myelin-basic-protein and Body-Weight

The therapeutic effects of mesenchymal stem cells-extracellular vesicles have been proved in many inflammatory animal models. In the current study, we aimed to investigate the effect of extracellular vesicles (EVs) derived from human umbilical cord-MSC (hUCSC-EV) on the clinical score and inflammatory/anti-inflammatory cytokines on the EAE mouse model. After induction of EAE in C57Bl/6 mice, they were treated intravenously with hUCSC-EV or vehicle. The clinical score and body weight of all mice was registered every day. On day 30, mice were sacrificed and splenocytes were isolated for cytokine assay by ELISA. Cytokine expression of pro-/anti-inflammatory cytokine by real-time PCR, leukocyte infiltration by hematoxylin and eosin (H&E) staining, and the percent of glial fibrillary acidic protein (GFAP) and myelin basic protein (MBP) positive cells by immunohistochemistry were assessed in the spinal cord. Our results showed that hUCSC-EV-treated mice have lower maximum mean clinical score (MMCS), pro-inflammatory cytokines, and inflammatory score in comparison to the control mice. We also showed that hUCSC-EV administration significantly improved body weight and increased the anti-inflammatory cytokines and the frequency of Treg cells in the spleen. There was no significant difference in the percent of GFAP and MBP positive cells in the spinal cord of experimental groups. Finally, we suggest that intravenous administration of hUCSC-EV alleviate induce-EAE by reducing the pro-inflammatory cytokines, such as IL-17a, TNF-α, and IFN-γ, and increasing the anti-inflammatory cytokines, IL-4 and IL-10, and also decrease the leukocyte infiltration in a model of MS. It seems that EVs from hUC-MSCs have the same therapeutic effects similar to EVs from other sources of MSCs, such as adipose or bone marrow MSCs.

Topics: Animals; Body Weight; Encephalomyelitis, Autoimmune, Experimental; Extracellular Vesicles; Female; Fetal Blood; Gene Expression Regulation; Glial Fibrillary Acidic Protein; Humans; Interferon-gamma; Interleukin-10; Interleukin-17; Interleukin-4; Mesenchymal Stem Cells; Mice; Mice, Inbred C57BL; Myelin Basic Protein; Spinal Cord; Spleen; T-Lymphocytes, Regulatory; Treatment Outcome; Tumor Necrosis Factor-alpha

Synthetic glucocorticoids (sGCs) such as dexamethasone (DEX), while used to mitigate inflammation and disease progression in premature infants with severe bronchopulmonary dysplasia (BPD), are also associated with significant adverse neurologic effects such as reductions in myelination and abnormalities in neuroanatomical development. Ciclesonide (CIC) is a sGC prodrug approved for asthma treatment that exhibits limited systemic side effects. Carboxylesterases enriched in the lower airways convert CIC to the glucocorticoid receptor (GR) agonist des-CIC. We therefore examined whether CIC would likewise activate GR in neonatal lung but have limited adverse extra-pulmonary effects, particularly in the developing brain. Neonatal rats were administered subcutaneous injections of CIC, DEX or vehicle from postnatal days 1-5 (PND1-PND5). Systemic effects linked to DEX exposure, including reduced body and brain weight, were not observed in CIC treated neonates. Furthermore, CIC did not trigger the long-lasting reduction in myelin basic protein expression in the cerebral cortex nor cerebellar size caused by neonatal DEX exposure. Conversely, DEX and CIC were both effective at inducing the expression of select GR target genes in neonatal lung, including those implicated in lung-protective and anti-inflammatory effects. Thus, CIC is a promising, novel candidate drug to treat or prevent BPD in neonates given its activation of GR in neonatal lung and limited adverse neurodevelopmental effects. Furthermore, since sGCs such as DEX administered to pregnant women in pre-term labor can adversely affect fetal brain development, the neurological-sparing properties of CIC, make it an attractive alternative for DEX to treat pregnant women severely ill with respiratory illness, such as with asthma exacerbations or COVID-19 infections.

Topics: Animals; Animals, Newborn; Anti-Inflammatory Agents; Body Weight; Brain; Cerebellum; Cerebral Cortex; COVID-19 Drug Treatment; Dexamethasone; Female; Glucocorticoids; Lung; Mice; Mice, Inbred C57BL; Myelin Basic Protein; Organ Size; Pregnancy; Pregnenediones; Prodrugs; Rats; Rats, Sprague-Dawley; Receptors, Glucocorticoid; Signal Transduction

Ketogenic diet (KD) is defined as a high-fat, low-carbohydrate diet with appropriate amounts of protein, which has broad neuroprotective effects. However, the mechanisms of ameliorating the demyelination and of the neuroprotective effects of KD have not yet been completely elucidated. Therefore, the present study investigated the protection mechanism of KD treatment in the cuprizone (bis-cyclohexanone oxalydihydrazone, CPZ)-induced demyelination mice model, with special emphasis on neuroinflammation. After the KD treatment, an increased ketone body level in the blood of mice was detected, and a significant increase in the distance traveled within the central area was observed in the open field test, which reflected the increased exploration and decreased anxiety of mice that received CPZ. The results of Luxol fast blue and myelin basic protein (MBP) immunohistochemistry staining for the evaluation of the myelin content within the corpus callosum revealed a noticeable increase in the number of myelinated fibers and myelin score after KD administration in these animals. Concomitant, the protein expressions of glial fibrillary acidic protein (GFAP, an astrocyte marker), ionized calcium-binding adaptor molecule 1 (Iba-1, a microglial marker), CD68 (an activated microglia marker) and CD16/32 (a M1 microglial marker) were down-regulated, while the expression of oligodendrocyte lineage transcription factor 2 (OLIG2, an oligodendrocyte precursor cells marker) was up-regulated by the KD treatment. In addition, the KD treatment not only reduced the level of the C-X-C motif chemokine 10 (CXCL10), which is correlated to the recruitment of activated microglia, but also inhibited the production of proinflammatory cytokines, including interleukin 1β (IL-1β) and tumor necrosis factor-α (TNF-α), which are closely correlated to the M1 phenotype microglia. It is noteworthy, that the expression levels of histone deacetylase 3 (HADC3) and nod-like receptor pyrin domain containing 3 (NLRP3) significantly decreased after KD administration. In conclusion, these data demonstrate that KD decreased the reactive astrocytes and activated the microglia in the corpus callosum, and that KD inhibited the HADC3 and NLRP3 inflammasome signaling pathway in CPZ-treated mice. This suggests that the inhibition of the HADC3 and NLRP3 signaling pathway may be a novel mechanism by which KD exerts its protective actions for the treatment of demyelinating diseases.

Topics: Alanine Transaminase; Animals; Aspartate Aminotransferases; Astrocytes; Body Weight; Brain; Chemokine CXCL10; Cuprizone; Cytokines; Demyelinating Diseases; Diet, Ketogenic; Disease Models, Animal; Down-Regulation; Glial Fibrillary Acidic Protein; Histone Deacetylases; Inflammasomes; Male; Mice; Mice, Inbred C57BL; Microglia; Myelin Basic Protein; NLR Family, Pyrin Domain-Containing 3 Protein; Signal Transduction; Weight Loss

Topics: Acrolein; Animals; Body Weight; Brain; Drinking Behavior; Feeding Behavior; Glial Fibrillary Acidic Protein; Intermediate Filaments; Male; Myelin Basic Protein; Rats, Wistar

Hypoxic-ischemic (HI) injury to the developing brain remains a major cause of morbidity. To date, few therapeutic strategies could provide complete neuroprotection. Erythropoietin (EPO) has been shown to be beneficial in several models of neonatal HI. This study examines the effect of treatment with erythropoietin on postnatal day 2 (P2) rats introduced with HI injury.. Rats at P2 were randomized into four groups: sham, bilateral carotid artery occlusion (BCAO), BCAO + early EPO, and BCAO + late EPO groups. Pups in each group were injected with either saline or EPO (5000U/kg) intraperitoneally once at immediately (early) or 48h (late) after HI induction. Body weight was assessed at P2 before and day 7 after HI. Mortality Rate was assessed at 24h, 48h and 72h after HI and brain water content was assessed at 72h. Brain weight and expression of myelin basic protein (MBP) were assessed at day 7 and day 14. At day 31 to 35 following HI insult, neurological behavior function was assessed via Morris water maze (MWM) test.. HI cause significant higher mortality in male than in female (P=0.0445). Among the surviving animal, HI affect significantly the body growth, brain growth, MBP expression, and neurological behavior. EPO treatments at both early and late time points significantly benefit the rats in injury recovery, in which they promoted weight gains, reduced brain edema, as well as improved spatial learning ability and memory.. We demonstrated a single dose of EPO at 5000U/kg immediately or 48h after HI injury had significant benefit for the P2 rats in injury recovery, and there was no adverse effect associated with either EPO treatment.

Topics: Age Factors; Analysis of Variance; Animals; Animals, Newborn; Body Weight; Brain Edema; Developmental Disabilities; Disease Models, Animal; Erythropoietin; Hypoxia-Ischemia, Brain; Maze Learning; Myelin Basic Protein; Neuroprotective Agents; Rats; Rats, Sprague-Dawley

T3D-959, a dual PPAR-δ/PPAR γ nuclear receptor agonist and former diabetes drug candidate, has been repositioned as an Alzheimer's disease (AD)-modifying therapy.. This study examines the effectiveness and mechanisms of T3D-959's therapeutic effects using in vivo and ex vivo rat models of sporadic AD.. A sporadic AD model was generated by intracerebral (i.c.) administration of streptozotocin (STZ). Control and i.c. STZ treated rats were gavaged with saline or T3D-959 (0.3 to 3.0 mg/kg/day) for 28 days. Spatial learning and memory were evaluated using the Morris water maze test. Frontal lobe slice cultures generated 24 hours after i.c. STZ or vehicle were used to study early effects of T3D-959 (0.5-1.0 μM) on viability and molecular markers of AD.. T3D-959 significantly improved spatial learning and memory in i.c STZ-treated rats. Mechanistically, T3D-959 significantly improved culture viability and brain morphology, reduced levels of oxidative stress and Aβ, and normalized expression of phospho-tau, choline acetyltransferase, and myelin-associated glycoprotein. Protective effects occurred even at the lowest tested dose of T3D-959.. Pre-clinical proof of concept has been demonstrated that T3D-959 can improve multiple pathologies of AD resulting in significant improvements in cognitive function and molecular and biochemical indices of neurodegeneration. These results support the theses that (1) effective disease modification in AD can be achieved by targeting relevant nuclear receptors, and (2) treating AD as a metabolic disease has the potential to be disease remedial. A Phase 2a trial of T3D-959 in mild-to-moderate AD patients has been initiated (ClinicalTrials.gov identifier NCT02560753).

