Active shrinkage protects neurons following axonal transection.

Trauma, vascular events, or neurodegenerative processes can lead to axonal injury and eventual transection (axotomy). Neurons can survive axotomy, yet the underlying mechanisms are not fully understood. Excessive water entry into injured neurons poses a particular risk due to swelling and subsequent death. Using in vitro and in vivo neurotrauma model systems based on laser transection and surgical nerve cut, we demonstrated that axotomy triggers actomyosin contraction coupled with calpain activity. As a consequence, neurons shrink acutely to force water out through aquaporin channels preventing swelling and bursting. Inhibiting shrinkage increased the probability of neuronal cell death by about 3-fold. These studies reveal a previously unrecognized cytoprotective response mechanism to neurotrauma and offer a fresh perspective on pathophysiological processes in the nervous system.

The electrophysiological and behavioral evaluation of the peptide hemopressin and cannabinoid CB1 receptor agonist and antagonist in pentylenetetrazol model of epilepsy in rats.

This study endeavoured to assess the effect of hemopressin (Hp), a nano peptide obtained from the alpha chain of hemoglobin, on chronic epileptic activity and its potential correlation with cannabinoid receptor type 1 (CB1). Male Wistar albino rats (230-260 g) were used. The kindling process was conducted by administering a sub-convulsant dose of pentylenetetrazol (PTZ) (35 mg/kg, i.p) three times a week for a maximum of 10 weeks. Tripolar electrodes and external cannula guides for intracerebroventricular (i.c.v) injections were surgically implanted in the skulls of kindled rats. On the day of the experiment, doses of Hp, AM-251, and ACEA were administered prior to the PTZ injections. Electroencephalography recordings and behavioural observations were conducted simultaneously for 30 min after the PTZ injection. The administration of Hp (0.6 µg, i.c.v) resulted in a decrease in epileptic activity. The CB1 receptor agonist ACEA (7.5 µg, i.c.v) showed an anticonvulsant effect, but the CB1 receptor antagonist AM-251 (0.5 µg, i.c.v) displayed a proconvulsant effect. The co-administration of Hp (0.6 µg, i.c.v) and ACEA (7.5 µg, i.c.v) and of Hp (0.6 µg, i.c.v) and AM-251 (0.5 µg, i.c.v) produced an anticonvulsant effect. However, when AM-251 was administered prior to Hp, it produced a proconvulsant impact that overrode Hp's intended anticonvulsant effect. Interestingly, the co-administration of Hp (0.03 µg) + AM-251 (0.125 µg) unexpectedly exhibited an anticonvulsant effect. Electrophysiological and behavioural evaluations demonstrated the anticonvulsant effect of Hp in the present model, highlighting the possibility that Hp may act as an agonist for the CB1 receptor.

Evaluation of the association between sexual dysfunction and demyelinating plaque location and number in female multiple sclerosis patients.

Purpose To investigate the frequency of sexual dysfunction (SD) in female multiple sclerosis (MS) patients and to explore its association with the location and number of demyelinating lesions. Material and Methods We evaluated 42 female patients and 41 healthy subjects. All patients underwent neurological examination and 1.5 T brain and full spinal MRI. All subjects completed the female sexual function index (FSFI), Beck Depression Inventory (BDI), Beck Anxiety Inventory (BAI), and Short-Form 36 Quality of Life Scale (SF-36). All participants were also evaluated for serum thyroid stimulating hormone (TSH), T4, estradiol, and total testosterone. Results No statistically significant differences between the MS and control groups were found for age, body mass index (BMI), serum TSH, T4, E2, and total testosterone level. MS patients had a statistically significantly lower FSFI and SF-36 scores and higher BDI and BAI scores compared with healthy subjects. The location and number of demyelinating lesions were not associated with SD. Conclusion In our cohort, this difference in SD appears unrelated to the location and number of demyelinating lesions. These findings highlight the importance of the assessment and treatment of psychiatric comorbidities, such as depression and anxiety, in MS patients reporting SD.

