Y. Ziya Ziylan

Changes in the permeability of the blood-brain barrier in acute hyperammonemia. Effect of dexamethasone.

This study was designed to determine the contribution of elevated plasma ammonia levels to blood-brain barrier (BBB) abnormalities in the presence of intact liver. The permeability changes of the BBB were investigated grossly with Evans blue (EB) and quantitatively by measuring the blood-to-brain transfer content for alpha-aminoisobutyric acid (AIB) in normal rats and rats subjected to sublethal doses of ammonium acetate (NH4OAc) (750 and 600 mg/kg ip; at 30-min intervals). Some rats were pretreated with dexamethasone (DXN). Injection of NH4OAc increased both plasma and brain ammonia concentrations about 16-and 5-fold, respectively, above the control level. In rats receiving NH4OAc injection, the blood-to-brain transfer constant (Ki) for AIB was increased 3- to 11-fold. The elevated Ki values were limited to certain gray matter areas and less pronounced permeability changes were detected in white matter. Extravasation sites of EB were more restricted and were especially observed in thalamus and cerebellum, whereas cortex and white matter were unaffected. Dexamethasone pretreatment for 3 d reduced both leakage of EB and the Ki for AIB in NH4OAc injected animals, whereas acute treatment appeared ineffective. Dexamethasone did not prevent the development of coma but slightly decreased the ammonia concentration in plasma and brain. The results obtained indicate that hyperammonemia may disrupt BBB integrity not only to AIB and EB but also enhance the transport of other solutes.

Age-related changes in regional patterns of blood-brain barrier breakdown during epileptiform seizures induced by pentylenetetrazol

Age related changes in blood-brain barrier (BBB) permeability to macromolecules were investigated during seizures induced by pentylenetetrazol in rats aged from 6 to 120 days. Evans blue was used as a visual indicator of BBB integrity. BBB leakage due to seizures was present only in animals in which the mean arterial blood pressure (BP) rose with the seizure onset. Although considerable BBB damage was found partly in similar areas in young and adult rat brains, in adults the leakage of Evans blue was most intense in preoptic area, colliculus inferior, hypothalamus and cerebellum whereas the BBB opening was comparatively rare, in the same areas of young brains. In 6- and 15-day-old rats which did not differ in BP changes from the adults, the leakage was extremely intense in hypothalamus and hippocampus and in contrast to 30- or 120-day-old rats there was no leakage of Evans blue in preoptic area and cerebral cortex. From the results obtained, the conclusion may be drawn that the brain regions which are vulnerable to seizures induced by pentylenetetrazol differ markedly in developing and adult rats. On the other hand, in adult animals, either certain brain areas are more vulnerable to pentylenetetrazol or the BBB has an increased fragility particular to seizure activity. These results indicate that the sensitivity of BBB mechanisms may depend on proliferation of capillaries and changes in their internal structure and may emphasize it.

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.

Pathophysiology of the opening of the blood-brain and blood-cerebrospinal fluid barriers in acute hypertension

Acute hypertension, produced by i.v. Aramine injection, opened the blood-brain and blood-cerebrospinal fluid (CSF) barriers to Evans blue-albumin. In rabbits the threshold for blood-brain barrier (B-BB) opening was approximately 160 mm Hg and for blood--CSF barrier opening 150 mm Hg. The blood-brain and blood-CSF barriers were not opened by blood pressure elevations less than 80 mm Hg. Multiple blue spots (1- to 10-mm diameter) which show Evans blue-albumin extravasation, were seen throughout the cerebral cortex, occasionally in the medulla-pons, and cerebellum. Diffuse extravasation was not seen and the extravasation was nearly symmetrical in the two hemispheres. The barrier permeability was increased when systemic blood pressure was elevated rapidly rather than gradually to the threshold level. Endothelial or epithelial cell destruction was never observed in light and electron microscopic studies. Arterial blood and CSF PCO2, PO2 and pH remained constant, which is indicative of the lack of significant metabolic effect caused by hypertension. Barrier opening in acute hypertension is postulated to be due primarily to the direct mechanical effect of increased intraluminal pressure in cerebral vessels, which may cause widening of the tight junctions between endothelial cells.

