[' ', '0124052', 'PRINCIPAL', 'INVESTIGATOR:', 'HILLARD,', 'CECILIA', 'J', 'ADDRESS:', 'MEDICAL', 'COLLEGE', 'OF', 'WISCONSIN', '@701', 'WATERTOWN', 'PLANK', 'RD', 'MILWAUKEE,', 'WI', '53226', 'PERFORMING', 'ORG.;', 'MEDICAL', 'COLLEGE', 'OF', 'WISCONSIN,', 'MILWAUKEE,', 'WISCONSIN', 'SPONSORING', 'ORG.:', 'NATIONAL', 'INSTITUTE', 'ON', 'DRUG', 'ABUSE', 'FY', ':', '94', 'FONDS:', '$177,002', 'TYPE', 'OF', 'AWARD:', 'Competing', 'Continuation', '(Type', '2)', 'SOMMARY:', 'Although', 'cigarette', 'smoking', 'continue', 'to', 'be', 'the', 'most', 'popular', 'and', 'persistent', 'method', 'of', 'recreational', 'drug', 'use', 'in', 'the', 'U.S.,', 'relatively', 'little', 'is', 'known', 'about', 'the', 'neurochemical', 'effects', 'of', 'nicotine', 'in', 'the', 'brain', 'or', 'about', 'the', 'neurochemical', 'consequences', 'of', 'chronic', 'nicotine', 'exposure.', 'The', 'overall', 'hypothesis', 'to', 'be', 'tested', 'by', 'the', 'studies', 'outlined', 'in', 'this', 'proposal', 'is', 'that', 'the', 'functioning', 'of', 'the', 'nicotinic', 'cholinergic', 'receptor', '(nAChR)', 'in', 'the', 'CNS', 'of', 'the', 'rat', 'is', 'altered', 'by', 'chronic', 'exposure', 'of', 'the', 'animal', 'to', 'nicotine.', 'We', 'will', 'carry', 'out', 'studies', 'to', 'explore', 'this', 'hypothesis', 'and', 'to', 'determine', 'the', 'mechanism(s)', 'involved', 'in', 'the', 'effects', 'of', 'nicotine', 'that', 'we', 'identify.', 'The', 'specific', 'aims', 'of', 'the', 'studies', 'outlined', 'in', 'this', 'proposal', 'are', '(1)', 'to', 'explore', 'using', 'neurochemical', 'methods,', 'the', 'ionic', 'events', 'that', 'follow', 'nAChR', 'activation', 'in', 'synaptosomes;', '(2)', 'to', 'determine', 'whether', 'the', 'number', 'of', 'nAChR', 'is', 'changed', 'following', 'chronic', 'exposure', 'of', 'rats', 'to', 'nicotine', 'using', 'an', 'approach', 'that', 'does', 'not', 'rely', 'on', 'agonist', 'binding', 'to', 'the', 'receptor;', '(3)', 'to', 'determine', 'whether', 'the', 'functional', 'status', 'of', 'the', 'nAChR', 'ionophore', 'is', 'altered', 'following', 'chronic', 'treatment', 'with', 'nicotine;', '(4)', 'to', 'explore', 'the', 'mechanism', 'involved', 'in', 'the', 'effecta', 'of', 'in', 'vivo', 'nicotine', 'on', 'synaptosomal', 'membrane', 'potential;', '(5)', 'to', 'explore', 'the', 'effects', 'of', 'chronic', 'nicotine', 'on', 'synaptosomal', 'calcium', 'influx', '(6)', 'to', 'separate', 'nAChR', 'containing', 'synaptoscme', 'and', 'determine', 'whether', 'they', 'are', 'selectively', 'affected', 'by', 'nicotine', 'exposure', 'or', 'if', 'a', 'large', 'pool', 'of', 'synaptosome', 'will', 'be', 'loaded', 'with', 'PTSA', 'and', 'the', 'entry', 'of', 'cations', 'in', 'response', 