Ranjan Bose

Effects of excitotoxins on free radical indices in mouse brain

Excitotoxins and free radicals individually have been implicated in several neurological disorders including those associated with aging. We observed that systemically administered domoic acid enhanced mouse brain superoxide dismutase activity with either an associated decrease or no change in mouse brain lipid peroxidation. These findings reflect a state of adequately compensated oxidative stress induced by excitotoxins. In homogenates containing disrupted cells from various regions of mouse brain, however, kainic acid produced a 2 to 5-fold increase in lipid peroxidation. This suggests that excitotoxins cause lipid peroxidation possibly by acting at intracellular loci which become more accessible following disruption of cells in vitro and by extrapolation, possibly in vivo due to cellular permeability changes during the edematous stage of ischemic and other related neuropathological conditions.

Effect of mannitol, nimodipine, and indomethacin singly or in combination on cerebral ischemia in rats.

The effects of mannitol, nimodipine, and indomethacin on ischemic neuronal injury were examined in 45 rats divided equally into nine groups subjected to 10 minutes of forebrain ischemia. Of two control groups, one received maintenance fluids while the other received a normal saline bolus. In the remaining seven groups, mannitol, nimodipine, and indomethacin were administered singly or in combination 5 minutes before forebrain ischemia. Seven days after ischemia, the brains were perfusion-fixed, sectioned coronally into 2.8-mm slices, and stained with hematoxylin and eosin. Ischemic neurons were directly counted on predetermined regions of standardized serial sections. Considerable amelioration of ischemic injury (ischemic neurons/total neurons) was observed with mannitol (ischemic injury, 7 +/- 5% in the hippocampal CA1/CA2 sectors and 28 +/- 17% in the CA3 sector). This is in contrast to control values of 64 +/- 11% and 80 +/- 6%, respectively, and those obtained in the normal saline group of 70 +/- 10% and 59 +/- 13%, respectively. The beneficial effect with nimodipine reached significance in only the hippocampal CA3 sector (ischemic injury, 35 +/- 21%). Indomethacin showed no significant benefit. Combining the agents resulted in significantly reduced neuronal injury compared with control groups, although the effect was not greater than that achieved with mannitol alone. The degree of ischemic injury was least when all three agents were used in combination (ischemic injury, 12 +/- 12% in the hippocampal CA1/CA2 sectors and 4 +/- 4% in the CA3 sector). Our data support the concept that successfully blocking the ischemic cascade with a single, diversely acting agent or multiple agents will evoke the best beneficial response.

Biological and methodological implications of prostaglandin involvement in mouse brain lipid peroxidation measurements

Enhanced cyclooxygenase-mediated prostaglandin (PG) turnover occurring during sacrifice and biochemical processing of tissues also generates malondialdehyde (MDA), a product of lipid peroxidation (LPO). Studies reporting on LPO estimated by thiobarbituric acid reactive substances (TBARS) have failed to consider such artefactual increases. This study reports the relative proportion of PG metabolism-derived MDA (PG-MDA) in mouse brain regions during the TBARS assay. The cyclooxygenase inhibitor indomethacin significantly lowered MDA in fronto-parietal cortex and corpus striatum. Indomethacin (50–800 μg/ml, in vitro) increased estimated TBARS in whole brain. Such enhancement was absent when indomethacin (20–80 μg/sample) was added to the MDA standard curve, reflecting its interaction with TBARS other than MDA. PG-MDA contributes as much as 15% to the total estimated value of MDA in fronto-parietal cortex and corpus striatum and must be corrected for in LPO studies.

Parenteral domoic acid impairs spatial learning in mice

The present study is the first to examine the effect of a single intraperitoneal injection of the neuroexcitotoxin domoic acid on learning in mice. Compared to saline controls, animals exposed to domoic acid (2.0 mg/kg) showed significant impairment on the acquisition of the place task in the Morris water maze. Observation of swim paths taken by mice searching for the underwater platform revealed a failure on the part of the domoic acid-exposed mice to select the appropriate problem solving strategies. The results, along with neuroanatomic work done here and elsewhere, suggest that impairment of acquisition and retention of this spatial navigation task by domoic acid, involves a neuropathology that includes not only the hippocampus, but other limbic, and possibly extralimbic brain regions.