Modulation of Excitability of Stellate Neurons in the Ventral Cochlear Nucleus of Mice by ATP-Sensitive Potassium Channels.

Major voltage-activated ionic channels of stellate cells in the ventral part of cochlear nucleus (CN) were largely characterized previously. However, it is not known if these cells are equipped with other ion channels apart from the voltage-sensitive ones. In the current study, it was aimed to study subunit composition and function of ATP-sensitive potassium channels (KATP) in stellate cells of the ventral cochlear nucleus. Subunits of KATP channels, Kir6.1, Kir6.2, SUR1, and SUR2, were expressed at the mRNA level and at the protein level in the mouse VCN tissue. The specific and clearly visible bands for all subunits but that for Kir6.1 were seen in Western blot. Using immunohistochemical staining technique, stellate cells were strongly labeled with SUR1 and Kir6.2 antibodies and moderately labeled with SUR2 antibody, whereas the labeling signals for Kir6.1 were too weak. In patch clamp recordings, KATP agonists including cromakalim (50 µM), diazoxide (0.2 mM), 3-Amino-1,2,4-triazole (ATZ) (1 mM), 2,2-Dithiobis (5-nitro pyridine) (DTNP) (330 µM), 6-Chloro-3-isopropylamino- 4H-thieno[3,2-e]-1,2,4-thiadiazine 1,1-dioxide (NNC 55-0118) (1 µM), 6-chloro-3-(methylcyclopropyl)amino-4H-thieno[3,2-e]-1,2,4-thiadiazine 1,1-dioxide (NN414) (1 µM), and H2O2 (0.88 mM) induced marked responses in stellate cells, characterized by membrane hyperpolarization which were blocked by KATP antagonists. Blockers of KATP channels, glibenclamide (0.2 mM), tolbutamide (0.1 mM) as well as 5-hydroxydecanoic acid (1 mM), and catalase (500 IU/ml) caused depolarization of stellate cells, increasing spontaneous action potential firing. In conclusion, KATP channels seemed to be composed dominantly of Kir 6.2 subunit and SUR1 and SUR2 and activation or inhibition of KATP channels regulates firing properties of stellate cells by means of influencing resting membrane potential and input resistance.

Proconvulsant Effect of Papaverine on Penicillin-Induced Epileptiform Activity in Rats.

AIM: Papaverine is a vasodilator agent that is an opium alkaloid. It exhibits its effects by inhibiting the phosphodiesterase enzyme. Papaverine administration is widely used to avoid symptomatic vasospasm after subarachnoid hemorrhage. We aimed, in this research, to study the effects of papaverine on the epileptic discharges stimulated by penicillin. MATERIAL AND METHODS: Adult female Wistar rats (220±30 g) were included in this research (n=30). Rats were anesthetized with urethane (1.25 g/kg) and then the left cerebral cortex was reached by opening a burr hole with a drill. Penicillin G sodium salt (500 IU)(200 IU/1 µl) was injected into the left lateral ventricle to produce epileptiform activity. Thirty minutes before penicillin G sodium injection, papaverine was administered at doses of 5, 10, 20 or 40 mg/kg intraperitoneally. RESULTS: There was no significant difference in spike frequency between the control group and the groups given 5 mg/kg, 10 mg/ kg or 40 mg/kg papaverine, while 20 mg/kg papaverine significantly increased the spike frequency (p < 0.05). CONCLUSION: Papaverine augments the epileptiform activity produced by penicillin injection. It is important to remember that papaverine might induce convulsions in patients who have epilepsy. More research is required to understand the mechanisms of the proconvulsant influence of papaverine in epilepsy.

