tetrodotoxin and Body-Weight

We tested whether tetrodotoxin (TTX) functions as a stress relieving substance in puffer fish. We orally administered TTX to the juveniles of hatchery-reared non-toxic tiger puffer Takifugu rubripes and measured the effects of TTX on brain corticotropin-releasing hormone (CRH) mRNA expression and plasma cortisol levels in comparison with effects in non-toxic juveniles. Firstly, the reciprocal connections of CRH and adrenocorticotropic hormone (ACTH) were confirmed by dual-label immunohistochemistry. CRH-immunoreactive (ir) cell bodies were detected in the hypothalamus and CRH-ir fibers were observed to project to ACTH-ir cells in the rostral pars distalis of the pituitary. Next, a TTX-containing diet (2.35 mouse units (517 ng)/g diet) or a non-toxic diet were fed to the fish for 28 days under a recirculating system. Standard length and body weight became significantly larger in the TTX-treated group. The degree of loss of the caudal fin, which is an indicator of the degree of agonistic interactions, where high values show a higher loss of caudal fin of a fish due to nipping by other individuals, was significantly lower in the TTX-treated group. Relative CRH mRNA expression levels in the brain and cortisol levels in the plasma were significantly lower in the TTX-treated group. These results indicate that TTX functions as a stress relieving substance by affecting the CRH-ACTH-cortisol axis and reducing agonistic interactions in tiger puffer juveniles.

Topics: Adrenocorticotropic Hormone; Animal Fins; Animals; Behavior, Animal; Body Weight; Brain Chemistry; Corticotropin-Releasing Hormone; Gene Expression; Hydrocortisone; Pituitary Gland; RNA, Messenger; Takifugu; Tetrodotoxin

Functional immobility of the diaphragm by mechanical ventilation impairs neuromuscular transmission and may result in ventilator-induced diaphragmatic dysfunction. We compared 3 diaphragmatic immobilization models with respect to their effects on expression of adult and fetal acetylcholine receptors (AChRs), muscle-specific receptor tyrosine kinase (MuSK), and muscle fiber morphology.. Diaphragms of rats were immobilized by either: (1) phrenicotomy; (2) presynaptic tetrodotoxin nerve blockade; or (3) postsynaptic polyethylene orthosis. AChR subtypes and MuSK were quantified by Western blot and immunohistochemistry. Muscle fiber morphology was evaluated by hematoxylin-eosin staining.. Adult AChRs remained unchanged, whereas fetal AChRs and MuSK were upregulated in all models. Denervation induced the strongest changes in muscle morphology.. Each diaphragm immobilization model led to severe morphologic and postsynaptic receptor changes. Postsynaptic polyethylene orthosis, a new model with an intact and functioning motor unit, best reflects the clinical picture of a functionally immobilized diaphragm. Muscle Nerve 55: 101-108, 2017.

Topics: Animals; Body Weight; Denervation; Diaphragm; Embryo, Mammalian; Gene Expression Regulation, Developmental; In Vitro Techniques; Male; Neuromuscular Junction; Protein Transport; Rats; Rats, Sprague-Dawley; Receptor Protein-Tyrosine Kinases; Receptors, Cholinergic; Tetrodotoxin

Maturation-associated changes in the internal distribution of tetrodotoxin (TTX) in the goby Yongeichthys criniger were investigated in 29 and 40 female specimens collected from Okinawa, Japan, from August 2008 to June 2009 (Group I), and from November 2009 to August 2010 (Group II), respectively. In Group I, based on changes in the gonadosomatic index (GSI) and histologic observation of the ovary, the period from October 2008 through January 2009 was estimated to be the 'previtelline-forming period', February through March 2009 the 'vitelline-forming period', April through June 2009 the 'spawning period', and August 2008 the 'end of spawning period' of the preceding year. The TTX content (mouse unit [MU] per gram tissue) of each Y. criniger tissue (skin, muscle, liver, and ovary) quantified by liquid chromatography/mass spectrometry (LC/MS) was generally high during the spawning period and continued to rise until the end of spawning period, especially in the ovary. Total TTX per individual increased considerably during the spawning period, most of which located in the ovary, indicating that Y. criniger obtains a high amount of TTX during the spawning period, and accumulates most of it in the ovary. In contrast, the TTX content of the skin was highest at the end of spawning period, and most of the total TTX located in the skin during this period as well as during the previtelline-forming period. In Group II, the maturation stage of the ovaries of all specimens was determined, and the specimens were grouped accordingly. In the perinucleolus stage, yolk vesicle stage, and yolk globule stage I, most of the TTX was localized in the skin, but the TTX in the ovary greatly increased as the maturation stage advanced from yolk globule stage I to yolk globule stage III.