PROTECTION OF BLOOD-BRAIN BARRIER BREAKDOWN BY NIFEDIPINE IN ADRENALINE-INDUCED ACUTE HYPERTENSION

The question of whether influxes of ionic Ca+2 into cerebral endothelium plays an important role in increased vascular permeability consequent to an acute hypertension is not accurately resolved. We tested the effect of nifedipine, a calcium entry blocker, on the cerebrovascular permeability for proteins in adrenalin-induced acute hypertension. The experiments were carried out on male Wistar rats. The experimental groups consisted of normotensive saline controls, adrenaline-induced hypertensive rats, and adrenalin-induced hypertensive rats as pre-treated or post-treated with a bolus of nifedipine. Brains of hypertensive rats showed increased permeability to Evans Blue-Albumin complex, when blood pressure elevated rapidly to more than 170 mmHg. The number and size of areas of Evans-Blue extravasation were smaller if an increase in blood pressure was prevented. The short lasting elevation of blood pressure did not result in protein extravasation in brains of hypertensive rats. The results suggest that nifedipine can modify the permeability disruptions observed in acutely hypertensive rats. The data also support the hypothesis that Ca+2 may be responsible for the changes in permeability of BBB in hypertension by mediating the contraction of vascular muscles

Effect of dimethyl sulfoxide on blood-to-brain transfer of α-aminoisobutyric acid: Examination of regional blood-brain barrier function

The effect of dimethyl sulfoxide (DMSO) on brain capillary permeability has been controversial. We have studied the effect of DMSO on unidirectional transport of alpha-aminoisobutyric acid (AIB) across the blood-brain barrier (BBB) in rats. Rats were treated with 15% DMSO intraperitoneally (i.p.), intravenously (i.v.) or by an i.p. injection in combination with an i.v. injection, or in some cases intra-arterially by rapid infusion into left external carotid artery. The unidirectional blood-to-brain transfer constant (Ki) for AIB was measured in each group after the animals were killed. DMSO administration did not significantly increase Ki as compared to control Kj. These results show that it is unlikely that DMSO increases the permeability of BBB and therefore do not support the proposal that DMSO can act as a carrier at the BBB for compounds with restricted vascular permeability.

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.

Hormonal influence on the permeability of the blood-brain barrier: Effect of an analog of adrenocorticotropic hormone, β 1–24 corticotrophin

Regional unidirectional transport of alpha-aminoisobutyric acid (AIB) (mol. wt.: 104) and sucrose (mol wt.: 342) which have a low permeability across the intact endothelium was investigated in brain of rats either treated with synacthène: an analog of ACTH, tetracosactide retard (beta-1-24 corticotrophin) or in brain of placebo-treated controls. Three days treatment with synacthène, reduced the rate of influx of AIB and sucrose in most of the brain regions studied especially in thalamus, hypothalamus, cortex, and caudate nucleus without affecting the vascular compartment. The brainstem, cerebellum and white matter were less affected. These experimental findings may suggest that ACTH exhibits significant influence on hormonal regulation of blood-brain barrier permeability. Thereby such a regulation may involve the entry of polar compounds into the CNS and may influence the central effects of diffusion-limited drugs.

Water and electrolyte content of rabbit brain after opening the blood-brain barrier by acute hypertension

Changes in water and electrolyte content of the brain and edema formation after acute, drug-induced hypertension were studied in albino rabbits. Blood-brain and blood-cerebrospinal fluid (CSF) barriers opened to Evans blue-albumin when systemic blood pressure was elevated abruptly to more than 160 mm Hg by i.v. injection of Aramin. No statistically significant changes in sodium and potassium content of brain, muscle, and CSF were observed. Measurable brain edema did not develop. The results suggest that short-lasting hypertensive barrier opening does not cause brain edema, but may enhance a tendency for brain edema.