['eee', ' ', '3®', 'BOWMAN', 'GRAY', 'SCHOOL', 'OF', 'MEDICINE', 'OF', 'WAKE', 'FOREST', 'UNIVERSITY,', 'PAGE', 'NO.', '2', 'purpose.', 'Drs.', 'Lunt', 'and', 'Wonnocott', 'have', 'indicated', 'their', 'interest', 'in', 'collaborating', 'on', 'examining', 'the', 'expression', 'from', 'mRNA', 'of', 'the', 'various', 'nicotine', 'receptor', 'subunits', 'in', 'the', 'oocyte', 'preparation.', 'These', 'would', 'be', 'extremely', 'interesting', 'studies', 'since,', 'as', 'they', 'indicate,', 'it', 'would', 'be', 'possible', 'to', 'obtain', 'the', 'mRNA', 'from', 'Dr.', 'James', 'Patrick', 'at', 'the', 'Salk', 'Institute,', 'San', 'Diego,', 'and', 'test', 'its', 'expression', 'in', 'the', 'oocyte', 'as', 'to', 'specificity', 'for', 'nicotine', 'receptor', 'binding', 'or', 'other', 'related', 'compounds.', 'This', 'technique', 'is', 'therefore', 'urgently', 'needed', 'in', 'order', 'to', 'functionally', 'classify', 'specific', 'mRNAs', 'as', 'capable', 'of', 'nicotinic', 'receptor', 'expression', 'in', 'the', 'oocyte', 'and', 'as', 'a', 'means', 'of', 'performing', 'in', 'situ', 'hybridization', 'studies', 'to', 'localize', 'critical', 'structural', 'features', 'of', 'the', 'receptor', 'in', 'the', 'mammalian', 'CNS.', 'The', 'above', 'studies', 'will', 'be', 'pursued', 'in', 'addition', 'to', 'others', 'outlined', 'in', 'the', 'original', 'proposal', 'concerned', 'with', 'in', 'vivo', 'experimentation', 'in', 'the', 'freely', 'moving', 'animal.', 'The', 'patch', 'clamp', 'experiments', 'will', 'essentially', 'take', 'the', 'place', 'of', 'the', 'on-going', 'conventional', 'intracellular', 'studies', 'of', 'in', 'vitro', 'hypothalamic', 'slice', 'since', 'much', 'of', 'the', 'same', 'information', 'can', 'now', 'be', 'obtained', 'in', 'the', 'patch', 'clamp', 'paradigm,', 'as', 'well', 'as', 'additional', 'voltage', 'clamp', 'and', 'single', 'channel', 'information', 'which', 'cannot', 'be', 'obtained', 'with', 'conventional', 'intracellular', 'recording', 'procedures.', 'The', 'cost', 'of', 'setting', 'up', 'the', 'facility', 'is', 'a', 'one', 'time', 'outlay', 'of', 'roughly', '$53,715', 'which', 'will', 'cover', 'the', 'costs', 'of', 'the', 'inverted', 'microscope,', 'patch', 'clamp', 'amplifier,', 'interface', 'electronics,', 'and', 'a', 'PC', 'computer', 'to', 'analyze', 'the', 'data', 'which', 'is', 'compatible', 'with', 'the', 'supplied', 'software.', 'These', 'items', 'are', 'listed', 'below', 'and', 'an', 'itemized', 'quote', 'for', 'the', 'microscope', 'is', 'included', 'as', 'an', 'appendix', 'to', 'this', 'letter.', 'The', 'personnel', 'costs', 'can', 'be', 'handled', 'in', 'much', 'the', 'same', 'manner', 'as', 'before', 'and', 'will', 'not', 'require', 'additional', 'support.', 'Nikon', 'Inverted', 'Microscope', '(Diaphot)', '$23,815', '(Includes', 'Epi-Fluorescence,', 'Namarski', 'Optics,', 'associated', 'stands', 'and', 'attach-', 'ments', 'for', 'micromanipulators)', '.', 'Nikon/Narishige', 'N-T', 'Micromanipulator', '13,898', 'system', '(quote', 'for', 'two', 'systems).', 'Patch-Clamp', 'Amplifier', '(Axon', 'Inst.)', '6,000', 'Axo-Lab', 'Accessories', '(A/D', 'converter,', '6,000', 'Interface', 'for', 'PC', '1/0', 'board,', 'P-clamp', 'software,', 'Axon', 'Inst.).', 'AT/PC', 'computer', 'for', 'running', 'P-clamp', '4,000', 'software.', 'Total', '$53,713', 'gi', 'o', 'Q)', 'w', 'wo', 'Q', 'Lie)', 'By', '16)', ' ']