Topics: Animals; Body Weight; Chromatography, High Pressure Liquid; Female; Mass Spectrometry; Organ Size; Ovary; Perciformes; Poisons; Reproduction; Seasons; Sexual Behavior, Animal; Sexual Maturation; Skin; Tetrodotoxin; Vitellins

In the present study, we analysed possible alterations in adrenergic, nitrergic and sensory functioning in mesenteric arteries from rats at 1 and 21 months after partial portal vein ligation, and the mechanisms involved in these alterations, if any. For this purpose, we analysed the vasoconstrictor response to EFS (electrical field stimulation) and the effect of the α-antagonist phentolamine, the NOS (nitric oxide synthase) inhibitor L-NAME (N(G)-nitro-L-arginine methyl ester) and the CGRP (calcitonin gene-related peptide) receptor antagonist CGRP-(8-37) in mesenteric segments from ST (short-term; 1 month) and LT (long-term; 21 months) SO (sham-operated) and pre-hepatic PH (portal hypertensive) rats. The vasomotor responses to NA (noradrenaline), the NO donor DEA-NO (diethylamine NONOate) and CGRP were analysed. NA, NO and CGRP releases were measured. Phospho-nNOS (neuronal NOS) expression was studied. The vasoconstrictor response to EFS was decreased in STPH animals. Phentolamine decreased this vasoconstrictor response more strongly in SO animals. Both L-NAME and CGRP-(8-37) increased vasoconstrictor response to EFS more strongly in PH than SO segments. PH did not modify vasomotor responses to NA, DEA-NO or CGRP, but it decreased NA release while increasing those of NO and CGRP. Phospho-nNOS expression was increased by PH. In LTPH, no differences were observed in vasoconstrictor response to EFS, vasomotor responses or neurotransmitter release when compared with age-matched SO animals. In conclusion, the mesenteric innervation may participate in the development of the characteristic hyperdynamic circulation observed in STPH through the joint action of decreased adrenergic influence, and increased nitrergic and sensory innervations influences. The participation of each innervation normalizes under conditions of LTPH.

Topics: Animals; Blood Pressure; Blotting, Western; Body Weight; Calcitonin Gene-Related Peptide; Electric Stimulation; Hypertension, Portal; In Vitro Techniques; Isometric Contraction; Male; Mesenteric Arteries; NG-Nitroarginine Methyl Ester; Nitric Oxide; Nitric Oxide Synthase Type I; Norepinephrine; Organ Size; Peptide Fragments; Phentolamine; Phosphorylation; Rats; Rats, Wistar; Sensory Receptor Cells; Tetrodotoxin; Time Factors; Vasodilator Agents

Peripheral nerve hyperexcitability (PNH) is one of the distal peripheral neuropathy phenotypes often present in patients affected by type 2 diabetes mellitus (T2DM). Through in vivo and ex vivo electrophysiological recordings in db/db mice, a model of T2DM, we observed that, in addition to reduced nerve conduction velocity, db/db mice also develop PNH. By using pharmacological inhibitors, we demonstrated that the PNH is mediated by the decreased activity of K(v)1-channels. In agreement with these data, we observed that the diabetic condition led to a reduced presence of the K(v)1.2-subunits in juxtaparanodal regions of peripheral nerves in db/db mice and in nerve biopsies from T2DM patients. Together, these observations indicate that the T2DM condition leads to potassium channel-mediated PNH, thus identifying them as a potential drug target to treat some of the DPN related symptoms.

Topics: Action Potentials; Age Factors; Animals; Blood Glucose; Body Weight; Diabetes Mellitus, Type 2; Disease Models, Animal; Electric Stimulation; Humans; Kv1.2 Potassium Channel; Male; Mice; Mice, Mutant Strains; Mutation; Neural Conduction; Peripheral Nerves; Peripheral Nervous System Diseases; Potassium Channel Blockers; Protein Subunits; Ranvier's Nodes; Receptors, Leptin; Sodium Channel Blockers; Sodium Channels; Tetrodotoxin

Previous research has demonstrated that diabetes induced learning and memory deficits. However, the mechanism of memory impairment induced by diabetes is poorly understood. Sprague-Dawley rats were used in the present study to investigate the effect of streptozotocin (STZ)-induced diabetes on spatial learning and memory with the Morris water maze. The excitability of CA1 pyramidal neurons in hippocampus was also examined. Diabetes impaired spatial learning and memory of rats. Diabetes decreased the membrane excitability of CA1 pyramidal neurons, effects which may contribute to the behavioral deficits. To investigate the further ionic mechanisms, the sodium currents and the potassium currents were detected. Diabetes decreased both transient and persistent sodium currents, and increased both transient and sustained potassium currents, which leads to the reduction of neuron excitability and to the increase of firing accommodation. The results of the present study suggested that sodium and potassium currents contributed to the inhibitory effect of diabetes on neuron excitability, further influencing learning and memory processing. Modulating the ion channels and increasing the membrane excitability are possible candidates for preventing the impairments of diabetes on hippocampal function.

