Allele-specific expression of the MAOA gene and X chromosome inactivation in in vitro produced bovine embryos
| Autor: | T. O. Diesel, Rodolfo Rumpf, J. O. Carvalho, A.R. Ferreira, Margot Alves Nunes Dode, G. M. Machado, Maurício Machaim Franco, E. O. Melo |
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| Rok vydání: | 2010 |
| Předmět: |
Male
animal structures Biology X-inactivation Andrology X Chromosome Inactivation Gene expression Genetics medicine Animals Blastocyst RNA Messenger Gene Monoamine Oxidase reproductive and urinary physiology X chromosome Alleles Cumulus Cells Reverse Transcriptase Polymerase Chain Reaction Gene Expression Profiling Embryogenesis Embryo Cell Biology Sequence Analysis DNA DNA Methylation Molecular biology Gene expression profiling medicine.anatomical_structure Phenotype embryonic structures Cattle Female Polymorphism Restriction Fragment Length Developmental Biology |
| Zdroj: | Molecular reproduction and development. 77(7) |
| ISSN: | 1098-2795 |
| Popis: | During embryogenesis, one of the two X chromosomes is inactivated in embryos. The production of embryos in vitro may affect epigenetic mechanisms that could alter the expression of genes related to embryo development and X chromosome inactivation (XCI). The aim of this study was to understand XCI during in vitro, pre-implantation bovine embryo development by characterizing the allele-specific expression pattern of the X chromosome-linked gene, monoamine oxidase A (MAOA). Two pools of ten embryos, comprised of the 4-, 8- to 16-cell, morula, blastocyst, and expanded blastocyst stages, were collected. Total RNA from embryos was isolated, and the RT-PCR-RFLP technique was used to observe expression of the MAOA gene. The DNA amplicons were also sequenced using the dideoxy sequencing method. MAOA mRNA was detected, and allele-specific expression was identified in each pool of embryos. We showed the presence of both the maternal and paternal alleles in the 4-, 8- to 16-cell, blastocyst and expanded blastocyst embryos, but only the maternal allele was present in the morula stage. Therefore, we can affirm that the paternal X chromosome is totally inactivated at the morula stage and reactivated at the blastocyst stage. To our knowledge, this is the first report of allele-specific expression of an X-linked gene that is subject to XCI in in vitro bovine embryos from the 4-cell to expanded blastocyst stages. We have established a pattern of XCI in our in vitro embryo production system that can be useful as a marker to assist the development of new protocols for in vitro embryo production. |
| Databáze: | OpenAIRE |
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