cAMP signaling pathways play crucial roles in photoreceptor cells and other retinal cell types. Previous studies demonstrated a circadian rhythm of cAMP level in chick photoreceptor cell cultures that drives the rhythm of activity of the melatonin synthesizing enzyme arylalkylamine N-acetyltransferase and the rhythm of affinity of the cyclic nucleotide-gated channel for cGMP. Here, we report that the photoreceptor circadian clock generates a rhythm in Ca(2+)/calmodulin-stimulated adenylyl cyclase activity, which accounts for the temporal changes in the cAMP levels in the photoreceptors. The circadian rhythm of cAMP in photoreceptor cell cultures is abolished by treatment with the l-type Ca(2+) channel antagonist nitrendipine, while the Ca(2+) channel agonist, Bay K 8644, increased cAMP levels with continued circadian rhythmicity in constant darkness. These results indicate that the circadian rhythm of cAMP is dependent, in part, on Ca(2+) influx. Photoreceptor cell cultures exhibit a circadian rhythm in Ca(2+)/calmodulin-stimulated adenylyl cyclase enzyme activity with high levels at night and low levels during the day, correlating with the temporal changes of cAMP in these cells. Transcripts encoding two of the Ca(2+)/calmodulin-stimulated adenylyl cyclases, type 1 and type 8 (Adcy1 and Adcy8), displayed significant daily rhythms of mRNA expression under a light-dark cycle, but only the Adcy1 transcript rhythm persisted in constant darkness. Similar rhythms of Adcy1 mRNA level and Ca(2+)/calmodulin-stimulated adenylyl cyclase activity were observed in retinas of 2-week-old chickens. These results indicate that a circadian clock controls the expression of Adcy1 mRNA and Ca(2+)/calmodulin-stimulated adenylyl cyclase activity; and calcium influx into these cells gates the circadian rhythm of cAMP, a key component in the regulation of photoreceptor function.

译文

cAMP信号通路在感光细胞和其他视网膜细胞类型中起着至关重要的作用。先前的研究表明,雏鸡感光细胞培养物中cAMP水平的昼夜节律驱动褪黑激素合成酶芳基烷基胺N-乙酰基转移酶的活性节律和环状核苷酸门控通道对cGMP的亲和力节律。在这里,我们报告了感光体昼夜节律时钟在Ca(2)/钙调蛋白刺激的腺苷酸环化酶活性中产生节律,这说明了感光体中cAMP水平的时间变化。通过用l型Ca(2) 通道拮抗剂尼群地平处理,消除了感光细胞培养物中cAMP的昼夜节律,而Ca(2) 通道激动剂Bay K 8644在持续的黑暗中增加了cAMP水平,并持续了昼夜节律。这些结果表明,cAMP的昼夜节律部分取决于Ca(2) 的流入。感光细胞培养物在Ca(2)/钙调蛋白刺激的腺苷酸环化酶活性中表现出昼夜节律,夜间水平高,白天水平低,与这些细胞中cAMP的时间变化相关。编码两个Ca(2)/钙调蛋白刺激的腺苷酸环化酶 (1型和8型) (Adcy1和Adcy8) 的转录本在明暗循环下显示出明显的每日mRNA表达节律,但只有Adcy1转录本节律持续持续黑暗。在2周龄鸡的视网膜中观察到Adcy1 mRNA水平和Ca(2)/钙调蛋白刺激的腺苷酸环化酶活性的相似节律。这些结果表明,生物钟控制Adcy1 mRNA的表达和Ca(2)/钙调蛋白刺激的腺苷酸环化酶活性; 钙流入这些细胞中,激活了cAMP的昼夜节律,cAMP是调节感光细胞功能的关键组成部分。

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