Circumventricular organ origin of domoic acid-induced neuropathology and toxicology

The neuroexcitotoxin, domoic acid, was responsible for an episode of mussel poisoning in Eastern Canada in 1987. Severe neurologic impairment and some deaths occurred. We have characterized the nature of domoate-induced neuropathology in the mouse brain. Domoic acid was administered intraperitoneally at doses of 2, 3 or 7 mg/kg to Swiss-Webster mice. Brains were examined at 0.5, 1, 24, 48 or 72 h postinjection for evidence of damage. Significant pathologic changes occurred only after the largest dose of domoic acid. Damage was confined to circumventricular organs lacking a blood-brain barrier and their environs, including the organon vasculosum of the lamina terminalis, subfornical organ, mediobasal hypothalamus and area postrema. The neural damage induced by domoic acid was evident at as early as 30 min after injection and increased by 60 min postinjection. The loci of domoic acid-induced neuropathological changes accounts for several central and peripheral effects and toxicities observed following systemic domoate treatment, these included gastroduodenal lesions, hypodipsia, analgesia, and blood pressure fluctuations.

Kynurenic acid protects against gastroduodenal ulceration in mice injected with extracts from poisonous atlantic shellfish

1. Mice were treated with an extract prepared from poisonous Atlantic mussels. 2. Gastric and duodenal ulcers, duodenal hyperemia and peritoneal ascites resulted from administration of the shellfish extract, with an LD84 of 1.0 ml. 3. Kynurenic acid, an antagonist at excitatory amino acid receptors, protected significantly against gastroduodenal ulcers, ascites and hyperemia when given at 60 or 75 min post-extract. 4. It is likely that the gastrointestinal damage evoked by this extract is due to its domoic acid content and that kynurenic acid may prove useful against domoic acid-induced gastropathy.

Domoic acid-induced neuro visceral toxic syndrome: Characterization of an animal model and putative antidotes

A rodent model of neurovisceral toxic syndrome induced by the neuroexcitant amino acid, domoic acid, is described, along with the activity of a putative antidote, the nonselective excitotoxin antagonist, kynurenic acid. Both an extract of contaminated mussels and pure domoic acid induced a characteristic syndrome including: sluggishness, scratching stereotypy, convulsions and death. Autopsy revealed gastric and duodenal lesions and peritoneal ascites. Kynurenic acid significantly obtunded these behavioral and physiological effects, particularly when given 60-75 min after the toxic insult. Probenecid, a blocker of organic acid transport, and tryptophan, a precursor of endogenous brain kynurenic acid, increased the time frame in which kynurenic acid exerted its protective effects. Kynurenic acid alone, in nontoxin-challenged animals significantly blocked cold-stress gastric lesions, significantly reduced basal gastric acid secretion and was protective to a lesser degree against ethanol-induced gastric mucosal injury. The murine model of domoate toxicity represents an inexpensive, reliable and sensitive biological assay for screening commercial shellfish for excitotoxin contamination. We are currently exploring kynurenic acid and other compounds for possible therapeutic use in both current and any future victims of neuroexcitant amino acid toxicosis.

Pharmacological and toxicological investigations of Cesium

Cesium, a mineral resource abundantly present in Manitoba with important existing and potential industrial applications was investigated to study its effects on biological systems. Several rodent models of pharmacological activities were utilized. The profile that emerged indicated that cesium is only moderately toxic and exerts salubrious effects which could be gainfully investigated for application in the treatment of certain psychological disorders and some tumors. Its conjunction with existing pharmacological agents for these two types of disorders could yield a pharmacologically active yet less toxic therapeutic combination.

Cesium in mammals: Acute toxicity, organ changes and tissue accumulation

Abstract The acute toxicity of cesium given intraperitoneally (IP) as CsCl in mice is characterized by autonomic upset and by a multiphasic excitant‐depressant action on the central nervous system. The predominantly depressant action can progress to respiratory embarassment, cyanosis, spinal convulsions and death. Light microscopy of mice treated with 2.0 mEq Cs+ kg‐1 IP for 14 days showed lymphoid hyperplasia in small intestine. Measurement of tissue cesium by the proton‐induced x‐ray emission (PIXE) technique is convenient and reproducible. Cesium displays a generalized bioactivity that merits further study.

Kynurenic acid protects against neurotoxicity and lethality of toxic extracts from contaminated atlantic coast mussels

1. An aqueous acidic extract from poisonous Atlantic coast mussels was injected i.p. into groups of Swiss-Webster mice at the LD84 value (= 40 ml of extract/kg). Body-scratching, clonic convulsions and death were consistently observed in response to XTRT injection. These results are consistent with the presumed presence of the neuroexcitant amino acid, domoic acid, in the contaminated mussels. 2. Kynurenic acid at 300 and 600 mg/kg i.p. protected powerfully and significantly against extract-induced neurotoxicity. Greatest protection was observed when the antidote was administered between 30 and 90 min after injection of the toxic extract.