Brain-derived neurotrophic factor (BDNF) acts primarily via the JAK/STAT pathway to promote neurite growth in the major pelvic ganglion of the rat: Part 2
Brain-derived neurotrophic factor (BDNF) acts primarily via the JAK/STAT pathway to promote neurite growth in the major pelvic ganglion of the rat: Part 2
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DOI：
10.1111/j.1743-6109.2006.00292.x
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发表时间：
2006-09-01
影响因子：
3.5
通讯作者：
Lin, Ching-Shwun
中科院分区：
文献类型：
--
作者：
Lin, Guiting;Bella, Anthony J.;Lin, Ching-Shwun
介绍。膀胱癌和前列腺癌的手术和放射治疗可能损害海绵状神经并引起勃起功能障碍（ED）。我们之前的研究表明，脑源性神经营养因子（BDNF）可以恢复神经源性ED大鼠的勃起功能。我们现在研究BDNF在大盆腔神经节（MPG）外植体中的信号传导机制。目的：探讨BDNF在mpg培养中的神经营养作用的信号传导机制。从雄性大鼠大盆腔神经节分离，免疫组化和免疫荧光染色定位BDNF受体、泛神经营养因子75 （p75）、原肌球蛋白相关激酶B （TrkB）和原肌球蛋白相关激酶C （TrkC）。用BDNF治疗MPG背尾区，以确定促进神经突起生长的最佳剂量。然后使用特异性激酶抑制剂AG490、KT5720、LY294002和U0126单独或在BDNF治疗之前治疗MPG。Western blot检测经处理的MPG的神经突生长情况，以及JAK2、STAT1和STAT3的表达和磷酸化情况。主要结果测量。测量MPG中神经突生长的长度，以量化BDNF的作用并确定特定的信号通路。在对照和处理过的MPG中，测定了JAK2、STAT1和STAT2磷酸化与未磷酸化蛋白的比值，证实了JAK/STAT是主要的信号通路。原肌球蛋白相关激酶B和TrkC定位于神经元，而p75定位于神经元周围卫星胶质细胞（SGC）。BDNF促进MPG神经突生长的最佳剂量为25 ~ 50 ng/mL。在四种特异性激酶抑制剂中，AG490在抑制MPG神经突生长以及bdnf诱导的JAK2、STAT1和stat3磷酸化方面的作用最强。大鼠MPG中TrkB和TrkC在神经元中表达，而p75在SGC中表达。促进MPG神经突生长的最佳BDNF剂量为25 ~ 50 ng/mL。BDNF主要通过激活JAK/STAT通路促进MPG神经突生长。
Introduction. Surgical and radiation therapies of bladder and prostate cancers may damage cavernous nerves and cause erectile dysfunction (ED). We previously showed that brain-derived neurotrophic factor (BDNF) could restore erectile function in a neurogenic ED rat model. We now investigated the signaling mechanism of BDNF in major pelvic ganglia (MPG) explants.Aim. To identify the signaling mechanism that mediates the neurotrophic effect of BDNF in cultured MPG.Methods. Major pelvic ganglia was isolated from male rats for immunohistochemistry and immunofluorescence staining to locate BDNF receptors, pan-neurotrophin 75 (p75), tropomyosin-related kinase B (TrkB), and tropomyosin-related kinase C (TrkC). The dorso-caudal region of MPG was treated with BDNF to determine the optimal dosage for promoting neurite growth. Specific kinase inhibitors AG490, KT5720, LY294002, and U0126 were then used to treat MPG either alone or prior to BDNF treatment. The treated MPG was examined for neurite growth and for expression and phosphorylation of JAK2, STAT1, and STAT3 by Western blot analysis.Main Outcome Measures. Lengths of neurite growth from MPG were measured to quantify the effects of BDNF and to identify specific signaling pathways. Ratios of phosphorylated vs. unphosphoryated proteins of JAK2, STAT1, and STAT2 in control and treated MPG were determined to confirm JAK/STAT as the principal signaling pathway.Results. Tropomyosin-related kinase B and TrkC were localized to neurons whereas p75 to perineuronal satellite glial cells (SGC). The optimal dosage of BDNF for promoting MPG neurite growth was between 25 and 50 ng/mL. Among the four specific kinase inhibitors, AG490 was the strongest in suppressing MPG neurite growth as well as BDNF-induced phosphorylation of JAK2, STAT1, and STAT3.Conclusions. In rat MPG, TrkB and TrkC were expressed in neurons, whereas p75 in SGC. Optimal BDNF dosage for promoting MPG neurite growth was between 25 and 50 ng/mL. BDNF promotes MPG neurite growth primarily by activating the JAK/STAT pathway.