'to', 'nicotine', 'and', 'other', 'agonist', 'will', 'be', 'measured', 'as', 'a', 'decrease', 'in', 'fluorescence', 'in', 'the', 'presence', 'of', 'thallium.', 'We', 'will', 'explore', 'the', 'effects', 'of', 'nAChR', 'activation', 'on', 'intrasynaptosomal', 'free', 'calcium', 'concentrations', 'using', 'either', 'Fura-2', 'and', 'conventional', 'spectrofluorometric', 'approaches', 'or', 'Indo-1', 'with', 'flow', 'cytometry.', 'Our', 'method', 'for', 'labeling,', 'quantifying', 'and', 'sorting', 'synaptosomes', 'with', 'nAchR', 'will', 'involva', 'coupling', 'a', 'fluorochrome', 'to', 'a', 'commercially', 'available', 'antibody', 'that', 'recognizes', 'the', 'Beta?', 'subunit', 'of', 'the', 'CNS', 'nAChR', '(mAb270).', 'We', 'will', 'use', 'the', 'technique', 'of', 'flow', 'cytometry', 'coupled', 'wit', 'fluorescence', 'activated', 'cell', 'sorting', 'to', 'determine', 'the', 'concentration', 'of', 'antibody', '(and', 'hence', 'the', 'concentration', 'of', 'nAChR)', 'in', 'synapto-somes', 'fram', 'nicotine', 'exposed', 'animals.', 'Furthermore,', 'we', 'will', 'use', 'the', 'cell', 'sorting', 'capabilities', 'of', 'the', 'flow', 'cytometer', 'to', 'isolate', 'a', 'synaptosomal', 'fraction', 'that', 'is', 'enriched', 'in', 'nACHR-contalning', 'terminals.', 'tt', 'is', 'anticipated', 'that', 'the', 'successful', 'completion', 'of', 'these', 'studies', 'will', 'enhance', 'our', 'understanding', 'of', 'the', 'mechanism', 'ef', 'action', 'of', 'nicotine,', 'give', 'insight', 'into', 'the', 'compensatory', 'mechanisms', 'inyolved', 'in', 'the', 'development', 'of', 'tolerance', 'and', 'provide', 'general', 'information', 'regarding', 'regulatory', 'proceases', 'that', 'operate', 'in', 'the', 'nerve', 'terminal.', '6/7/466', 'DIALOG', '(R)File', '266:Fed.', 'Rea.', 'in', 'Progress', '{c)', 'format', 'only', '1995', 'Dialog', 'Info.Svcs.', 'All', 'rts.', 'reserv.', '0224021', 'IDENTIFYING', 'NO.:', 'SRO1DA04208-06', 'AGENCY', 'CODE:', 'CRISP', 'MECHANISM', 'OF', 'CIGARETTE', 'SMOKE-INDUCED', 'IMMUNOSUPPRESSION', 'PRINCIPAL', 'INVESTIGATOR:', 'SOPORI,', 'MOHAN', 'L', 'ADDRESS:', 'LOVELACE', 'MEDICAL', 'FOUNDATION', '2425', 'RIDGECREST', 'DR,', 'SE', 'ALBUQUERQUE,', 'NM', '87108-5127', 'PERFORMING', 'ORG.:', 'LOVELACE', 'INSTITUTES,', 'ALBUQUERQUE,', 'NEW', 'MEXICO', 'SPONSORING', 'ORG.:', 'NATIONAL', 'INSTITUTE', 'ON', 'DRUG', 'ABUSE', 'Fr', ':', '94', 'FUNDS:', '$157,513', 'TYPE', 'OF', 'AWARD:', 'Noncompeting', 'Continuation', '(Type', '5)', 'SUMMARY:', 'Cigarette', 'smoking', 'is', 'associated', 'with', 'elevated', 'risks', 'of', 'upper', 'vespiratory', 'tract', 'infections,', 'cancers', 'of', 'several', 'organs,', 'and', 'cardiovascular', '§114@', '2814', '+', ' ', ' ']