X-Message-Number: 27621 Date: Wed, 15 Feb 2006 20:31:13 -0800 (PST) From: Doug Skrecky <> Subject: nicotine for vitrification solutions? [Complete block of ethanol effects was achieved by the highest dose of nicotine (20 microM).] Neurotox Res. 2005;7(4):319-22. Nicotine blocks ethanol-induced apoptosis in primary cultures of rat cerebral cortical and cerebellar granule cells. Nicotine's counteraction of adverse effects of ethanol on cognitive function and motor coordination may play a major role in the observed high incidence of smoking among alcoholics. Previously, we have observed protective effects of nicotine against ethanol-induced neurotoxicity in cultured cortical and cerebellar granule cells as determined by lactate dehydrogenase assay. Ethanol-induced apoptosis may be a contributory mechanism to its neuronal toxicity. In this study we sought to determine whether ethanol induces formation of caspase 3 (reflective of apoptosis) in these cells and whether these effects may be blocked by nicotine pretreatment. Primary cultures of cerebral cortical and cerebellar granule cells were prepared from the brains of 20 day old Sprague-Dawley fetuses. Exposure of cells to ethanol (10-100 mM) for 3 days resulted in a dose-dependent increase in caspase 3 activity and cytotoxicity. Pretreatment with nicotine (5-20 microM) dose dependently attenuated these effects of ethanol. Complete block of ethanol effects was achieved by the highest dose of nicotine (20 microM). Nicotine, at concentrations administered, did not affect caspase activity or neuronal viability. These results suggest that at least some of the neurotoxic effects of ethanol may be mediated by apoptosis and that pretreatment with nicotine can prevent these effects of ethanol. Anti-apoptotic effects of nicotine in this model may be suggestive of potential use of nicotinic agonists in neurotoxic insults and/or neurodegenerative disorders. Neurotox Res. 2004;6(4):311-6. Nicotine inhibits ethanol-induced toxicity in cultured cerebral cortical cells. The high incidence of smoking among alcoholics may be partially due to nicotine's ability to counteract some of the adverse effects of ethanol on motor coordination and/or cognitive functions. Neuroprotective effects of nicotine on ethanol-induced toxicity in cerebellar granular cells have been observed. In this study, we sought to determine whether similar protection is observed in neocortical cells and if so, what specific nicotinic receptor subtypes may be mediating the actions of nicotine. Primary cultures of neocortical cells were prepared from 20-day embryos obtained from time-pregnant Sprague-Dawley rats. Cells were cultured for 10 days and were then exposed for 3 days to various concentrations of ethanol with and without pretreatment with nicotine and nicotinic antagonists. Cellular toxicity was evaluated by measuring the lactate dehydrogenase level. Administration of ethanol (10-100 mM) resulted in a dose-dependent toxicity. Pretreatment with nicotine 5-20 microM resulted in a dose-dependent protection against ethanol-induced toxicity. The effects of nicotine were blocked by pretreatment with nicotinic antagonists such as mecamylamine (1-20 microM), dihydro-beta-erythroidine (DHBE) 50 nM-1.0 microM and methyllycaconitine (MLA) 5 nM-1 microM in a dose-dependent manner. Compared to previous studies, higher ethanol concentrations were required to induce toxicity in neocortical vs cerebellar granule cells. Moreover, the effects of nicotine in the neocortical cells were blocked by lower concentrations of MLA, but higher concentrations of DHBE compared to cerebellar cells. Collectively, the results suggest differential sensitivity of various neuronal populations to the toxic effect of ethanol. Furthermore, protective effects of nicotine against alcohol in various regions appear to be mediated by different nicotinic receptor subtypes. The neuroprotective effect of nicotine against ethanol-induced toxicity may be a contributing factor to the high incidence of smoking among alcoholics. Alcohol Clin Exp Res. 2000 Oct;24(10):1583-92. Chronic, but not acute, nicotine exposure attenuates ethanol withdrawal-induced hippocampal damage in vitro. BACKGROUND: Long-term ethanol use and long-term tobacco use frequently occur together, which suggests concurrent dependence on ethanol and nicotine. Consequences of this form of polydrug dependence are not well understood, however. Previous evidence suggests detrimental effects of long-term ethanol and beneficial effects of nicotine exposure on neuronal viability. Thus, the present study was designed to use an organotypic hippocampal slice culture model to examine the ability of chronic and acute nicotine exposure to reduce neurotoxicity associated with withdrawal from long-term ethanol exposure. METHODS AND RESULTS: Twenty-four hours of withdrawal after continuous 10 day ethanol exposure (50 or 100 mM in culture medium) resulted in cytotoxicity in hippocampal slice explants obtained from neonatal rat, most notably in pyramidal cell layers of the CA1 region. Exposure of slices to the N-methyl-D-aspartate receptor blocker MK-801 during ethanol withdrawal significantly reduced this toxicity. Exposure of slices to nicotine (0.1-10.0 microM) during the 24 hr withdrawal period did not reduce hippocampal damage. However, treatment of slices with nicotine (0.1-10.0 microM) during 10 days of ethanol exposure was associated with significant reductions in subsequent withdrawal-induced cytotoxicity, an effect reduced by mecamylamine coexposure with nicotine and ethanol. CONCLUSIONS: These findings indicate the development of marked hippocampal neurotoxicity during withdrawal from long-term ethanol exposure that is mediated, in part, by overactivation of N-methyl-D-aspartate receptors. Furthermore, these data suggest that one consequence of concurrent dependence on ethanol and nicotine may be reduced neurological damage associated with ethanol withdrawal. Dig Dis Sci. 1990 Jan;35(1):106-12. The influence of acute or chronic nicotine treatment on ethanol-induced gastric mucosal damage in rats. The influences of acute or chronic nicotine pretreatment on ethanol-induced changes on gastric secretion, mucosal blood flow (GMBF), and glandular mucosal damage were studied in anesthetized rats. Ethanol administration decreased gastric acid secretion and GMBF, which were accompanied by a marked increase in gastric mucosal damage. Acute nicotine incubation 2 or 4 mg dose-dependently elevated both the titratable acid in the luminal solution and the gastric secretory volume; it also prevented the depressive action on GMBF and gastric mucosal damage in ethanol-treated animals. Chronic nicotine treatment for 10 days reduced the inhibitory action of ethanol on gastric acid secretion; the higher dose (25 micrograms/ml drinking water) potentiated the decrease of GMBF and the ulcerogenic property of ethanol. However, chronic treatment with the lower dose (5 micrograms/ml drinking water) had the opposite effects; it also markedly increased the gastric secretory volume. It is concluded that acute nicotine pretreatment elevates, whereas chronic nicotine pretreatment differentially affects GMBF. These effects could account for their protective or preventive actions on ethanol ulceration. The increase in nonacid gastric secretory volume by nicotine could partially explain its antiulcer effect. Furthermore, the acid secretory state of the stomach appears unrelated to the ulcerogenic property of ethanol. Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=27621