BACKGROUND<\/p>\n
The cystic fibrosis transmembrane conductance regulator (CFTR) plays a critical role in electrolyte and fluid transport in epithelial cells, and women with cystic fibrosis (CF), caused by CFTR gene mutations, have a higher incidence of infertility.<\/p>\n
METHODS<\/p>\n
In the present study, we investigated the expression of CFTR in porcine oviduct and its functional role in oviductal HCO3<\/SUB>–<\/sup> secretion and embryo development with RT–PCR, western blot, patch-clamp, short-circuit current (I<\/I>sc<\/SUB>), pH measurement and embryo culture.<\/p>\n RESULTS<\/p>\n RT–PCR and western blot analysis showed the expression of CFTR mRNA and protein in the oviduct with its localization demonstrated by immunohistochemstry. The whole-cell patch-clamp recording revealed a forskolin (FSK)-activated current with electrophysiological and pharmacological characteristics of CFTR. The I<\/I>sc<\/SUB> measurement showed that FSK-stimulated an increase in the I<\/I>sc<\/SUB>, which could be significantly reduced by CFTR inhibitor or removal of both CO2<\/SUB> and HCO3<\/SUB>–<\/sup>. pH measurement showed a FSK stimulated alkalization at the apical surface, which could be inhibited by CFTR inhibitor, indicating CFTR-mediated HCO3<\/SUB>–<\/sup> secretion. Mouse embryo development from 2-cell to morula or blastocyst stage was significantly inhibited in the absence of HCO3<\/SUB>–<\/sup> or when co-cultured with HCO3<\/SUB>–<\/sup> secretion-deficient CFTR mutant cells as compared with the wild-type. RT–PCR, western blot and immunostaining showed the expression of soluble adenylate cyclase (sAC), the known HCO3<\/SUB>–<\/sup> sensor, in embryos. Treatment with its inhibitors, 2-hydroxyestradiol and KH7, prevented the HCO3<\/SUB>–<\/sup> dependent embryo development.<\/p>\n CONCLUSION<\/p>\n The present results suggest that CFTR-mediated oviductal HCO3<\/SUB>–<\/sup> secretion may be vital for sAC-dependent early embryo development, a defect of which may contribute to the reduced fertility seen in women with CF.<\/p>\n","protected":false},"excerpt":{"rendered":" BACKGROUND The cystic fibrosis transmembrane conductance regulator (CFTR) plays a critical role in electrolyte and fluid transport in epithelial cells, and women with cystic fibrosis (CF), caused by CFTR gene mutations, have a higher incidence of infertility. METHODS In the … Continue reading