Tyrosine phosphorylation is a novel pathway for regulation of chloride secretion in shark rectal gland.
Tyrosine phosphorylation is a novel pathway for regulation of chloride secretion in shark rectal gland.
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酪氨酸磷酸化是调节鲨鱼直肠腺氯离子分泌的新途径。
DOI：
10.1152/ajprenal.1995.269.4.f594
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
ForrestJr,JN
中科院分区：
文献类型：
--
作者：
Lehrich,RW;ForrestJr,JN
我们使用特定的酪氨酸激酶抑制剂金雀异黄素来定义酪氨酸磷酸化在角鲨直肠腺中氯离子转运调节中的作用，这是一种通过囊性纤维化跨膜电导调节器（CFTR）样通道分泌氯离子的模型。在灌注的腺体中，金雀异黄素 (100 microM) 迅速将氯离子分泌量从基础值 159 +/- 36 增加到 966 +/- 49 mueq.h-1.g-1 (P < 0.0001)。布门他尼完全逆转金雀异黄素诱导的分泌。在直肠腺管细胞的原代培养单层中，染料木黄酮（而非无活性的 7-葡萄糖苷形式染料木黄酮）以剂量依赖性方式增加短路电流，从基础值 2.7 +/- 4.3 增加到 104 +/- 10 microA/cm2 (P < 0.0001)。顶部施用金雀异黄素比基底外侧添加显着诱导更多的氯离子分泌。金雀异黄素不会增加灌注腺体或培养单层细胞的腺苷 3',5'-环单磷酸 (cAMP) 含量。使用抗磷酸酪氨酸抗体，我们观察到多种蛋白质的磷酸化。分子量分别为 250、210、55 和 53 kDa 的四种肽对金雀异黄素处理做出反应，酪氨酸磷酸化降低。这些数据证明了以下内容：1) 金雀异黄素在灌注的直肠腺和培养的肾小管细胞中诱导布美他尼敏感的氯化物分泌； 2) 这些作用并不伴随组织cAMP的升高，表明金雀异黄素诱导的分泌不是由cAMP-蛋白激酶A途径介导的； 3) 金雀异黄素敏感肽存在于直肠腺细胞中，是参与调节氯离子分泌的候选肽。（摘要截断为 250 字）
We used the specific tyrosine kinase inhibitor genistein to define the involvement of tyrosine phosphorylation in the regulation of chloride transport in the rectal gland of the dogfish shark, a model for chloride secretion via a cystic fibrosis transmembrane conductance regulator (CFTR)-like channel. In the perfused gland, genistein (100 microM) promptly increased chloride secretion from basal values of 159 +/- 36 to 966 +/- 49 mueq.h-1.g-1 (P < 0.0001). Bumentanide fully reversed genistein-induced secretion. In primary culture monolayers of rectal gland tubular cells, genistein, but not the inactive 7-glucoside form, genistin, increased short-circuit current in a dose-dependent manner, from basal values of 2.7 +/- 4.3 to 104 +/- 10 microA/cm2 (P < 0.0001). Apically applied genistein induced significantly greater chloride secretion than basolateral addition. Genistein did not increase the adenosine 3',5'-cyclic monophosphate (cAMP) content of either perfused glands or cultured monolayers. Using an anti-phosphotyrosine antibody, we observed phosphorylation of multiple proteins. Four peptides, with molecular masses of 250, 210, 55, and 53 kDa, responded to genistein treatment with a decrease in tyrosine phosphorylation. These data demonstrate the following: 1) genistein induces bumetanide-sensitive chloride secretion in both perfused rectal glands and cultured tubular cells; 2) these effects are not accompanied by an elevation of tissue cAMP, indicating that genistein-induced secretion is not mediated by the cAMP-protein kinase A pathway; and 3) genistein-sensitive peptides are present in the rectal gland cell and are candidates for involvement in the regulation of chloride secretion.(ABSTRACT TRUNCATED AT 250 WORDS)