title: Differential effects of culture and nuclear transfer on relative transcript levels of genes with key roles during preimplantation.
abstract: It is well known that the preimplantation culture environment to which embryos are exposed influences the expression of developmentally important genes. Recently, it has been reported that MEMalpha, a culture medium commonly used for somatic cells, allows high rates of preimplantation development and development to term of mouse somatic cell nuclear transfer (SCNT) embryos. The objective of this study was to compare the differential effects of this medium and of the nuclear transfer procedure on the relative mRNA abundance of several genes with key roles during preimplantation. The relative mRNA levels of nine genes (Glut 1, Glut 5, G6PDH, Bax, Survivin, Gpx 1, Oct4, mTert and IGF2bp1) were quantified at blastocyst stage on cumulus cell cloned embryos cultured in MEMalpha, as well as on in vivo cultured and MEMalpha cultured controls. Only three of the nine transcripts analysed (Glut 5, Gpx 1 and Igf2bp1) were significantly down-regulated at blastocyst stage in in vitro produced controls. However, most genes analysed in our MEMalpha cultured cloned embryos showed altered transcription levels. Interestingly, between cloned and in vitro produced controls only the transcription levels measured for Glut 1 were significantly different. This result suggests that Glut 1 may be a good marker for embryo quality after cumulus cell nuclear transfer.
authors.affiliation: Dpto. de Reproducción Animal y Conservación de Recursos Zoogenéticos, INIA, Madrid, Spain.
authors.forename: P N
authors.initials: PN
authors.lastname: Moreira
authors.forename: R
authors.initials: R
authors.lastname: Fernández-Gonzalez
authors.forename: M A
authors.initials: MA
authors.lastname: Ramirez
authors.forename: M
authors.initials: M
authors.lastname: Pérez-Crespo
authors.forename: D
authors.initials: D
authors.lastname: Rizos
authors.forename: B
authors.initials: B
authors.lastname: Pintado
authors.forename: A
authors.initials: A
authors.lastname: Gutiérrez-Adán
mesh_terms: D000818:Animals; D001755:Blastocyst; D019976:Cloning, Organism; D003470:Culture Media; D046149:Embryo Culture Techniques / Q000379:methods*; D047108:Embryonic Development / Q000235:genetics; D005260:Female; D018507:Gene Expression Regulation, Developmental; D005819:Genetic Markers; D051272:Glucose Transporter Type 1 / Q000378:metabolism; D008297:Male; D051379:Mice; D053652:Nuclear Transfer Techniques; D012333:RNA, Messenger / Q000378:metabolism; D014158:Transcription, Genetic / Q000502:physiology
chemical_list: D003470:Culture Media; D005819:Genetic Markers; D051272:Glucose Transporter Type 1; D012333:RNA, Messenger