Cumulus-specific genes are transcriptionally silent following somatic cell nuclear transfer in a mouse model.
This study investigated whether four cumulus-specific genes: follicular stimulating hormone receptor (FSHr), hyaluronan synthase 2 (Has2), prostaglandin synthase 2 (Ptgs2) and steroidogenic acute regulator protein (Star), were correctly reprogrammed to be transcriptionally silent following somatic cell nuclear transfer (SCNT) in a murine model. Cumulus cells of C57xCBA F1 female mouse were injected into enucleated oocytes, followed by activation in 10 micromol/L strontium chloride for 5 h and subsequent in vitro culture up to the blastocyst stage. Expression of cumulus-specific genes in SCNT-derived embryos at 2-cell, 4-cell and day 4.5 blastocyst stages was compared with corresponding in vivo fertilized embryos by real-time PCR. It was demonstrated that immediately after the first cell cycle, SCNT-derived 2-cell stage embryos did not express all four cumulus-specific genes, which continually remained silent at the 4-cell and blastocyst stages. It is therefore concluded that all four cumulus-specific genes were correctly reprogrammed to be silent following nuclear transfer with cumulus donor cells in the mouse model. This would imply that the poor preimplantation developmental competence of SCNT embryos derived from cumulus cells is due to incomplete reprogramming of other embryonic genes, rather than cumulus-specific genes.
Tong
Guo-qing
GQ
Nuclear Reprogramming and Stem Cell Laboratory, Department of Obstetrics and Gynecology, Yong Loo Lin School of Medicine, National University of Singapore, 119074 Singapore.
Heng
Boon-chin
BC
Ng
Soon-chye
SC
D000818:Animals; D051546:Cyclooxygenase 2 / Q000235:genetics; D004622:Embryo, Mammalian / Q000378:metabolism; D005260:Female; D020868:Gene Silencing; D014453:Glucuronosyltransferase / Q000235:genetics; D000076002:Hyaluronan Synthases; D008297:Male; D051379:Mice; D023421:Models, Animal; D053652:Nuclear Transfer Techniques; D010750:Phosphoproteins / Q000235:genetics; D011962:Receptors, FSH / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics*; D000098682:Steroidogenic Acute Regulatory Protein
D010750:Phosphoproteins; D011962:Receptors, FSH; D000098682:Steroidogenic Acute Regulatory Protein; C077623:Ptgs2 protein, mouse; D051546:Cyclooxygenase 2; D014453:Glucuronosyltransferase; C491099:Has2 protein, mouse; D000076002:Hyaluronan Synthases