Biochemical and developmental evidence that ooplasmic maturation of prepubertal bovine oocytes is compromised.
Our previous studies have shown that oocytes collected from prepubertal calves lack developmental competence. The overall objective of this study was to assess causes by comparing biochemical and physiologic changes during in vitro maturation of oocytes collected from ovaries of adult cattle at slaughter and from superstimulated calves (<6 mo old) by either laporotomy or ultrasound-guided follicular aspiration. Activity and/or concentrations of maturation-promoting factor (MPF), mitogen-activated protein kinase (MAPK), and inositol 1,4,5-trisphosphate receptor (IP(3)R) were determined by measuring phosphorylation of histone H-1 kinase, phosphorylation of myelin basic protein, or Western blotting, respectively, and were compared between oocytes collected from calves and for those collected from cows. The activities of MPF and MAPK and the relative amount of IP(3)R were significantly lower in calf oocytes. The physiologic significance of these observations was determined by assessing the developmental potential of embryos derived by reciprocal transfer of metaphase II (M-II) chromosomes between cow and calf ooplasts and transfer of adult cumulus cells (G0/G1) into cow and calf ooplasts. Procedural controls consisted of transfer of M-II between adult oocytes and parthenogenic activation of adult and calf oocytes. Adult parthenogenically activated oocytes cleaved and developed to blastocysts at a higher rate than did similarly activated calf oocytes (42.1% vs. 3.4%, P < 0.05). Cleavage was also higher in reciprocal M-II transfer embryos containing adult ooplasm (46.2% vs. 12.0%, P < 0.05). Cleavage (66.7% vs. 21.9%, P < 0.05) and development to blastocyst (20.1% vs. 4.8%, P < 0.05) of nuclear transfer embryos reconstructed from adult cumulus cells was higher after transfer to adult ooplasts. Collectively, these results support the hypothesis that lack of developmental competence of calf oocytes is due to their failure or inability to complete ooplasmic maturation.
Salamone
D F
DF
Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, Massachusetts 01003, USA.
Damiani
P
P
Fissore
R A
RA
Robl
J M
JM
Duby
R T
RT
D000375:Aging; D000818:Animals; D001755:Blastocyst / Q000502:physiology; D015153:Blotting, Western; D015220:Calcium Channels / Q000032:analysis; D002417:Cattle / Q000254:growth & development*; D002970:Cleavage Stage, Ovum; D003593:Cytoplasm / Q000502:physiology*; D005260:Female; D053496:Inositol 1,4,5-Trisphosphate Receptors; D016200:Maturation-Promoting Factor / Q000032:analysis; D020928:Mitogen-Activated Protein Kinases / Q000032:analysis; D004676:Myelin Basic Protein / Q000378:metabolism; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure*; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000032:analysis; D012741:Sexual Maturation
D015220:Calcium Channels; D053496:Inositol 1,4,5-Trisphosphate Receptors; D004676:Myelin Basic Protein; D018160:Receptors, Cytoplasmic and Nuclear; D011494:Protein Kinases; C102333:histone H1 kinase; D016200:Maturation-Promoting Factor; D020928:Mitogen-Activated Protein Kinases