GPR68 Contributes to Persistent Acidosis-Induced Activation of AGC Kinases and Tyrosine Phosphorylation in Organotypic Hippocampal Slices.
GPR68 Contributes to Persistent Acidosis-Induced Activation of AGC Kinases and Tyrosine Phosphorylation in Organotypic Hippocampal Slices.
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DOI：
10.3389/fnins.2021.692217
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发表时间：
2021
影响因子：
4.3
通讯作者：
Zha XM
中科院分区：
文献类型：
--
作者：
Zhou G;Zha XM
持续性酸中毒发生在缺血和多种神经系统疾病中。在以前的研究中，酸性刺激导致神经元细胞内钙离子迅速增加。然而，长期酸中毒如何改变神经元信号传导在很大程度上仍不清楚。在我们以前的研究中，我们发现GPR 68介导的PKC活性对酸中毒诱导的皮质切片损伤具有保护作用。在这里，我们首先询问是否同样的原则适用于器官型海马切片。我们的数据表明，1小时pH 6诱导PKC磷酸化的GPR 68依赖的方式。Go 6983是一种PKC抑制剂，它使野生型（WT）中酸中毒诱导的神经元损伤恶化，但对GPR 68 −/−切片没有影响。接下来，为了更深入地了解脑组织中的酸信号传导，我们用pH 6处理器官型海马切片1小时，并通过Western印迹进行激酶组分析。酸中毒对细胞周期蛋白依赖性激酶（CDK）或酪蛋白激酶2活性（CMGC家族的两个成员）或共济失调毛细血管扩张突变（ATM）/ATM和RAD 3相关（ATR）活性影响不大，但降低了MAPK/CDK底物的磷酸化。相反，酸中毒诱导CaMKIIα、PKA和Akt的激活。除了这些丝氨酸/苏氨酸激酶，酸中毒也诱导酪氨酸磷酸化。由于GPR 68在脑神经元中广泛表达，我们询问GPR 68是否有助于酸中毒诱导的信号传导。删除GPR 68对酸中毒诱导的CaMKII磷酸化没有影响，减弱了磷酸化Akt和磷酸化PKA底物，同时消除了酸中毒诱导的酪氨酸磷酸化。这些数据表明，长期酸中毒激活了由AGC激酶、CaMKII和酪氨酸激酶介导的信号级联网络。GPR 68是酸中毒诱导的PKC激活和酪氨酸磷酸化的主要介质，而GPR 68依赖性和非依赖性机制都有助于PKA和Akt的激活。
Persistent acidosis occurs in ischemia and multiple neurological diseases. In previous studies, acidic stimulation leads to rapid increase in intracellular calcium in neurons. However, it remains largely unclear how a prolonged acidosis alters neuronal signaling. In our previous study, we found that GPR68-mediated PKC activities are protective against acidosis-induced injury in cortical slices. Here, we first asked whether the same principle holds true in organotypic hippocampal slices. Our data showed that 1-h pH 6 induced PKC phosphorylation in a GPR68-dependent manner. Go6983, a PKC inhibitor worsened acidosis-induced neuronal injury in wild type (WT) but had no effect in GPR68−/− slices. Next, to gain greater insights into acid signaling in brain tissue, we treated organotypic hippocampal slices with pH 6 for 1-h and performed a kinome profiling analysis by Western blot. Acidosis had little effect on cyclin-dependent kinase (CDK) or casein kinase 2 activity, two members of the CMGC family, or Ataxia telangiectasia mutated (ATM)/ATM and RAD3-related (ATR) activity, but reduced the phosphorylation of MAPK/CDK substrates. In contrast, acidosis induced the activation of CaMKIIα, PKA, and Akt. Besides these serine/threonine kinases, acidosis also induced tyrosine phosphorylation. Since GPR68 is widely expressed in brain neurons, we asked whether GPR68 contributes to acidosis-induced signaling. Deleting GPR68 had no effect on acidosis-induced CaMKII phosphorylation, attenuated that of phospho-Akt and phospho-PKA substrates, while abolishing acidosis-induced tyrosine phosphorylation. These data demonstrate that prolonged acidosis activates a network of signaling cascades, mediated by AGC kinases, CaMKII, and tyrosine kinases. GPR68 is the primary mediator for acidosis-induced activation of PKC and tyrosine phosphorylation, while both GPR68-dependent and -independent mechanisms contribute to the activation of PKA and Akt.
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DOI：
10.1523/jneurosci.5021-11.2012
发表时间：
2012-03-21
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Jing L;Chu XP;Jiang YQ;Collier DM;Wang B;Jiang Q;Snyder PM;Zha XM
通讯作者：
Zha XM
影响因子：
16.6
作者：
Bi M;Gladbach A;van Eersel J;Ittner A;Przybyla M;van Hummel A;Chua SW;van der Hoven J;Lee WS;Müller J;Parmar J;Jonquieres GV;Stefen H;Guccione E;Fath T;Housley GD;Klugmann M;Ke YD;Ittner LM
通讯作者：
Ittner LM
影响因子：
3.7
作者：
Li H;Wang D;Singh LS;Berk M;Tan H;Zhao Z;Steinmetz R;Kirmani K;Wei G;Xu Y
通讯作者：
Xu Y
影响因子：
4.7
作者：
Hosford PS;Mosienko V;Kishi K;Jurisic G;Seuwen K;Kinzel B;Ludwig MG;Wells JA;Christie IN;Koolen L;Abdala AP;Liu BH;Gourine AV;Teschemacher AG;Kasparov S
通讯作者：
Kasparov S
影响因子：
5.5
作者：
Hutter-Schmid, Bianca;Kniewallner, Kathrin M.;Humpel, Christian
通讯作者：
Humpel, Christian