Comparative proteome analysis of porcine follicular fluid and serum reveals that excessive alpha(2)-macroglobulin in serum hampers successful expansion of cumulus-oocyte complexes.
Porcine follicular fluid (pFF) constitutes the micro-environment of the maturing oocyte. Although pFF is a transudate of serum, in pigs, it is superior to serum in promoting in vitro expansion of the cumulus cells, a specialized cell population surrounding the oocyte. A comparative proteome analysis of autologous serum and pFF was performed to investigate proteins involved in successful cumulus expansion of porcine oocytes. iTRAQ labeling followed by 2-D LC ESI-Q-TOF MS/MS revealed 63 proteins common to both fluids of which the abundance of 13 proteins was significantly different (p<0.05). Seven proteins were more concentrated in serum whereas six proteins were more abundant in pFF. To investigate the importance of these proteins, the cumulus matrices of COCs were collected after in vitro maturation in media supplemented with either of both biologically fluids and then subjected to 2-D PAGE analysis. alpha(2)-Macroglobulin and CH4 and secrete domains of swine IgM, which were both less abundant in pFF, were absent from cumulus matrix extracts after in vitro maturation in pFF. Although both proteins were incorporated in the matrices of cumulus-oocyte complexes matured in serum, depletion of alpha(2)-macroglobulin from serum could significantly compensate for the impaired cumulus expansion of oocytes matured in serum.
Bijttebier
Jo
J
Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, 9820 Merelbeke, Belgium. jo.bijttebier@ugent.be
Tilleman
Kelly
K
Dhaenens
Maarten
M
Deforce
Dieter
D
Van Soom
Ann
A
Maes
Dominiek
D
D000818:Animals; D001798:Blood Proteins / Q000032:analysis*; D054885:Cumulus Cells / Q000166:cytology / Q000378:metabolism*; D005260:Female; D015571:Follicular Fluid / Q000737:chemistry*; D006801:Humans; D008970:Molecular Weight; D009865:Oocytes / Q000166:cytology / Q000378:metabolism*; D040081:Protein Array Analysis; D020543:Proteome / Q000032:analysis*; D013552:Swine; D000511:alpha-Macroglobulins / Q000378:metabolism*
D001798:Blood Proteins; D020543:Proteome; D000511:alpha-Macroglobulins