Nagy James I, Ionescu Andrei V, Lynn Bruce D, Rash John E
Department of Physiology, Faculty of Medicine, University of Manitoba, Winnipeg, Manitoba R3E 3J7, Canada.
J Comp Neurol. 2003 Sep 22;464(3):356-70. doi: 10.1002/cne.10797.
The cellular localization, relation to other glial connexins (Cx30, Cx32, and Cx43), and developmental expression of Cx29 were investigated in the mouse central nervous system (CNS) with an anti-Cx29 antibody. Cx29 was enriched in subcellular fractions of myelin, and immunofluorescence for Cx29 was localized to oligodendrocytes and myelinated fibers throughout the brain and spinal cord. Oligodendrocyte somata displayed minute Cx29-immunopositive puncta around their periphery and intracellularly. In developing brain, Cx29 levels increased during the first few postnatal weeks and were highest in the adult brain. Immunofluorescence labeling for Cx29 in oligodendrocyte somata was intense at young ages and was dramatically shifted in localization primarily to myelinated fibers in mature CNS. Labeling for Cx32 also was localized to oligodendrocyte somata and myelin and absent in Cx32 knockout mice. Cx29 and Cx32 were minimally colocalized on oligodendrocytes somata and partly colocalized along myelinated fibers. At gap junctions on oligodendrocyte somata, Cx43/Cx32 and Cx30/Cx32 were strongly associated, but there was minimal association of Cx29 and Cx43. Cx32 was very sparsely associated with astrocytic connexins along myelinated fibers. With Cx26, Cx30, and Cx43 expressed in astrocytes and Cx29, Cx32, and Cx47 expressed in oligodendrocytes, the number of connexins localized to gap junctions of glial cells is increased to six. The results suggested that Cx29 in mature CNS contributes minimally to gap junctional intercellular communication in oligodendrocyte cell bodies but rather is targeted to myelin, where it, with Cx32, may contribute to connexin-mediated communication between adjacent layers of uncompacted myelin.
利用抗Cx29抗体在小鼠中枢神经系统(CNS)中研究了Cx29的细胞定位、与其他胶质连接蛋白(Cx30、Cx32和Cx43)的关系以及发育表达情况。Cx29在髓磷脂的亚细胞组分中富集,Cx29的免疫荧光定位于整个脑和脊髓的少突胶质细胞和有髓纤维。少突胶质细胞胞体在其周边和细胞内显示出微小的Cx29免疫阳性斑点。在发育中的脑内,Cx29水平在出生后的头几周内升高,在成年脑中最高。少突胶质细胞胞体中Cx29的免疫荧光标记在幼年时强烈,而在成熟CNS中其定位主要显著转移至有髓纤维。Cx32的标记也定位于少突胶质细胞胞体和髓磷脂,且在Cx32基因敲除小鼠中不存在。Cx29和Cx32在少突胶质细胞胞体上极少共定位,而在有髓纤维上部分共定位。在少突胶质细胞胞体的缝隙连接处,Cx43/Cx32和Cx30/Cx32强烈相关,但Cx29和Cx43的相关性极小。沿着有髓纤维,Cx32与星形胶质细胞连接蛋白的关联非常稀少。星形胶质细胞表达Cx26、Cx30和Cx43,少突胶质细胞表达Cx29、Cx32和Cx47,定位于胶质细胞缝隙连接处的连接蛋白数量增加到了六种。结果表明,成熟CNS中的Cx29对少突胶质细胞胞体中的缝隙连接细胞间通讯贡献极小,而是靶向至髓磷脂,在那里它与Cx32一起可能有助于未压实髓磷脂相邻层之间的连接蛋白介导的通讯。