myelin-basic-protein and Optic-Nerve-Diseases

This study evaluated the function of mouse optic nerves after transient retinal ischemia using in vitro electrophysiologic recordings of compound action potentials (CAPs) correlated with diffusion tensor imaging (DTI) injury markers with confirmation by immunohistochemistry-determined pathology.. Retinal ischemia was induced in 7- to 8-week-old female C57BL/6 mice by elevating intraocular pressure to 110 mm Hg for 60 minutes. At 3 and 7 days after retinal ischemia, optic nerves were removed for CAP measurements. The CAP amplitude was recorded using suction electrodes in isolated control and injured optic nerves followed by ex vivo DTI evaluation. After DTI, optic nerves were embedded in paraffin and cut for immunohistochemical analyses.. Consistent with previous in vivo DTI measurements, a 25% decrease in axial diffusivity with normal radial diffusivity was seen at 3 days after retinal ischemia, suggesting axonal injury without myelin damage. At 7 days, there was no additional change in axial diffusivity compared with that at 3 days, but radial diffusivity significantly increased by 50%, suggestive of significant myelin damage due to sustained axonal injury. The relative anisotropy (RA) progressively decreased after retinal ischemia when compared with that of the controls. The CAP amplitude in injured nerves also progressively decreased after retinal ischemia, which correlated with the reduced RA (r = 0.80).. This study suggests that CAP amplitude reflects both axonal and myelin integrity and RA is an optimal parameter for functional assessment compared with axial or radial diffusivity alone in murine optic nerves after retinal ischemia.

Topics: Action Potentials; Animals; Axons; Diffusion Tensor Imaging; Disease Models, Animal; Electrophysiology; Female; Mice; Mice, Inbred C57BL; Myelin Basic Protein; Myelin Sheath; Nerve Degeneration; Optic Nerve Diseases; Reperfusion Injury; Retinal Diseases; Retinal Ganglion Cells

To examine chronic changes occurring at 6 months following partial optic nerve (ON) transection, assessing optic axons, myelin, and visual function.. Dorsal ON axons were transected, leaving ventral optic axons vulnerable to secondary degeneration. At 3 and 6 months following partial transection, toluidine-blue stained sections were used to assess dimensions of the ON injury site. Transmission electron microscopy (TEM) images of ventral ON were used to quantify numbers, diameter, area, and myelin thickness of optic axons. Immunohistochemistry and fluoromyelin staining were used to assess semiquantitatively myelin protein, lipids in ventral ON, and retinal ganglion cells (RGCs) in midventral retina. Visuomotor function was assessed using optokinetic nystagmus.. Following partial ON transection, optic axons and function remained disrupted at 6 months. Although ventral ON swelling observed at 3 months (P ≤ 0.05) receded to normal by 6 months, ultrastructurally, myelinated axons remained swollen (P ≥ 0.05), and myelin thickness increased (P ≤ 0.05) due to loosening of lamellae and an increase in the number of intraperiodic lines. Axons with decompacted myelin persisted and were distinguished as having large axonal calibers and thicker myelin sheaths. Nevertheless, progressive loss of myelin lipid staining with fluoromyelin was seen at 6 months. Despite no further loss of ventral optic axons between 3 and 6 months (P ≥ 0.05), visuomotor function progressively declined at 6 months following partial transection (P ≤ 0.05).. Continued decompaction of myelin, altered myelin structure, and swelling of myelinated axons are persistent features of the chronic phases of secondary degeneration and likely contribute to progressive loss of visual function.

Topics: Animals; Axons; Chronic Disease; Female; Fluorescent Antibody Technique, Indirect; Microscopy, Electron, Transmission; Myelin Basic Protein; Myelin Sheath; Nerve Degeneration; Nystagmus, Optokinetic; Optic Nerve; Optic Nerve Diseases; Optic Nerve Injuries; Rats; Retinal Ganglion Cells; Tubulin

