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The Expression Analysis Of Ube2e3 During Mouse Embryonic Development

Posted on:2012-05-09Degree:MasterType:Thesis
Country:ChinaCandidate:Y J XingFull Text:PDF
GTID:2210330362951367Subject:Biochemistry and Molecular Biology
Abstract/Summary:PDF Full Text Request
Ube2e3, encoding a ubiquitin conjugating enzyme Ube2e3 in mouse, is identified as a growth-inhibitory gene. Ube2e3 involves in the ubiquitin/proteasome system and is ubquitiously expressed among diverse tissues of adult mouse. In this study, the expression patterns of Ube2e3 were examined using in situ hybridization and quantitative reverse transcription-PCR (qRT-PCR) at mouse mid-late embryonic stage. The expression variation of Ube2e3 in NIH3T3 cells treated by the inhibitors of DNA methyltransferase and histone deacetylases was analyzed to investigate the primary regulation mechanism.Bioinformatics analysis indicated that Ube2e3 was conserved evolutionarily. Its protein was highly conserved among vertebrates, with the amino-acid sequences of Homo sapiens and Mus musculus exhibiting 100% identity. The core domain of Ube2e3 was conserved in the phylogenetic evolution, whereas its amino terminal extension revealed high specificity. In situ hybridization illustrated that Ube2e3 was prominently expressed in brain, neural tube, limb buds and branchial arch at E10.5 (Embryonic day 10.5), while mainly detected in brain, limb buds and somite at E11.5. At late development period, the expression of Ube2e3 was mainly distributed in the central nervous system and skeletal system, less in tongue, heart, lung and liver. Quantitative analysis showed that Ube2e3 was continuously expressed during mid-later-gestation period. At E15.5, Ube2e3 was expressed abundantly in brain, weakly in lung, liver and kidney, accompanied by the lowest level in tongue and heart. During the brain development, the expression level of Ube2e3 first ascended and then decreased from E12.5 to E18.5, the peak of which sustained starting at E14.5 till E16.5. What's more, the expression of Ube2e3 had no change between the control cells and those treated by 5-aza-CdR and 4-PBA respectively or in combination.Together, these results suggest that Ube2e3 may relate to the development of nervous system and skeletal system during mouse embryonic development, exempt from the regulation of DNA methylation and histone acetylation in NIH3T3 cells.
Keywords/Search Tags:Ube2e3, in situ hybridization, embryonic development, epigenetic regulation
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