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Day/Night Differences in the Stimulation of Adenylate Cyclase Activity by Calcium/Calmodulin in Chick Pineal Cell Cultures: Evidence for Circadian Regulation of Cyclic AMP

Joseph S. Takahashi

Northwestern University Institute for Neuroscience and Howard Hughes Medical Institute, Department of Neurobiology and Physiology, Northwestern University, Evanston, IL 60208; Departmentof Biology, Central Missouri State University, Warrensburg, MO 64093.

In chick pineal cell culture, stimulation of adenylate cyclase with the diterpene forskolin was greater during the subjective night than during the subjective day. This rhythm of cyclic AMP (cAMP) stimulation mimicked the rhythm of unstimulated cAMP measured previously during LD cycles from flow-through culture. Direct measurement of adenylate cyclase activity in permeabilized cells revealed an adenylate cyclase activity activated by Ca2+/calmodulin during the night but not during the day. However, this difference in adenylate cyclase activity at twotimes of the circadian cycle is apparent only when permeabilized cells were prewashed with buffer containing GTP. When cAMP was measured from flow-through cultures maintained in continuous darkness to determine whether a circadian clock may regulate cAMP, a low-amplitude rhythm was measured. The circadian rhythm of cAMP was similar to the cAMP rhythm previously measured on LD cycles except that the rhythm in darkness had a lower amplitude. Similar to the suppression of melatonin, cAMP was suppressed by light presented during the middle of the night. LD differencesin nocturnal cAMP levels were abolished with dipyridamole, an inhibitor of cyclic GMP (cGMP) phosphodiesterase. These results suggest that the rhythm of cAMP in chick pineal cells involves the stimulation of adenylate cyclase by Ca2+/calmodulin during the night and a GTP-dependent suppression of adenylate cyclase activity during the day. The photic suppression of cAMP at night involves the activation of a dipyridamole-sensitive, cGMP phosphodiesterase.

Key Words: diurnal • circadian rhythm • melatonin • pineal • G protein • GTP • phosphodiesterase

Journal of Biological Rhythms, Vol. 13, No. 6, 479-493 (1998)
DOI: 10.1177/074873098129000318


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