Sex differences in modulating blood brain barrier permeability by NO in pentylenetetrazol-induced epileptic seizures.

Susceptibility to epilepsy as well as BBB dysfunction in some pathological conditions varies depending on sex difference. It has recently been shown that systemically given NO donor and antagonists modify the nature of seizures induced by PTZ (pentylenetetrazol) differently in male and female rats. This study investigates the role of NO on BBB permeability in PTZ seizures with sex differences using NO donor, sodium nitroprusside (SNP), and NOS inhibitor, NG-nitro-L-arginine methyl ester (L-NAME). Nitrite+nitrate levels as indices of NO generation in the brain were also assessed. L-NAME prolonged seizure latency in male rats, seizure intensity and seizure duration were lessened. L-NAME depicted opposite effects in seizure nature in female rats. SNP prolonged seizure latency, while seizure intensity and duration were lessened only in female rats. L-NAME in male rats increased L-NAME use in female rats (not in male rats) which resulted in a more leaky BBB especially in midbrain, thalamus, hippocampus, corpus striatum and cerebellum whereas SNP use in male rats and not in female rats resulted in pronounced BBB opening in all brain regions studied than PTZ per se. L-NAME while decreasing nitrite+nitrate levels in male rat brains, acted in an opposite fashion in females. SNP use depicted an inverse picture with respect to L-NAME, with an opposite action in different sexes. This study reveals that NO effect on BBB in PTZ-induced seizures depends unequivocally on sex difference. The sex-dependent action of NO in seizures and in CNS pathologies warrants further investigation.

The relationship between erythropoietin pretreatment with blood-brain barrier and lipid peroxidation after ischemia/reperfusion in rats.

Blood-brain barrier (BBB) leakage plays a role in the pathogenesis of many pathological states of the brain including ischemia and some neurodegenerative disorders. In recent years, erythropoietin (EPO) has been shown to exert neuroprotection in many pathological conditions including ischemia in the brain. This study aimed to investigate the effects of EPO on BBB integrity, infarct size and lipid peroxidation following global brain ischemia/reperfusion in rats. Wistar male rats were divided into four groups (each group n=8); Group I; control group (sham-operated), Group II; ischemia/reperfusion group, Group III; EPO treated group (24 h before decapitation--000 U/kg r-Hu EPO i.p.), Group IV; EPO+ ischemia/reperfusion group (24 h before ischemia/reperfusion--3000 U/kg r-Hu EPO i.p.). Global brain ischemia was produced by the combination of bilateral common carotid arteries occlusion and hemorrhagic hypotension. Macroscopical and spectrophotometrical measurement of Evans Blue (EB) leakage was observed for BBB integrity. Infarct size was calculated based on 2,3,5-triphenyltetrazolium chloride (TTC) staining. Lipid peroxidation in the brain tissue was determined as the concentration of thiobarbituric acid-reactive substances (TBARS) for each group. Ischemic insult caused bilateral and regional BBB breakdown (hippocampus, cortex, corpus striatum, midbrain, brain stem and thalamus). EPO pretreatment reduced BBB disruption, infarct size and lipid peroxide levels in brain tissue with 20 min ischemia and 20 min reperfusion. These results suggest that EPO plays an important role in protecting against brain ischemia/reperfusion through inhibiting lipid peroxidation and decreasing BBB disruption.

Evidence for ascorbic acid transport system in rat brain capillaries.

Although ascorbic acid (AA) crosses the choroid plexus and may enter the brain at an appreciable rate, it is not clearly established that there exist transport system(s) carrying this vitamin from blood into the brain cells across the brain capillaries. Thus the rate of its uptake by choroid plexus and cerebral capillaries were evaluated in vitro in this study. Choroid plexus and brain capillaries were isolated from two-month-old male Sprague-Dawley rats. Time course of AA incorporation in micro vessels and choroid plexus was studied up to 30 min. After stopping the incorporation with the excess of cold isotonic saline, micro vessels were filtered and sonicated. The intracellular incorporated AA radioactivity was measured by liquid scintillation counting. AA uptake by micro vessel was tested for Na+-dependence and saturability. The time course studies showed linear increase in total uptake and accumulation of AA by choroid plexus and endothelial cells up to 30 min. Treatment with oubain or replacement with sodium chloride showed that uptake is an Na+- independent process. Transport of AA to cerebrospinal fluid and brain was also shown to be readily saturated by increasing the level of cold AA. These results document that the brain capillary endothelial cells are able to transport and accumulate AA, and may have a critical role in the homeostasis and regulation of cerebral ascorbic acid concentration.

Nitric oxide involvement in seizures elicited by pentylentetrazol and sex dependence.

