Soluble adenylyl cyclase coordinates intracellular pH homeostasis and biomineralization in calcifying cells of a marine animal
Soluble adenylyl cyclase coordinates intracellular pH homeostasis and biomineralization in calcifying cells of a marine animal
复制标题
DOI：
10.1152/ajpcell.00524.2022
复制
发表时间：
2023-03-01
影响因子：
5.5
通讯作者：
Hu，Marian Y.
中科院分区：
文献类型：
--
作者：
Chang，William Weijen;Thies，Angus B.;Hu，Marian Y.
生物矿化细胞浓缩溶解的无机碳（DIC）并从矿物沉淀部位去除质子。然而，协调pH稳态和钙化细胞的生物矿化的分子调控机制知之甚少。在这里，我们报告的酸碱敏感酶可溶性腺苷酸环化酶（SAC）协调细胞内pH值（pHi）的调节在钙化的初级间充质细胞（PMC）的海胆幼虫。单细胞转录组学、原位杂交和免疫细胞化学阐明了骨骼发生过程中sAC的时空表达。PMCs的实时pHi成像显示，用两种结构不相关的小分子下调sAC活性抑制了PMCs的pHi调节，这种作用通过加入细胞可渗透的cAMP来挽救。药理学sAC抑制也显著降低了正常骨针生长和骨针再生，建立了PMC pH调节和生物矿化之间的联系。最后，增加的sAC mRNA的表达，检测到在骨骼矿化和暴露于CO2诱导的酸化。这些发现表明，需要转录调控的sAC，以促进磷酸化和补偿酸性胁迫。这项工作突出了中央的作用，sAC在协调酸碱调节和生物矿化的钙化细胞的海洋动物。
Biomineralizing cells concentrate dissolved inorganic carbon (DIC) and remove protons from the site of mineral precipitation. However, the molecular regulatory mechanisms that orchestrate pH homeostasis and biomineralization of calcifying cells are poorly understood. Here, we report that the acid-base sensing enzyme soluble adenylyl cyclase (sAC) coordinates intracellular pH (pHi) regulation in the calcifying primary mesenchyme cells (PMCs) of sea urchin larvae. Single-cell transcriptomics, in situ hybridization, and immunocytochemistry elucidated the spatiotemporal expression of sAC during skeletogenesis. Live pHiimaging of PMCs revealed that the downregulation of sAC activity with two structurally unrelated small molecules inhibited pHiregulation of PMCs, an effect that was rescued by the addition of cell-permeable cAMP. Pharmacological sAC inhibition also significantly reduced normal spicule growth and spicule regeneration, establishing a link between PMC pHiregulation and biomineralization. Finally, increased expression of sAC mRNA was detected during skeleton remineralization and exposure to CO2-induced acidification. These findings suggest that transcriptional regulation of sAC is required to promote remineralization and to compensate for acidic stress. This work highlights the central role of sAC in coordinating acid-base regulation and biomineralization in calcifying cells of a marine animal.