Pituitary adenylate cyclase-activating peptide induces neurite outgrowth in cultured monkey trigeminal ganglion cells: Involvement of receptor PAC1
Pituitary adenylate cyclase-activating peptide induces neurite outgrowth in cultured monkey trigeminal ganglion cells: Involvement of receptor PAC1
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发表时间：
2013-01
期刊：
影响因子：
2.2
通讯作者：
E. Nakajima;R. D. Walkup;A. Fujii;T. Shearer;M. Azuma
中科院分区：
文献类型：
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作者：
E. Nakajima;R. D. Walkup;A. Fujii;T. Shearer;M. Azuma
目的我们以往对兔三叉神经(TGN)的研究表明，垂体腺苷环化酶激活肽(PACAP)可促进神经元突起的延长和角膜敏感性的恢复。本研究的目的是：1)建立猴三叉神经(TGN)细胞的培养方法；2)检测PACAP在我们的培养体系中是否诱导轴突的萌发和延长；3)研究PACAP诱导轴突延长的信号机制；4)检测PACAP对猴泪腺泡细胞分泌泪液蛋白的作用。方法建立恒河猴TGN细胞原代培养体系。免疫组织化学染色检测PACAP受体PAC1的细胞分布。用神经丝抗体染色测定培养的TGN神经节细胞的轴突长度。用实时定量聚合酶链式反应(QPCR)检测mRNA的表达。免疫印迹法检测培养的腺泡细胞分泌泪液蛋白。结果分离培养的TGN细胞含有神经节细胞和雪旺细胞，PAC1受体在这两种细胞中均有表达。PACAP-27可显著诱导神经突起生长，PACAP 6-27可抑制PACAP-27的作用。腺苷环化酶和磷脂酶C的抑制剂也抑制了轴突的生长。在培养过程中，卵泡抑素被PACAP-27上调。PACAP可促进泪液蛋白的分泌。结论PAC1的激活参与了TGN突起的生长。
Purpose Our previous studies in the rabbit trigeminal nerve (TgN) showed that pituitary adenylate cyclase-activating peptide (PACAP) accelerated the extension of neuronal processes and recovery of corneal sensitivity. The purposes of the present study were 1) develop a procedure to culture trigeminal nerve (TgN) cells from monkeys, 2) test whether PACAP induces sprouting and elongation of axons in our culture system, 3) investigate the signaling mechanisms producing axon elongation induced by PACAP, and 4) test the action of PACAP on tear protein secretion by monkey lacrimal acinar cells. Methods Primary cultures of TgN cells were established from rhesus monkeys. Cellular distribution of the PACAP receptor, PAC1, was determined with immunostaining. Axonal length in cultured TgN ganglion cells was evaluated with staining by antibody for neurofilament. mRNA expression was determined with quantitative real-time polymerase chain reaction (qPCR). Secretion of tear protein from cultured acinar cells was measured with immunoblotting. Results Our results showed that dissociated, cultured TgN cells contained neuronal ganglion and Schwann cells, and the PAC1 receptor was expressed in both cell types. PACAP-27 significantly induced neurite outgrowth, which was inhibited by PACAP 6–27. Inhibitors for adenylate cyclase and phospholipase C also inhibited neurite outgrowth. Follistatin was upregulated by PACAP-27 during the culture period. PACAP enhanced secretion of tear proteins. Conclusions Our data suggested PAC1 activation is involved in TgN neurite outgrowth.