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Role of cAMP-dependent protein kinase A activity in endothelial cell cytoskeleton rearrangement.

机译:cAMP依赖性蛋白激酶A活性在内皮细胞骨架重排中的作用。

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To examine signaling mechanisms relevant to cAMP/protein kinase A (PKA)-dependent endothelial cell barrier regulation, we investigated the impact of the cAMP/PKA inhibitors Rp diastereomer of adenosine 3',5'-cyclic monophosphorothioate (Rp-cAMPS) and PKA inhibitor (PKI) on bovine pulmonary artery and bovine lung microvascular endothelial cell cytoskeleton reorganization. Rp-cAMPS as well as PKI significantly increased the formation of actin stress fibers and intercellular gaps but did not alter myosin light chain (MLC) phosphorylation, suggesting that the Rp-cAMPS-induced contractile phenotype evolves in an MLC-independent fashion. We next examined the role of extracellular signal-regulated kinases (ERKs) in Rp-cAMPS- and PKI-induced actin rearrangement. The activities of both ERK1/2 and its upstream activator Raf-1 were transiently enhanced by Rp-cAMPS and linked to the phosphorylation of the well-known ERK cytoskeletal target caldesmon. Inhibition of the Raf-1 target ERK kinase (MEK) either attenuated or abolished Rp-cAMPS- and PKI-induced ERK activation, caldesmon phosphorylation, and stress fiber formation. In summary, our data elucidate the involvement of the p42/44 ERK pathway in cytoskeletal rearrangement evoked by reductions in PKA activity and suggest the involvement of significant cross talk between cAMP- and ERK-dependent signaling pathways in endothelial cell cytoskeletal organization and barrier regulation.
机译:为了检查与cAMP /蛋白激酶A(PKA)依赖性内皮细胞屏障调节相关的信号传导机制,我们研究了cAMP / PKA抑制剂腺苷3',5'-环一硫代磷酸酯(Rp-cAMPS)和PKA的Rp非对映异构体的影响抑制剂(PKI)对牛肺动脉和牛肺微血管内皮细胞的细胞骨架重组。 Rp-cAMPS以及PKI显着增加了肌动蛋白应激纤维和细胞间间隙的形成,但并未改变肌球蛋白轻链(MLC)的磷酸化,表明Rp-cAMPS诱导的收缩表型以MLC独立的方式进化。接下来,我们研究了细胞外信号调节激酶(ERK)在Rp-cAMPS和PKI诱导的肌动蛋白重排中的作用。 Rp-cAMPS瞬时增强了ERK1 / 2及其上游激活因子Raf-1的活性,并与众所周知的ERK细胞骨架靶分子caldesmon的磷酸化有关。 Raf-1目标ERK激酶(MEK)的抑制作用减弱或消除了Rp-cAMPS和PKI诱导的ERK活化,卡尔德斯蒙磷酸化和应力纤维形成。总而言之,我们的数据阐明了p42 / 44 ERK通路参与了PKA活性降低引起的细胞骨架重排,并提示在内皮细胞的细胞骨架组织和屏障调节中依赖cAMP和ERK的信号通路之间存在明显的串扰。

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