Topics: Alzheimer Disease; Analysis of Variance; Animals; Animals, Newborn; Antibiotics, Antineoplastic; Antipsychotic Agents; Blood Glucose; Body Weight; Brain; Choline O-Acetyltransferase; Disease Models, Animal; Dose-Response Relationship, Drug; Glial Fibrillary Acidic Protein; Glucosephosphate Dehydrogenase; In Vitro Techniques; Maze Learning; Myelin Basic Protein; Neurotoxicity Syndromes; Organ Culture Techniques; PPAR delta; PPAR gamma; Rats; Rats, Long-Evans; Spatial Learning; Streptozocin

The aim of this study was to evaluate the effects of orexin-A (OX-A) on behavioral and pathological parameters and on gene expression of some multiple sclerosis-related peptides in a model of experimental autoimmune encephalomyelitis (EAE). EAE was induced by subcutaneous administration of MOG 35-55. Following immunization, the treatment was initiated by using SB.334867 (orexin-1 receptor antagonist) and/or OX-A. Locomotor activity and exploratory behaviors were monitored using open field and T-maze continuous alternation task (T-CAT) respectively. Pain sensitivity was assessed by hot-plate test. Histopathological assessments were performed by H&E staining. The expression of TGF-β, MBP, MMP-9, IL-12, iNOS and MCP-1 were measured using real-time PCR method in lumbar spinal cord. OX-A administration in EAE mice remarkably attenuated the clinical symptoms, increased latency response in hot plate test, inhibited infiltration of inflammatory cells, up-regulated mRNA expression of TGF-β as well as MBP and down-regulated mRNA expression of iNOS, MMP-9 and IL-12. In contrast SB.334867 administration in EAE mice deteriorated the clinical symptoms, decreased the alternation in T-CAT, increased infiltration of inflammatory cells, down-regulated mRNA expression of TGF-β and MBP and up-regulated mRNA expression of iNOS. Results of this study suggest that the orexinergic system might be involved in pathological development of EAE. These findings suggest orexinergic system as a potential target for treatment of multiple sclerosis.

Topics: Animals; Attention; Avoidance Learning; Benzoxazoles; Body Weight; Cytokines; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Exploratory Behavior; Female; Matrix Metalloproteinase 9; Maze Learning; Mice; Mice, Inbred C57BL; Myelin Basic Protein; Myelin-Oligodendrocyte Glycoprotein; Naphthyridines; Nitric Oxide Synthase Type II; Orexin Receptor Antagonists; Orexins; Peptide Fragments; Severity of Illness Index; Spinal Cord; Time Factors; Urea

Preclinical Research The aim of the present study was to evaluate the neuroprotective benefits of rhGLP-1 in diabetic rats subjected to acute cerebral ischemia/reperfusion injury induced by middle cerebral artery occlusion/reperfusion (MCAO/R). Streptozotocin (STZ)-induced diabetic rats were pretreated with rhGLP-1 (10, 20, or 40 μg/kg ip, tid) for 14 days. During this time, body weight and fasting blood glucose levels were assessed. Rats were then subjected to MCAO 90 min/R 24 h. At 2 and 24 h of reperfusion, rats were evaluated for neurological deficits and blood samples were collected to analyze markers of brain injury. Rats were then sacrificed to assess the infarction volume. rhGLP-1 pretreatment lowered blood glucose levels, improved neurological scores, attenuated infarct volumes, and reduced the blood levels of S100 calcium-binding protein B (S100B), neuron-specific enolase (NSE), and myelin basic protein (MBP). rhGLP-1 has neuroprotective benefits in diabetic rats with cerebral ischemia/reperfusion injury and could potentially be used as a prophylatic neuroprotectant in diabetic patients at high risk of ischemic stroke. Drug Dev Res 77 : 124-133, 2016.   © 2016 Wiley Periodicals, Inc.

Topics: Animals; Body Weight; Brain Ischemia; Diabetes Mellitus, Experimental; Disease Models, Animal; Glucagon-Like Peptide 1; Hypoglycemic Agents; Male; Myelin Basic Protein; Neuroprotective Agents; Phosphopyruvate Hydratase; Rats; Rats, Sprague-Dawley; Recombinant Proteins; Reperfusion Injury; S100 Calcium Binding Protein beta Subunit; Streptozocin; Treatment Outcome

Infections are common following stroke and associated with worse outcome. Using an animal model of pneumonia, we assessed the effect of infection and its treatment on the immune response and stroke outcome.. Lewis rats were subjected to transient cerebral ischemia and survived for 4weeks. One day after stroke animals were exposed to aerosolized Staphylococcus aureus, Pseudomonas aeruginosa or saline. Antibiotics (ceftiofur or enrofloxacin) were started immediately after exposure or delayed for 3days. Behavioral tests were performed weekly. ELISPOT assays were done on lymphocytes from spleen and brain to assess autoimmune responses to myelin basic protein (MBP).. Among animals that received immediate antibiotic therapy, infection was associated with worse outcome in ceftiofur but not enrofloxacin treated animals. (The outcome with immediate enrofloxacin therapy was so impaired that further worsening may have been difficult to detect.) A delay in antibiotic therapy was associated with better outcomes in both ceftiofur and enrofloxacin treated animals. Infection was associated with an increased likelihood of developing Th1(+) responses to MBP in non-infarcted brain (OR=2.94 [1.07, 8.12]; P=0.04), and Th1(+) responses to MBP in spleen and non-infarcted brain were independently associated with a decreased likelihood of stroke recovery (OR=0.16 [0.05, 0.51; P=0.002 and OR=0.32 [0.12, 0.84]; P=0.02, respectively).. Infection worsens stroke outcome in ceftiofur treated animals and increases Th1 responses to MBP. These data may help explain how infection worsens stroke outcome and suggest that treatment of infection may contribute to this outcome.

Topics: Animals; Anti-Bacterial Agents; Antineoplastic Agents; Body Temperature; Body Weight; Cephalosporins; Cytokines; Disease Models, Animal; Enrofloxacin; Fluoroquinolones; Lymphocytes; Male; Myelin Basic Protein; Nervous System Diseases; Pneumonia; Rats; Rats, Inbred Lew; Staphylococcus aureus; Statistics, Nonparametric; Stroke

Multiple sclerosis is a chronic inflammatory, demyelinating degenerative disease of the central nervous system. Current treatments target pathological immune responses to counteract the inflammatory processes. However, these drugs do not restrain the long-term progression of clinical disability. For this reason, new therapeutic approaches and identification of novel target molecules are needed to prevent demyelination or promote repair mechanisms. Transient Receptor Potential Ankyrin 1 (TRPA1) is a nonselective cation channel with relatively high Ca

Topics: Adenomatous Polyposis Coli; Animals; Apoptosis; bcl-2 Homologous Antagonist-Killer Protein; Body Weight; Brain; Cuprizone; Demyelinating Diseases; Disease Models, Animal; Fibroblast Growth Factor 2; Gene Expression Regulation; Gliosis; Mice; Mice, Knockout; Monoamine Oxidase Inhibitors; Myelin Basic Protein; Nerve Tissue Proteins; Oligodendroglia; Signal Transduction; TRPA1 Cation Channel

The inflammatory mediator lipopolysaccharide (LPS) has been shown to induce acute gliosis in neonatal mice. However, the progressive effects on the murine neurodevelopmental program over the week that follows systemic inflammation are not known. Thus, we investigated the effects of repeated LPS administration in the first postnatal week in mice, a condition mimicking sepsis in late preterm infants, on the developing central nervous system (CNS).. Systemic inflammation was induced by daily intraperitoneal administration (i.p.) of LPS (6 mg/kg) in newborn mice from postnatal day (PND) 4 to PND6. The effects on neurodevelopment were examined by staining the white matter and neurons with Luxol Fast Blue and Cresyl Violet, respectively. The inflammatory response was assessed by quantifying the expression/activity of matrix metalloproteinases (MMP), toll-like receptor (TLR)-4, high mobility group box (HMGB)-1, and autotaxin (ATX). In addition, B6 CX3CR1(gfp/+) mice combined with cryo-immunofluorescence were used to determine the acute, delayed, and lasting effects on myelination, microglia, and astrocytes.. LPS administration led to acute body and brain weight loss as well as overt structural changes in the brain such as cerebellar hypoplasia, neuronal loss/shrinkage, and delayed myelination. The impaired myelination was associated with alterations in the proliferation and differentiation of NG2 progenitor cells early after LPS administration, rather than with excessive phagocytosis by CNS myeloid cells. In addition to disruptions in brain architecture, a robust inflammatory response to LPS was observed. Quantification of inflammatory biomarkers revealed decreased expression of ATX with concurrent increases in HMGB1, TLR-4, and MMP-9 expression levels. Acute astrogliosis (GFAP(+) cells) in the brain parenchyma and at the microvasculature interface together with parenchymal microgliosis (CX3CR1(+) cells) were also observed. These changes preceded the migration/proliferation of CX3CR1(+) cells around the vessels at later time points and the subsequent loss of GFAP(+) astrocytes.. Collectively, our study has uncovered a complex innate inflammatory reaction and associated structural changes in the brains of neonatal mice challenged peripherally with LPS. These findings may explain some of the neurobehavioral abnormalities that develop following neonatal sepsis.

Topics: Age Factors; Anethole Trithione; Animals; Animals, Newborn; Body Weight; Cerebellum; CX3C Chemokine Receptor 1; Demyelinating Diseases; Developmental Disabilities; Gene Expression Regulation; Green Fluorescent Proteins; HMGB1 Protein; Inflammation; Lipopolysaccharides; Mice; Mice, Inbred C57BL; Mice, Transgenic; Microglia; Myelin Basic Protein; Nervous System Malformations; Neurodegenerative Diseases; Receptors, Chemokine; Time Factors; Toll-Like Receptor 4

There is increasing evidence that poor growth of preterm infants is a risk factor for poor long-term development, while the effects of early postnatal growth restriction are not well known. We utilized a rat model to examine the consequences of different patterns of postnatal growth and hypothesized that early growth failure leads to impaired development and insulin resistance. Rat pups were separated at birth into normal (N, n = 10) or restricted intake (R, n = 16) litters. At d11, R pups were re-randomized into litters of 6 (R-6), 10 (R-10) or 16 (R-16) pups/dam. N pups remained in litters of 10 pups/dam (N-10). Memory and learning were examined through T-maze test. Insulin sensitivity was measured by i.p. insulin tolerance test and glucose tolerance test.. By d10, N pups weighed 20% more than R pups (p < 0.001). By d15, the R-6 group caught up to the N-10 group in weight, the R-10 group showed partial catch-up growth and the R-16 group showed no catch-up growth. All R groups showed poorer scores in developmental testing when compared with the N-10 group during T-Maze test (p < 0.05). Although R-16 were more insulin sensitive than R-6 and R-10, all R groups were more glucose tolerant than N-10.. In rats, differences in postnatal growth restriction leads to changes in development and in insulin sensitivity. These results may contribute to better elucidating the causes of poor developmental outcomes in human preterm infants.

Topics: Animals; Animals, Newborn; Blood Glucose; Body Composition; Body Weight; Brain; Eating; Female; Homeostasis; Insulin; Male; Maze Learning; Myelin Basic Protein; Rats

Diffusion tensor imaging (DTI) suggests the presence of white matter abnormality at the prodromal stage in human Alzheimer's disease (AD).. To use a mouse model of AD to determine whether the white matter abnormality detected by in vivo DTI is associated with functional deficits and axon damage.. Amyloid-β1-42 (Aβ1-42) was injected into the left lateral ventricle in mice. Two months after the injection, in vivo DTI and visual evoked potential (VEP) recordings were performed, followed by immunohistochemistry of phosphorylated neurofilament and myelin basic protein.. DTI of Aβ1-42-treated mice showed a significant increase of radial diffusivity in white matter including the optic nerves and tracts. The abnormality was associated with decreased amplitude and increased latency of VEP. Immunohistochemistry confirmed a significant loss of axons and myelin integrity.. White matter damage induced by Aβ1-42 in mice can be detected non-invasively by DTI.

