Department of Anatomy & Cell Biology, University of North Dakota School of Medicine and Health Sciences, Grand Forks, ND 58203, USA.
Exp Neurol. 2013 Feb;240:75-87. doi: 10.1016/j.expneurol.2012.10.023. Epub 2012 Nov 1.
Previous studies have demonstrated that ciliary neurotrophic factor (CNTF) enhances survival and process outgrowth from magnocellular neurons in the paraventricular (PVN) and the supraoptic (SON) nuclei. However, the mechanisms by which CNTF facilitates these processes remain to be determined. Therefore, the aim of this study was to identify the immediate signal transduction events that occur within the rat SON following administration of exogenous rat recombinant CNTF (rrCNTF) and to determine the contribution of those intracellular signaling pathway(s) to neuronal survival and process outgrowth, respectively. Immunohistochemical and Western blot analyses demonstrated that axonal injury and acute unilateral pressure injection of 100 ng/μl of rrCNTF directly over the rat SON resulted in a rapid and transient increase in phosphorylated-STAT3 (pSTAT3) in astrocytes but not neurons in the SON in vivo. Utilizing rat hypothalamic organotypic explant cultures, we then demonstrated that administration of 25 ng/ml rrCNTF for 14days significantly increased the survival and process outgrowth of OT magnocellular neurons. In addition, pharmacological inhibition of the Jak-STAT pathway via AG490 and cucurbitacin I significantly reduced the survival of OT magnocellular neurons in the SON and PVN; however, the contribution of the Jak-STAT pathway to CNTF-mediated process outgrowth remains to be determined. Together, these data indicate that CNTF-induced survival of OT magnocellular neurons is mediated indirectly through astrocytes via the Jak-STAT signaling pathway.
先前的研究表明,睫状神经营养因子(CNTF)可增强室旁核(PVN)和视上核(SON)中的大细胞神经元的存活和突起生长。然而,CNTF 促进这些过程的机制仍有待确定。因此,本研究旨在确定外源性大鼠重组 CNTF(rrCNTF)给药后大鼠 SON 内发生的即刻信号转导事件,并确定这些细胞内信号通路分别对神经元存活和突起生长的贡献。免疫组织化学和 Western blot 分析表明,轴突损伤和急性单侧压力注射 100ng/μl rrCNTF 直接作用于大鼠 SON,导致星形胶质细胞中磷酸化 STAT3(pSTAT3)的快速和短暂增加,但 SON 中的神经元没有增加。利用大鼠下丘脑器官培养物,我们随后证明,给予 25ng/ml rrCNTF 14 天可显著增加 OT 大细胞神经元的存活和突起生长。此外,通过 AG490 和葫芦素 I 抑制 Jak-STAT 途径可显著减少 SON 和 PVN 中 OT 大细胞神经元的存活;然而,Jak-STAT 途径对 CNTF 介导的突起生长的贡献仍有待确定。总之,这些数据表明,CNTF 诱导的 OT 大细胞神经元存活是通过星形胶质细胞间接介导的,通过 Jak-STAT 信号通路。