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Single cell expression analysis of genes with potential mrna gradient in mouse oocytes
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SYSNO ASEP 0463008 Document Type K - Proceedings Paper (Czech conf.) R&D Document Type The record was not marked in the RIV Title Single cell expression analysis of genes with potential mrna gradient in mouse oocytes Author(s) Dorosh, Andriy (BTO-N) RID
Margaryan, Hasmik (BTO-N) RID
Vodička, Martin (FGU-C) RID, ORCID, SAI
Ergang, Peter (FGU-C) RID, ORCID
Šídová, Monika (BTO-N) RID
Dvořáková-Hortová, Kateřina (BTO-N)Number of authors 6 Source Title Book of abstracts of XXIInd Symposium of biology and immunology of reproduction. - Vestec u Prahy : Biotechnologický ústav AVČR, 2016 / Pěknicová Jana ; Elzeinová Fatima ; Kubátová Alena
S. 34-34Number of pages 1 s. Action XXIInd Symposium of biology and immunology of reproduction with international participation Event date 26.05.2016-28.05.2016 VEvent location Třešť Country CZ - Czech Republic Event type WRD Language eng - English Country CZ - Czech Republic Keywords single cell expresion analysis ; RT-qPCR ; oocytes ; Laser capture microdissection ; in situ hybridization Subject RIV EB - Genetics ; Molecular Biology R&D Projects GA 14-05547S GA ČR - Czech Science Foundation (CSF) Institutional support BTO-N - RVO:86652036 Annotation In frogs, there are clearly visible differently pigmented animal and vegetal poles of the egg determined before fertilization and leading to asymmetrical divisions. Mammalian egg does not show any comparable differentiation and it has been generally accepted that even the individual blastomeres in 2-cell and 4-cell embryos are homogenous. However, recent findings suggest that those blastomeres display different gene expression patterns and might already possess some inclinations to specific cell lineages. We therefore raised a question, whether there could be any mRNA or protein gradients in pre-fertilization oocytes similar to a previously described amphibian egg one. In mammalian eggs, there is a membrane region that is poor in microvilli, cortical granules are absent beneath plasma membrane and sperm cells generally do not bind to this location. This microvilli free region also covers the egg nucleus, and cytoskeleton localization differs markedly to the rest of the cortical space, forming actin –myosin II cortical cap/ring and is considered as animal pole. The purpose of this study was to determine gene products that can be detected at single cell level using qPCR and display gradient like distribution in mature oocytes. We checked expression of 12 selected genes in a pool of 10 oocytes and single mature oocytes. Then, we analysed gene expression in fixed intact oocytes and those undergoing laser capture microdissection procedure (LCMD). Eventually, we have determined six candidate genes for the study of intracellular spatial gene expression in mature mammalian oocytes by subcellular qPCR and in situ hybridization. Workplace Institute of Biotechnology Contact Monika Kopřivová, Monika.Koprivova@ibt.cas.cz, Tel.: 325 873 700 Year of Publishing 2017
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