Silva Ana, Naia Luana, Dominguez Alejandro, Ribeiro Márcio, Rodrigues Joana, Vieira Otília V, Lessmann Volkmar, Rego Ana Cristina
CNC - Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal.
Neurodegener Dis. 2015;15(4):207-18. doi: 10.1159/000375447. Epub 2015 Apr 21.
Several cellular mechanisms have been proposed to explain the pathogenesis of Huntington's disease (HD), including the lack of striatal brain-derived neurotrophic factor (BDNF). Thus, by preferentially binding to tropomyosin receptor kinase B (TrkB) receptor, BDNF is an important neurotrophin implicated in striatal neuronal survival.
To study the influence of BDNF and TrkB receptors in intracellular signaling pathways and caspase-3 activation in HD striatal cells.
HD mutant knockin and wild-type striatal cells were transduced with preproBDNF or full-length TrkB receptors to analyze BDNF processing, AKT and extracellular signal-regulated kinase (ERK) activation and the activity of caspase-3 in the absence or presence of staurosporine (STS).
HD mutant cells transduced with preproBDNF-mCherry (mCh) expressed similar levels of pro- and mature BDNF compared to WT cells, but HD cells released lower levels of pro- and mature BDNF. Despite this, BDNF-mCh overexpression rescued decreased AKT phosphorylation and reduced the caspase-3 activation observed in HD cells. Activated ERK was also enhanced in HD BDNF-mCh/TrkB-eGFP receptor co-cultures. Of relevance, overexpression of TrkB-eGFP in HD cells decreased caspase-3 activation, and stimulation of TrkB-eGFP-transduced mutant cells with recombinant human BDNF reduced both basal and STS-induced caspase-3 activation.
The results highlight the importance of BDNF-induced TrkB receptor signaling in rescuing HD-mediated apoptotic features in striatal cells.
已经提出了几种细胞机制来解释亨廷顿舞蹈病(HD)的发病机制,包括纹状体脑源性神经营养因子(BDNF)的缺乏。因此,BDNF通过优先结合原肌球蛋白受体激酶B(TrkB)受体,是一种参与纹状体神经元存活的重要神经营养因子。
研究BDNF和TrkB受体对HD纹状体细胞内信号通路和半胱天冬酶-3激活的影响。
用前体BDNF或全长TrkB受体转导HD突变体敲入和野生型纹状体细胞,以分析在不存在或存在星形孢菌素(STS)的情况下BDNF的加工、AKT和细胞外信号调节激酶(ERK)的激活以及半胱天冬酶-3的活性。
与野生型细胞相比,用前体BDNF-樱桃红(mCh)转导的HD突变体细胞表达的前体BDNF和成熟BDNF水平相似,但HD细胞释放的前体BDNF和成熟BDNF水平较低。尽管如此,BDNF-mCh的过表达挽救了HD细胞中AKT磷酸化的降低,并减少了观察到的半胱天冬酶-3激活。在HD BDNF-mCh/TrkB-eGFP受体共培养物中,激活的ERK也增强。相关的是,HD细胞中TrkB-eGFP的过表达降低了半胱天冬酶-3的激活,用重组人BDNF刺激TrkB-eGFP转导的突变体细胞减少了基础和STS诱导的半胱天冬酶-3激活。
结果强调了BDNF诱导的TrkB受体信号在挽救纹状体细胞中HD介导的凋亡特征方面的重要性。