Src family kinase inhibitors blunt PACAP-induced PAC1 receptor endocytosis, phosphorylation of ERK, and the increase in cardiac neuron excitability.
Src family kinase inhibitors blunt PACAP-induced PAC1 receptor endocytosis, phosphorylation of ERK, and the increase in cardiac neuron excitability.
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Src 家族激酶抑制剂会减弱 PACAP 诱导的 PAC1 受体内吞作用、ERK 磷酸化以及心脏神经元兴奋性的增加。
DOI：
10.1152/ajpcell.00223.2017
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Parsons,RodneyL
中科院分区：
文献类型：
--
作者：
Tompkins,JohnD;Clason,ToddA;Buttolph,ThomasR;Girard,BeatriceM;Linden,AnneK;Hardwick,JeanC;Merriam,LauraA;May,Victor;Parsons,RodneyL
垂体腺苷环化酶激活多肽(PACAP，ADCYAP1)激活PAC1受体(Adcyap1r1)显著增加豚鼠心肌神经元兴奋性。这种兴奋性的调节在一定程度上是通过质膜G蛋白依赖的腺苷酸环化酶激活和下游信号级联来调节的。然而，在PAC1受体和内体信号内化之后，导致兴奋性增强的其他机制被激活。SRC家族在许多营养因子和G蛋白偶联受体的内吞过程中起着重要作用。本研究探讨了Src家族激酶是否也支持PACAP诱导的PAC1受体内化、ERK的磷酸化以及增强神经元的兴奋性。使用稳定表达绿色荧光蛋白标记的PAC1受体的人胚胎肾细胞，用Src家族激酶抑制剂PP2(10µM)处理后，PACAP诱导的PAC1受体内化显著减少，Western印迹分析表明，PP2和Src抑制物1(Src-1，2µM)同时降低了ERK的激活。相反，Src家族的激酶抑制剂并不能消除PACAP诱导的钙升高，这种升高是由肌醇(1，4，5)-三磷酸诱导的内质网钙释放引起的。从磷酸化ERK免疫染色的共聚焦分析来看，PP2处理显著减弱了心脏神经节整体贴壁标本内神经元中ERK的PACAP活性。细胞内记录显示，PP2还显著钝化了PACAP诱导的心脏神经元兴奋性的增加。这些研究表明，在PAC1受体内化、MEK/ERK信号的激活和神经元兴奋性的调节中，Src相关的激酶活性。目前的结果进一步支持了PAC1受体内体信号作为调节细胞功能的关键机制的重要性。
Pituitary adenylate cyclase activating polypeptide (PACAP,Adcyap1) activation of PAC1 receptors (Adcyap1r1) significantly increases excitability of guinea pig cardiac neurons. This modulation of excitability is mediated in part by plasma membrane G protein-dependent activation of adenylyl cyclase and downstream signaling cascades. However, additional mechanisms responsible for the enhanced excitability are activated following internalization of the PAC1 receptor and endosomal signaling. Src family kinases play critical roles mediating endocytosis of many trophic factor and G protein-coupled receptors. The present study investigated whether Src family kinases also support the PACAP-induced PAC1 receptor internalization, phosphorylation of ERK, and enhanced neuronal excitability. Using human embryonic kidney cells stably expressing a green fluorescent protein-tagged PAC1 receptor, treatment with the Src family kinase inhibitor PP2 (10 µM) markedly reduced the PACAP-induced PAC1 receptor internalization, and in parallel, both PP2 and Src inhibitor 1 (Src-1, 2 µM) reduced ERK activation determined by Western blot analysis. In contrast, Src family kinase inhibitors did not eliminate a PACAP-induced rise in global calcium generated by inositol (1,4,5)-trisphosphate-induced release of calcium from endoplasmic reticulum stores. From confocal analysis of phosphorylated ERK immunostaining, PP2 treatment significantly attenuated PACAP activation of ERK in neurons within cardiac ganglia whole mount preparations. Intracellular recordings demonstrated that PP2 also significantly blunted a PACAP-induced increase in cardiac neuron excitability. These studies demonstrate Src-related kinase activity in PAC1 receptor internalization, activation of MEK/ERK signaling, and regulation of neuronal excitability. The present results provide further support for the importance of PAC1 receptor endosomal signaling as a key mechanism regulating cellular function.
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多种机制有助于 PAC1 调节副交感心脏神经元兴奋性
DOI：
--
发表时间：
2016
期刊：
影响因子：
--
作者：
R. Parsons;J. Tompkins;J. Hardwick;L. A. Merriam;B. Girard;V. May
通讯作者：
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DOI：
10.1152/ajpcell.00164.2016
发表时间：
2016-08
期刊：
American journal of physiology. Cell physiology
影响因子：
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作者：
J. Tompkins;Todd A. Clason;J. Hardwick;B. Girard;L. A. Merriam;V. May;R. Parsons
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J. Tompkins;Todd A. Clason;J. Hardwick;B. Girard;L. A. Merriam;V. May;R. Parsons
影响因子：
5.5
作者：
May, Victor;Buttolph, Thomas R.;Parsons, Rodney L.
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Parsons, Rodney L.
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Brandon Zimmerman;M. Simaan;Mi‐Hye Lee;L. Luttrell;S. Laporte
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S. Laporte
影响因子：
11.8
作者：
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