PKC and PKA, but not PKG mediate LPS-induced CGRP release and [Ca2+]i elevation in DRG neurons of neonatal rats
PKC and PKA, but not PKG mediate LPS-induced CGRP release and [Ca2+]i elevation in DRG neurons of neonatal rats
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DOI：
10.1002/jnr.1249
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发表时间：
2001-11-15
影响因子：
4.2
通讯作者：
Wang, X
中科院分区：
文献类型：
--
作者：
Hou, LF;Wang, X
降钙素基因相关肽 (CGRP) 在背根神经节 (DRG) 神经元中产生，并从初级传入神经元释放，以介导血流动力学效应和神经源性炎症。在这项工作中，我们确定了脂多糖（LPS）（一种炎症刺激剂）是否可以触发培养的 DRG 神经元释放 CGRP，如果可以，则确定哪种细胞信号通路参与了这种反应。钙 ([Ca2+](i)) 的细胞质浓度在神经递质释放中起着关键作用，因此还使用 Fluo-3/AM 在培养的 DRG 细胞中测定 [Ca2+](i)。结果表明，LPS (0.1-10 mug/ml) 会以时间和浓度依赖性方式从 DRG 神经元中诱发 CGRP 释放。 LPS 还以浓度依赖性方式增加 [Ca2+](i)。蛋白激酶 C (PKC) 抑制剂 Calphostin C 0.5 muM 或 RO-31-8220 0.1 muM，以及 cAMP 依赖性蛋白激酶 (PKA) 特异性抑制剂 RP-CAMPS 30 muM 或非特异性抑制剂 H8 1 muM 抑制 1 mug/ml LPS 诱发的 CGRP 释放和 DRG 神经元的 [Ca2+](i) 增加。 cGMP依赖性蛋白激酶(PKG)抑制剂Rp-8-pCPT-cGMPS 30 muM不会阻断LPS反应。这些数据表明，LPS可能通过PKC和PKA而不是PKG信号通路刺激新生大鼠DRG神经元中的CGRP释放和[Ca2+](i)升高。 J.神经科学。资源。 66:592-600, 2001。(C) 2001 Wiley-Liss, Inc.
Calcitonin gene-related peptide (CGRP), is produced in dorsal root ganglia (DRG) neurons and released from primary afferent neurons to mediate hemodynamic effects and neurogenic inflammation. In this work, we determined whether lipopolysaccharide (LPS), an inflammatory stimulator, could trigger CGRP release from cultured DRG neurons and if so, which cellular signaling pathway was involved in this response. Cytoplasmic concentration of calcium ([Ca2+](i)) plays a key role in neurotransmitter release, therefore [Ca2+](i) was also determined in cultured DRG cells using fluo-3/AM. The results showed that LPS (0.1-10 mug/ml) evoked CGRP release in a time- and concentration-dependent manner from DRG neurons. LPS also increased [Ca2+](i) in a concentration-dependent manner. The protein kinase C (PKC) inhibitors, calphostin C 0.5 muM or RO-31-8220 0.1 muM, and the cAMP-dependent protein kinase (PKA) specific inhibitor RP-CAMPS 30 muM or nonspecific inhibitor H8 1 muM inhibited 1 mug/ml LPS-evoked CGRP release and [Ca2+](i) increase from DRG neurons. The cGMP-dependent protein kinase (PKG) inhibitor Rp-8-pCPT-cGMPS 30 muM did not block the LPS response. These data suggest that LPS may stimulate CGRP release and [Ca2+](i) elevation through PKC and PKA but not PKG signaling pathway in DRG neurons of neonatal rats. J. Neurosci. Res. 66:592-600, 2001. (C) 2001 Wiley-Liss, Inc.