Nagy J I, Rash J E
Department of Physiology and Pathophysiology, Faculty of Medicine, University of Manitoba, 745 Bannatyne Ave, Winnipeg, MB, R3E 0J9, Canada.
Department of Biomedical Sciences, and Program in Molecular, Cellular and Integrative Neurosciences, Colorado State University, Fort Collins, CO, USA.
Eur J Neurosci. 2017 Jul;46(2):1790-1804. doi: 10.1111/ejn.13614. Epub 2017 Jun 27.
Electrical synapses formed by connexin36 (Cx36)-containing gap junctions between interneurons in the cerebellar cortex have been well characterized, including those formed between basket cells and between Golgi cells, and there is gene reporter-based evidence for the expression of connexin45 (Cx45) in the cerebellar molecular layer. Here, we used immunofluorescence approaches to further investigate expression patterns of Cx36 and Cx45 in this layer and to examine localization relationships of these connexins with each other and with glial connexin43 (Cx43). In mice, strain differences were found, such that punctate labelling for Cx36 was differentially distributed in the molecular layer of C57BL/6 vs. CD1 mice. In mice with EGFP reporter representing Cx36 expression, Cx36-puncta were localized to processes of stellate cells and other cerebellar interneurons. Punctate labelling of Cx45 was faint in the molecular layer of wild-type mice and was increased in intensity in mice with Cx36 gene ablation. The vast majority of Cx36-puncta co-localized with Cx45-puncta, which in turn was associated with the scaffolding protein zonula occludens-1. In rats, Cx45-puncta were also co-localized with Cx36-puncta and additionally occurred along Bergmann glial processes adjacent to Cx43-puncta. The results indicate strain and species differences in Cx36 as well as Cx45 expression, possible compensatory processes after loss of Cx36 expression and localization of Cx45 to both neuronal and Bergmann glial gap junctions. Further, expression of both Cx43 and Cx45 in Bergmann glia of rat may contribute to the complex properties of junctional coupling between these cells and perhaps to their reported coupling with Purkinje cells.
小脑皮质中间神经元之间由含连接蛋白36(Cx36)的缝隙连接形成的电突触已得到充分表征,包括篮状细胞之间以及高尔基细胞之间形成的电突触,并且有基于基因报告的证据表明连接蛋白45(Cx45)在小脑分子层中表达。在此,我们使用免疫荧光方法进一步研究该层中Cx36和Cx45的表达模式,并检查这些连接蛋白彼此之间以及与胶质连接蛋白43(Cx43)的定位关系。在小鼠中,发现了品系差异,使得Cx36的点状标记在C57BL/6小鼠与CD1小鼠的分子层中分布不同。在具有代表Cx36表达的EGFP报告基因的小鼠中,Cx36点状结构定位于星状细胞和其他小脑中间神经元的突起。Cx45的点状标记在野生型小鼠的分子层中很微弱,而在Cx36基因敲除的小鼠中强度增加。绝大多数Cx36点状结构与Cx45点状结构共定位,而Cx45点状结构又与支架蛋白闭合蛋白-1相关。在大鼠中,Cx45点状结构也与Cx36点状结构共定位,并且还沿着与Cx43点状结构相邻的伯格曼胶质细胞突起出现。结果表明Cx36以及Cx45表达存在品系和物种差异,Cx36表达缺失后可能存在补偿过程,以及Cx45定位于神经元和伯格曼胶质细胞的缝隙连接。此外,大鼠伯格曼胶质细胞中Cx43和Cx45的表达可能有助于这些细胞之间连接偶联的复杂特性,也许还有助于它们与浦肯野细胞的报道偶联。