Topics: Animals; Blood Glucose; Body Weight; CA1 Region, Hippocampal; Cell Membrane; Data Interpretation, Statistical; Diabetes Mellitus, Experimental; Electrophysiological Phenomena; Hippocampus; Male; Maze Learning; Memory; Neurons; Patch-Clamp Techniques; Potassium Channel Blockers; Potassium Channels, Voltage-Gated; Pyramidal Cells; Rats; Rats, Sprague-Dawley; Sodium Channels; Tetrodotoxin

Changes in tetrodotoxin (TTX) content of the puffer fish Takifugu rubripes during seed production were examined. Two mature female puffer fish T. rubripes (samples 1 and 2) that contained TTX were used. The toxic eggs were artificially fertilized, and hatchlings were reared in an indoor tank for 50 days and then in a netcage at sea for an additional 48 or 38 days. The TTX content of the fertilized eggs of sample 1 was initially 13.0 microg TTX/g, transiently increased to 67.6 microg TTX/g at 4 days after hatching, and then gradually decreased to 0.28 microg TTX/g at 98 days. In contrast, the total TTX content in an individual was 0.016 microg TTX at the fertilization stage and 0.01-0.03 microg TTX at the larval stage until 30 days after hatching. Thereafter, the total TTX content increased remarkably during culture in the netcage at sea, reaching 4.80 microg TTX at 98 days. Change in the TTX content of sample 2 showed a similar tendency to that of sample 1. The present study showed that the TTX content per gram of puffer fish body weight decreased during progression from fertilized eggs to juveniles, whereas the total TTX content increased.

Topics: Animals; Body Weight; Female; Takifugu; Tetrodotoxin; Time Factors; Tissue Distribution; Zygote

In this study, we investigated the hepatic uptake clearance (CL(uptake)) of tetrodotoxin (TTX) in the marine puffer fish Takifugu rubripes by integration plot analysis after a single bolus injection of 0.25mg TTX/kg body weight into the hepatic vein at 20 degrees C. The blood concentration of TTX decreased over time after the injection, from 1451+/-45 ng/mL at 10 min to 364+/-59 ng/mL at 60 min. TTX concentrations in the spleen and kidney decreased in parallel with the blood concentrations, whereas those in the muscle and skin remained almost the same throughout the experiment. In contrast, the TTX concentration in the liver gradually increased, reaching 1240+/-90 ng/g liver at 60 min after injection. The amount of TTX that had accumulated in the liver 60 min after injection accounted for 63+/-5% of the administered dose. Integration plot analysis indicated a CL(uptake) of 3.1 mL/min/kg body weight in the liver for TTX, a rate far below that of the hepatic portal vein blood flow rate (at most, 9%). This finding is consistent with negligible extraction of TTX by the liver. The results demonstrated conclusively that the liver-specific distribution of TTX in T. rubripes is achieved by removal from the systemic circulation, but not by the hepatic first-pass effect.

Topics: Animals; Body Weight; Injections, Intravenous; Liver; Organ Size; Poisons; Takifugu; Tetrodotoxin; Tissue Distribution

Marine puffer fish accumulates tetrodotoxin (TTX) in the liver and ovary. In this study, we examined the pharmacokinetics of TTX in Takifugu rubripes by a single administration under general anesthesia at 20 degrees C for 300 min. The blood concentration-time profile showed multiple distinct phases after injection into hepatic portal vein. The area under the blood concentration-time curve (AUC) increased linearly at the dosage of 0.25-0.75 mg TTX/kg body weight, and the total body clearance was 2.06+/-0.17 mL/min/kg body weight. The AUCs following administration into the hepatic portal vein and hepatic vein were closely similar (147+/-33 versus 141+/-1 ng.min/microL), indicating negligible hepatic first-pass effect. Comparison of the AUCs following an administration to the hepatic vein and gastrointestinal tract (0.25 mg TTX/kg body weight) elucidated the bioavailability of TTX to be 62%. There was no significant increase in the AUCs following direct injection into the gastrointestinal tract (0.50 versus 1.0 mg TTX/kg body weight). At the dosage of 0.25 mg TTX/kg body weight into the hepatic vein, hepatic portal vein or gastrointestinal tract, TTX amount in the liver accounted for 84+/-6%, 70+/-9% or 49+/-17% of the total TTX amount applied, respectively. These results demonstrate that TTX is absorbed into the systemic circulation from the gastrointestinal tract by saturable mechanism and finally accumulated in the liver within 300 min.

Topics: Animals; Biological Availability; Body Weight; Dose-Response Relationship, Drug; Female; Gastrointestinal Tract; Hepatic Veins; Injections, Intravenous; Portal Vein; Takifugu; Tetraodontiformes; Tetrodotoxin; Time Factors; Tissue Distribution

Functional changes induced by inflammation persist following recovery from the inflammatory response, but the mechanisms underlying these changes are not well understood. Our aim was to investigate whether the excitability and synaptic properties of submucosal neurons remained altered 8 wk post-trinitrobenzene sulfonic acid (TNBS) treatment and to determine whether these changes were accompanied by alterations in secretory function in submucosal preparations voltage clamped in Ussing chambers. Mucosal serotonin (5-HT) release measurements and 5-HT reuptake transporter (SERT) immunohistochemistry were also performed. Eight weeks after TNBS treatment, colonic inflammation resolved, as assessed macroscopically and by myeloperoxidase assay. However, fast excitatory postsynaptic potential (fEPSP) amplitude was significantly increased in submucosal S neurons from previously inflamed colons relative to those in control tissue. In addition, fEPSPs from previously inflamed colons had a hexamethonium-insensitive component that was not evident in age-matched controls. AH neurons were hyperexcitable, had shorter action potential durations, and decreased afterhyperpolarization 8 wk following TNBS adminstration. Neuronally mediated colonic secretory function was significantly reduced after TNBS treatment, although epithelial cell signaling, as measured by responsiveness to both forskolin and bethanecol in the presence of tetrodotoxin, was comparable with control tissue. 5-HT levels and SERT immunoreactivity were comparable to controls 8 wk after the induction of inflammation, but there was an increase in glucagon-like peptide 2-immunoreactive L cells. In conclusion, sustained alterations in enteric neural signaling occur following the resolution of colitis, which are accompanied by functional changes in the absence of active inflammation.

