Apoptosis in cumulus cells, but not in oocytes, may influence bovine embryonic developmental competence.
Aim of our study was to clarify if the occurrence of apoptosis in oocytes and cumulus cells is correlated to bovine oocyte developmental competence. The cumulus-oocyte complexes (COCs) were selected according to cumulus status: G1 with more than five layers of compact cumulus cells, G2 with one to five layers of compact cumulus cells and G3 with expanded cumulus cells. The degree of apoptosis in cumulus cells and oocytes measured by caspase staining and TUNEL assay before and after maturation, and 24 h post-insemination was compared to the cleavage, blastocyst formation and hatching rates of each group. Highest cleavage, blastocyst and hatching rates were found in cumulus-oocyte complexes with more than five layers of compact cumulus cells, but no apoptosis was detected in immature or in vitro matured oocytes, regardless of the cumulus status. Many cumulus cells contained active caspases before maturation, but caspase activity declined dramatically after maturation. TUNEL positive cells were rarely observed in each cumulus-oocyte complex upon oocyte recovery, but a huge increase of them was seen after in vitro maturation. Significantly more TUNEL and caspase positive cells were found in G2 cumulus-oocyte complexes. Our results suggest that: (i) oocyte apoptosis does not account for the inferior oocyte quality of G2 and G3; (ii) apoptosis occurs in cumulus cells regardless of the number and compactness of cumulus cells; and (iii) the degree of apoptosis in the compact cumulus-oocyte complexes (G1 and G2) is negatively correlated to the developmental competence of oocyte.
Yuan
Y Q
YQ
Laboratory of In Vitro Technology, Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium.
Van Soom
A
A
Leroy
J L M R
JL
Dewulf
J
J
Van Zeveren
A
A
de Kruif
A
A
Peelman
L J
LJ
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D001562:Benzimidazoles; D020169:Caspases / Q000032:analysis; D002417:Cattle / Q000196:embryology*; D002478:Cells, Cultured; D047108:Embryonic Development; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D005456:Fluorescent Dyes; D020287:In Situ Nick-End Labeling; D009865:Oocytes / Q000166:cytology* / Q000502:physiology; D006080:Ovarian Follicle / Q000166:cytology*
D001562:Benzimidazoles; D005456:Fluorescent Dyes; D020169:Caspases; C017807:bisbenzimide ethoxide trihydrochloride