Topics: Amyloid beta-Peptides; Animals; Body Weight; Diffusion Magnetic Resonance Imaging; Disease Models, Animal; Evoked Potentials, Visual; Female; Functional Laterality; Leukoencephalopathies; Male; Mice; Mice, Inbred C57BL; Myelin Basic Protein; Neurofilament Proteins; Optic Nerve; Peptide Fragments

The etiology of schizophrenia (SZ) is complex and largely unknown. Neuroimaging and postmortem studies have suggested white matter disturbances in SZ. In the present study, we tested the white matter deficits hypothesis of SZ using a mouse model of SZ induced by NMDA receptor antagonist MK-801. We found that mice with repeated chronic MK-801 administration showed increased locomotor activity in the open field test, less exploration of a novel environment in the hole-board test, and increased anxiety in the elevated plus maze but no impairments were observed in coordination or motor function on accelerating rota-rod. The total white matter volume and corpus callosum volume in mice treated with MK-801 were significantly decreased compared to control mice treated with saline. Myelin basic protein and 2', 3'-cyclic nucleotide 3'-phosphodiesterase were also significantly decreased in the mouse model of SZ. Furthermore, we observed degenerative changes of myelin sheaths in the mouse model of SZ. These results provide further evidence of white matter deficits in SZ and indicate that the animal model of SZ induced by MK-801 is a useful model to investigate mechanisms underlying white matter abnormalities in SZ.

Topics: 2',3'-Cyclic-Nucleotide Phosphodiesterases; Animals; Behavior, Animal; Blotting, Western; Body Weight; Disease Models, Animal; Dizocilpine Maleate; Excitatory Amino Acid Antagonists; Immunoenzyme Techniques; Male; Mice; Mice, Inbred C57BL; Microscopy, Electron, Transmission; Motor Activity; Myelin Basic Protein; Nerve Fibers, Myelinated; Real-Time Polymerase Chain Reaction; Receptors, N-Methyl-D-Aspartate; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Schizophrenia; White Matter

To study the effects of perinatal recurrent infection on the brain development in immature mice.. Six pregnant C57BL6 mice were randomly assigned to three groups: intrauterine infection, perinatal recurrent infection and control. The intrauterine infection group was intraperitoneally injected with LPS (0.5 mg/kg) on the 18th day of pregnancy. The perinatal recurrent infection group was injected with LPS (0.5 mg/kg) on the 18th day of pregnancy and their offsprings were intraperitoneally injected with the same dose of LPS daily from postnatal day 3 to 12. The control group was administered with normal saline at the same time points as the recurrent infection group. The short-time neurobehaviors were assessed on postnatal day 13. The mice were then sacrificed to measure brain weights and neuropathological changes using cresyl violet staining. Western blot was used to evaluate the expression of TNF-α, Caspase-3 and myelin basic protein (MBP).. The brain weights of the recurrent infection group were significantly lower than the control and intrauterine infection groups (P<0.05) and the recurrent infection group displayed significant neuropathological changes. Perinatal recurrent infection resulted in increased expression levels of TNF-α and Caspase-3, and decreased expression level of MBP compared with the intrauterine infection and control groups (P<0.01). The neurobehavior test showed that the recurrent infection group used longer time in gait reflex, right reflex and geotaxis reflex compared with the control and intrauterine infection groups on postnatal day 13 (P<0.05).. Perinatal recurrent infection may exacerbate inflammatory response and cell death in the immature brain, which may be one of the important factors for perinatal brain injury.

Topics: Animals; Animals, Newborn; Bacterial Infections; Body Weight; Brain; Caspase 3; Female; Mice; Mice, Inbred C57BL; Myelin Basic Protein; Pregnancy; Recurrence; Reflex

Multiple sclerosis (MS) is an autoimmune disease characterized by inflammatory cell infiltration of the central nervous system (CNS) and multifocal demyelination. Clinical data and clinical indicators demonstrate that estrogen improves the relapse-remittance of MS patients. This study aimed to investigate the anti-inflammatory effects and the underlying mechanism(s) of action of estrogen and estrogen receptor α (ERα) in an experimental autoimmune encephalomyelitis (EAE) mouse model of MS. An ERα recombinant lentivirus was constructed. Mouse neurons were cultured in serum-free culture medium, and ERα recombinant lentivirus with a multiplicity of infection (MOI) of 5 was used to infect the neurons. Furthermore, neuronal ERα mRNA and protein expression were detected using real-time quantitative PCR and western blot analysis. We sterotaxically injected ERα recombinant lentivirus into the lateral ventricle of mouse brains, and successfully identified infected neurons using Flag immunofluorescence staining to determine the optimal dose. A total of 75 C57BL/6 mice were ovariectomized. After 2 weeks, EAE was induced with myelin oligodendrocyte glycoprotein (MOG) 35-55 peptide. The EAE mice were divided into 5 groups: the estrogen group (treatment with estradiol), the ERα agonist group (treatment with raloxifene), the ERα recombinant lentivirus group (ERα group, treatment with ERα recombinant lentivirus), the empty virus group and the normal saline (NS) group; clinical symptoms and body weight were compared among the groups. We assessed EAE-related parameters, detected pathological changes with immunohistochemistry and quantified the expression of myelin basic protein (MBP), matrix metalloproteinase-9 (MMP-9), and a subset of EAE-related cytokines using enzyme-linked immunosorbent assay (ELISA). We successfully constructed an ERα recombinant lentivirus. C57BL/6 mouse neurons can survive in culture for at least 8 weeks. During that period, the recombinant lentivirus was able to infect the neurons, while sustaining green fluorescence protein (GFP) expression. ERα recombinant lentivirus also infected the neurons at a MOI of 5. The ERα mRNA and protein expression levels were higher in the infected neurons compared to the uninfected ones. We successfully infected the CNS of C57BL/6 mice by stereotaxically injecting ERα recombinant lentivirus into the lateral ventricle of the mouse brains and induced EAE. The lentivirus-mediated overexpression of ERα reduced the inciden

Topics: Animals; Body Weight; Brain; Central Nervous System; Cytokines; Demyelinating Diseases; Encephalomyelitis, Autoimmune, Experimental; Estrogen Receptor alpha; Green Fluorescent Proteins; HEK293 Cells; Humans; Immunohistochemistry; Inflammation; Lentivirus; Matrix Metalloproteinase 9; Mice; Mice, Inbred C57BL; Myelin Basic Protein; Nerve Fibers; Recombination, Genetic; RNA, Messenger; Spinal Cord; Transfection

Brain weight and size are known to be reduced in adult leptin-deficient Lep/Lep (OB) mice when compared with the wild-type (+/+) mice (C57BL/6: B6). We here analyzed leptin's effects on myelination by examining morphometrically the myelin sheath (MS) in the cerebrum of postnatal day (P) 14 and P28 OB that had received leptin 1 nmol/capita/day from P7 to P14 or P28 (OB+lep), in comparison with OB and B6. We examined myelin basic protein (MBP) mRNA levels and the differentiation of oligodendrocytes by comparing the number of oligodendrocyte precursor cells (OPCs) and the mature oligodendrocytes in the cerebrum between OB, OB+lep, and B6 on P14 and P28. MBP-mRNA expression was lower in OB than in B6 on P14 and P28. On P14, it was higher in OB+lep than in OB but was still lower than in B6, whereas on P28 it was even higher in OB+lep than in B6. On P28, the radii of myelinated axons were larger in OB than in B6 and OB+lep. The MS on P28 was significantly thinner in OB than in B6, but there was no significant difference between OB and OB+lep. There were significantly fewer mature oligodendrocytes in OB and OB+lep than in B6 on P28, whereas on P14 there were significantly fewer OPCs in OB and OB+lep than in B6. Our results suggested that leptin regulates the myelination of oligodendrocytes and that the replenishment of leptin in OB recovered myelination but did not affect the differentiation of OPCs from P7 to P28.

Topics: Age Factors; Animals; Animals, Newborn; Antigens; Body Weight; Cell Differentiation; Cerebrum; Gangliosides; Gene Expression Regulation, Developmental; Leptin; Mice; Mice, Inbred C57BL; Mice, Neurologic Mutants; Microscopy, Electron, Transmission; Myelin Basic Protein; Myelin Sheath; O Antigens; Oligodendroglia; Proteoglycans; Receptor, Platelet-Derived Growth Factor alpha; RNA, Messenger

A peripheral indicator of the presence and magnitude of brain injury has been a sought-after tool by clinicians. We measured neuron-specific enolase (NSE), myelin basic protein (MBP), and S100B, prior to and after scaled cortical impact in immature pigs, to determine if these purported markers increase after injury, correlate with the resulting lesion volume, and if these relationships vary with maturation. Scaled cortical impact resulted in increased lesion volume with increasing age. Concentrations of NSE, but not S100B or MBP, increased after injury in all age groups. The high variability of S100B concentrations prior to injury may have precluded detection of an increase due to injury. Total serum markers were estimated, accounting for the allometric growth of blood volume, and resulted in a positive correlation of both NSE and S100B with lesion volume. Even with allometric scaling of blood volume and a uniform mechanism of injury, NSE had only a fair to poor predictive value. In a clinical setting, where the types of injuries are varied, more investigation is required to yield a panel of serum markers that can reliably predict the extent of injury. Allometric scaling may improve estimation of serum marker release in pediatric populations.

Topics: Aging; Algorithms; Animals; Biomarkers; Blood Volume; Body Weight; Brain Injuries; Cerebral Cortex; Enzyme-Linked Immunosorbent Assay; Female; Male; Myelin Basic Protein; Nerve Growth Factors; Phosphopyruvate Hydratase; Predictive Value of Tests; S100 Calcium Binding Protein beta Subunit; S100 Proteins; Swine

In rats, iron deficiency produces an alteration in myelin formation. However, there is limited information on the effects of this condition on oligodendroglial cell (OLGc) proliferation and maturation. In the present study, we further analyzed the hypomyelination associated with iron deficiency by studying the dynamics of oligodendrogenesis. Rats were fed control (40 mg Fe/kg) or iron-deficient (4 mg Fe/kg) diets from gestation day 5 until postnatal day 3 (P3) or 11 (P11). OLGc proliferation, migration and differentiation were investigated before and after an intracranial injection of apotransferrin at 3 days of age (P3). The proliferating cell population was evaluated at P3. Iron-deficient (ID) animals showed an increase in the oligodendrocyte precursors cell (OPC) population in comparison with controls. The overall pattern of migration of cells labeled with BrdU was investigated at P11. Iron deficiency increased the amount of BrdU(+) cells in the corpus callosum (CC) and decreased OLGc maturation and myelin formation. Changes in nerve conduction were analyzed by measuring visual evoked potentials. Latency and amplitude were significantly disturbed in ID rats compared with controls. Both parameters were substantially normalized when animals were treated with a single intracranial injection of 350 ng apotransferrin (aTf). The current results give support to the idea that iron deficiency increases the number of proliferating and undifferentiated cells in the CC compared with the control. Treatment with aTf almost completely reverted the effects of iron deficiency, both changing the migration pattern and increasing the number of mature cells in the CC and myelin formation.

