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小鼠视网膜出生后发育过程中连接蛋白36、43和45的表达

Expression of connexins 36, 43, and 45 during postnatal development of the mouse retina.

作者信息

Kihara Alexandre Hiroaki, Mantovani de Castro Leandro, Belmonte Mônica Aparecida, Yan Chao Yun Irene, Moriscot Anselmo Sigari, Hamassaki Dânia Emi

机构信息

Department of Cell & Developmental Biology, University of São Paulo, São Paulo, SP, Brazil.

出版信息

J Neurobiol. 2006 Nov;66(13):1397-410. doi: 10.1002/neu.20299.

Abstract

Gap junction channels formed by connexins (Cx) may play essential roles in some processes that occur during retinal development, such as apoptosis and calcium wave spread. The present study was undertaken to determine the distribution pattern of Cx36, Cx43, and Cx45 by immunofluorescence, as well as their gene expression levels by quantitative PCR during postnatal development of the mouse retina. Our results showed an increased expression of neuronal Cx36 from P1 until P10, when this Cx reached adult levels, and it was mainly distributed in the outer and inner plexiform layers. In turn, Cx43 was almost absent in retinal progenitor cells at P1, it became more prominent in glial cell processes about P10, and did not change until adulthood. Double-labeling studies in situ and in vitro with antivimentin, a Müller cell marker, confirmed that Cx43 was expressed by these cells. In addition, quantitative PCR showed that Cx43 and vimentin shared very similar temporal expression patterns. Finally, in contrast to Cx36 and Cx43, Cx45 mRNA was strongly down-regulated during development. In early postnatal days, Cx45 was seen ubiquitously distributed throughout the retina in cells undergoing proliferation and differentiation, as well in differentiated neurons. In adult retina, this protein had a more restricted distribution both in neurons and glial cells, as confirmed in situ and in vitro. In conclusion, we observed a distinct temporal expression pattern for Cx36, Cx43, and Cx45, which is probably related to particular roles in retinal function and maintenance of homeostasis during development of the mouse retina.

摘要

由连接蛋白(Cx)形成的间隙连接通道可能在视网膜发育过程中发生的某些过程中发挥重要作用,如细胞凋亡和钙波传播。本研究旨在通过免疫荧光法确定Cx36、Cx43和Cx45的分布模式,以及通过定量PCR确定它们在小鼠视网膜出生后发育过程中的基因表达水平。我们的结果显示,神经元Cx36从出生后第1天(P1)到第10天(P10)表达增加,此时该连接蛋白达到成年水平,并且主要分布在外丛状层和内丛状层。相应地,Cx43在P1时视网膜祖细胞中几乎不存在,在大约P10时在神经胶质细胞突起中变得更加突出,并且直到成年都没有变化。用穆勒细胞标志物波形蛋白进行的原位和体外双标记研究证实,Cx43由这些细胞表达。此外,定量PCR显示Cx43和波形蛋白具有非常相似的时间表达模式。最后,与Cx36和Cx43相反,Cx45 mRNA在发育过程中强烈下调。在出生后早期,Cx45在经历增殖和分化的细胞以及分化的神经元中普遍分布于整个视网膜。在成年视网膜中,这种蛋白在神经元和神经胶质细胞中的分布更受限制,原位和体外研究均证实了这一点。总之,我们观察到Cx36、Cx43和Cx45有明显的时间表达模式,这可能与它们在小鼠视网膜发育过程中的视网膜功能和内环境稳态维持中的特定作用有关。

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