269 EXPRESSION OF TANDEM-PORE DOMAIN K⁺ CHANNELS IN BOVINE OOCYTES AND PRE-IMPLANTATION EMBRYOS

Abstract

Tandem-pore domain K⁺ (K_2P) channels that contribute to setting the resting membrane potential of excitable and nonexcitable cells are expressed in many kinds of cells and tissues. Recent studies have shown that TASK [TWIK (Tandem of P domains in Weak Inward rectifying K⁺ channels)-related acid-sensitive K⁺ channels] and TREK (TWIK-Related K⁺ channels), members of K_2P channel family that are involved in a variety of cellular functions, are expressed in human myometrium, placenta, and cytotrophoblast cells. However, their expression in bovine oocytes and embryos has not yet been reported. In this study, we examined whether TASK and TREK channels are expressed in bovine immature (germinal vesicle-stage) and mature (metaphase II-stage) oocytes and in pre-implantation (2-cell- and 16-cell-stage) embryos using RT-PCR and immunocytochemistry. RT-PCR data showed that TASK-1, TASK-3, TREK-1, TREK-2, and TRAAK channels were expressed in bovine immature and mature oocytes. Interestingly, the expression levels of TREK channels were 2-fold higher than those of TASK channels as judged by semiquantitative RT-PCR and real-time PCR with cDNA synthesized from 50 individual immature and mature oocytes (P < 0.05, n = 4). Intensity of genes was normalized with respect to that of GAPDH. Consistent with RT-PCR data, immunocytochemical data showed that TASK-1, TASK-3, TREK-1, TREK-2, and TRAAK channels were expressed in bovine immature and mature oocytes. The fluorescence intensity of TREK channels was higher than that of TASK channels (P < 0.05, n = 5). TASK and TREK channels were also expressed in pre-implantation embryos. Of TREK channels, the TREK-2 channel was strongly expressed in immature and mature oocytes and in pre-implantation embryos (P < 0.05, n = 5). For statistics, Student's t-test was used, with P < 0.05 as the criterion for significance. Our results show that TASK-1, TASK-3, TREK-1, TREK-2, and TRAAK channels were expressed in bovine immature and mature oocytes and pre-implantation embryos. These results suggest that TASK and TREK channels could be involved in various physiological processes in mammalian oocytes and embryos.