Diffusion tensor imaging (DTI) measures the random motion of water molecules reflecting central nervous system tissue integrity and pathology. Glaucoma damages retinal ganglion cells (RGCs) and their axons. The authors hypothesized that DTI-derived axonal and myelin injury biomarkers may be used to detect early axonal damage and may be correlated with RGC loss in the mouse model of optic nerve crush (ONC).. The progression of RGC axon degeneration was quantitatively assessed with DTI in vivo, corroborated with axon/myelin immunohistochemical staining and retrograde fluorogold labeling in mice after ONC.. Decreased axial diffusivity (λ(‖)) and relative anisotropy (RA) of damaged axons were observed from 6 hours to 14 days, reflecting axonal injury. DTI detected axonal injury at 6 hours after ONC when SMI-31 did not detect axonal abnormality. Both decreased λ(‖), and SMI-31 identified axon damage at 3 days after ONC. Decreased λ(‖) correlated with reduced SMI-31-positive axon counts from 3 days after ONC. In contrast, the increased λ(⊥) was seen only in the distal segment of optic nerve whereas decreased myelin basic protein-positive axon counts were seen in all segments 3 days after ONC. The number of retrograde-labeled RGCs did not decline significantly until 7 days after ONC. There was a significant correlation between RGC loss and optic nerve axon damage.. The authors demonstrated that in vivo DTI detected axonal injury earlier than SMI-31. Results suggest that in vivo DTI of optic nerve injury may be used as a noninvasive tool for assessing the pathogenesis of RGC axonal injury.

Topics: Animals; Axons; Biomarkers; Cell Count; Diffusion Tensor Imaging; Disease Models, Animal; Female; Mice; Mice, Inbred C57BL; Myelin Basic Protein; Nerve Crush; Nerve Degeneration; Neurofilament Proteins; Optic Nerve Diseases; Retinal Ganglion Cells; Stilbamidines

The neurotrophin nerve growth factor (NGF) is produced by different cell types in the anterior and posterior eye, exerting a neuroprotective role in the adult life. The visual system is highly sensitive to NGF and the retina and optic nerve provides suitable subjects for the study of central nervous system degeneration. The model of bilateral carotid occlusion (two-vessel occlusion, 2VO) is a well-established model for chronic brain hypoperfusion leading to brain capillary pathology, to retina and optic nerve degeneration. In order to study if a single intravitreal injection of NGF protects the retina and the optic nerve from degeneration during systemic circulatory diseases, we investigated morphological and molecular changes occurring in the retina and optic nerve of adult rats at different time-points (8, 30 and 75 days) after bilateral carotid occlusion.. We demonstrated that a single intravitreal injection of NGF (5 microg/3 microl performed 24 hours after 2VO ligation) has a long-lasting protective effect on retina and optic nerve degeneration. NGF counteracts retinal ganglion cells degeneration by early affecting Bax/Bcl-2 balance- and c-jun- expression (at 8 days after 2VO). A single intravitreal NGF injection regulates the demyelination/remyelination balance after ischemic injury in the optic nerve toward remyelination (at 75 days after 2VO), as indicated by the MBP expression regulation, thus preventing optic nerve atrophy and ganglion cells degeneration. At 8 days, NGF does not modify 2VO-induced alteration in VEFG and related receptors mRNA expression.. The protective effect of exogenous NGF during this systemic circulatory disease seems to occur also by strengthening the effect of endogenous NGF, the synthesis of which is increased by vascular defect and also by the mechanical lesion associated with NGF or even vehicle intraocular delivery.

Topics: Analysis of Variance; Animals; bcl-2-Associated X Protein; Carotid Artery Diseases; Disease Models, Animal; Gene Expression Regulation; Injections, Intraventricular; Male; Myelin Basic Protein; Nerve Growth Factor; Optic Nerve Diseases; Proto-Oncogene Proteins c-bcl-2; Proto-Oncogene Proteins c-jun; Rats; Rats, Sprague-Dawley; Receptor, Nerve Growth Factor; Receptor, trkA; Reflex, Pupillary; Retinal Degeneration; RNA, Messenger; Tubulin

T-cell autoimmunity to myelin basic protein was recently shown to be neuroprotective in injured rat optic nerves. In the present study, using the mouse optic nerve, we examined whether active immunization rather than passive transfer of T-cells can be beneficial in protecting retinal ganglion cells (RGCs) from post-traumatic death. Before severe crush injury of the optic nerve, SJL/J and C3H.SW mice were actively immunized with encephalitogenic or nonencephalitogenic peptides of proteolipid protein (PLP) or myelin oligodendrocyte glycoprotein (MOG), respectively. At different times after the injury, the numbers of surviving RGCs in both strains immunized with the nonencephalitogenic peptides pPLP 190-209 or pMOG 1-22 were significantly higher than in injured controls treated with the non-self-antigen ovalbumin or with a peptide derived from beta-amyloid, a non-myelin-associated protein. Immunization with the encephalitogenic myelin peptide pPLP 139-151 was beneficial only when the disease it induced, experimental autoimmune encephalomyelitis, was mild. The results of this study show that survival of RGCs after axonal injury can be enhanced by vaccination with an appropriate self-antigen. Furthermore, the use of nonencephalitogenic myelin peptides for immunization apparently allows neuroprotection without incurring the risk of an autoimmune disease. Application of these findings might lead to a promising new approach for treating optic neuropathies such as glaucoma.

