Bovine cumulus cell expansion does not depend on the presence of an oocyte secreted factor.
Communication between the oocyte and its somatic cells has been shown to be important in oocyte development. Here we examined how the oocyte may be involved in bovine cumulus cell expansion. Intact bovine cumulus oocyte complexes (COC) were obtained by puncturing antral follicles. From the intact COC, oocytectomised complexes (OOX) were produced by micro surgical removal of the oocyte. Clumps of cumulus cells (CC) were obtained by micro-dissection. Intact or OOX complexes or CC were matured in the presence of fetal calf serum and hFSH (6 mlU/ml) for 24 hr and the degree of expansion measured. The presence of the oocyte is not essential to allow bovine cumulus expansion to occur as expansion occurred in all groups. Murine OOX complexes from eCG primed 35-40-day-old C57BL6/CBA F1 hybrids (known to require the presence of an oocyte secreted factor for cumulus expansion) were cultured with or without denuded bovine oocytes (1 oocyte/microliter). Murine OOX complexes expanded only in the presence of denuded bovine oocytes. Thus some factor produced by bovine oocytes enabled expansion of murine OOX complexes. To determine whether the factor is secreted by bovine oocytes, murine OOX were cultured with or without media conditioned by bovine oocytes (1 oocyte/microliter for 4 hr). Significant expansion of murine OOX occurred in media conditioned by bovine oocytes. This shows that the cumulus expansion enabling effect of bovine oocytes is released into the surrounding media. Media conditioned by bovine oocytes and then frozen for up to 1 month showed that the activity by the factor can withstand freezing.
Ralph
J H
JH
Division of Development and Reproduction, Roslin Institute, Midlothian, United Kingdom.
Telfer
E E
EE
Wilmut
I
I
D000818:Animals; D002417:Cattle; D002450:Cell Communication / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002462:Cell Membrane / Q000502:physiology; D017077:Culture Media, Conditioned; D003593:Cytoplasm / Q000502:physiology; D005260:Female; D066298:In Vitro Techniques; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D009865:Oocytes / Q000166:cytology* / Q000254:growth & development / Q000378:metabolism*; D006080:Ovarian Follicle / Q000166:cytology*; D015044:Zona Pellucida / Q000502:physiology
D017077:Culture Media, Conditioned