Connexin36 localization to pinealocytes in the pineal gland of mouse and rat.
Connexin36 localization to pinealocytes in the pineal gland of mouse and rat.
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Connexin36 定位于小鼠和大鼠松果体中的松果体细胞。
DOI：
10.1111/ejn.13602
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Nagy,JI
中科院分区：
文献类型：
--
作者：
Wang,SG;Tsao,DD;Vanderpool,KG;Yasumura,T;Rash,JE;Nagy,JI
已知松果体中的几种细胞类型可以建立细胞间间隙连接，但这些连接的连接蛋白成分尚未完全表征。具体来说，已经在松果体中检查了连接蛋白 36 (Cx36) 蛋白和 mRNA 的表达，但产生 Cx36 和形成含 Cx36 间隙连接的细胞的身份尚未确定。我们使用 Cx36 的免疫荧光和冷冻断裂复制品免疫金标记 (FRIL) 来研究 Cx36 在成年小鼠和大鼠松果体中的细胞和亚细胞定位。 Cx36 的免疫荧光标记仅可视化为分布在整个松果体中的点或短免疫阳性链，并且在 Cx36 缺失小鼠的松果体切片中不存在。通过双重免疫荧光标记，Cx36 定位于色氨酸羟化酶阳性和 5-羟色胺阳性松果体细胞细胞体及其大的初始过程，包括这些过程的交叉点和显示过程汇合的位点。细胞连接标记物闭合小带-1 (ZO-1) 的标记与 Cx36 的标记重叠或密切相关。因此，松果体细胞形成含有 Cx36 的间隙连接，其中还包含支架蛋白 ZO-1。 FRIL揭示了Cx36标记在松果体细胞之间超微结构限定的间隙连接处，其中大部分位于具有网状、带状或串状结构的间隙连接处。结果表明，松果体细胞的内分泌功能及其褪黑激素的分泌是通过含有 Cx36 的间隙连接的细胞间通讯来支持的，现在可以使用 Cx36 缺失小鼠进行测试。
Several cell types in the pineal gland are known to establish intercellular gap junctions, but the connexin constituents of those junctions have not been fully characterized. Specifically, the expression of connexin36 (Cx36) protein and mRNA has been examined in the pineal, but the identity of cells that produce Cx36 and that form Cx36‐containing gap junctions has not been determined. We used immunofluorescence and freeze fracture replica immunogold labelling (FRIL) of Cx36 to investigate the cellular and subcellular localization of Cx36 in the pineal gland of adult mouse and rat. Immunofluorescence labelling of Cx36 was visualized exclusively as puncta or short immunopositive strands that were distributed throughout the pineal, and which were absent in pineal sections from Cx36 null mice. By double immunofluorescence labelling, Cx36 was localized to tryptophan hydroxylase‐positive and 5‐hydroxytryptamine‐positive pinealocyte cell bodies and their large initial processes, including at intersections of those processes and at sites displaying a confluence of processes. Labelling for the cell junction marker zonula occludens‐1 (ZO‐1) either overlapped or was closely associated with labelling for Cx36. Pinealocytes thus form Cx36‐containing gap junctions that also incorporate the scaffolding protein ZO‐1. FRIL revealed labelling of Cx36 at ultrastructurally defined gap junctions between pinealocytes, most of which was at gap junctions having reticular, ribbon or string configurations. The results suggest that the endocrine functions of pinealocytes and their secretion of melatonin is supported by their intercellular communication via Cx36‐containing gap junctions, which may now be tested by the use of Cx36 null mice.
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