Ripoll Léa, von Zastrow Mark
bioRxiv. 2023 Dec 6:2023.12.06.570478. doi: 10.1101/2023.12.06.570478.
The cAMP cascade is widely recognized to transduce its physiological effects locally through spatially limited cAMP gradients. However, little is known about how the adenylyl cyclase enzymes, which initiate cAMP gradients, are localized. Here we answer this question in physiologically relevant striatal neurons and delineate how AC localization impacts downstream signaling functions. We show that the major striatal AC isoforms are differentially sorted between ciliary and extraciliary domains of the plasma membrane, and that AC9 is uniquely targeted to endosomes. We identify key sorting determinants in the N-terminal cytoplasmic domain responsible for isoform-specific localization. We also show that AC9-containing endosomes accumulate activated dopamine receptors and form an elaborately intertwined network with juxtanuclear PKA stores bound to Golgi membranes. Finally, we show that endosomal localization is critical for AC9 to selectively elevate PKA activity in the nucleus relative to the cytoplasm. These results reveal a precise spatial landscape of the cAMP cascade in neurons and a key role of AC localization in directing downstream signal transduction to the nucleus.
环磷酸腺苷(cAMP)级联反应通过空间上有限的cAMP梯度在局部转导其生理效应,这一点已得到广泛认可。然而,关于启动cAMP梯度的腺苷酸环化酶如何定位,人们所知甚少。在这里,我们在生理相关的纹状体神经元中回答了这个问题,并描述了腺苷酸环化酶(AC)的定位如何影响下游信号功能。我们发现,主要的纹状体AC亚型在质膜的纤毛和非纤毛区域之间进行差异分选,并且AC9独特地靶向内体。我们确定了N端胞质结构域中负责亚型特异性定位的关键分选决定因素。我们还表明,含有AC9的内体积累活化的多巴胺受体,并与结合在高尔基体膜上的近核蛋白激酶A(PKA)储存形成一个错综复杂的交织网络。最后,我们表明内体定位对于AC9选择性提高细胞核相对于细胞质中的PKA活性至关重要。这些结果揭示了神经元中cAMP级联反应精确的空间格局,以及AC定位在引导下游信号转导至细胞核中的关键作用。