Topics: Action Potentials; Animals; Bethanechol; Body Weight; Cell Count; Colforsin; Colitis; Colon; Enteric Nervous System; Enteroendocrine Cells; Excitatory Postsynaptic Potentials; Glucagon-Like Peptide 2; Guinea Pigs; Male; Membrane Potentials; Neurons; Peptide YY; Peroxidase; Serotonin; Serotonin Plasma Membrane Transport Proteins; Submucous Plexus; Tetrodotoxin; Trinitrobenzenesulfonic Acid; Veratridine

Intense or chronic stress can produce pathophysiological alterations in the systems involved in the stress response. The amygdala is a key component of the brain's neuronal network that processes and assigns emotional value to life's experiences, consolidates the memory of emotionally significant events, and organizes the behavioral response to these events. Clinical evidence indicates that certain stress-related affective disorders are associated with changes in the amygdala's excitability, implicating a possible dysfunction of the GABAergic system. An important modulator of the GABAergic synaptic transmission, and one that is also central to the stress response is norepinephrine (NE). In the present study, we examined the hypothesis that stress impairs the noradrenergic modulation of GABAergic transmission in the basolateral amygdala (BLA). In control rats, NE (10 microM) facilitated spontaneous, evoked, and miniature IPSCs in the presence of beta and alpha(2) adrenoceptor antagonists. The effects of NE were not blocked by alpha(1D) and alpha(1B) adrenoceptor antagonists, and were mimicked by the alpha(1A) agonist, A61603 (1 microM). In restrain/tail-shock stressed rats, NE or A61603 had no significant effects on GABAergic transmission. Thus, in the BLA, NE acting via presynaptic alpha(1A) adrenoceptors facilitates GABAergic inhibition, and this effect is severely impaired by stress. This is the first direct evidence of stress-induced impairment in the modulation of GABAergic synaptic transmission. The present findings provide an insight into possible mechanisms underlying the antiepileptogenic effects of NE in temporal lobe epilepsy, the hyperexcitability and hyper-responsiveness of the amygdala in certain stress-related affective disorders, and the stress-induced exacerbation of seizure activity in epileptic patients.

Topics: 2-Amino-5-phosphonovalerate; 6-Cyano-7-nitroquinoxaline-2,3-dione; Adrenergic alpha-Antagonists; Adrenergic beta-Antagonists; Aging; Amygdala; Anesthetics, Local; Animals; Animals, Newborn; Bicuculline; Body Weight; Dose-Response Relationship, Drug; Drug Interactions; Estrenes; Excitatory Amino Acid Antagonists; GABA Antagonists; gamma-Aminobutyric Acid; Imidazoles; In Vitro Techniques; Isoquinolines; Male; Membrane Potentials; Morpholines; Neural Inhibition; Neurons; Norepinephrine; Patch-Clamp Techniques; Phosphodiesterase Inhibitors; Propranolol; Pyrrolidinones; Rats; Rats, Sprague-Dawley; Receptors, Adrenergic, alpha-1; Restraint, Physical; Stress, Physiological; Tetrahydronaphthalenes; Tetrodotoxin; Time Factors

The fat-derived hormone leptin regulates energy balance in part by modulating the activity of neuropeptide Y and proopiomelanocortin neurons in the hypothalamic arcuate nucleus. To study the intrinsic activity of these neurons and their responses to leptin, we generated mice that express distinct green fluorescent proteins in these two neuronal types. Leptin-deficient (ob/ob) mice differed from wild-type mice in the numbers of excitatory and inhibitory synapses and postsynaptic currents onto neuropeptide Y and proopiomelanocortin neurons. When leptin was delivered systemically to ob/ob mice, the synaptic density rapidly normalized, an effect detectable within 6 hours, several hours before leptin's effect on food intake. These data suggest that leptin-mediated plasticity in the ob/ob hypothalamus may underlie some of the hormone's behavioral effects.

Topics: Animals; Arcuate Nucleus of Hypothalamus; Body Weight; Eating; Evoked Potentials; Excitatory Postsynaptic Potentials; Feeding Behavior; gamma-Aminobutyric Acid; Ghrelin; Glutamic Acid; Green Fluorescent Proteins; In Vitro Techniques; Leptin; Luminescent Proteins; Mice; Mice, Obese; Mice, Transgenic; Neuronal Plasticity; Neurons; Neuropeptide Y; Patch-Clamp Techniques; Peptide Hormones; Pro-Opiomelanocortin; Recombinant Fusion Proteins; Synapses; Tetrodotoxin; Transgenes

The aims of this study were two-fold: first, to simplify the method for creating a recently described neuropathic pain model in the rat, and second, to evaluate the effects of a number of drugs with analgesic or antihyperalgesic properties, in this model. Continuous intravenous vincristine infusion (1-100 microg kg(-1) day (-1)) for 14 days resulted in a dose dependent tactile allodynia (as measured by von Frey filaments) by 7 days at doses between 30 - 100 microg kg(-1) day (-1), with a hindlimb motor deficit observed at doses greater than 50 microg kg(-1) day (-1). No thermal hyperalgesia was observed. Systemic morphine, lidocaine, mexiletine and pregabalin (given intraperitoneally) produced significant reduction of the allodynia, while tetrodotoxin was without effect. Continuous intravenous infusion of vincristine in rats thus provides a reliable model of chemotherapy induced neuropathy which may be used in defining the mechanism and pharmacology of this clinically relevant condition.

