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Differential Expression of Embryonic Stem Cell Genes in the Central and Peripheral Lens Epithelial Cells

Author: ZengXiaoMing
Tutor: LiWanCheng
School: Hunan Normal University
Course: Cell Biology
Keywords: Lens Stem cells CD9 SOX2 DLAD Polyclonal antibody
CLC: Q343
Type: Master's thesis
Year: 2010
Downloads: 23
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Abstract

Throughout the life course, the lens epithelial cells (Lens epithelial cells, LECs) almost all of the ongoing split its daughter cells continue to migrate from the genital area of ??the lens to the equatorial region, extending, and finally differentiate into lens fibers. Our previous studies have shown that in the central region of rabbit lens epithelial stem cells may be present in the lens, one of the aims of this study is to further confirm our previous conclusions. Of 6-month-old rabbits as experimental material, separating the central lens area and the surrounding area epithelial cells, and cultured in vitro for. Cells cultured in vitro analysis by RT-PCR and immunofluorescence technology, its main results are as follows: 1, different regions of the lens epithelial cells can be cultured in vitro success; 2, RT-PCR and immunofluorescence results show that embryonic stem cell gene CD9 and SOX2 in primary cultures of central lens epithelial cells significantly stronger than in the surrounding area were significantly different. This result is further supported by the center of the lens may exist lens stem cells in this reasoning. On the other hand, it is known lens epithelial cells through a self-elimination of their cells and a series of processes and differentiate into lens fibers. The DNA restriction endonuclease DLAD (DNase Ⅱ-like acid DNase, like type II DNA enzyme) in lens epithelial cell differentiation process of the denuclearization of lens fibers play an important role. DLAD protein coding genes have been cloned, but no DLAD antibody was sold on the market, thus impeding the functions on DLAD study. In order to be able to smoothly follow-up experiments, another part of the contents of this paper was prepared for the study of lens epithelial cell differentiation antibody - mouse DLAD (mouse DLAD, mDLAD) polyclonal antibodies. To this end, I first prepared and purified in this laboratory is actively studying the protein plays an important role in the vertebrate eye and other organizations - serine / threonine protein phosphatase 2A catalytic subunit p (PP-2Acβ ) of the GST fusion protein. This fusion protein as antigen recognition successfully prepared retinal pigment epithelial cells (ARPE) endogenous PP-2A antiserum. In this antibody successfully prepared based on the experience, I am actively preparing polyclonal antibody preparation DLAD. However, through a series of research I found that existing technologies and methods are almost impossible to induce in prokaryotic gene expression mDLAD. Part of the reason may be a DLAD encoded endonuclease, exogenous expression of the enzyme it may degrade the host DNA, so the host is not yet clear through some mechanism to prevent the expression of the exogenous enzyme to protect the host's own .

Differential Expression of Embryonic Stem Cell Genes in the Central and Peripheral Lens Epithelial Cells

Abstract

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