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Expression and localization of two-pore domain K⁺ channels in bovine germ cells

Department of Physiology, College of Medicine and Institute of Health Sciences, Medical Research Center for Neural Dysfunction, Gyeongsang National University, 90 Chilam-dong, Jinju, Gyeongnam 660-751, South Korea¹ CHO-A Biotechnology Research Institute, CHO-A Pharmaceutical Company Ltd, Seoul 150-992, South Korea² Animal Genetic Resources Station, National Institute of Animal Science, RDA, Namwon 590-832, South Korea

Correspondence should be addressed to D Kang; Email: dawon{at}gnu.ac.kr

Two-pore domain K⁺ (K_2P) channels that help set the resting membrane potential of excitable and nonexcitable cells are expressed in many kinds of cells and tissues. However, the expression of K_2P channels has not yet been reported in bovine germ cells. In this study, we demonstrate for the first time that K_2P channels are expressed in the reproductive organs and germ cells of Korean cattle. RT-PCR data showed that members of the K_2P channel family, specifically KCNK3, KCNK9, KCNK2, KCNK10, and KCNK4, were expressed in the ovary, testis, oocytes, embryo, and sperm. Out of these channels, KCNK2 and KCNK4 mRNAs were abundantly expressed in the mature oocytes, eight-cell stage embryos, and blastocysts compared with immature oocytes. KCNK4 and KCNK3 were significantly increased in eight-cell stage embryos. Immunocytochemical data showed that KCNK2, KCNK10, KCNK4, KCNK3, and KCNK9 channel proteins were expressed at the membrane of oocytes and blastocysts. KCNK10 and KCNK4 were strongly expressed and distributed in oocyte membranes. These channel proteins were also localized to the acrosome sperm cap. In particular, KCNK3 and KCNK4 were strongly localized to the post-acrosomal region of the sperm head and the equatorial band within the sperm head respectively. These results suggest that K_2P channels might contribute to the background K⁺ conductance of germ cells and regulate various physiological processes, such as maturation, fertilization, and development.