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Aquaporin-9 is a regulator of antral growth and ovulation
Antral formation and growth during folliculogenesis are fundamental step for accomplishment both the development and extrusion of competent oocyte. Although it has been suggested that aquaproteins (AQPs) may mediate in fluid pass into the antral cavity of the follicle, many parts are still unmasked. In this study, we examined the role of AQP9 in ovulation using in vitro follicle culture system and siRNA. Expression level of AQP 9 mRNA was dramatically increased from 24 hr and kept until 56 hr (8 hr post hCG injection) after eCG injection. AQP9 mRNA level was mostly high in the theca cells of tertiary follicles and preovulatory follicles compared with granulosa cells. The healthy follicles were isolated using sharp needles at 12 hr post eCG injection (3 wks old CD-1 mice). AQP9 specific siRNAs were transfected into the theca cells with lipofectamin 2000 for 24 hr and 36 hr. After 6 hr of hCG administration into the medium, we measure the diameter of the follicles. The number of ovulated oocytes was counted at 16 hr of hCG. The follicle size was increased more than 30% in control compared with that of AQP9 siRNA treatment group. On the other hand, the ovulation rate was dramatically decreased by suppression of AQP expression (41% vs 15% respectively in control and siRNA). At eCG 48 hr, we cannot find any ovulation marker changed in siRNA treatment group. From these results, we know that AQP9 accelerates the antral growth and ovulation.
