The potential role of gap junction communication between cumulus cells and bovine oocytes during in vitro maturation.
Preliminary studies in our laboratory have indicated that modulating cumulus expansion early or late during culture has a profound influence on the subsequent development of cumulus-enclosed oocytes. Our objectives were to evaluate the effect of short term exposure to recombinant human follicle-stimulating hormone (r-hFSH) during in vitro maturation (IVM) on cumulus expansion and developmental competence of bovine oocytes. A highly significant (P < 0.0001) improvement in blastocyst development rate as a proportion of cleaved oocytes after IVM of oocytes was observed in the presence of r-hFSH for the first 6 hr of culture. To demonstrate the importance of the functional coupling between the oocyte and the cumulus compartment during that period of 6 hr, cumulus-oocyte complexes (COCs) were matured with r-hFSH for the first 6 hr followed by 18 hr in presence of 1-heptanol or 1-octanol (gap junction inhibitors) to block the communication between the two. With the coupling inhibitors, the blastocyst yield was significantly decreased (P < 0.05). A brief treatment (30 min) with the weak base methylamine, known to reverse the gap junction inhibitors effect, significantly (P < 0.05) reversed the inhibitory action of these agents on the blastocyst rate. Gap junction communication between the oocyte and surrounding cumulus cells was further studied using microinjection of the fluorescent dye Lucifer Yellow. Morphological evidences (dye transfer) were obtained that support the presence of functional coupling for a longer period with the FSH short exposure. In conclusion, high developmental rates of bovine oocytes can be achieved with a short exposure to r-hFSH. This effect is believed to be mediated through gap junctions as developmental competence of oocytes is compromised by the inhibition of their function.
Atef
Ali
A
Centre de Recherche en Biologie de la Reproduction (CRBR), Département des Sciences Animales, Ste-Foy, Québec, Canada.
François
Paradis
P
Christian
Vigneault
V
Marc-André
Sirard
S
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D046149:Embryo Culture Techniques; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D017629:Gap Junctions / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D009865:Oocytes / Q000166:cytology / Q000502:physiology*; D006080:Ovarian Follicle / Q000166:cytology / Q000502:physiology*
D005640:Follicle Stimulating Hormone