Involvement of CFTR in oviductal HCO3- secretion and its effect on soluble adenylate cyclase-dependent early embryo development
Authors: Chen, Min Hui; Chen, Hui; Zhou, Zhen; Ruan, Ye Chun; Wong, Hau Yan; Lu, Yong Chao; Guo, Jing Hui; Chung, Yiu Wa; Huang, Ping Bo; Huang, He Feng; Zhou, Wen Liang; Chan, Hsiao Chang
Source: Human Reproduction, Volume 25, Number 7, 19 July 2010 , pp. 1744-1754(11)
Publisher: Oxford University Press
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.
In the present study, we investigated the expression of CFTR in porcine oviduct and its functional role in oviductal HCO3 secretion and embryo development with RTPCR, western blot, patch-clamp, short-circuit current (Isc), pH measurement and embryo culture.
RTPCR 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 Isc measurement showed that FSK-stimulated an increase in the Isc, which could be significantly reduced by CFTR inhibitor or removal of both CO2 and HCO3. pH measurement showed a FSK stimulated alkalization at the apical surface, which could be inhibited by CFTR inhibitor, indicating CFTR-mediated HCO3 secretion. Mouse embryo development from 2-cell to morula or blastocyst stage was significantly inhibited in the absence of HCO3 or when co-cultured with HCO3 secretion-deficient CFTR mutant cells as compared with the wild-type. RTPCR, western blot and immunostaining showed the expression of soluble adenylate cyclase (sAC), the known HCO3 sensor, in embryos. Treatment with its inhibitors, 2-hydroxyestradiol and KH7, prevented the HCO3 dependent embryo development.
The present results suggest that CFTR-mediated oviductal HCO3 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.
Document Type: Research Article
Publication date: 2010-07-19
- Human Reproduction features full-length, peer-reviewed papers reporting original research, clinical case histories, as well as opinions and debates on topical issues. Papers published cover the scientific and medical aspects of reproductive physiology and pathology, endocrinology, andrology, gonad function, gametogenesis, fertilization, embryo development, implantation, pregnancy, genetics, genetic diagnosis, oncology, infectious disease, surgery, contraception, infertility treatment, psychology, ethics and social issues. The highest scientific and editorial standard is maintained throughout the journal along with a rapid rate of publication.
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- In this Subject: Anatomy & Physiology , Obstetrics & Gynecology
- By this author: Chen, Min Hui ; Chen, Hui ; Zhou, Zhen ; Ruan, Ye Chun ; Wong, Hau Yan ; Lu, Yong Chao ; Guo, Jing Hui ; Chung, Yiu Wa ; Huang, Ping Bo ; Huang, He Feng ; Zhou, Wen Liang ; Chan, Hsiao Chang