Reduced expression and function of aquaporin-3 in mouse metaphase-II oocytes induced by controlled ovarian hyperstimulation were associated with subsequent low fertilization rate.
ABSTRACT Aquaporin-3 (AQP3), one isoform of water channel family, has been found to be expressed in mouse oocytes. The present study aimed to investigate whether functional AQP3 was expressed in oocytes induced by controlled ovarian hyperstimulation (COH), and whether altered oocyte AQP3 expression was associated with changes in fertilization rate.
Sixty ICR female mice were divided into two groups: COH and control. AQP3 mRNA expression of mouse metaphase II (MII) oocytes was quantified by real-time RT-PCR. The water permeability of oocytes was assessed with cell swelling test. The fertilization profiles of oocytes were generated via in vitro fertilization.
AQP3 mRNA was expressed in both natural and COH-induced mouse oocytes. COH significantly reduced AQP3 mRNA expression. The volume of oocytes was significantly increased after exposure to hypotonic medium and pretreatment with HgCl(2) attenuated hypotonic medium-induced increase in oocyte volume and water permeability coefficient (Pf). Furthermore, the expression of AQP3, Pf and the fertilization rate were significantly lower in COH oocytes than those in control.
AQP3 might play an important role in controlling oocyte quality and a low in vitro fertilization rate of COH mice might, in part, result from reduced AQP3 expression and water permeability in mouse oocytes.
- SourceAvailable from: Mariusz Skowronski[show abstract] [hide abstract]
ABSTRACT: Aquaporins (AQPs) are a family of membrane channel proteins that facilitate bulk water transport. To date, 11 isoforms of AQPs have been reported to be expressed in the female and male reproductive systems. The purpose of our study was to determine the localization and quantitative changes in the expression of AQP1, 5 and 9 within the pig uterus during different stages of the estrous cycle and early pregnancy. Immunoperoxidase and semi-quantitative immunoblotting techniques were used to examine the distribution and changes in amounts of AQP1, AQP5 and AQP9 in uteral cells of pigs at the early (Days 2-4), middle (10-12), late (14-16) stage of the luteal phase and late (18-20) stage of the follicular phase of the estrous cycle as well as on Days 14-16 and 30-32 of gestation (the onset and the end of implantation process). The results demonstrated that AQP1, 5, and 9 were clearly detected in all studied stages of the estrous cycle and pregnancy. AQP1 was localized within uterine blood vessels. In cyclic gilts, endometrial and myometrial expression of AQP1 protein did not change significantly but increased during gestation. AQP5 was localized in smooth muscle cells and uterine epithelial cells. Endometrial expression of AQP5 protein did not change significantly between Days 2-4 and 10-12 of the estrous cycle but increased on Days 14-16 and 18-20 as well as during early pregnancy. Myometrial expression of AQP5 did not differ significantly during the estrous cycle but increased in the pregnancy. The anti-AQP9 antibody labeled uterine epithelial cells of uterus. Endometrial expression of AQP9 did not change significantly between Days 2-4 and 10-12 of the estrous cycle but increased on Days 14-16 and 18-20 as well as during early pregnancy. The results suggest that a functional and distinctive collaboration exists among diverse AQPs in water handling during the different uterine phases in the estrous cycle and early pregnancy.Reproductive Biology and Endocrinology 01/2010; 8:109. · 2.14 Impact Factor
- [show abstract] [hide abstract]
ABSTRACT: To investigate whether the embryotoxic effect of peritoneal fluid (PF) from infertile women with mild endometriosis on mouse oocytes and embryos is associated with changes in embryonic epidermal growth factor (EGF), insulin-like growth factor-I (IGF-I), and their receptors. Experimental animal study. University-based research laboratory. Adult ICR mice. Peritoneal fluid was obtained from fertile women with no endometriosis (PF-NE) and infertile women with mild endometriosis (PF-E). In vitro fertilization was performed, and mouse two-cell stage embryos were cultured in human tubal fluid medium with or without PF. Rates of fertilization, cleavage, and blastulation. The embryonic EGF and IGF-I levels in culture medium were analyzed by enzyme-linked immunosorbent assay, and EGF receptor, IGF-I receptor, and phosphorylated extracellular signal-regulated protein kinases (p-ERK) expression was determined by immunofluorescence and confocal microscopy. When oocytes and embryos were cultured in media with PF-E, the fertilization capability of oocytes and the development potential of embryos were decreased. The levels of embryonic EGF, IGF-I, and their receptors were increased. However, p-ERK of the postreceptor signal transduction pathway was down-regulated. Endometriotic PF may attenuate oocyte and embryo development by impairing embryonic growth factor/receptor/signal transduction, resulting in endometriotic infertility.Fertility and sterility 05/2010; 93(8):2538-44. · 3.97 Impact Factor
- [show abstract] [hide abstract]
ABSTRACT: This study aimed to investigate whether aquaporin 3 (Aqp3) mRNAs are expressed in immature oocytes and altered during in vitro maturation process. Five- to 6-week-old female ICR mice were primed by gonadotropin for 24 and 48 h. Immature oocytes obtained 48 h after priming were also matured in vitro for 17 to 18 h. In vivo matured oocytes were obtained after 48 h priming followed by hCG injection. Total RNAs were extracted from 80 to 150 oocytes in each experimental group, and the levels of Aqp3 mRNA were quantified by real-time reverse transcriptase polymerase chain reaction. The experiments were repeated twice using different oocytes. The Aqp3 mRNA was expressed in immature oocytes, as well as in in vitro and in vivo matured oocytes. The expression level was higher in immature oocytes obtained 48 h after priming (17.2 ± 8.6, mean ± SD) than those with no priming (5.7 ± 0.8) or obtained 24 h after priming (2.5 ± 0.8). The expression of Aqp3 mRNA decreased after in vitro maturation (1.2 ± 0.5), which was similar to in vivo matured oocytes (1.0 ± 0.0). Our work demonstrated that Aqp3 mRNA expression increased during the development of immature oocyte but decreased after completion of in vitro maturation. The results indicate that AQP3 is certainly needed for the acquisition of immature oocytes' full growing potential within antral follicles.Zygote 02/2011; 19(1):9-14. · 1.50 Impact Factor