Maternal housekeeping proteins translated during bovine oocyte maturation and early embryo development.
Protein synthesis from maternal mRNA is needed to sustain oocyte maturation and embryo development prior to the maternal-embryonic transition (MET). Therefore, proteins that are expressed throughout this time are important and may be considered as maternal housekeeping proteins (MHKP). Our objectives were first, identify the translated protein patterns of bovine embryo development and secondly, determine the MHKP. Proteins synthesized during oocyte maturation and embryo development (2, 4 and 8-cell stages) were labeled using [S(35)]-Met and [S(35)]-Cys, and visualized by 2-DE. Embryos were cultured with alpha-amanitine to inhibit new transcription. Only 46 proteins were present throughout all stages. Ten spots were identified by MALDI-TOF and MS/MS: HSC71; HSP70; CypA; UCH-L1; GSTM5; Cct5; E-FABP; 2,3-BPGM, ubiquitin-conjugating enzyme E2D3; and beta-actin/gamma-actin. A new method called in silico protein identification confirmation was developed using EST databases. This method is a promising approach for use in rare tissue or from species with an incomplete protein database. This study has revealed that the translated protein patterns show a transition that brings the embryo to the MET. The needs in translated proteins between oocyte maturation and embryo development are different. In summary, this study represents the bases for future proteomics studies on bovine oocytes and embryos.
Massicotte
Lyne
L
Centre de Recherche en Biologie de la Reproduction, Département des Sciences Animales, Université Laval, Ste-Foy, Québec, Canada.
Coenen
Karine
K
Mourot
Marina
M
Sirard
Marc-André
MA
D000818:Animals; D002417:Cattle; D015180:Electrophoresis, Gel, Two-Dimensional; D047108:Embryonic Development; D009865:Oocytes / Q000166:cytology*; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D011506:Proteins; D012333:RNA, Messenger