Topics: Acetates; Amines; Analgesics; Animals; Antineoplastic Agents, Phytogenic; Body Weight; Cyclohexanecarboxylic Acids; Excitatory Amino Acid Antagonists; Gabapentin; gamma-Aminobutyric Acid; Hot Temperature; Ion Channel Gating; Male; Pain; Pain Measurement; Pain Threshold; Rats; Rats, Sprague-Dawley; Tetrodotoxin; Vincristine

The expression of five myosin heavy chain (MHC) isoforms was analyzed in the rat soleus (Sol) and the deep and superficial medial gastrocnemius (dGM, sGM) muscle after 2 and 4 wk of TTX paralysis by using immunohistochemical techniques. In Sol, after 4 wk of paralysis, fibers containing type I MHC were either pure type I (14%) or also contained developmental (D; 76%), IIa (26%), or IIx (18%) MHC. Values for corresponding fibers in dGM were 8.5, 65, 38, and 22%. Also, by 4 wk an increase was seen in the proportions of fibers expressing IIa MHC in Sol (from 16 to 38%) and dGM (from 24 to 74%). In a region of sGM in control muscles containing pure IIb fibers, a major proportion (86%) remained pure after 4 wk of paralysis, with the remainder coexpressing IIb and IIx. The results indicate that TTX-induced muscle paralysis results in an increase in fibers containing multiple MHC isoforms and that the D isoform appears in a major proportion of these hybrid fibers.

Topics: Animals; Body Weight; Female; Immunohistochemistry; Muscle Fibers, Skeletal; Muscle, Skeletal; Muscular Atrophy; Myosin Heavy Chains; Organ Size; Paralysis; Protein Isoforms; Rats; Rats, Sprague-Dawley; Tetrodotoxin; Time Factors

1. In the present study, we investigated the effect of GABA and selective GABA agonists and antagonists on neurally induced tracheal contractions in streptozotocin (STZ) diabetic rats. 2. Contractile responses to electrical field stimulation (EFS) in rat tracheal rings were completely abolished by atropine and tetrodotoxin, but were unaffected by the ganglion blocker hexamethonium, indicating that they were mediated via neuronal release of acetylcholine (ACh). 3. Contractions induced by EFS, but not by exogenous ACh, were inhibited by GABA and the selective GABA(B) receptor agonist baclofen, but not by the selective GABA(A) receptor agonist 3-aminopropane sulphonic acid. The inhibitory effects of GABA or baclofen were not affected by the GABA(A) antagonist bicuculline, but were significantly reversed by the GABA(B) antagonist phaclofen. 4. The inhibitory effects of both GABA and baclofen were found to be significantly greater in trachea from control rats compared with tissues from diabetic rats. 5. Non-adrenergic, non-cholinergic relaxation responses elicited by EFS in precontracted tracheal rings from diabetic and control rats were similar in magnitude and were unaffected by GABA or GABA analogues. 6. These results suggest that GABA decreases the response to EFS by directly inhibiting the evoked release of ACh through GABA(B) receptors in rat trachea and that STZ-induced diabetes causes an impairment in the inhibitory effect of GABA on neurally induced contractions in this tissue.

Topics: Acetylcholine; Animals; Baclofen; Bicuculline; Blood Glucose; Body Weight; Diabetes Mellitus, Experimental; Dose-Response Relationship, Drug; Electric Stimulation; GABA Agonists; GABA Antagonists; gamma-Aminobutyric Acid; In Vitro Techniques; Male; Muscle Contraction; Rats; Rats, Wistar; Taurine; Tetrodotoxin; Trachea; Vasodilator Agents

This study examined the effect of fasting on the neural control of ion transport and paracellular permeability in piglet jejunum. Muscle-stripped tissues from fed or 48-h fasted piglets were mounted in Ussing chambers. Neural blockade with tetrodotoxin (TTX) or antagonists of muscarinic or nicotinic receptors caused reductions in basal short-circuit current that were approximately threefold greater in fasted piglets. The TTX-induced reduction in short-circuit current in fasted piglets was due to a decrease in residual ion flux and was abolished in the absence of HCO(-)(3). Intestinal paracellular permeability, as indicated by tissue conductance (G(t)) and fluxes of inulin and mannitol, was significantly increased by fasting. TTX increased inulin flux and G(t) in fed but not fasted piglets. In fasted piglets, carbachol reduced G(t) by 29% and mannitol flux by 27% but had no effect on these parameters in the fed state. We conclude that fasting enhances enteric neural control of basal ion transport and increases paracellular permeability in piglet jejunum. Tonic release of enteric neurotransmitters regulates paracellular permeability in the fed state, and cholinergic stimulation restores fasting-induced elevations in paracellular permeability to fed levels.