Isolation and culture of adult mouse vestibular nucleus neurons

Background/aim: Isolated cell cultures are widely used to study neuronal properties due to their advantages. Although embryonic animals are preferred for culturing, their morphological or electrophysiological properties may not reflect adult neurons, which may be important in neurodegenerative diseases. This paper aims to develop a method for preparing isolated cell cultures of medial vestibular nucleus (MVN) from adult mice and describe its morphological and electrophysiological properties.Materials and methods: Vestibular nucleus neurons were mechanically and enzymatically isolated and cultured using a defined medium with known growth factors. Cell survival was measured with propidium iodide, and electrophysiological properties were investigated with current-clamp recording.Results: Vestibular neurons grew neurites in cultures, gaining adult-like morphological properties, and stayed viable for 3 days in culture. Adding bovine calf serum, nerve growth factor, or insulin-like growth factor into the culture medium enhanced neuronal viability. Current-clamp recording of the cultured neurons revealed tonic and phasic-type neurons with similar input resistance, resting membrane potential, action potential amplitude, and duration. Conclusion: Vestibular neurons from adult mice can be cultured, and regenerate axons in a medium containing appropriate growth factors. Culturing adult vestibular neurons provides a new method to study age-related pathologies of the vestibular system.

The Effects of Phrenic Nerve Degeneration by Axotomy and Crush on the Electrical Activities of Diaphragm Muscles of Rats.

The aim of this study was to investigate the effect of axotomy and crush-related degeneration on the electrical activities of diaphragm muscle strips of experimental rats. In the present study, twenty-one male Wistar-albino rats were used and divided into three groups. The animals in the first group were not crushed or axotomized and served as controls. Phrenic nerves of the rats in the second and third groups were crushed or axotomized in the diaphragm muscle. Resting membrane potential (RMP) was decreased significantly in both crush and axotomy of diaphragm muscle strips of experimental rats (p < 0.05). Depolarization time (T DEP) and half-repolarization (1/2 RT) time were significantly prolonged in crush and axotomy rats (p < 0.05). Crushing or axotomizing the phrenic nerves may produce electrical activities in the diaphragm muscle of the rat by depolarization time and half-repolarization time prolonged in crush and axotomy rats.

Smoking-related Alterations in Serum Levels of Thyroid Hormones and Insulin in Female and Male Students.

CONTEXT: Cigarette smoking has large-scale and complex effects on the endocrine system. Various studies related to cigarette smoking have provided differing results. Therefore, more research is needed to determine the effects on the body that are created by cigarette smoking. OBJECTIVES: The study was designed to investigate the effects of cigarette smoking, primarily on thyroid hormones in serum, such as on levels of total triiodothyronine (tT3), free triiodothyronine (fT3), total thyroxine (tT4), free thyroxine (fT4), thyroid-stimulating hormone (TSH) (ie, thyrotropin), and insulin of young students aged 18-25 y. DESIGN: This study was a randomized, controlled trial. SETTING: The study was performed in the Department of Physiology, School of Medicine, Yuzuncu Yil University (Van, Turkey). PARTICIPANTS: Eighty healthy students, 40 females and 40 males, were included in the study. INTERVENTION: Of the 40 female participants, 25 were smokers, and 15 were nonsmokers. Of the 40 male participants, 25 were smokers, and 15 were nonsmokers. The intervention (smoking) group, therefore, consisted of 50 participants, and the control (nonsmoking) group consisted of 30 participants. OUTCOME MEASURES: Serum concentrations of thyroid hormones and insulin were determined by enzyme-linked immunesorbent assays (ELISAs), using monoclonal antibodies; and by measurement of blood glucose, using a glucometer. RESULTS: The study found that both female and male smokers had higher levels of serum tT3 and insulin hormone than nonsmokers had. A positive correlation was found between age and insulin resistance in male smokers. The study also found that male smokers had higher levels of serum tT3 and fT4 hormone than female smokers had. CONCLUSIONS: Smoking may be associated with an increased secretion of thyroid hormones and the development of insulin resistance. With aging, insulin resistance may increase more in male smokers than in female smokers.

Investigation of the hepatoprotective effects of Sesame (Sesamum indicum L.) in carbon tetrachloride-induced liver toxicity.