It has been known that susceptibility to some types of epilepsy is affected by sex. In addition, the role of NO in epileptogenesis is still unclear; NO has been suggested to be either an anticonvulsive or a proconvulsive agent. In an attempt to elucidate both the role of NO and sex differences in sensitivity to seizures, male and female Wistar rats were treated intraperitoneally (i.p.) by pentylentetrazol (PTZ)(80 mg/kg) and by a nitric oxide synthase(NOS) inhibitor N-omega-nitro-L-arginine-mthylester(L-NAME)(50mg/kg) and a NO precursor sodium-nitroprusside(SNP)(2.5mg/kg)- applied 15 min. before PTZ injection. Latency, frequency, severity, and duration of generalized clonic and clonic-tonic convulsions were recorded. Furthermore, alterations in severity, latency, frequency, and duration of convulsions were observed to correlate with NO. Both sexes, injected with PTZ, showed repetitive seizure patterns. Seizures were found to be more severe in females. L-NAME and SNP pretreatment produced paradoxical effects on PTZ-induced seizures in both sexes. L-NAME completely prevented PTZ-induced seizures in male rats, whereas increased severity, frequency, duration, and significantly shortened the latency in female rats. Unexpectedly, SNP increased convulsion severity, frequency, duration, and shortened latencies in male, whereas it decreased convulsion severity, frequency, and duration and prolonged latency in females. These results indicate that endogenous NO is involved in the regulation of convulsive action suggesting a role depending on sex.

Nicotine improves learning and memory in rats: morphological evidence for acetylcholine involvement.

It has been suggested that nicotine improves rapid information processing (learning and memory) tasks. However, it is not clear which aspects of cognition actually underlie these improvements because relatively less attention has been given to nicotinic cholinergic systems compared to muscarinic systems. The authors therefore studied the effects of nicotine on the learning and memory performance by a step-through passive avoidance task. Nicotine (0.4 mg/kg) was administered s.c. single dose (acute group), once a day for 3 days (subchronic group) or 21 days (chronic group). Nicotine treated and control rats were trained in one trial learning step-through passive avoidance task, where retention latencies were carried out 1 h, 24 h, and 3 days after learning trial. Treatment with nicotine before training session prolonged the latencies significantly (p < .01). Control group, acute, subacute and chronic nicotine treatment groups showed latencies 4.75 +/- 0.6, 69.4 +/- 14, 116.2 +/- 30, and 118.5 +/- 23 s, respectively. In addition, to prove the actual contribution of nicotinic cholinergic system in improvement of learning and memory processing, histological methods that permit the visualization and quantification of ACh levels were used. Electron microscopic evaluation revealed increased numbers of Ach-containing vesicles especially in hippocampus in chronic nicotine-treated rats; although frontal and temporal cortex in addition to hippocampus showed increment in Ach vesicles in a lesser extent in all nicotine treatment groups. These results indicate that long-term nicotine treatment can be important for improving cognitive function in regard to increased cholinergic activity.

Tolerance to pentylentetrazol-induced convulsions and protection of cerebrovascular integrity by chronic nicotine.

The authors' previous studies have shown that in nicotine-induced seizures sensitivity was decreased and blood-brain barrier (BBB) disruption was prevented as a consequence of nicotine pretreatment. This study aimed to investigate the possible protective actions of nicotine on cerebrovascular permeability and seizures induced by pentylentetrazol (PTZ) injection. Cerebrovascular effects of nicotine were evaluated by measuring the permeability changes of BBB using Evans-Blue (EB) dye and specific gravity (SG), which indicates brain water and protein content. The experiments were carried out on Wistar rats. Animals were randomly divided into two groups. Convulsions were induced by injection of PTZ (80 mg/kg i.v.) in rats either pretreated with nicotine daily with a low dose of 0.8 mg/kg day for 21 days or injected with a single dose of 6 mg/kg mecamylamine. The same procedures were followed in control rats with the exception that they were injected only with saline. PTZ injection caused tonic-clonic convulsions and increased the EB dye leakage and specific gravity values in saline-injected control rat brains. Daily injection of nicotine lessened the intensity of seizures. These were accompanied by marked decreases in both the leakage of EB and brain water content. Acute administration of a nAChR antagonist mecamylamine significantly increased seizure latency and decreased the duration of seizures. Thereby, mecamylamine reduced the EB leakage and water content in most brain regions. These results indicate that development of tolerance to PTZ convulsions can be produced by chronic nicotine administration in rats. The mechanism for this effect currently needs clarification. Moreover, the data also suggest that cholinergic activity may account for occurrence of PTZ-induced convulsions.