Topics: Age Factors; Analysis of Variance; Animals; Animals, Newborn; Apoproteins; Basic Helix-Loop-Helix Transcription Factors; Body Weight; Brain; Bromodeoxyuridine; Cell Count; Cell Differentiation; Cell Proliferation; Corpus Callosum; Demyelinating Diseases; Electroencephalography; Evoked Potentials, Visual; Female; Gene Expression Regulation, Developmental; Hematocrit; Iron Deficiencies; Myelin Basic Protein; Nerve Tissue Proteins; Neural Cell Adhesion Molecule L1; Oligodendroglia; Photic Stimulation; Pregnancy; Prenatal Exposure Delayed Effects; Proliferating Cell Nuclear Antigen; Rats; Sialic Acids; Transferrin

Simvastatin, the most widely used cholesterol-lowering drug, has been reported to protect the adult brain from ischemia. Nevertheless, little is known about its action on developing brain after stroke. Although a few reports have found recently that simvastatin displays anti-inflammation and anti-apoptosis properties and improves the cognitive and morphological consequences in the neonatal rats after hypoxia-ischemia (HI) damage, to our best knowledge, there has been no study of the effect of it on myelin formation after neonatal brain damage. Therefore, we investigated whether simvastatin could promote the myelination of oligodendrocytes in the neonatal rats after HI and explored the possible role of microglial responses in this process.. Postnatal day 7 Sprague-Dawley rats were subjected to HI. White matter integrity and myelination were evaluated by the densitometry of myelin basic protein (MBP) immunostaining. OX-42 immunoreactivity and nissl staining were used for identifying microglial responses and the structure changes of white matter and adjacent gray matter after HI. Simvastatin was administrated prophylactically to rats.. HI induced serious hypomyelination especially in external and internal capsules 3 and 7 days after HI, accompanying with microglial activation remarkably. Simvastatin treatment greatly increased the densities of MBP immunostaining, inhibited microglial activation and reduced the numbers of pyknotic cells and neuronal loss.. The present study shows that simvastatin treatment in neonatal rats attenuates HI-induced developing oligodendrocytes injury and hypomyelination. Its anti-inflammatory properties via suppression of microglial activation are likely to contribute to this action. It provides experimental evidence to support the neuroprotective effects of statins in neonatal ischemic stroke.

Topics: Age Factors; Analysis of Variance; Animals; Animals, Newborn; Body Weight; CD11b Antigen; Demyelinating Diseases; Disease Models, Animal; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Hypoxia-Ischemia, Brain; Myelin Basic Protein; Rats; Rats, Sprague-Dawley; Simvastatin

Peripheral neurotoxicity is a frequent complication limiting docetaxel chemotherapy in patients with cancer. We developed an experimental model that closely mimics the course of neuropathy in patients, aiming to investigate both the mechanisms of neurotoxicity at biochemical, functional and morphological levels and the potential neuroprotective role of neuroactive steroids. We demonstrated that treatment with dihydroprogesterone (DHP) or progesterone (P) counteracts docetaxel-induced neuropathy, preventing nerve conduction and thermal threshold changes, and degeneration of skin nerves in the foodpad. Neuroactive steroids also counteract the changes in gene expression of several myelin proteins and calcitonin gene-related peptide induced by docetaxel in sciatic nerve and lumbar spinal cord, respectively. Most nerve abnormalities observed during the treatment with docetaxel spontaneously recovered after drug withdrawal, similarly to what occurs in patients. However, results of midterm follow-up experiments indicated that animals cotreated with DHP or P have a faster recovery of the neuropathy compared with docetaxel-treated rats. Our study confirmed that neuroactive steroids exert a protective effect on peripheral nerves at different levels, suggesting that they might represent a new therapeutic frontier for patients with chemotherapy-induced neuropathy.

Topics: 20-alpha-Dihydroprogesterone; Analysis of Variance; Animals; Body Weight; Disease Models, Animal; Docetaxel; Gene Expression Regulation; Male; Myelin Basic Protein; Myelin P0 Protein; Myelin Proteins; Nerve Fibers; Neural Conduction; Neuroprotective Agents; Pain Threshold; Peripheral Nervous System Diseases; Progesterone; Rats; Rats, Inbred F344; Sciatic Nerve; Taxoids

Human diabetic patients often lose touch and vibratory sensations, but to date, most studies on diabetes-induced sensory nerve degeneration have focused on epidermal C-fibers. Here, we explored the effects of diabetes on cutaneous myelinated fibers in relation to the behavioral responses to tactile stimuli from diabetic mice. Weekly behavioral testing began prior to streptozotocin (STZ) administration and continued until 8 weeks, at which time myelinated fiber innervation was examined in the footpad by immunohistochemistry using antiserum to neurofilament heavy chain (NF-H) and myelin basic protein (MBP). Diabetic mice developed reduced behavioral responses to non-noxious (monofilaments) and noxious (pinprick) stimuli. In addition, diabetic mice displayed a 50% reduction in NF-H-positive myelinated innervation of the dermal footpad compared with non-diabetic mice. To test whether two neurotrophins nerve growth factor (NGF) and/or neurotrophin-3 (NT-3) known to support myelinated cutaneous fibers could influence myelinated innervation, diabetic mice were treated intrathecally for 2 weeks with NGF, NT-3, NGF and NT-3. Neurotrophin-treated mice were then compared with diabetic mice treated with insulin for 2 weeks. NGF and insulin treatment both increased paw withdrawal to mechanical stimulation in diabetic mice, whereas NT-3 or a combination of NGF and NT-3 failed to alter paw withdrawal responses. Surprisingly, all treatments significantly increased myelinated innervation compared with control-treated diabetic mice, demonstrating that myelinated cutaneous fibers damaged by hyperglycemia respond to intrathecal administration of neurotrophins. Moreover, NT-3 treatment increased epidermal Merkel cell numbers associated with nerve fibers, consistent with increased numbers of NT-3-responsive slowly adapting A-fibers. These studies suggest that myelinated fiber loss may contribute as significantly as unmyelinated epidermal loss in diabetic neuropathy, and the contradiction between neurotrophin-induced increases in dermal innervation and behavior emphasizes the need for multiple approaches to accurately assess sensory improvements in diabetic neuropathy.

Topics: Animals; Blood Glucose; Body Weight; Diabetes Mellitus, Experimental; Dose-Response Relationship, Drug; Drug Interactions; Immunohistochemistry; Insulin; Male; Mice; Mice, Inbred C57BL; Myelin Basic Protein; Nerve Fibers, Myelinated; Nerve Growth Factors; Neurofilament Proteins; Pain Measurement; Physical Stimulation; Sensation Disorders; Skin; Time Factors

Clinical and experimental evidence indicate that the presence of intrauterine inflammation in pregnancy is not only a cause of preterm birth but is also associated with perinatal brain damage and long-term neurological handicap. In the present study, the neuropathological outcome was investigated in surviving pups in a model of inflammation-induced preterm delivery. C57BL/6 mice were subjected to intrauterine injection of lipopolysaccharide (LPS) or saline, at a time corresponding to 79% of average gestation (gestational day 15). Fetuses that survived after LPS administration were sacrificed on postnatal day 14 (PND 14). At PND 14, the brain weight of LPS-exposed pups was significantly lower than that of saline-exposed. A high proportion of LPS-exposed brains were found affected and exhibited hypomyelination, enlarged ventricles, and in some cortical gray matter lesions were evident. None of these pathologies were detected in sham-treated animals.. Intrauterine inflammation impaired brain development and various brain lesions were produced in both the white and gray matter after intrauterine LPS administration in mice.

Topics: Animals; Animals, Newborn; Body Weight; Brain; Female; Inflammation; Lipopolysaccharides; Mice; Mice, Inbred C57BL; Myelin Basic Protein; Organ Size; Pregnancy; Prenatal Exposure Delayed Effects

Chronic exposure to the copper-chelating agent, cuprizone (CPZ), is an increasingly popular model for producing demyelination. More importantly, cessation of cuprizone exposure allows for full remyelination, which represents a window of opportunity for determining the influence of environmental factors on regenerative processes. In the present study, CPZ-treated animals were assessed for functional status of systemic and central cytokine responsiveness to LPS, as well as assessment for signs of body weight changes. Exposure of male C57BL/6J mice to 5 weeks of 0.2% CPZ in the diet was optimal in producing demyelination and microglial activation, as measured by myelin basic protein, CD11b, and CD45 immunohistochemistry. Acute challenge with LPS at the end of 5 weeks CPZ treatment did not alter IL-1beta, IL-6, nor TNFalpha responses in the spleen and corpus callosum. Similarly, repeated exposure to LPS during the remyelination phase (CPZ removal) did not influence these measures to LPS. Plasma corticosterone was unaffected following acute challenge of CPZ-pretreated animals, but after repeated LPS treatment, there was a significant augmentation of the corticosterone response in CPZ-pretreated mice. Interestingly, the basal concentration of IL-1beta in the corpus callosum of CPZ treated animals was significantly increased, which was in keeping with the increase in activated microglial cells. In conclusion, the cuprizone model of demyelination and remyelination does not appear to influence the systemic nor central IL-1, IL-6, and TNF responses to acute nor repeated LPS. This opens up the possibility for studying the contribution of systemic inflammatory processes on remyelination after cessation of CPZ treatment.

Topics: Acute Disease; Animals; Body Weight; CD11b Antigen; Chelating Agents; Chronic Disease; Corpus Callosum; Corticosterone; Cuprizone; Cytokines; Demyelinating Diseases; Disease Models, Animal; Leukocyte Common Antigens; Lipopolysaccharides; Male; Mice; Mice, Inbred C57BL; Microglia; Myelin Basic Protein; Myelin Sheath; Nerve Regeneration; Spleen

Transferrin (Tf), the iron transport glycoprotein found in biological fluids of vertebrates, is synthesized mainly by hepatocytes. Tf is also synthesized by oligodendrocytes (Ol), and several lines of evidence indicate that brain Tf could be involved in myelinogenesis. Because Tf is postnatally expressed in the brain, we sought to investigate whether Tf could intervene in Ol differentiation. For this purpose, we analyzed transgenic mice overexpressing the complete human Tf gene in Ol. We show that the hTf transgene was expressed only from 5 days postpartum onward. In the brain of 14-day-old transgenic mice, the DM-20 mRNA level was decreased, whereas the PLP, MBP, CNP, and MAG mRNA levels were increased. We counted a higher proportion of Ol expressing the O4 (Ol-specific antigens) and PLP in brain cells cultured from transgenic mice. These results support the idea that overexpressing Tf in the brain accelerates the oligodendrocyte lineage maturation. Accordingly, by NMR imaging acquisition of diffusion tensor in hTf transgenic mice, we observed early maturation of the cerebellum and spinal cord and more myelination in the corpus callosum. In addition, hTf overexpression led to an increase in Sox10 mRNA and protein. Increases in Sox10 and in Tf expression occur simultaneously during brain development. The Olig1 mRNA level also increased, but long after the rise of hTf and Sox10. The Olig2 mRNA level remained unchanged in the brain of transgenic mice. Our findings suggest that Tf could influence oligodendrocyte progenitor differentiation in the CNS.

