Rapid automated high-performance liquid chromatographic analysis of cyclic adenosine 3',5'-monophosphate. Synthesis in brain tissues.
A rapid and sensitive automated system for measuring cyclic adenosine 3',5'-monophosphate (cAMP) synthesized from either radiolabelled adenine or adenosine 5'-triphosphate (ATP) in intact and broken cell tissue preparations, respectively, is described. After incubation with radiolabelled precursor, tissue samples are deproteinized and then injected directly onto a reversed-phase high-performance liquid chromatographic column. The column effluent fraction which contains cAMP is collected into scintillation vials and assayed for tritium by liquid scintillation spectrometry. Since the high-performance liquid chromatographic and fraction collection procedures are automated, over fifty samples can be analyzed in duplicate in a single day. The utility of this assay is illustrated by investigations of the effects of beta-adrenergic receptor stimulation on cAMP synthesis in tissue slices prepared from rat cerebral cortex and dopamine on cAMP synthesis in striatal membrane preparations.
Lin
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Saller
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Salama
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D000225:Adenine / Q000378:metabolism; D000818:Animals; D001322:Autoanalysis; D001923:Brain Chemistry / Q000187:drug effects; D002540:Cerebral Cortex / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003342:Corpus Striatum / Q000378:metabolism; D000242:Cyclic AMP / Q000032:analysis* / Q000096:biosynthesis / Q000302:isolation & purification; D004298:Dopamine / Q000494:pharmacology; D018492:Dopamine Antagonists; D006220:Haloperidol / Q000494:pharmacology; D066298:In Vitro Techniques; D007545:Isoproterenol / Q000494:pharmacology; D008297:Male; D011433:Propranolol / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D013056:Spectrophotometry, Ultraviolet
D018492:Dopamine Antagonists; D011433:Propranolol; D000242:Cyclic AMP; D006220:Haloperidol; D000225:Adenine; D007545:Isoproterenol; D004298:Dopamine