Ponsioen Theodorus L, van Luyn Marja J A, van der Worp Roelofje J, Pas Hendri H, Hooymans Johanna M M, Los Leonoor I
University Medical Center Groningen and University of Groningen, Department of Ophthalmology, Groningen, The Netherlands.
Mol Vis. 2008 Mar 26;14:652-60.
To investigate the capacity of cultured Müller cells to synthesize collagens, since previous studies indicated that Müller cells could be involved in collagen remodeling at the vitreoretinal border in adult human eyes.
Spontaneously immortalized cultured human Müller cells were analyzed for the presence of mRNA of types I-VII, IX, XI, and XVII collagen by RT-PCR. Furthermore, Müller cells were immunocytochemically stained for light microscopic (LM) evaluation of these collagens and their main characteristics. Finally, cell extracts and culture medium were evaluated by western blot (WB) analysis using anticollagen antibodies.
Cultured Müller cells contained mRNA for types I-VII, IX, and XI collagen, but not for type XVII collagen. LM and WB confirmed the intracellular expression of all the above-mentioned collagens with the exception of type XVII. Collagen secretion into the medium was established for types I-VII, IX, and XI collagen.
Cultured Müller cells can synthesize internal limiting lamina and vitreous collagens. Possible collagen production by Müller cells could explain and expand on previous in vivo morphological findings in the embryonic and postnatal period and in pathologic conditions.
研究培养的 Müller 细胞合成胶原蛋白的能力,因为先前的研究表明 Müller 细胞可能参与成人眼睛玻璃体视网膜边界处的胶原重塑。
通过逆转录聚合酶链反应(RT-PCR)分析自发永生化的培养人 Müller 细胞中 I-VII、IX、XI 和 XVII 型胶原蛋白的 mRNA 存在情况。此外,对 Müller 细胞进行免疫细胞化学染色,以通过光学显微镜(LM)评估这些胶原蛋白及其主要特征。最后,使用抗胶原蛋白抗体通过蛋白质印迹(WB)分析评估细胞提取物和培养基。
培养的 Müller 细胞含有 I-VII、IX 和 XI 型胶原蛋白的 mRNA,但不含有 XVII 型胶原蛋白的 mRNA。光学显微镜和蛋白质印迹证实除 XVII 型外上述所有胶原蛋白的细胞内表达。已确定 I-VII、IX 和 XI 型胶原蛋白分泌到培养基中。
培养的 Müller 细胞可以合成内界膜和玻璃体胶原蛋白。Müller 细胞可能产生的胶原蛋白可以解释并扩展先前在胚胎期、出生后时期以及病理状况下的体内形态学发现。