Freidin Mona, Asche-Godin Samantha, Abrams Charles K
Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Department of Neurology, State University of New York, Downstate Medical Center, Brooklyn, NY 11203, USA.
Exp Neurol. 2015 Jan;263:339-49. doi: 10.1016/j.expneurol.2014.10.014. Epub 2014 Oct 23.
X-linked Charcot-Marie-Tooth disease (CMT1X) is an inherited peripheral neuropathy caused by mutations in GJB1, the human gene for Connexin32 (Cx32). This present study uses Ilumina Ref8-v2 BeadArray to examine the expression profiles of injured and uninjured sciatic nerves at 5, 7, and 14 days post-crush injury (dpi) from Wild Type (WT) and Cx32-knockout (Cx32KO) mice to identify the genes and signaling pathways that are dysregulated in the absence of Schwann cell Cx32. Given the assumption that loss of Schwann cell Cx32 disrupts the regeneration and maintenance of myelinated nerve leading to a demyelinating neuropathy in CMT1X, we initially hypothesized that nerve crush injury would result in significant increases in differential gene expression in Cx32KO mice relative to WT nerves. However, microarray analysis revealed a striking collapse in the number of differentially expressed genes at 5 and 7 dpi in Cx32KO nerves relative to WT, while uninjured and 14 dpi time points showed large numbers of differentially regulated genes. Further comparisons within each genotype showed limited changes in Cx32KO gene expression following crush injury when compared to uninjured Cx32KO nerves. By contrast, WT nerves exhibited robust changes in gene expression at 5 and 7 dpi with no significant differences in gene expression by 14dpi relative to uninjured WT nerve samples. Taken together, these data suggest that the gene expression profile in uninjured Cx32KO sciatic nerve strongly resembles that of a WT nerve following injury and that loss of Schwann cell Cx32 leads to a basal state of gene expression similar to that of an injured WT nerve. These findings support a role for Cx32 in non-myelinating and regenerating populations of Schwann cells in normal axonal maintenance in re-myelination, and regeneration of peripheral nerve following injury. Disruption of Schwann cell-axonal communication in CMT1X may cause dysregulation of signaling pathways that are essential for the maintenance of intact myelinated peripheral nerves and to establish the necessary conditions for successful regeneration and remyelination following nerve injury.
X连锁型夏科-马里-图思病(CMT1X)是一种遗传性周围神经病,由连接蛋白32(Cx32)的人类基因GJB1发生突变引起。本研究使用Illumina Ref8-v2 BeadArray检测野生型(WT)和Cx32基因敲除(Cx32KO)小鼠在坐骨神经挤压伤后5天、7天和14天(dpi)时受伤和未受伤坐骨神经的表达谱,以确定在施万细胞Cx32缺失时失调的基因和信号通路。鉴于施万细胞Cx32的缺失会破坏有髓神经的再生和维持,导致CMT1X出现脱髓鞘性神经病这一假设,我们最初推测,相对于WT神经,神经挤压伤会导致Cx32KO小鼠的差异基因表达显著增加。然而,微阵列分析显示,相对于WT,Cx32KO神经在5 dpi和7 dpi时差异表达基因的数量显著减少,而未受伤和14 dpi时间点则显示出大量差异调节基因。每种基因型内的进一步比较表明,与未受伤的Cx32KO神经相比,挤压伤后Cx32KO基因表达的变化有限。相比之下,WT神经在5 dpi和7 dpi时基因表达有显著变化,而在14 dpi时相对于未受伤的WT神经样本,基因表达无显著差异。综上所述,这些数据表明,未受伤的Cx32KO坐骨神经中的基因表达谱与受伤后的WT神经非常相似,施万细胞Cx32的缺失导致基因表达的基础状态类似于受伤的WT神经。这些发现支持了Cx32在施万细胞的非髓鞘形成和再生群体中对正常轴突维持、再髓鞘形成以及损伤后周围神经再生的作用。CMT1X中施万细胞与轴突通讯的破坏可能导致对维持完整有髓周围神经至关重要的信号通路失调,并为神经损伤后成功再生和再髓鞘形成建立必要条件。