Topics: Animals; Cell Survival; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Female; Fluorescent Dyes; Mice; Mice, Inbred Strains; Myelin Basic Protein; Myelin Proteins; Myelin Proteolipid Protein; Myelin-Associated Glycoprotein; Myelin-Oligodendrocyte Glycoprotein; Nerve Crush; Optic Nerve Diseases; Optic Nerve Injuries; Peptide Fragments; Retinal Ganglion Cells; Stilbamidines; T-Lymphocytes; Vaccination

Eighteen normal human eye-bank eyes (age: 18-81 years), five fetal eyes (16-24 weeks), 11 primary open-angle glaucoma (POAG) eyes (age: 76-89 years), and two Schnabel's cavernous optic atrophy eyes were examined using a biotinylated-hyaluronan binding protein to study the changes in the distribution of hyaluronic acid (HA) in the fetal, adult and glaucomatous optic nerve head. The vitreous body served as a positive control. Sections treated with Streptomyces hyaluronidase were used to confirm specificity. Monoclonal antibodies to myelin basic protein (MBP) and glial fibrillary acidic protein (GFAP) were used as additional controls. In fetal optic nerve, HA was localized in blood vessels, peripapillary sclera and the pial septae in the retrolaminar nerve. No staining was associated with axons. Staining for MBP was negative. In adults, HA was found surrounding the myelin sheaths in the retrolaminar nerve; staining decreased with age. In contrast, HA staining in myelinated peripheral nerves (e.g. ciliaries) remained unchanged with age. HA also was localized to the adventitia of arteries and veins throughout the posterior segment. Compared to age-matched normal eyes, HA staining was virtually absent around myelin sheaths of the retrolaminar nerve in POAG eyes. Similar changes were not found in other HA positive structures. In Schnabel's cavernous optic atrophy. HA was present in increased amount in the atrophic area, but virtually absent in the remaining retrolaminar nerve. HA staining was invariably positive in vitreous, and Streptomyces hyaluronidase treated sections were negative. In adults, staining of MBP was associated with the myelin sheath in the retrolaminar nerve. In contrast to HA, staining of MBP was unchanged with age and in POAG. In Schnabel's atrophy, MBP staining disappeared only in the atrophic area. HA in the retrolaminar optic nerve appears to be associate with the space-filling matrix between myelin sheaths. HA is not present in the axon bundles prior to myelination of the optic nerve. HA in the retrolaminar optic nerve appears to decrease with age and is further reduced in POAG; however, corresponding changes are not found in MBP or in peripheral nerves. Perhaps, decreased amounts of HA is related to a higher susceptibility to elevated intraocular pressure or to optic nerve atrophy. In Schnabel's cavernous optic atrophy, HA is present in increased amount only in the atrophic area while MBP is markedly decreased, suggesting in situ product

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Aging; Fetus; Glaucoma, Open-Angle; Glial Fibrillary Acidic Protein; Histocytochemistry; Humans; Hyaluronic Acid; Middle Aged; Myelin Basic Protein; Myelin Sheath; Optic Nerve; Optic Nerve Diseases; Vitreous Body

Regarding immunopathogenesis of the Vogt-Koyanagi-Harada syndrome (VKHS), peripheral blood lymphocytes of a female patient with this disease were tested for sensitization against several antigens such as myelinic basic protein (BP) and uveoretinal, brain and spleen tissue homogenates. For comparison, cells of seven patients with chorioretinitis only and cells of six patients with encephalitis of unknown etiology were also tested. Cells of healthy donors served as controls. The electromobility test and the leucocyte migration test were used as in vitro test systems for cell-mediated immunological reactivity. The results show a strong sensitivity against BP and uveoretinal and brain tissue homogenates in VKHS. In the patients suffering from chorioretinitis, a strong reactivity against uveoretinal homogenate only was found, while in patients with encephalitis, responses to brain antigens only were observed. Reactions towards spleen tissue homogenate were negative in all experiments. The extent to which this tissue-specific immunological reactivity, which is different from the monoreactivity in chorioretinitis or encephalitis, might be a characteristic phenomenon of the VKHS is discussed.

Topics: Eye Proteins; Female; Humans; Immunity, Cellular; Immunologic Techniques; In Vitro Techniques; Lymphocytes; Middle Aged; Myelin Basic Protein; Neurologic Manifestations; Optic Nerve Diseases; Skin Diseases; Syndrome; Uveitis, Anterior