Topics: Animal Nutritional Physiological Phenomena; Animals; Atropine; Bicarbonates; Biological Transport; Body Weight; Eating; Enteric Nervous System; Fasting; Ganglionic Blockers; Intestinal Absorption; Intestinal Mucosa; Jejunum; Mecamylamine; Membrane Potentials; Neurotransmitter Agents; Parasympatholytics; Swine; Tetrodotoxin

The aim of the study was to address discrepant findings in the literature regarding coupling between decreased functional demand during disuse and reduced capillarity. We previously reported [K. Tyml, O. Mathieu-Costello, and E. Noble. Microvasc. Res. 49: 17-32, 1995] that severe disuse of rat extensor digitorum longus (EDL) muscle caused by a 2-wk application of tetrodotoxin (TTX) on the sciatic nerve is not accompanied by capillary loss. Using the same animal model, the present study examined whether this absence of coupling could be explained in terms of 1) too short a duration of disuse and 2) muscle-specific response to disuse. Fischer 344 rats were exposed to either no treatment (control) or to 2- or 8-wk TTX applications. Fiber size, capillary density per fiber cross-sectional area, and capillary-to-fiber (C/F) ratio were determined by morphometry in the EDL muscle (control, 2- and 8-wk groups) and in the superficial portion of medial gastrocnemius (Gas) muscle (control, 2 wk). In both muscles, microvascular blood flow was evaluated by intravital microscopy [red blood cell velocity in capillaries (V(RBC))] and by laser Doppler flowmetry (LDF). Regardless of duration of TTX application or muscle type, TTX-induced disuse resulted in a significant reduction of fiber area (44-71%). However, capillary density increased in EDL muscle (both at 2 and 8 wk) but not in Gas muscle. C/F ratio decreased in EDL muscle at 8 wk (18%) and in Gas muscle (39%). This indicates that the effect on capillarity depended on duration of disuse and on muscle type. V(RBC) and LDF signal were significantly larger in EDL than in Gas muscle. Analysis of change in capillarity vs. V(RBC) suggested that the outcome of disuse may be modulated by blood flow. We conclude that the duration of skeletal muscle disuse per se does not dictate capillary loss, and we hypothesize that discrepant findings of coupling between functional demand and capillarity could be due to the presence/absence of flow-related angiogenesis superimposed on the capillary removal process during disuse.

Topics: Animals; Blood Volume; Body Weight; Capillaries; Hemodynamics; Hindlimb; Ischemia; Male; Microcirculation; Muscle, Skeletal; Muscular Atrophy; Rats; Rats, Inbred F344; Sciatic Nerve; Tetrodotoxin; Time Factors

Portal-systemic encephalopathy (PSE) is associated with an increased brain tissue turnover of serotonin (5-HT). Despite increased 5-HT metabolism, brain 5-HT release in rats with a portacaval shunt (PCS) seems to be unaltered. Although this may indicate that the overall 5-HT output is unaltered in PSE, it is also possible that the 5-HT release pattern might be altered in some way. In the present study, the potassium-evoked frontal neocortical release of 5-HT was studied in experimental chronic PSE. KCI (60 mM) produced marked increases in the 5-HT output compared with basal values both in PCS and sham rats. Simultaneously, the KCI challenge resulted in significant elevations in the 5-HT release of PCS compared with sham. In Ca2+-free medium, the difference between PCS and sham rats in the KCl-evoked release of 5-HT was abolished. In the presence of TTX (1 mM), both groups displayed increased extracellular 5-HT levels. Again, a difference with higher amplitude of the 5-HT release in PCS compared with sham was evident. It is concluded that in experimental chronic PSE an augmented neocortical 5-HT release compared with the normal in vivo situation is available. The possible mechanism(s) responsible for the difference in neocortical 5-HT output between PCS and sham-operated rats in response to the KCl-challenge is discussed.

Topics: Animals; Body Weight; Calcium; Chronic Disease; Extracellular Space; Frontal Lobe; Hepatic Encephalopathy; Hydroxyindoleacetic Acid; Liver; Male; Microdialysis; Neurons; Organ Size; Potassium Chloride; Rats; Rats, Sprague-Dawley; Serotonin; Tetrodotoxin

Although a link between the hypothalamic-pituitary-thyroid (HPT) axis and affective disorder has been established, the mechanism underlying this relationship remains unclear. Since the serotonin (5-HT) system appears to be involved in the pathophysiology of mood disorders, the time course of the effects of thyroidectomy (TXT) with or without thyroxine (T4) replacement on 5-HT1A receptors and 5-HT uptake sites was examined. TXT caused a significant increase in 3H-8-hydroxy-2-(di-n-propylamino)-tetralin (3H-DPAT) binding to 5-HT1A receptors in the cortex and hippocampus at 7 days and this increase was also evident at 35 days following TXT. By contrast, TXT did not have a significant effect on 3H-DPAT binding in the hypothalamus or in the dorsal raphe nucleus. TXT did not affect the binding of 3H-cyanoimipramine (3H-CN-IMI) to 5-HT uptake sites in any of the brain regions analyzed, or at any of the time points studied. Administration of high-dose T4 for 28 days caused the binding of 3H-DPAT to recover to sham levels in the cortex, to increase in the hippocampus and hypothalamus, and had no effect in the dorsal raphe nucleus. Replacement with high-dose T4 had no effect on 3H-CN-IMI binding to 5-HT uptake sites when compared to sham-operated animals at all time points examined. These results suggest that a neuromodulatory link may exist between the HPT axis and 5-HT1A receptors in the limbic regions of the rat brain. Depending on the brain region examined, a differential response to circulating levels of thyroid hormone was observed.

