Proteomic analysis of parthenogenetic and in vitro fertilized porcine embryos.
Proteomic data from embryos are essential for the completion of whole proteome catalog due to embryo-specific expression of certain proteins. In this study, using reverse phase LC-MS/MS combined with 1-D SDS-PAGE, we identified 1625 mammalian and 735 Sus scrofa proteins from porcine zygotes that included both cytosolic and membranous proteins. We also found that the global protein profiles of parthenogenetically activated (PA) and in vitro fertilized (IVF) zygotes were similar but differences in expression of individual proteins were also evident. These differences were not due to culture conditions, polyspermy or non-activation of oocytes, as the same culture method was used in both groups, the frequency of polyspermy was 24.3+/-3.0% and the rates of oocyte activation did not differ (p>0.05) between PA and IVF embryos. Consistent with proteomic data, fluorescent Hoechst 33 342 staining and terminal deoxynucleotidyl transferase dUTP nick end labeling assay also revealed that PA embryos were of poor quality as they contained less cells per blastocyst and were more predisposed to apoptosis (p<0.05), although their in vitro development rates were similar. To our knowledge, this is the first report on global peptide sequencing and quantification of protein in PA and IVF embryos by LC-MS/MS that may be useful as a reference map for future studies.
Gupta
Mukesh Kumar
MK
Department of Animal Biotechnology, Bio-Organ Research Center, Konkuk University, Hwayang-dong, Gwangjin-Gu, Seoul, South Korea.
Jang
Jung Mi
JM
Jung
Jin Woo
JW
Uhm
Sang Jun
SJ
Kim
Kwang Pyo
KP
Lee
Hoon Taek
HT
D000818:Animals; D001755:Blastocyst / Q000378:metabolism; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004622:Embryo, Mammalian / Q000378:metabolism*; D005306:Fertilization; D005307:Fertilization in Vitro; D010312:Parthenogenesis; D011506:Proteins / Q000032:analysis; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics; D015203:Reproducibility of Results; D013552:Swine / Q000196:embryology*; D053719:Tandem Mass Spectrometry
D011506:Proteins; D020543:Proteome