Soluble adenylyl cyclase is localized to cilia and contributes to ciliary beat frequency regulation via production of cAMP.
Soluble adenylyl cyclase is localized to cilia and contributes to ciliary beat frequency regulation via production of cAMP.
复制标题
DOI：
10.1085/jgp.200709784
复制
发表时间：
2007-07
期刊：
影响因子：
--
通讯作者：
Salathe M
中科院分区：
文献类型：
--
作者：
Schmid A;Sutto Z;Nlend MC;Horvath G;Schmid N;Buck J;Levin LR;Conner GE;Fregien N;Salathe M
纤毛气道上皮细胞在气道疾病恶化期间会受到细胞内CO2/HCO 3 −的持续变化，但CO2/HCO 3 −敏感性可溶性腺苷酸环化酶（sAC）在纤毛搏动调节中的作用尚不清楚。我们现在不仅显示了sAC在人气道上皮中的表达（通过RT-PCR、Western印迹和免疫荧光），而且还显示了其在轴丝中的特异性定位（Western印迹和免疫荧光）。利用荧光标记的PKA亚基（仅在纤毛细胞中由foxj 1启动子表达）之间的FRET，对纤毛细胞中[cAMP]变化的真实的时间估计揭示了CO2/HCO 3 −介导的cAMP产生。这种cAMP的产生被sAC抑制剂特异性阻断，但不被跨膜腺苷酸环化酶（tmAC）抑制剂阻断。此外，这种cAMP的产生刺激了不依赖于细胞内pH的纤毛搏动频率（CBF），因为PKA和sAC抑制剂能够独特地阻断CO2/HCO 3 −介导的CBF变化（而tmAC抑制剂没有影响）。因此，sAC定位于运动的气道纤毛，并且其有助于人气道CBF的调节。此外，CO2/HCO 3 −增加确实可逆地刺激完整细胞中sAC的细胞内cAMP产生。
Ciliated airway epithelial cells are subject to sustained changes in intracellular CO2/HCO3 − during exacerbations of airway diseases, but the role of CO2/HCO3 −-sensitive soluble adenylyl cyclase (sAC) in ciliary beat regulation is unknown. We now show not only sAC expression in human airway epithelia (by RT-PCR, Western blotting, and immunofluorescence) but also its specific localization to the axoneme (Western blotting and immunofluorescence). Real time estimations of [cAMP] changes in ciliated cells, using FRET between fluorescently tagged PKA subunits (expressed under the foxj1 promoter solely in ciliated cells), revealed CO2/HCO3 −-mediated cAMP production. This cAMP production was specifically blocked by sAC inhibitors but not by transmembrane adenylyl cyclase (tmAC) inhibitors. In addition, this cAMP production stimulated ciliary beat frequency (CBF) independently of intracellular pH because PKA and sAC inhibitors were uniquely able to block CO2/HCO3 −-mediated changes in CBF (while tmAC inhibitors had no effect). Thus, sAC is localized to motile airway cilia and it contributes to the regulation of human airway CBF. In addition, CO2/HCO3 − increases indeed reversibly stimulate intracellular cAMP production by sAC in intact cells.
登录
查看更多内容
DOI：
10.1073/pnas.0400050101
发表时间：
2004-03-02
影响因子：
11.1
作者：
Esposito, G;Jaiswal, BS;Gossen, JA
通讯作者：
Gossen, JA
影响因子：
4.8
作者：
Litvin, TN;Kamenetsky, M;Levin, LR
通讯作者：
Levin, LR
影响因子：
5.8
作者：
Reed, BY;Gitomer, WL;Pak, CYC
通讯作者：
Pak, CYC
影响因子：
5.5
作者：
Fragoso, MA;Fernandez, V;Conner, GE
通讯作者：
Conner, GE
DOI：
10.1084/jem.20050778
发表时间：
2005-08-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Han H;Stessin A;Roberts J;Hess K;Gautam N;Kamenetsky M;Lou O;Hyde E;Nathan N;Muller WA;Buck J;Levin LR;Nathan C
通讯作者：
Nathan C