Topics: 2',3'-Cyclic-Nucleotide Phosphodiesterases; Age Factors; Analysis of Variance; Animals; Animals, Newborn; Blotting, Northern; Blotting, Western; Body Weight; Brain; Cell Count; Cell Differentiation; Cells, Cultured; Enzyme-Linked Immunosorbent Assay; Gene Expression Regulation, Developmental; Humans; Immunohistochemistry; Magnetic Resonance Imaging; Mice; Mice, Inbred C57BL; Mice, Transgenic; Myelin Basic Protein; Myelin Proteolipid Protein; Myelin-Associated Glycoprotein; Nerve Tissue Proteins; Oligodendroglia; Radioimmunoassay; Receptors, Cell Surface; RNA, Messenger; Transferrin

Neonatal and congenital hydrocephalus are common problems in humans. Hydrocephalus was induced in 1-day-old rats by injection of kaolin into the cisterna magna. At 7 and 21 days, magnetic resonance (MR) imaging was used to assess ventricle size, then brains were subjected to histopathological and biochemical analyses. Hydrocephalic pups did not exhibit delays in righting or negative geotaxis reflexes during the first week. At 7 days, there was variable ventricular enlargement with periventricular white matter edema, axon damage, reactive astrogliosis, and accumulation of macrophages in severe but not mild hydrocephalus. Cellular proliferation in the subependymal zone was significantly reduced. The cortical subplate neuron layer was disrupted. In rats allowed to survive to 21 days, weight was significantly lower in severely hydrocephalic rats. They also exhibited impaired memory in the Morris water maze test. Despite abnormal posture, there was minimal quantitative impairment of walking ability on a rotating cylinder. At 21 days, histological studies showed reduced corpus callosum thickness, fewer mature oligodendrocytes, damaged axons, and astroglial/microglial reaction. Reduced myelin basic protein, increased glial fibrillary acidic protein, and stable synaptophysin content were demonstrated by immunochemical methods. In conclusion, impairment in cognition and motor skills corresponds to ventricular enlargement and white matter destruction. Quantitative measures of weight, memory, ventricle size, and myelin, and glial proteins in this neonatal model of hydrocephalus will be useful tools for assessment of experimental therapeutic interventions.

Topics: Age Factors; Animals; Animals, Newborn; Antidiarrheals; Behavior, Animal; Blotting, Western; Body Weight; Brain Injuries; Cerebral Ventricles; Enzyme-Linked Immunosorbent Assay; Hydrocephalus; Immunohistochemistry; Kaolin; Ki-67 Antigen; Magnetic Resonance Imaging; Myelin Basic Protein; Nerve Tissue Proteins; Rats; Rats, Sprague-Dawley; Time Factors; Walking

In sera from normal rats and from rats injected with whole myelin in complete Freund adjuvant to induce EAE we study the presence of antibodies capable to inhibit the reactivity of autoantibodies directed to myelin basic protein (MBP).. Sera from rats that developed or not clinical signs of EAE were obtained previously to immunization, at acute stage of the disease and when the animals were completely recuperated, and chromatographied on a protein G-Sepharose column to obtain the retained (IgG) fractions. Then these fractions were depleted of anti-MBP reactivity by affinity chromatography and the ability of these depleted sera to block the reactivity of anti-MBP IgG antibodies was analyzed by an immunoblot technique.. IgG fractions from preimmune sera inhibited the anti-MBP IgG reactivity associated to EAE. The analysis of sick EAE animals showed that the inhibitory activity faded away with the onset of the clinical signs but returned at its maximum value during the spontaneous remission. Animals that never developed clinical EAE did not show changes in the level of inhibitory activity that was similar to that observed in the preimmune sera.. The presence of IgG antibodies blocking the anti-MBP IgG reactivity correlates with the development of the clinical signs of EAE.

Topics: Animals; Autoantibodies; Blotting, Western; Body Weight; Chromatography, Affinity; Dose-Response Relationship, Immunologic; Encephalomyelitis, Autoimmune, Experimental; Immunization, Passive; Immunoglobulin G; Immunoglobulin M; Male; Myelin Basic Protein; Rats; Rats, Wistar; Severity of Illness Index

Considering that sex steroids can influence the immune system, we studied the development of experimental autoimmune encephalomyelitis (EAE), a T-cell-mediated autoimmune disease of the central nervous system, and the concomitant cell-mediated immunity in gonadally intact and gonadectomized male Wistar rats given testosterone supplementation.. Sham-operated rats and surgically castrated animals were orally self-administered with vehicle or testosterone added in the water bottle for 20 days before EAE induction. The androgenic effect of oral testosterone self-administration was evidenced by changes in body weight, and in the weights of androgen-dependent testes and seminal vesicles. Testosterone administration reduced the incidence of clinical signs of EAE in sham-operated animals and reversed the clinical symptoms of the disease associated with castrated EAE animals. The clinical signs observed in the different groups correlated with changes in delayed-type hypersensitivity and mononuclear cell-proliferative responses to the encephalitogenic myelin basic protein. Moreover, testosterone but not cholesterol supplementation in vitro suppressed the proliferative response of mononuclear cells to myelin basic protein suggesting that testosterone may affect specific immune functions through direct actions on immune cells. Finally, self-administration of testosterone induced also elevated corticosterone levels that in sham-operated rats correlated with the low incidence of the disease and in gonadectomized animals could be involved in the remission of clinical symptoms of EAE.. These results suggest that orally self-administered testosterone can modulate specific cellular immune responses and serum corticosterone levels leading to changes in the development of EAE.

Topics: Administration, Oral; Animals; Body Weight; Cell Proliferation; Cholesterol; Corticosterone; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Immune Tolerance; Immunity, Cellular; Immunosuppressive Agents; Leukocytes, Mononuclear; Lymphocyte Activation; Male; Myelin Basic Protein; Orchiectomy; Organ Size; Rats; Rats, Wistar; Self Administration; T-Lymphocytes; Testis; Testosterone

One hemisphere of postnatal day 8 (P8) rats or P10 mice was irradiated with a single dose of 4-12 Gy, and animals were killed from 2 h to 8 weeks after irradiation (IR). In the subventricular zone (SVZ) and the granular cell layer (GCL) of the dentate gyrus, harboring neural and other progenitor cells, nitrosylation and p53 peaked 2-12 h after IR, followed by markers for active caspase-3, apoptosis-inducing factor and TUNEL (6-24 h). Ki67-positive (proliferating) cells had disappeared by 12 h and partly reappeared by 7 days post-IR. The SVZ and GCL areas decreased approximately 50% 7 days after IR. The development of white matter was hampered, resulting in 50-70% less myelin basic protein staining. Pretreatment with erythropoietin did not confer protection against IR. Caspase inhibition by overexpression of XIAP prevented caspase-9 and caspase-3 activation but not cell death, presumably because of increased caspase-independent cell death.

Topics: Active Transport, Cell Nucleus; Animals; Apoptosis; Body Weight; Brain; Caspase 3; Caspase 9; Caspase Inhibitors; Caspases; Cell Death; Cell Proliferation; DNA Fragmentation; Dose-Response Relationship, Radiation; Enzyme Activation; Enzyme Inhibitors; Erythropoietin; Hippocampus; Immunohistochemistry; In Situ Nick-End Labeling; Ki-67 Antigen; Mice; Mice, Inbred C57BL; Myelin Basic Protein; Proteins; Rats; Rats, Wistar; Stem Cells; Time Factors; Tumor Suppressor Protein p53; X-Linked Inhibitor of Apoptosis Protein

In the present work we show morphological data of the in vivo susceptibility of CNS myelin to sodium metavanadate [V(+5)] in adult rats. The possible role of vanadium in behavioral alterations and in brain lipid peroxidation was also investigated. Animals were injected intraperitoneally (i.p.) with 3 mg/kg body weight (bw) of sodium metavanadate [1.25 V/kg bw/day] for 5 consecutive days. Open field and rotarod tests were performed the day after the last dose had been administered and then animals were sacrificed by different methods for histological and lipid peroxidation studies. The present results show that intraperitoneal administration of V(+5) to adult rats resulted in changes in locomotor activity, specific myelin stainings and lipid peroxidation in some brain areas. They support the notion that CNS myelin could be a preferential target of V(+5)-mediated lipid peroxidation in adult rats. The mechanisms underlying this action could affect the myelin sheath leading to behavioral perturbations.

Topics: Animals; Behavior, Animal; Body Weight; Brain Chemistry; Central Nervous System; Gold; Immunohistochemistry; Lipid Peroxidation; Male; Motor Activity; Myelin Basic Protein; Myelin Sheath; Organ Size; Postural Balance; Rats; Vanadium

A1 adenosine receptors (A1ARs) are widely expressed in the brain during development. To examine whether A1AR activation can alter postnatal brain formation, neonatal rats from postnatal days 3 to 14 were treated with the A1AR agonist N6-cyclopentyladenosine (CPA) in the presence or absence of the peripheral A1AR antagonist 8-(p-sulfophenyl)-theophylline (8SPT). CPA or CPA + 8SPT treatment resulted in reductions in white matter volume, ventriculomegaly, and neuronal loss. Quantitative electron microscopy revealed reductions in total axon volume following A1AR agonist treatment. We also observed reduced expression of myelin basic protein in treated animals. Showing that functional A1ARs were present over the ranges of ages studies, high levels of specific [3H]CCPA binding were observed at PD 4, 7 and 14, and receptor-G protein coupling was present at each age. These observations show that activation of A1ARs with doses of CPA that mimic the effects of high adenosine levels results in damage to the developing brain.

Topics: Adenosine; Animals; Animals, Newborn; Body Weight; Cell Count; Cerebral Cortex; Cerebral Ventricles; Drug Combinations; Drug Interactions; GTP-Binding Proteins; Guanosine 5'-O-(3-Thiotriphosphate); Hippocampus; Microscopy, Electron; Myelin Basic Protein; Nerve Degeneration; Nerve Fibers, Myelinated; Neuroglia; Neurons; Presynaptic Terminals; Purinergic P1 Receptor Agonists; Purinergic P1 Receptor Antagonists; Rats; Rats, Sprague-Dawley; Receptors, Purinergic P1; Telencephalon; Theophylline

Globoid cell leukodystrophy is an autosomal recessive disease with progressive demyelination caused by a deficiency of the lysosomal enzyme galactosylceramidase. Bone marrow transplantation (BMT) is a therapeutic option for patients with late-onset disease and for patients with early onset disease that had an early diagnosis owing to an affected sibling. This therapy, however, typically is not effective for early onset disease when the diagnosis occurs after several months of life. In an effort to enable a broader range of patients to benefit from BMT, we tested whether combining substrate-reduction therapy with BMT would result in a greater benefit than either treatment alone in the twitcher mouse model of globoid cell leukodystrophy. Twitcher mice treated with L-cycloserine, an inhibitor of 3-ketodyhydrosphingosine synthase, and transplanted with 50 +/- 5 x 10(6) bone marrow cells on d 10 had a mean life-span of 112 d compared with 51 d for BMT alone (p < 0.001) or L-cycloserine alone, which was previously reported to be 56 d. L-Cycloserine treatment also was initiated neonatally to determine whether it would allow for a delayed BMT to have therapeutic value. Twitcher mice given only BMT at 18 d or only a short course of L-cycloserine died at 36 and 37 d, respectively. Twitcher mice given a short course of L-cycloserine + BMT at 18 d lived to 58 d (p = 0.0006). In conclusion, substrate-reduction therapy enhanced the value of BMT in twitcher mice, suggesting that this combination strategy might benefit patients with globoid cell leukodystrophy.

