Unsain N, Nuñez N, Anastasía A, Mascó D H
Laboratorio de Neurobiología, Centro de Biología Celular y Molecular, Cátedra de Biología Celular, Facultad de Ciencias Exactas, Físicas y Naturales, Universidad Nacional de Córdoba, Av. Vélez Sarsfield 1611, X5016 GCA, Córdoba, Argentina.
Neuroscience. 2008 Jun 26;154(3):978-93. doi: 10.1016/j.neuroscience.2008.04.038. Epub 2008 May 2.
In neuronal cultures it has been demonstrated that neurotrophins can elicit neuronal death through the p75 neurotrophic receptor (p75ntr) in the absence of concomitant Trk signaling. However, it was suggested that p75ntr induces neuronal death after status epilepticus (SE) in neuronal populations that express relatively high quantities of tropomyosin receptor kinase B (TrkB). Here, using Western blot and immunohistochemistry analyses in the hippocampus, we found that 3-h SE caused a remarkable decrease in TrkB expression and phosphorylation, and a significant increase in p75ntr. TrkB modification occurs before the overexpression of the tumor suppressor protein p53, accompanies the cell damage taking place in the dentate gyrus, and precedes the CA1 neuronal injury as assessed by Fluoro-Jade B and Nissl staining. Co-immunoprecipitation of brain-derived neurotrophic factor (BDNF) or its immature form proBDNF showed increased interaction with p75ntr after its binding to TrkB was reduced. Interestingly, proBDNF also increases its binding with p75ntr after seizures that do not cause neuronal death (animals injected with pilocarpine that fail to enter SE). However, in those animals, TrkB protein levels remained unchanged and its phosphorylation increased. Our results indicate an intrinsic capacity of neurons in vivo to modify final neurotrophin output by changing the proportion of their receptors' expression and the receptors' interaction with their ligands. These early events support the idea that neurotrophins may be involved in the induction of neuronal death in vivo under pathological conditions.
在神经元培养中已证明,在没有伴随的Trk信号传导的情况下,神经营养因子可通过p75神经营养因子受体(p75ntr)引发神经元死亡。然而,有人提出,在表达相对大量原肌球蛋白受体激酶B(TrkB)的神经元群体中,p75ntr在癫痫持续状态(SE)后诱导神经元死亡。在这里,我们使用海马体中的蛋白质印迹和免疫组织化学分析发现,3小时的SE导致TrkB表达和磷酸化显著降低,p75ntr显著增加。TrkB修饰发生在肿瘤抑制蛋白p53过表达之前,伴随着齿状回中发生的细胞损伤,并先于通过Fluoro-Jade B和尼氏染色评估的CA1神经元损伤。脑源性神经营养因子(BDNF)或其未成熟形式proBDNF的免疫共沉淀显示,在其与TrkB的结合减少后,与p75ntr的相互作用增加。有趣的是,在未导致神经元死亡的癫痫发作后(注射匹鲁卡品但未进入SE的动物),proBDNF与p75ntr的结合也增加。然而,在这些动物中,TrkB蛋白水平保持不变,其磷酸化增加。我们的结果表明,体内神经元具有通过改变其受体表达比例以及受体与配体的相互作用来改变最终神经营养因子输出的内在能力。这些早期事件支持了神经营养因子可能在病理条件下参与体内神经元死亡诱导过程的观点。