Quantitative expression of pluripotency-related genes in parthenogenetically produced buffalo (Bubalus bubalis) embryos and in putative embryonic stem cells derived from them

• Published in: Gene Expression Patterns 2014-09, Vol.16 (1), p.23-30
• Main Authors: Singh, K.P., Kaushik, R., Mohapatra, S.K., Garg, V., Rameshbabu, K., Singh, M.K., Palta, P., Manik, R.S., Singla, S.K., Chauhan, M.S.
• Format: Article
• Language: English
• Subjects: Animals; Blastocyst - metabolism; Bubalus bubalis; Buffaloes - embryology; Buffaloes - genetics; Buffaloes - metabolism; Cells, Cultured; Embryonic Development; Embryonic stem cells; Embryonic Stem Cells - cytology; Embryonic Stem Cells - metabolism; Embryonic Stem Cells - physiology; Embryos; Gene Expression Regulation, Developmental; Morula - metabolism; Oocytes - metabolism; Parthenogenesis; Pluripotent Stem Cells - cytology; Pluripotent Stem Cells - metabolism; Pluripotent Stem Cells - physiology; Real time PCR
• ISSN: 1567-133X; 1872-7298
• DOI: 10.1016/j.gep.2014.06.004

•We have successfully produced the parthenogenetic buffalo embryos of different developmental stages.•We have isolated the inner cell mass of blastocysts to culture parthenogenetically derived embryonic stem cells (pES).•To our knowledge this is the first report to quantify the pluripotency genes in different stages of embryos and different passages of embryonic stem cells in farm animals. Parthenogenetically produced embryos and embryonic stem (ES) cells derived from them offer a unique model for investigating the role of transcription factors in embryonic genome activation (EGA), pluripotent lineage specification and in pluripotency and self-renewal of ES cells because of the unique nature of these embryos. There is little information on the quantitative expression of important genes in parthenogenetically produced embryos and in ES cells derived from them. The present study examined the quantitative expression of some important genes in parthenogenetically produced buffalo embryos and in putative parthenogenetic ES cells (pES) cells. The quantitative expression of OCT-4, SOX-2, NANOG, REX-1, FOXD-3 and NUCLEOSTEMIN, which is very low in immature and mature oocytes, and in embryos at 2-, 4- and 8- to 16-cell stage, increases significantly at morula and blastocyst stage. The expression level of TELOMERASE, c-MYC and STAT-3, which is high in immature oocytes decreases during embryonic development followed by either an increase at the morula stage (TELOMERASE) or a low expression level maintained throughout development till blastocyst stage (c-MYC and STAT-3). There is a progressive decline in the expression level of OCT-4, SOX-2, c-MYC, REX-1, NUCLEOSTEMIN, TELOMERASE and STAT-3 during long term culture of pES cells.