Topics: 8-Hydroxy-2-(di-n-propylamino)tetralin; Animals; Autoradiography; Body Weight; Brain Chemistry; Hyperthyroidism; Imipramine; Kinetics; Male; Radioimmunoassay; Raphe Nuclei; Rats; Rats, Sprague-Dawley; Receptors, Serotonin; Serotonin Antagonists; Tetrodotoxin; Thyroid Gland; Thyroidectomy; Thyroxine; Triiodothyronine

The role of oxygen radicals in isocapnic hyperpnea-induced bronchoconstriction (HIB) of guinea pigs was investigated using scavengers of the radicals. In series 1, 50 young guinea pigs were randomly divided into seven groups: control 1, control 2, chlorisondamine, tetrodotoxin (TTX), acute dimethylthiourea (DMTU), tachykinin depletion, and 5% CO2 in air. Animals of the control 2 group received vehicle (saline) infusion while those of the control 1 group did not. Chlorisondamine was used to block ganglionic transmission, TTX to interrupt nerve conduction, DMTU to scavenge hydroxyl radicals, and chronic capsaicin pretreatment to deplete tachykinins. The animals in the last group were ventilated with dry 5% CO2 in air during hyperpnea. In series 2, 13 additional animals were used to test the effects of intratracheal administration of superoxide dismutase and catalase (SOD + CAT) on HIB. Each animal was anesthetized with pentobarbital sodium, cannulated with a tracheal cannula and venous catheter, paralyzed with gallamine triethiodide, and mechanically ventilated. During the baseline period, each animal was ventilated normally with humidified air. Then it was hyperventilated 15 min with a dry gas mixture of 95% O2-5% CO2, except animals in the last group of series 1. Subsequently, all animals returned to normal ventilation with humidified air for 45 min (recovery period). The maximal expiratory flow and dynamic compliance were obtained periodically during the recovery period. The isocapnic hyperpnea using 95% O2-5% CO2, but not 5% CO2 in air, caused bronchoconstriction that was significantly blocked by acute DMTU, acute SOD + CAT, and tachykinin depletion. In an additional group of six animals, acute DMTU did not significantly alter acetylcholine-induced airway constriction.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Axons; Body Weight; Bronchoconstriction; Carbon Dioxide; Catalase; Free Radical Scavengers; Free Radicals; Guinea Pigs; Lung Compliance; Male; Oxygen Consumption; Reflex; Respiratory Function Tests; Respiratory Mechanics; Superoxide Dismutase; Tachykinins; Tetrodotoxin

1. Previous reports of the effects of disuse induced by tetrodotoxin (TTX) have demonstrated alterations in muscle function suggesting changes in the quality of contractile proteins. 2. We extended these studies to the effects of TTX-induced disuse on the Ca2+ activation characteristics of myofibrillar ATPase of the rat gastrocnemius. 3. Atrophic responses were as previously reported (St-Pierre, D.M.M. and Gardiner P.F. (1985) Effect of disuse on mammalian fast-twitch muscle: joint fixation compared with neurally applied tetrodotoxin. Exp. Neurol. 90, 635-651; St-Pierre, D.M.M. et al. (1987). Recovery of muscle from tetrodotoxin-induced disuse and the influence of daily exercise; 1. Contractile properties. Exp. Neurol. 98, 472-488.) with a significant decrease in left gastrocnemius weight compared to control (C) (1.25 +/- 0.06 for C vs 0.72 +/- 0.04 for TTX, X +/- SEM, P less than or equal to 0.01). 4. Myofibrillar protein yield (mg/g wet weight) was also depressed (92.8 +/- 4.5 for C vs 70.3 +/- 3.7 for TTX; P less than or equal to 0.01). 5. Maximum ATPase of myofibrils (nmol Pi/mg/min) was decreased (441 +/- 28 for C vs 181 +/- 30 for TTX, P less than or equal to 0.01). 6. Furthermore, the Hill n which reflects the cooperative aspects of Ca2+ activation of the myofibrillar ATPase was depressed (1.58 +/- 0.07 for C vs 1.29 +/- 0.09 for TTX; P less than or equal to 0.01). 7. The results suggest that muscle perturbations resulting from disuse are partially related to changes in the myofibril.

