NPKCε, a P2Y2-R downstream effector in regulated mucin secretion from airway goblet cells
NPKCε, a P2Y2-R downstream effector in regulated mucin secretion from airway goblet cells
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DOI：
10.1152/ajpcell.00051.2007
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发表时间：
2007-11-01
影响因子：
5.5
通讯作者：
Davis, C. William
中科院分区：
文献类型：
--
作者：
Ehre, Camille;Zhu, Yunxiang;Davis, C. William
气道杯状细胞粘蛋白分泌受P2Y(2)嘌呤受体激动剂激活控制，通过Gq/PLC、肌醇-1,4,5-三磷酸（IP3）、二酰基甘油、Ca2+和蛋白激酶C （PKC）起作用。之前，我们发现SPOC1细胞表达cPKC α、nPKC delta、nPKC epsilon和nPKC eta；其中，只有nPKC δ移位到膜上与粘蛋白分泌有关（Abdullah LH, Bundy JT, Ehre C, Davis CW）。[J] .中国生物医学工程学报，2016,31(2):444 - 444。我们已经验证了这些结果，并通过使用SPOC1细胞和小鼠模型测试PKC表达改变对调节粘蛋白释放的影响，来追求PKC效应异构体的身份。过表达cPKC α、nPKC δ和nPKC eta的SPOC1细胞与空逆转录病毒载体表达细胞的ATP γ S-和phorpol -1,2- myriate -13-acetate （PMA）刺激的粘蛋白分泌水平相同。分泌激素诱导的粘蛋白分泌仅在过表达nPKCe的细胞中升高（ATP γ S和PMA分别为14.6%和23.5%）。同样，只有感染了激酶缺陷型nPKCe的SPOC1细胞，相对于野生型（WT）亚型的过表达，表现出预期的受刺激的粘蛋白分泌减少。与WT小鼠相比，P2Y(2)-R缺失小鼠的ATP γ s刺激的离体小鼠气管粘蛋白分泌减少了82%，证明了小鼠模型在研究调节粘蛋白分泌方面的实用性。同窝的WT和nPKC δ敲除（KO）小鼠具有几乎相同水平的受刺激的粘蛋白分泌，而nPKCe KO小鼠的粘蛋白释放相对于其WT同窝的小鼠几乎被消除。我们得出结论，nPKCe是杯状细胞分泌反应中P2Y(2)-R激活下游的效应异构体。在活化细胞中观察到的nPKC δ易位可能与粘蛋白分泌无关，而是与杯状细胞生物学的其他方面有关。
Air-way goblet cell mucin secretion is controlled by agonist activation of P2Y(2) purinoceptors, acting through Gq/PLC, inositol-1,4,5-trisphosphate (IP3), diacylglycerol, Ca2+ and protein kinase C (PKC). Previously, we showed that SPOC1 cells express cPKC alpha, nPKC delta, nPKC epsilon, and nPKC eta; of these, only nPKC delta translocated to the membrane in correlation with mucin secretion (Abdullah LH, Bundy JT, Ehre C, Davis CW. Am J Physiol Lung Physiol 285: L149-L160, 2003). We have verified these results and pursued the identity of the PKC effector isoform by testing the effects of altered PKC expression on regulated mucin release using SPOC1 cell and mouse models. SPOC1 cells overexpressing cPKC alpha, nPKC delta, and nPKC eta had the same levels of ATP gamma S- and phorbol-1,2-myristate-13-acetate (PMA)-stimulated mucin secretion as the levels in empty retroviral vector expressing cells. Secretagogue-induced mucin secretion was elevated only in cells overexpressing nPKCe (14.6 and 23.5%, for ATP gamma S and PMA). Similarly, only SPOC1 cells infected with a kinase-deficient nPKCe exhibited the expected diminution of stimulated mucin secretion, relative to wild-type (WT) isoform overexpression. ATP gamma S-stimulated mucin secretion from isolated, perfused mouse tracheas was diminished in P2Y(2)-R null mice by 82% relative to WT mice, demonstrating the utility of mouse models in studies of regulated mucin secretion. Littermate WT and nPKC delta knockout (KO) mice had nearly identical levels of stimulated mucin secretion, whereas mucin release was nearly abolished in nPKCe KO mice relative to its WT littermates. We conclude that nPKCe is the effector isoform downstream of P2Y(2)-R activation in the goblet cell secretory response. The translocation of nPKC delta observed in activated cells is likely not related to mucin secretion but to some other aspect of goblet cell biology.