Oocyte maturation-related gene expression in the canine oviduct, cumulus cells, and oocytes and effect of co-culture with oviduct cells on in vitro maturation of oocytes.

Abstract

In contrast to most other mammals, canine oocytes are ovulated in an immature state and undergo oocyte maturation within the oviduct during the estrus stage. The aim of the study was to investigate whether oviduct cells from the estrus stage affect the maturation of oocytes and show gene expression patterns related to oocyte maturation. We analyzed MAPK1/3, SMAD2/3, and BMP6/15 expression in oviduct cells, cumulus cells, and oocytes from anestrus, estrus, and diestrus stages. Next, we investigated the effect of co-culture with oviduct cells derived from the estrus stage upon in vitro maturation (IVM) of canine oocytes. There was significantly higher MAPK1/3 (1.42±0.02 and 2.23±0.06), SMAD2/3 (0.77±0.03 and 2.39±0.07), and BMP15 (2.21±0.16) expression in oviduct cells at the estrus stage (P<0.05). In cumulus cells, MAPK1 (1.26±0.07), SMAD2/3 (0.82±0.01, 1.04±0.01), and BMP6 (13.09±0.11) expression was significantly higher in the estrus stage (P<0.05). In oocytes, significant upregulation of MAPK1/3 (14,960±3121 and 1668±253.4), SMAD3 (774.6±79.62), and BMP6 (8500±895.4) expression was found in the estrus stage (P<0.05). After 72h of IVM culture, a significantly higher maturation rate was observed in oocytes co-cultured with oviduct cells (10.0 ± 1.5%) than in the control group (3.2 ± 1.4%). We demonstrate that oviduct cells at the estrus stage highly expressed MAPK1/3, SMAD2/3, and BMP15. Furthermore, canine oviduct cells from the estrus stage enhance the culture environment for canine oocyte maturation.