Topics: Animals; Body Weight; Bone Marrow Transplantation; Cycloserine; Glial Fibrillary Acidic Protein; Immunohistochemistry; Leukodystrophy, Globoid Cell; Mice; Mice, Inbred C57BL; Myelin Basic Protein; Rhombencephalon

Bone marrow transplantation (BMT) has therapeutic value for twitcher (globoid cell leukodystrophy) mice, which suffer from a genetic deficiency of the lysosomal enzyme galactosylceramidase that leads to progressive demyelination and early death. Preliminary investigations indicated that a semiallogeneic BMT resulted in graft vs. host disease (GVHD) in twitcher mice but not normal mice. Increased production of the cytokine IL-6 has been demonstrated in twitcher mice, and it has been linked with induction of GVHD. We investigated the effects of BMT in twitcher/IL-6 deficient mice and compared these findings with those from transplanted twitcher and control mice. After a semiallogeneic BMT, 11.4% of controls died within few weeks while the rest survived >100 days without GVHD. In contrast, 85% of the transplanted twitcher mice died by 70 days and 65% developed clinical signs of GVHD, e.g., alopecia and weight loss. In transplanted twitcher/IL-6 deficient mice, only 21% died by Day 70, none had alopecia, and 23% had weight loss. There was no difference in the onset day and severity of twitching between twitcher and twitcher/IL-6 deficient mice after BMT. In transplanted twitcher/IL-6 deficient mice, there was improvement of BBB integrity and a decrease in globoid cell number compared with nontransplanted twitcher/IL-6 deficient mice. In summary, these results demonstrate that an underlying pathology like globoid cell leukodystrophy leads to activation of GVHD responses in a donor-host combination that would not normally induce GVHD. Furthermore, IL-6 seems to play a key role because a deficiency of IL-6 results in a better prognosis.

Topics: Animals; Astrocytes; beta-Galactosidase; Blood-Brain Barrier; Body Weight; Bone Marrow Transplantation; Brain; Demyelinating Diseases; Female; Gliosis; Graft vs Host Disease; Immunohistochemistry; Interleukin-6; Lectins; Leukodystrophy, Globoid Cell; Macrophages; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Mice, Neurologic Mutants; Minor Histocompatibility Antigens; Myelin Basic Protein; Serum Albumin; Survival Rate

Polychlorinated biphenyls (PCBs) are a class of industrial compounds consisting of paired phenyl rings with various degrees of chlorination. They are now ubiquitous, persistent environmental contaminants that are routinely found in samples of human and animal tissues and are known to affect brain development. The effects of PCBs on brain development may be attributable, at least in part, to their ability to reduce circulating levels of thyroid hormone. However, the developmental effects of PCB exposure are not fully consistent with hypothyroidism. Because some individual PCB congeners interact strongly with various thyroid hormone binding proteins, several investigators have speculated that these congeners may be producing thyroid hormone-like effects on brain development. Therefore, we tested whether a mixture of PCBs, Aroclor 1254 (A1254), would produce an antithyroid or thyromimetic effect on the expression of known thyroid hormone-responsive genes in the developing brain. Pregnant female rats were fed various doses of A1254 (0, 1, 4, and 8 mg/kg) from gestational day 6 to weaning on postnatal day (P) 21. Pups derived from these dams were sampled on P5, P15, and P30. Total T4 was reduced by A1254 in a dose-dependent manner, but body weight of the pups or dams was not affected. The expression of RC3/Neurogranin and myelin basic protein was not affected by A1254 on P5 or P30. However, on P15, RC3/Neurogranin was elevated by A1254 in a dose-dependent manner, and myelin basic protein expression followed this general pattern. These data clearly demonstrate that the developmental effects of PCB exposure are not simply a function of PCB-induced hypothyroidism.

Topics: Animals; Autoradiography; Body Weight; Brain; Calmodulin-Binding Proteins; Female; Gene Expression Regulation; In Situ Hybridization; Myelin Basic Protein; Nerve Tissue Proteins; Neurogranin; Polychlorinated Biphenyls; Pregnancy; Radioimmunoassay; Rats; Rats, Sprague-Dawley; Receptors, Thyroid Hormone; RNA Probes; RNA, Messenger; Signal Transduction; Thyroid Hormones

We tested the hypotheses that an episode of hypoxemia near mid-gestation in fetal sheep has long-term effects on brain development and that the extent and type of damage is related to the stage of development within a particular brain structure at the time of the hypoxemia. Fetal sheep (n = 8) were made hypoxemic at 90 +/- 2 days (term approximately 147 days) by restricting the maternal blood supply to the placenta for 12 hours (h) using a vascular clamp so as to reduce fetal arterial O2 saturation by 50%-60%. Fetuses were killed 35 days later and the brains analysed histologically and immunohistochemically. Age-matched fetuses (n = 8) were used as controls. Gross brain damage was observed in only 1 fetus, the most acidemic during the period of hypoxemia. There was a reduction of 12% (p < 0.05) in the cross-sectional area of the cerebral cortex in hypoxemic fetuses compared with controls. In lobule 6 of the cerebella of hypoxemic fetuses, significant reductions were seen in (a) the volume density of Purkinje cells (33%), (b) the width of the molecular layer (13%), (c) the area of the inner granule cell layer (13%), (d) the area of the white matter (18%), and (e) the total cross-sectional area (15%). There were also significant reductions in the area of arborization of Purkinje cell dendritic trees (50%), in the branching density (25%), and in the number of dendritic spines (31%). In the ventral hippocampi of hypoxemic fetuses, there was a 36% reduction (p < 0.05) in the volume density of CA1 pyramidal cells and a 50% increase (p < 0.05%) in the number of astrocytes. We conclude that an episode of hypoxemia near mid-gestation reduces neuronal numbers in the hippocampus and cerebellum and probably also in the cerebral cortex. The growth of neural processes in a particular region will be significantly retarded if the hypoxemia occurs at an early stage of the growth of neural processes (e.g. cerebellum) but not if development is well advanced at the time of the insult (e.g. hippocampus). Damage is sustained in the white matter of the cerebral hemispheres if the insult is particularly severe. Together, these deficits could affect neural connectivity and impair postnatal brain function.

Topics: Animals; Blood Pressure; Body Weight; Brain; Embryonic and Fetal Development; Female; Fetus; Gases; Gestational Age; Glial Fibrillary Acidic Protein; Hypoxia; Immunohistochemistry; Microtubule-Associated Proteins; Myelin Basic Protein; Organ Size; Pregnancy; Pregnancy Complications; Pregnancy, Animal; Sheep; Time Factors

The postnatal development of the central nervous system (CNS) in house musk shrew in the early stage of maturation was studied. The electroencephalogram (EEG) and visual evoked potential (VEP) in association with catecholamine contents and myelin basic protein (MBP) immunoreactivity were carried out from the 1st to the 20th day of postnatal age. Different EEG patterns which were specific to behavioral states (awake and drowsy) were first recorded on the 5th day, and the total power which was obtained by power spectrum analysis increased after this stage. The latencies of all peaks in VEP markedly shortened between the 5th and the 7th day. Noradrenalin (NA) content of the brain showed a slight increase after the 3rd day, and reached maximum levels on the 7th day, which was delayed a few days compared to dopamine (DA). In hyperthyroidism, the peak latency of VEP was shortened and biosynthesis of NA in cerebral cortex and DA in hippocampus was accelerated. The most obvious change in MBP-immunoreactivity of the telencephalon occurred from the 7th to the 10th day. These morphological changes in the brain advanced at the identical time-course to those in the electrophysiological development and increment of DA and NA contents.

Topics: Aging; Animals; Behavior, Animal; Body Weight; Brain; Brain Chemistry; Catecholamines; Electrodes, Implanted; Electroencephalography; Evoked Potentials, Visual; Female; Hyperthyroidism; Immunohistochemistry; Male; Myelin Basic Protein; Myelin Sheath; Organ Size; Shrews

Selenium deficiency causes further impairment of thyroid hormone metabolism in iodine-deficient rats and therefore could have a role in the aetiology of both myxoedematous and neurological cretinism in humans. Thyroidal type I iodothyronine deiodinase (ID-I), cytosolic glutathione peroxidase and phospholipid hydroperoxide glutathione peroxidase activities were increased in iodine-deficient adult rats and their offspring at 11 days of age. Thyroidal ID-I activity was unchanged and thyroidal cytosolic glutathione peroxidase activity was decreased by more than 75% by combined selenium and iodine deficiency in 11-day-old rats, indicating that, while the thyroid retained an ability to produce 3,3',5-triiodothyronine (T3), the gland was probably more susceptible to peroxidative damage caused by increased hydrogen peroxide concentrations driven by increased thyrotrophin. Thyroidal atrophy, common in myxoedematous cretinism, did not occur in iodine- or selenium and iodine-deficient rat pups. Iodine deficiency increased brain type II iodothyronine deiodinase activity 1.5-fold in 4-day-old rats and 3-fold in 11-day-old rats, regardless of selenium status. Thus rats were able to activate compensatory mechanisms in brain that would maintain T3 concentrations in selenium and iodine deficiencies. Surprisingly, however, selenium deficiency had a greater effect than iodine deficiency on markers of brain development in rat pups. Expression of the brain-derived neurotrophic factor (BDNF) mRNA was decreased in selenium deficiency in 4- and 11-day-old pups and in combined selenium and iodine deficiency in 4-day-old pups. Iodine deficiency caused an increase in BDNF expression in 11-day-old pups but had no effect on 4-day-old pups. Myelin basic protein mRNA expression in brain was decreased by combined selenium and iodine deficiency in 11-day-old rats.

Topics: Animals; Animals, Suckling; Body Weight; Brain; Brain-Derived Neurotrophic Factor; Deficiency Diseases; Female; Glutathione Peroxidase; Iodine; Liver; Myelin Basic Protein; Organ Size; Proteins; Rats; RNA, Messenger; Selenium; Selenoproteins; Thyroid Gland; Thyroid Hormones

Chronic relapsing experimental autoimmune encephalomyelitis (crEAE), a model for multiple sclerosis, was used to test 2 regimens of insulin-like growth factor-I (IGF-I) treatment. We induced crEAE by injecting 3x10(7) myelin basic protein-(MBP) sensitized lymph node cells into adult female SJL/J mice. Fifty-one mice, divided randomly into 4 groups, were used in the first trial. Two groups received IGF-I (a gift of Cephalon, Inc.) 0.6 mg/kg/d subcutaneously from day 7 to day 16 and the other two groups received placebo injections. IGF-I treatment reduced clinical deficits during the first attack and during 2 subsequent relapses. Image analysis of immunostained and histological sections showed that IGF-I treatment reduced BBB defects and both the numbers and sizes of inflammatory, demyelinating, and demyelinated lesions. Twelve mice that had recovered from their first attack were used in our second trial to evaluate possible adverse effects of prolonged treatment with a higher dose of IGF-I. Six received 1.2 mg/kg/d for 6 weeks (days 19-63). No adverse effects of IGF-I treatment were identified. The eyes, hearts, livers, and kidneys of IGF-I-treated mice were normal histologically and their spleens also appeared normal except for mild to moderate microscopic increases in lymphopoesis. Our results suggest that prolonged IGF-I treatment is well tolerated and that the anti-inflammatory effects of IGF-I have a major role in reducing clinical deficits and lesion severity in crEAE. These effects, if present in multiple sclerosis, may benefit patients with this disease.

