Influence of pub gene Expression on Differentiation of Mouse Embryonic Stem Cells into Derivatives of Ecto-, Meso-, and Endoderm in vitro

E V Novosadova; Novosadova E V; E S Manuilova; Manuilova E S; E L Arsenieva; Arsenieva E L; A N Lebedev; Lebedev A N; N V Khaidarova; Khaidarova N V; V Z Tarantul; Tarantul V Z; I A Grivennikov; Grivennikov I A

doi:10.32607/20758251-2009-1-2-93-97

Influence of pub gene Expression on Differentiation of Mouse Embryonic Stem Cells into Derivatives of Ecto-, Meso-, and Endoderm in vitro

Authors: Novosadova EV¹, Manuilova ES¹, Arsenieva EL¹, Lebedev AN¹, Khaidarova NV¹, Tarantul VZ¹, Grivennikov IA¹
Affiliations:
1. Institute of Molecular Genetics, Russian Academy of Sciences
Issue: Vol 1, No 2 (2009)
Pages: 93-97
Section: Articles
URL: http://actanaturae.ru/2075-8251/article/view/10823
DOI: https://doi.org/10.32607/20758251-2009-1-2-93-97
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Abstract

The influence of low and high pub gene expression on the initial stages of the differentiation of mouse embryonic stem cells into derivatives of ecto-, meso-, and endoderm in vitro was investigated. As follows from the results of a RT -PCR analysis, the expression of the vimentin, somatostatin, GATA 4, and GATA 6 genes, being the markers of endodermal differentiation, does not vary in both the cells with high pub gene expression and the cells with low pub gene expression, as well as in the corresponding control lines. The cells with high pub gene expression are characterized by an increase in the expression of mesodermal differentiation gene-markers ( trI card, trI skel, c-kit, and IL-7), whereas the cells with low pub gene expression are specified by a decrease in their expression. According to the analyses carried out, the reverse is characteristic of the expression of ectodermal differentiation gene-markers ( nestin, β- III tubulin, gfap, and th). Expression of these genes decreases in cell lines with high pub gene expression, whereas their expression increases with the decrease in pub gene expression. Hence, it is suggested that the variations in the pub gene expression in the embryonic stem cells influence significantly the mesodermal and ectodermal differentiation of these cells.

Keywords

embryonic stem cells, differentiation, polymerase chain reaction, pub gene, mesoderm, endoderm, and ectoderm.

An embryonic stem (ES) cell is a unique model to use for the investigation of the processes underway at the early stages of embryogenesis [1]. It is well known that in the course of in vivo embryo development, ES cells are able to form all three embryonic layers in culture – endoderm, mesoderm, and ectoderm – and, thus, all cell types developing from them. Analysis of gene expression in the process of ES cell differentiation into specialized cell types shows that the succession and efficiency of gene expression in the course of in vitro differentiation corresponds, as a whole, to the sequence of these processes in vivo [10]. Hence, ES cells may be used as an adequate experimental model for the investigation of molecular mechanisms at the initial stages of differentiation. Moreover, the investigations of ES cell differentiation in this or other directions in response to the action of specific inducers (growth factors, cytokines) or to direct the genetic modification of these cells make it possible to understand the functions of the investigated substances and different genes in this process [1]. In the previous investigations, we obtained and described cDNA clones characterized by intensive transcription in the HIV-associated immunoblastic lymphomas using the method of subtractive hybridization [16, 17]. An analysis of those cDNA allowed us to detect among them, along with the previously described genes (set, сalpain, etc.), several cDNA coding genes with previously unknown functions. One of such lymphoma-specific genes was then termed pub. The protein product of the human pub gene (hPub) is highly homological to the mouse Pub protein (mPub) [5].