Topics: Adenosine Triphosphatases; Animals; Atrophy; Body Weight; Calcium; Enzyme Activation; Female; Muscles; Myofibrils; Organ Size; Rats; Rats, Inbred Strains; Tetrodotoxin

1. We measured the rate of occurrence of miniature endplate potentials (MEPPs) at identified endplates in frog cutaneous pectoris muscles treated with crude black widow spider venom (BWSV) or purified alpha-latrotoxin (alpha-LTX) in calcium-free solutions, and we examined the relationship between the length of the nerve terminal and the total number of quanta secreted, and the relationship between the number of quanta secreted and the number of vesicles remaining at different times. 2. The venom, or toxin, was applied in a modified Ringer solution with tetrodotoxin, 1 mM-EGTA and no divalent cations, and quantal secretion was started by applying Ca2(+)-free solutions with Mg2+. This was done to synchronize the quantal discharge at the various junctions in a muscle. Ringer solution was applied after the MEPP rate had declined to low levels, and then the muscle fibre was injected with Lucifer Yellow, the endplate stained for acetylcholinesterase and the length of the nerve terminal and the length of a sarcomere were measured on the fluorescent fibre. 3. The total number of quanta secreted by a terminal was measured under a wide variety of experimental conditions: the weights of the frogs ranged from 13 to 68 g, the temperature from 9 to 28 degrees C, and the concentration of Mg2+ from 2 to 10 mM. In one series of experiments the Mg2+ was withdrawn after 3-4 min and reapplied 35-40 min later in order to divide the total output of quanta into two approximately equal bouts of secretion that were well separated in time. 4. The total number of MEPPs recorded at a junction was loosely correlated with the length of its nerve terminal, but it was not affected by the temperature, the concentration of Mg2+ or the division of secretion into well-separated bouts of quantal release. The average total secretion per unit length was about 3700 quanta/sarcomere or about 1200 quanta/microns. 5. The average time course of quantal secretion per micrometre of terminal was determined at single junctions in muscles held at 22-23 degrees C or at 9-10 degrees C. Other muscles were fixed at various times during the course of secretion at each temperature and the number of synaptic vesicles remaining in cross-sections of the terminals were counted on electron micrographs. The number of vesicles remaining per micrometre of terminal was determined from the number per cross-section and the section thickness.(ABSTRACT TRUNCATED AT 400 WORDS)

Topics: Animals; Body Weight; Egtazic Acid; Female; Kinetics; Magnesium; Male; Membrane Potentials; Microscopy, Electron; Motor Endplate; Neuromuscular Junction; Rana pipiens; Spider Venoms; Synaptic Vesicles; Temperature; Tetrodotoxin

The extent to which the remaining active or passive components of muscle mechanical stress not associated with weightbearing are involved in preserving muscle morphological and functional characteristics in the rodent hindlimb suspension model is not known. Such information would be relevant to the construction of appropriate countermeasures for the disuse atrophy associated with muscle unloading. This question was addressed by superimposing 2 weeks of hindlimb suspension and neuromuscular quiescence, achieved by the chronic neural application of the sodium channel blocker tetrodotoxin. A major portion of the muscle size characteristics of the fast anti-gravity gastrocnemius and plantaris, and the functional characteristics of the plantaris, were maintained by the full range voluntary activity remaining after suspension. Muscle mass of the slow soleus was compromised regardless of this residual activity. Indeed, for fast ankle extensors, hindlimb unloading resembles more closely a model of normal usage than of disuse, but for slow extensors this condition appears to be extremely detrimental.

Topics: Action Potentials; Animals; Body Weight; Electromyography; Female; Hindlimb; Models, Biological; Muscle Contraction; Muscle Denervation; Muscles; Muscular Atrophy; Neuromuscular Junction; Rats; Rats, Inbred Strains; Tetrodotoxin; Weightlessness

1. Electrophysiological properties of soleus motoneurones in adult cats were examined with intracellular electrodes following alterations of activity of the soleus muscle induced by transection of the thoracic spinal cord or by conduction block of the muscle nerve with tetrodotoxin (TTX) cuffs. Attempts were also made to maintain muscle activity by daily stimulation of the maintain muscle activity by daily stimulation of the peripheral nerve. 2. Within 8 days after transection of the thoracic cord, soleus motoneurones showed a significant decrease in the duration of afterhyperpolarization following action potentials. This change in motoneurone properties induced by cord transection was prevented by daily stimulation of the sciatic nerve. 3. Soleus motoneurones showed a significant decrease in the duration of after-hyperpolarization within 8 days after conduction block of the soleus nerve with TTX. This change in montoneurone properties was prevented by daily stimulation of the nerve peripheral to the TTX cuff but not central to the cuff. 4. The soleus muscle showed a significant decrease in weight relative to body weight within 8 days after transection of the thoracic cord. This decrease in muscle weight following cord transection was prevented by daily stimulation of the sciatic nerve. 5. No fibrillation was detected in the soleus muscle 8 days after conduction block of the soleus nerve with TTX. The maximum twitch tension of the soleus muscle evoked by nerve stimulation showed no significant difference between the two sides treated and untreated with TTX. Fast axoplasmic transport measured with cholinesterase as a marker was not affected by TTX. Thus, there was no sign of functional although morphological abnormalities were found in some nerve fibres. 6. It is concluded that motoneurone properties in an adult depend partly upon some factors associated with activity of the innervated muscles and that such trophic signals are retrogradely carried by the motor axons.

Topics: Action Potentials; Animals; Body Weight; Cats; Cordotomy; Female; Motor Neurons; Muscles; Neural Conduction; Organ Size; Tetrodotoxin

Topics: Animals; Atropine; Body Weight; Female; Hexamethonium Compounds; Histamine; Injections; Male; Mice; Picrotoxin; Strychnine; Tetrodotoxin; Toxicology; Toxins, Biological; Tubocurarine