More than 600 chemicals can cause damage in liver, one of which is carbon tetrachloride (CCl4). Hepatoprotective agents could prevent tissue damage and reduce morbidity and mortality rates; such agents may include alternative or folkloric treatments. We investigated sesame (Sesamum indicum L.) for its hepatoprotective effect in CCl4-induced experimental liver damage. To this end, 0.8 mg/kg of sesame fixed oil was provided intraperitoneally to rats whose livers were damaged by CCl4. Tissue and blood samples were taken at the end of the experiments and evaluated histologically and biochemically. Ballooning degenerations and an increase in lipid droplets in liver parenchyma and increases in serum alanine transaminase, aspartate transaminase, and bilirubin were found in the CCl4 group. Biochemical and histopathological findings in the sesame fixed oil treated group were not significantly different from the CCl4 group. Sesame did not show a hepatoprotective effect in CCl4-induced liver toxicity.

The susceptibility of erythrocytes to oxidation during storage of blood: effects of melatonin and propofol.

OBJECTIVES: We investigated the effects of melatonin and propofol in lipid peroxidation and antioxidant capacity of erythrocytes in stored bloods. DESIGN AND METHODS: Donated blood was taken into three citrate-phosphate-dextrose containing blood bags. One bag was used as control, the others were added either melatonin or propofol. Erythrocyte lipid peroxidation and antioxidant capacity and their sensitivity to in vitro oxidation were measured on days 0, 7, 14, 21 and 28. RESULTS: In control group, erythrocyte malondialdehyde levels and sensitivity to in vitro oxidation were increased whereas glutathione, glutathione peroxidase and superoxide dismutase levels were decreased. Melatonin prevented malondialdehyde accumulation and preserved glutathione, glutathione peroxidase and superoxide dismutase levels. Propofol preserved glutathione and glutathione peroxidase levels but did not affect catalase and superoxide dismutase activities. CONCLUSIONS: We showed that melatonin in stored blood could prevent lipid peroxidation and increase the resistance of erythrocytes to in vitro oxidation while propofol did not show such effects.

Consequences of neurite transection in vitro.

In order to quantify degenerative and regenerative changes and analyze the contribution of multiple factors to the outcome after neurite transection, we cultured adult mouse dorsal root ganglion neurons, and with a precise laser beam, we transected the nerve fibers they extended. Cell preparations were continuously visualized for 24 h with time-lapse microscopy. More distal cuts caused a more elongated field of degeneration, while thicker neurites degenerated faster than thinner ones. Transected neurites degenerated more if the uncut neurites of the same neuron simultaneously degenerated. If any of these uncut processes regenerated, the transected neurites underwent less degeneration. Regeneration of neurites was limited to distal cuts. Unipolar neurons had shorter regeneration than multipolar ones. Branching slowed the regenerative process, while simultaneous degeneration of uncut neurites increased it. Proximal lesions, small neuronal size, and extensive and rapid neurite degeneration were predictive of death of an injured neuron, which typically displayed necrotic rather than apoptotic form. In conclusion, this in vitro model proved useful in unmasking many new aspects and correlates of mechanically-induced neurite injury.

Glutamate responsiveness of medial vestibular nucleus neurons in aged rats.

Disequilibrium, dizziness, vertigo and falls are vestibular system-related problems which are very common especially in older people. In order to clarify these age-related disorders one must understand first the age-related changes in the properties of vestibular neurons that are responsible for equilibrium. The responsiveness of medial vestibular nucleus (MVN) neurons to the NMDA and AMPA/kainate receptor agonists was investigated in slices prepared from young and aged rats using extracellular single cell recording techniques. In both young and aged rats bath application of NMDA and AMPA caused a reversible, dose-dependant increase in the spontaneous discharge of the MVN neurons. The excitatory effects of both NMDA and AMPA on the spontaneous activity of aged MVN neurons were similar to those of young MVN neurons. The spontaneous firing rates of the MVN cells were also similar in young and aged rats. These results suggest that the responsiveness of the NMDA and AMPA/kainate receptors and the excitability of the MVN neurons do not change with age.