Topics: Animals; Blood-Brain Barrier; Body Weight; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Female; Immunohistochemistry; In Situ Hybridization; Insulin-Like Growth Factor I; Macrophages; Magnetic Resonance Imaging; Mice; Mice, Inbred Strains; Myelin Basic Protein; Myelin Sheath; Nerve Regeneration; Neuroglia; Recurrence; RNA Probes

The mechanism of action underlying the beneficial effect of IFN-beta in multiple sclerosis (MS) is not understood. To date, little information is available on the effects of IFN-beta in experimental autoimmune encephalomyelitis (EAE), the animal correlate of the human disease MS. Therefore, we investigated the effects of recombinant rat IFN-beta (rrIFN-beta) on EAE in Lewis rats with emphasis on a treatment regimen during the paralytic phase of the disease. The results indicated that rrIFN-beta dose-dependently inhibited disease activity with complete prevention at a s.c. dose of 300,000 U/day, provided that treatment was continued for 3 wk. Discontinuation of treatment on day 17 postimmunization resulted in a protracted and relapsing disease course with strongly enhanced clinical severity. Detailed immunohistology of central nervous system (CNS) tissue of protected animals revealed an almost complete absence of CNS lesions and a >90% reduction in the number of infiltrating leukocytes. Accordingly, isolation of mononuclear cells from spinal cord tissue of successfully treated EAE rats revealed a reduction of approximately 95% in the number of cells that produce IFN-gamma in response to the encephalitogenic peptide MBP63-88. Furthermore, rrIFN-beta significantly enhanced serum corticosterone levels, which showed an inverse relationship with disease activity. We show that rrIFN-beta can have both beneficial and detrimental effects on disease activity dependent on the timing and the duration of treatment. Beneficial effects on EAE are associated with inhibition of the extravasation of blood-derived mononuclear cells in the CNS.

Topics: Animals; Autoantibodies; Body Weight; Corticosterone; Dose-Response Relationship, Drug; Drug Administration Schedule; Encephalomyelitis, Autoimmune, Experimental; Interferon-beta; Interferon-gamma; Lymphoid Tissue; Myelin Basic Protein; Rats; Rats, Inbred Lew; Spinal Cord; Time Factors

The present experiment deals with the effect of maternal deprivation (MD) and early weaning (EW) on the development and course of experimental allergic encephalomyelitis (EAE) in Dark August (DA) rats. Five litters (five to nine pups per liter) were subjected to MD (4 h daily) from Day 1 until Day 28. EW rats were weaned on Day 15 (EW-15, five litters) or Day 21 (EW-21, four litters). Control rats and MD rats were weaned on Day 28. At the age of 8 weeks, rats were immunized with guinea pig spinal cord in complete Freund's adjuvant and clinical signs of EAE were recorded daily. On Day 18 after immunization, rats were bled and sacrificed. Brain and spinal cord were examined histologically for EAE lesions. Serum anti-rat myelin basic protein (MBP) antibodies were detected by ELISA. MD female rats exhibited suppression of neurological and histological signs of EAE in comparison with control rats. MD and control females showed elevated anti-MBP antibody level compared to MD and control males. EW-15 female rats demonstrated potentiation of neurological signs of EAE compared to control females. EW-21 females developed more severe clinical signs and histological lesions compared to control females. These results show that neonatal experiences, such as maternal deprivation and early weaning, influence the development of EAE in adult DA rats.

Topics: Age Factors; Animals; Autoimmune Diseases; Body Weight; Brain; Encephalomyelitis, Autoimmune, Experimental; Female; Guinea Pigs; Immunity, Maternally-Acquired; Male; Maternal Deprivation; Myelin Basic Protein; Psychoneuroimmunology; Rats; Sex Factors; Spinal Cord; Stress, Psychological; Weaning

A Northern blot analysis of P0 and MBP myelin protein transcripts in the sciatic nerve from rats with alloxan-induced diabetes at two different time points is described. After 5 weeks of diabetes induction, only P0 mRNA is significantly increased by 39%, while at 14 weeks both P0 and MBP mRNA contents are markedly higher than controls. Insulin treatment normalizes glycemia levels, partially counteracts P0 mRNA increase at both stages of diabetes and delays MBP mRNA increase present only in chronic animals. We suggest that increased transcript levels of P0 and MBP in Schwann cells may represent a higher turnover of myelin sheath specific proteins in diabetic syndrome, as attempt to repair the hyperglycemia-induced nerve damage, which is partially prevented by insulin treatment.

Topics: Animals; Blood Glucose; Blotting, Northern; Body Weight; Densitometry; Diabetes Mellitus, Experimental; DNA, Complementary; Image Processing, Computer-Assisted; Insulin; Male; Myelin Basic Protein; Myelin Proteins; Rats; Rats, Sprague-Dawley; RNA, Messenger; Sciatic Nerve; Transcription, Genetic; Up-Regulation

The expression of myelin-specific protein genes, i.e. myelin proteolipid (PLP), basic (BP), and myelin associated glycoproteins (MAG) was studied in normal and severely undernourished 20-day-old rats. The undernutrition paradigm resulted in reductions of approximately 50, 25 and 65% in body weight, brain weight and brain myelin yield, respectively. The amount of total brain RNA was not significantly altered, although the amount of cyclophilin (CYC) mRNA was increased. In contrast, the steady-state levels of myelin specific mRNAs were significantly decreased by approximately 40, 20 and 40% for PLP, BP and MAG, respectively. In addition, polyadenylation of the PLP transcript was altered, producing an abnormal ratio of the 1.6 kb to the 3.2 kb PLP mRNAs. The results indicate that down-regulation of myelin-specific gene expression is involved in the mechanisms of hypomyelination in hunger disease, although the individual genes are differently altered. Furthermore, undernutrition may have additional effects on the posttranscriptional processing of the transcripts as indicated by the abnormal size distribution of PLP messages.

Topics: Animals; Blotting, Northern; Body Weight; Brain; Demyelinating Diseases; DNA Probes; Down-Regulation; Female; Myelin Basic Protein; Myelin Proteins; Myelin Proteolipid Protein; Myelin Sheath; Myelin-Associated Glycoprotein; Nucleic Acid Hybridization; Nutrition Disorders; Organ Size; Pregnancy; Rats; RNA, Messenger

The in vitro potency of the immunosuppressants Cyclosporin A (CsA), FK-506 and Prednisolone was assessed using the adoptive transfer model of EAE in the Lewis rat. Co-culture of encephalitogen-sensitised splenic leukocytes with Prednisolone did not inhibit the transfer of disease to naive histocompatible recipients despite significant suppression of neuroantigen-stimulated leukocyte proliferation by the drug. The addition of CsA (100 nM) to cultures inhibited the induction of adoptive EAE but a lower dose of the agent (10 nM) did not prevent the development of clinico-histopathological signs of disease. FK-506 (1 nM) was 100 times more effective than CsA at suppressing adoptive EAE thus emphasising the usefulness of the model in determining the relative efficacy of compounds to modify cell-dependent autoimmune disease.

Topics: Animals; Body Weight; Cell Division; Cells, Cultured; Cyclosporine; Encephalomyelitis, Autoimmune, Experimental; Immunization, Passive; Leukocyte Transfusion; Leukocytes; Male; Myelin Basic Protein; Prednisolone; Rats; Rats, Inbred Lew; Spleen; Tacrolimus

Effects of dexamethasone (DEX) on the relative abundance of myelin basic protein (MBP), proteolipid protein (PLP) and glial fibrillary acidic protein (GFAP) mRNAs in the developing rat brain were examined. After DEX (1.0 mg/kg body weight) or saline was administered intraperitoneally to 3-day-old rats for 7 consecutive days, wet weight, DNA content and the relative abundance of the glia-specific mRNAs in cerebrum and cerebellum were analyzed at postnatal days (P) 10, 20 and 30. DEX decreased both wet weight and DNA content in cerebellum more profoundly than in cerebrum. The appearance of MBP, PLP and GFAP mRNAs in cerebellum preceded that in cerebrum in the control group. In cerebrum, the relative abundance of MBP and PLP mRNAs was significantly less in the DEX group than that in the control group at P20 and P30. The relative abundance of the GFAP mRNA was significantly less in the DEX group than in the control group at P10 and P20, but there was no significant difference at P30. In cerebellum, a significant decrease in the abundance of MBP, PLP and GFAP mRNAs in the DEX group was observed only at P10 but not at P20 and P30. Our findings indicate that DEX suppresses expression of genes related to glial functions, especially myelination when administered in the early postnatal period.

Topics: Aging; Animals; Body Weight; Brain; Cerebellum; Dexamethasone; DNA; DNA Probes; Gene Expression; Glial Fibrillary Acidic Protein; Myelin Basic Protein; Myelin Proteins; Myelin Proteolipid Protein; Organ Size; Rats; Rats, Inbred Strains; RNA, Messenger

Selective depletion of central nervous system norepinephrine (NE) by the neurotoxin 6-hydroxydopamine (6-OHDA) in rats subsequently inoculated with myelin basic protein (MBP) and complete Freund's adjuvant (CFA) produced experimental autoimmune encephalomyelitis (EAE) without the usual expected degree of weakness. The preservation of strength occurred in spite of continued weight loss. Post-decapitation myoclonic convulsive kick latency and kick number, which are known to depend on spinal cord NE, agreed well with the degree of weakness through the clinical disease course. The only difference between EAE groups was that the stronger 6-OHDA pretreated EAE animals did not have an elevated pons-medulla NE compared to saline intracisternal-ventricular (i.c.v.) pretreated controls. We conclude that 6-OHDA can influence the clinical course of weakness by interfering with central noradrenergic activity independent of other features associated with disease in EAE. This effect of 6-OHDA may be exerted through alteration of the blood-spinal cord barrier function and/or central nervous system blood flow.

Topics: Animals; Body Weight; Brain; Catecholamines; Cerebral Ventricles; Cisterna Magna; Decerebrate State; Encephalomyelitis, Autoimmune, Experimental; Freund's Adjuvant; Hydroxydopamines; Injections, Intraventricular; Male; Myelin Basic Protein; Norepinephrine; Oxidopamine; Rats; Rats, Inbred Lew; Seizures

Postnatal exposure of rats to lead has been shown previously to cause CNS hypo-myelination. Since rats intoxicated with lead often show retarded growth, the superimposed malnutrition, which as such can cause hypomyelination, may contribute to myelin deficit. In the present study control rats and lead exposed rats which did not have any retardation of growth were examined by radioimmunological assay of myelin basic protein (MBP) of homogenates of cerebrum and cerebellum at 30, 60 and 120 days of age. Lead was administered on postnatal days 1-15 by daily intraperitoneal injections of 10 mg lead nitrate/kg body weight. This lead dose results in light microscopically discernible hemorrhagic encephalopathy in the cerebellum of 15-day old rats, but does not induce growth retardation (Sundström et al. 1983). The controls were injected with vehicle only. The amount of lead in the blood and brain homogenates of lead-exposed and control rats 15-200 days old was estimated by atomic absorption spectrophotometry. Significant differences between the lead-exposed and control rats were not found in the cerebral or cerebellar content of MBP. Considering the results of previous investigations, the findings do not exclude a hypo-myelinating effect of lead, but they suggest that exposure to lead without concomitant malnutrition does not cause hypo-myelination in the cerebrum and cerebellum of the developing rat.

