缝隙连接蛋白26缺陷聋的神经细胞中缝隙连接蛋白的表达差异
发布时间：2018-12-12 14:25
【摘要】：目的:通过诱导多能干细胞(iPSCs)技术获得缝隙连接蛋白26(Cx26)缺陷聋患儿的神经细胞,研究Cx26缺陷对神经细胞发育和基因表达的影响。方法:从3例Cx26缺陷造成的极重度耳聋患儿取得成纤维细胞,将之诱导为数个非整合iPSC细胞系,鉴定这些细胞系的形态和内源性、外源性基因表达。然后将这些细胞系向神经细胞方向分化,检测整个过程中形态、多能性基因、神经标记物、缝隙连接蛋白基因表达变化。结果:能够成功建立Cx26缺陷的3个iPSC细胞系,并且可以在体外分化为神经前体细胞和神经元细胞;这些细胞的形态、增殖、内源性、外源性基因表达与人胚胎干细胞基本一致;iPSC细胞系分化出的神经元细胞中,Cx32的表达明显上调,Cx36的表达略微上调,Cx26的表达没有明显变化。结论:Cx26缺陷不影响诱导多能干细胞的神经分化,但其过程中Cx32和Cx36表达上调,提示Cx32可能对Cx26缺陷发挥了代偿作用。
[Abstract]:Aim: to obtain nerve cells from deafness children with gap junction protein 26 (Cx26) deficiency by inducing pluripotent stem cell (iPSCs), and to study the effect of Cx26 deficiency on neuronal development and gene expression. Methods: fibroblasts were obtained from 3 children with extremely severe deafness caused by Cx26 deficiency. The fibroblasts were induced into several unintegrated iPSC cell lines and their morphology, endogenous and exogenous gene expression were identified. Then these cell lines were differentiated into nerve cells, and the expression changes of morphology, pluripotent genes, neural markers and gap junction protein genes in the whole process were detected. Results: three iPSC cell lines with Cx26 deficiency could be successfully established and differentiated into neural precursor cells and neuron cells in vitro, and the morphology, proliferation, endogenous and exogenous gene expression of these cells were basically consistent with those of human embryonic stem cells. In the differentiated neurons of iPSC cell line, the expression of Cx32 was up-regulated, the expression of Cx36 was slightly up-regulated, and the expression of Cx26 was not changed. Conclusion: Cx26 deficiency does not affect the neural differentiation of pluripotent stem cells, but the expression of Cx32 and Cx36 is up-regulated, suggesting that Cx32 may play a compensatory role on Cx26 deficiency.
【作者单位】： 中国医学科学院北京协和医院耳鼻咽喉科;中国医学科学院基础医学研究所;
【分类号】：R764.43
本文编号：2374737
[Abstract]:Aim: to obtain nerve cells from deafness children with gap junction protein 26 (Cx26) deficiency by inducing pluripotent stem cell (iPSCs), and to study the effect of Cx26 deficiency on neuronal development and gene expression. Methods: fibroblasts were obtained from 3 children with extremely severe deafness caused by Cx26 deficiency. The fibroblasts were induced into several unintegrated iPSC cell lines and their morphology, endogenous and exogenous gene expression were identified. Then these cell lines were differentiated into nerve cells, and the expression changes of morphology, pluripotent genes, neural markers and gap junction protein genes in the whole process were detected. Results: three iPSC cell lines with Cx26 deficiency could be successfully established and differentiated into neural precursor cells and neuron cells in vitro, and the morphology, proliferation, endogenous and exogenous gene expression of these cells were basically consistent with those of human embryonic stem cells. In the differentiated neurons of iPSC cell line, the expression of Cx32 was up-regulated, the expression of Cx36 was slightly up-regulated, and the expression of Cx26 was not changed. Conclusion: Cx26 deficiency does not affect the neural differentiation of pluripotent stem cells, but the expression of Cx32 and Cx36 is up-regulated, suggesting that Cx32 may play a compensatory role on Cx26 deficiency.
【作者单位】： 中国医学科学院北京协和医院耳鼻咽喉科;中国医学科学院基础医学研究所;
【分类号】：R764.43
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