Proteomic analysis of the major cellular proteins of bovine trophectoderm cell lines derived from IVP, parthenogenetic and nuclear transfer embryos: Reduced expression of annexins I and II in nuclear transfer-derived cell lines.
Trophectoderm cell lines were established from 8-day in vitro-cultured embryos of cattle derived from fertilization (IVF), somatic cell nuclear transfer (NT), or parthenogenetic activation (P) of in vitro-matured oocytes and from five 8-day-old in vivo (V) embryos. The most abundant cellular proteins of 5 V-, 16 NT-, 12 P-, and 16 IVF-derived cell lines were compared by 2D-gel electrophoresis and mass spectrometry; that is, the unaltered thiourea/urea extract of each cell culture was analyzed. Common protein spots (n=118) were examined, and 95% were identified with significant scores from protein and gene database searches. Of the proteins detected and identified, actin and cytokeratin-8 were found to be the most abundant. Other prominent cellular proteins were metabolic enzymes such as aldose reductase, phosphoglycerate mutase, enolase, triosephosphate isomerase, cytoskeletal interacting proteins transgelin and stratifin, anti-oxidant proteins peroxiredoxin 1 and anti-oxidant protein 2, and the calcium-dependent lipid-binding proteins annexins I and II. In comparative analysis of the 2D-gels, the NT-derived trophectoderm had less annexins I and II in comparison to the IVF- and P-derived trophectoderm. Because annexins I and II are abundant in the placenta and have functions important to the maintenance of placentation, the down-regulation of the annexin genes in the cultured NT trophectoderm may be related to the frequent failures of NT pregnancies.
Talbot
Neil C
NC
U.S. Department of Agriculture, Agricultural Research Service USDA, ARS, Animal and Natural Resources Institute, Animal Biosciences and Biotechnology Laboratory, Beltsville Agricultural Research Center, Beltsville, MD 20705-2350, United States. neil.talbot@ars.usda.gov
Powell
Anne M
AM
Caperna
Thomas J
TJ
Garrett
Wesley M
WM
D000818:Animals; D017305:Annexin A1 / Q000378:metabolism; D017306:Annexin A2 / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D015536:Down-Regulation; D004475:Ectoderm / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D046149:Embryo Culture Techniques; D004622:Embryo, Mammalian; D005307:Fertilization in Vitro / Q000662:veterinary; D053652:Nuclear Transfer Techniques / Q000662:veterinary; D010312:Parthenogenesis / Q000502:physiology*; D011506:Proteins / Q000032:analysis* / Q000378:metabolism; D020543:Proteome / Q000032:analysis; D040901:Proteomics; D014327:Trophoblasts / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*
D017305:Annexin A1; D017306:Annexin A2; D011506:Proteins; D020543:Proteome