Topics: Aging; Animals; Body Weight; Brain; Brain Chemistry; Electrophoresis, Polyacrylamide Gel; Immunosorbent Techniques; Lead; Lead Poisoning; Male; Myelin Basic Protein; Organ Size; Radioimmunoassay; Rats; Rats, Inbred Strains; Time Factors

Guinea pigs inoculated with a mixture of myelin basic protein and the myelin-specific ganglioside, sialosylgalactosylceramide (GM4), do not develop the signs or neuropathology of experimental allergic encephalomyelitis. The results indicate that GM4 cloaks the basic protein molecule so that it is no longer immunopathogenic in these animals. The interaction of basic protein with GM4, previously shown in vitro, appears to be relevant to the pathogenesis of experimental allergic encephalomyelitis in guinea pigs.

Topics: Animals; Body Weight; Brain; Encephalomyelitis, Autoimmune, Experimental; Gangliosides; Guinea Pigs; Myelin Basic Protein; Spinal Cord

Intravenous administration of mouse myelin basic protein covalently coupled with chromic chloride to syngeneic spleen cells (MBP-SC) prevents the subsequent induction of experimental allergic encephalitis (EAE). Whereas 1 in 28 mice receiving MBP-SC developed EAE after immunization with mouse spinal cord homogenate (MSCH) and adjuvants, 16 out of 25 mice receiving ovalbumin-coupled spleen cells (OA-SC) had EAE following encephalitogenic challenge. The effect of administration of antigen-coupled spleen cells on in vitro proliferation responses is shown.

Topics: Animals; Antigens; Body Weight; Encephalomyelitis, Autoimmune, Experimental; Female; Lymphocyte Activation; Mice; Mice, Inbred Strains; Myelin Basic Protein; Ovalbumin; Spleen

Lewis rats were maintained on diets which provided either an adequate or deficient concentration of biotin. Biotin deficiency produced a marked reduction in thymus size and cellularity, a depressed immune response to sheep erythrocytes and prevented the development of experimental allergic encephalomyelitis following immunization with guinea pig myelin basic protein. Total T cells, T-helper and T-suppressor cells were quantitatively the same in the spleens of rats fed biotin-adequate or deficient diets. By using an adoptive lymphocyte transfer procedure, it was determined that the afferent immune response to myelin basic protein was impaired in biotin deficiency. These results indicate that experimentally induced autoimmune disease is susceptible to the nutritional influence of dietary biotin.

Topics: Animals; Autoantibodies; Autoimmune Diseases; Biotin; Body Weight; Encephalomyelitis, Autoimmune, Experimental; Erythrocytes; Female; Immunization; Lymphocytes; Myelin Basic Protein; Organ Size; Rats; Rats, Inbred Lew; Sheep

The effects of maternal treatment with ritodrine, a beta 2-adrenergic agonist, on the biochemical development of fetal brain were studied in an animal model. Pregnant rats were treated with long and short dosage schedules. Fetuses were delivered by hysterotomy 4 h after the last dose. No differences in the fetal brain content of protein, DNA, glycogen, cholesterol or beta-adrenergic receptors were found. In this animal model, using relatively high maternal doses of ritodrine, there were no apparent effects on the fetal brain biochemical indices measured, suggesting a relatively high efficacy to toxicity ratio of ritodrine for brain development.

Topics: Animals; Body Weight; Brain; Brain Chemistry; Cholesterol; DNA; Female; Fetus; Glycogen; Myelin Basic Protein; Nerve Tissue Proteins; Organ Size; Pregnancy; Propanolamines; Rats; Receptors, Adrenergic, beta; Ritodrine

Topics: Animals; Body Weight; Disease Susceptibility; Female; Hypersensitivity; Immunity, Cellular; Immunization; Lymphocyte Activation; Male; Myelin Basic Protein; Myelin P2 Protein; Neuritis; Rats

Experimental allergic encephalomyelitis (EAE) was induced in guinea pigs with bovine myelin basic protein (BP) with adjuvant of either synthetic muramyl dipeptide (Mdp) or Mycobacterium tuberculosis (Tbc). The following results were obtained: (1) The body temperature of the animals was studied serially after sensitization and its elevation was shown to be an early sign of EAE. (2) Several animals developed the clinical and histological signs of hyperacute EAE. (3) An optimal combined dosage of BP and adjuvant was found for induction of clinical EAE and for the the production of complement fixing (CF) antibodies. (4) Little passive hemagglutinating (PH) antibody was produced by single immunization. These results displayed no essential difference in EAE induced by either adjuvant. (5) Detectable PH antibodies developed later in addition to CF antibodies in a few animals immunized with Tbc adjuvant. These animals were skin-tested to BP, and had recovered from body weight loss or limb weakness. The results suggest that humoral antibodies play a role in modifying the disease process, even if they are not essential in production of EAE.

Topics: Acetylmuramyl-Alanyl-Isoglutamine; Animals; Antibodies; Antibody Formation; Body Temperature; Body Weight; Encephalomyelitis, Autoimmune, Experimental; Female; Freund's Adjuvant; Glycopeptides; Guinea Pigs; Mycobacterium tuberculosis; Myelin Basic Protein; Skin

It was confirmed that experimental autoimmune encephalomyelitis EAE, could be induced in SJL/J mice with mouse spinal cord homogenate. It was shown that induction of EAE in mice was critically dependent on the concentration of pertussis vaccine. The encephalitogen present in mouse brain was the basic protein of myelin. The smaller form of the mouse and rat basic proteins induced EAE; thus the mouse like the rat responds to determinants other than the "tryptophan region," which induced EAE in guinea-pigs. Mice with EAE developed a cell-mediated immune response to myelin basic protein, as judged by inhibition of peritoneal cell migration. However, levels of antibody to mouse basic protein were low, as judged by radioimmunoassay. The establishment of this autoimmune disease model in the mouse will allow the application of well established techniques for the analysis of the immunologic mechanisms leading to disease manifestation.

Topics: Animals; Antibodies; Ascitic Fluid; Body Weight; Brain; Cell Migration Inhibition; Chemical Fractionation; Electrophoresis; Encephalomyelitis, Autoimmune, Experimental; Female; Immunity, Cellular; Immunization; Iodine Radioisotopes; Macrophages; Male; Mice; Mycobacterium; Myelin Basic Protein; Myelin Sheath; Peptides; Pertussis Vaccine; Radioimmunoassay; Rats; Spinal Cord; Tissue Extracts

The resistance of Strain 2 guinea pigs to experimental allergic encephalomyelitis (EAE) induced by inoculation with whole CNS tissue in complete Freund's adjuvant (CFA) has been confirmed. The resistance is even more pronounced when myelin basic protein (BP) is used in attempts to induce EAE. Strain 2 guinea pigs are also resistant to an immunization schedule (multiple injections with BP in IFA followed by a single injection of BP in CFA) known to induce significant levels of antibody in susceptible strains. The poor response of Strain 2 guinea pigs to BP is not the result of lack of specific B cells--antibody equivalent to that produced by Strain 13 animals is obtained when the inoculum contains 0.5 mg BP and 2.5 mycobacteria.

Topics: Amino Acid Sequence; Animals; Antibody Formation; Antigen-Antibody Complex; B-Lymphocytes; Binding Sites, Antibody; Body Weight; Dose-Response Relationship, Drug; Encephalomyelitis, Autoimmune, Experimental; Freund's Adjuvant; Guinea Pigs; Hypersensitivity, Delayed; Immunization Schedule; Male; Mycobacterium; Myelin Basic Protein; Skin Tests; T-Lymphocytes

The encephalitogenic activity of different parts of bovine encephalitogenic protein (BEP) has been tested in guinea-pigs, mainly at a dose equimolar to 32 mug of intact protein. A high activity was found with complete protein and with region 89-169; both peptide 43-115, tyr (modified at the tyrosine residue 67) and HNB-89-169 (modified at the tryptophan residue 115) showed a moderate but clear-cut activity, whereas that of peptide 1-42 was very faint. No qualitative differences in the disease induced by the different peptides could be found; in only one animal out of 139 were clinical signs observed without evidence of histological disease. Slight histological changes, similar to those seen at EAE, were found in four out of 19 animals injected with Freund's complete adjuvant only.

Topics: Animals; Antigens; Body Weight; Cattle; Encephalomyelitis, Autoimmune, Experimental; Guinea Pigs; Myelin Basic Protein; Peptides; Time Factors

The condition of experimental allergic encephalomyelitis (EAE) induction was investigated in several mice strains. SJL and C3H/He strains were found to be susceptible. A single immunization with mouse spinal cord, complete Freund's adjuvant (CFA) and pertussis vaccine produced clinical signs of EAE in SJL and C3H/He strains after 11 to 18 days. Isogenic spinal cord produced EAE in C3H/He strain. A single immunization with myelin basic protein from bovine spinal cord in CFA and pertussis vaccine produced EAE in SJL strain. EAE susceptibility of SJL strain correlated with the amount of mycobacteria used for sensitization. It was necessary to give pertussis vaccine intravenously in all cases.

Topics: Animals; Antigens; Body Weight; Brain; Encephalomyelitis, Autoimmune, Experimental; Female; Male; Mice; Mice, Inbred C3H; Myelin Basic Protein; Pertussis Vaccine; Spinal Cord

Topics: Animals; Animals, Newborn; Body Weight; Brain; Diet Therapy; Dietary Proteins; Encephalomyelitis, Autoimmune, Experimental; Female; Myelin Basic Protein; Myelin Sheath; Nerve Tissue Proteins; Organ Size; Pregnancy; Protein Deficiency; Rats; Rats, Inbred Strains; Spinal Cord

Topics: Animals; Antigen-Antibody Reactions; Antigens, Bacterial; Body Weight; Bordetella pertussis; Cross Reactions; Encephalomyelitis, Autoimmune, Experimental; Freund's Adjuvant; Histamine Release; Hot Temperature; Immunoglobulin E; Male; Mycobacterium; Myelin Basic Protein; Passive Cutaneous Anaphylaxis; Proteins; Rats; Rats, Inbred Strains; Reagins; Skin; Spinal Cord; Time Factors

Topics: Adrenal Glands; Animals; Body Weight; Bordetella pertussis; Cyproheptadine; Drug Therapy, Combination; Encephalomyelitis, Autoimmune, Experimental; Guinea Pigs; Histamine; Histamine H1 Antagonists; Histamine Release; Immunoglobulin E; Male; Methysergide; Mycobacterium; Myelin Basic Protein; Organ Size; Passive Cutaneous Anaphylaxis; Reagins; Serotonin; Serotonin Antagonists; Skin; Skin Tests; Spinal Cord; Stress, Physiological; Thymus Gland

Topics: Animals; Antigens; Ascitic Fluid; Body Weight; Cattle; Cell Migration Inhibition; Cells, Cultured; Encephalomyelitis, Autoimmune, Experimental; Guinea Pigs; Hypersensitivity, Delayed; Immunity, Cellular; Immunodiffusion; Iodine Radioisotopes; Lymphocyte Activation; Macrophages; Male; Myelin Basic Protein; Skin Tests