Huang Zhi, Petitto John M
J Neurol Disord. 2013;1(1). doi: 10.4172/2329-6895.1000117.
Our lab showed previously that whereas a substantial portion of chronically resected facial motor neurons reside in an atrophied state that can be reversed at 14 days following reinjury in wild-type (WT) mice, atrophy reversal was altered in immunodeficient mice. It was unclear, however, if the abnormal response at day 14 post-reinjury in immunodeficient mice might be due to differences in the kinetics of the reversal response or impaired regeneration. We sought to address this question, and test our working hypothesis that the normal regeneration of atrophied motor neurons is dependent on normal adaptive immunity, by comparing WT and immunodeficient recombination activating gene-2 knockout (RAG2-KO) mice that lack a mature T and B lymphocytes, at 3 and 28 days following reinjury. In WT mice, facial motor neurons that were resected for 10 weeks and subsequently reinjured for 3 days were able to regain fully an apparent 40% loss of countable neurons, and nearly 45% of that robust increase in neurons was sustained at 28 days post-reinjury in the WT mice. By contrast, at both 3 and 28 days post-reinjury RAG2-KO mice failed to show any increase in neuronal number. Size measurements showed that the surviving neurons of WT and RAG2-KO mice exhibited substantial motor neuron hypertrophy at 3 days post-reinjury, and similar levels of normal size motor neurons by 28 days post-reinjury. Among the WT mice, small numbers of T lymphocytes where found in the reinjured facial motor nucleus (FMN), and were significantly higher at 3 days, but not 28 days, in the reinjury compared to sham-reinjury groups. No differences were seen between the WT and RAG2-KO mice in overall microglial cell activity using CD11b expression following reinjury. These data suggest that many resected motor neurons did not survive the initial resection in RAG2-KO mice, whereas in WT mice they atrophied and could be restimulated by reinjury to regenerate their phenotype. Moreover, they indicate that normal T cell function, or some yet unknown function of the RAG2 gene in the brain, is essential for activating regeneration programs of atrophied motor neurons - programs with therapeutic potential for modifying neuroplasticity.
我们实验室之前的研究表明,在野生型(WT)小鼠中,虽然大部分长期切除的面部运动神经元处于萎缩状态,且在再次损伤后14天可逆转,但免疫缺陷小鼠的萎缩逆转情况有所改变。然而,尚不清楚免疫缺陷小鼠在再次损伤后第14天的异常反应是由于逆转反应动力学的差异还是再生受损。我们试图解决这个问题,并通过比较再次损伤后3天和28天的WT小鼠与缺乏成熟T和B淋巴细胞的免疫缺陷重组激活基因2敲除(RAG2-KO)小鼠,来检验我们的工作假设,即萎缩运动神经元的正常再生依赖于正常的适应性免疫。在WT小鼠中,切除10周后再损伤3天的面部运动神经元能够完全恢复明显40%的可计数神经元损失,并且在WT小鼠再次损伤后28天,神经元的强劲增加中有近45%得以维持。相比之下,在再次损伤后3天和28天,RAG2-KO小鼠的神经元数量均未显示出任何增加。大小测量显示,WT和RAG2-KO小鼠存活的神经元在再次损伤后3天表现出明显的运动神经元肥大,到再次损伤后28天,正常大小运动神经元的水平相似。在WT小鼠中,在再次损伤的面部运动核(FMN)中发现了少量T淋巴细胞,与假再次损伤组相比,再次损伤后3天的T淋巴细胞数量显著更高,但28天时并非如此。再次损伤后,使用CD11b表达评估,WT和RAG2-KO小鼠在总体小胶质细胞活性方面没有差异。这些数据表明,在RAG2-KO小鼠中,许多切除的运动神经元在初次切除后未能存活,而在WT小鼠中,它们发生萎缩,并可通过再次损伤重新刺激以再生其表型。此外,这些数据表明正常的T细胞功能,或RAG2基因在大脑中某些未知的功能,对于激活萎缩运动神经元的再生程序至关重要——这些程序具有调节神经可塑性的治疗潜力。