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| Effects of LH and E2 on the Expression of Oviductin in Bos grunniens |
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Abstract The aim of this study is to verify whether involved in mammalian embryonic development by Somatic cell. This paper discusses the effects of luteinizing hormone (LH) and 17β-estradiol (E2) on oviductin (Ovn) by oviduct epithelial cells. We culture yak (Bos grunniens) oviduct epithelial cells in vitro with different concentration LH and E2 (10, 25, 50, 100 and 200 ng/mL). The relative expression level of Ovn gene and protein in different groups were detected by qRT-PCR and immunofluorescence, and location of ovn protein was detected by immunofluorescence after 72 h. The result obtained in this study was as follows that: (1) LH improved the relative levels of Ovn mRNA; These improvements were highest in the 10 ng/mL LH group (P＜0.05), followed by 25, 50, 100 and 200 ng/mL groups, the relative levels of Ovn mRNA were the lowest in control group (0 ng/mL LH); E2 improved the relative levels of Ovn mRNA; These improvements were the highest in the 25 ng/mL E2 group (P＜0.05), followed by 50, 10, 100 and 200 ng/mL groups, the relative levels of Ovn mRNA were the lowest in control group (0 ng/mL E2). (2) Detection by immunofluorescence showed that Ovn protein could be observed in yak cumulus cells cytoplasm as well as within nuclear membrane, and the expression of nuclear membrane was higher than the cytoplasm. LH and E2 could improve the relative expression levels of Ovn protein in yak oviduct epithelial cells; The improvements were the highest in the 10 ng/mL LH group (P＜0.05). However, the Ovn protein expression was decreased when LH concentration over 25 ng /mL. The relative levels of Ovn protein were the lowest in control group (0 ng/mL LH); The effective range of E2 (10~25 ng/mL), and the highest Ovn protein copy numbers at 25 ng/mL(P＜0.05), and reduced protein expression during all others. However, the Ovn protein expression was decreased when E2 concentration over 50 ng/mL. The relative levels of Ovn protein were the lowest in control group (0 ng/mL E2). These results suggestted that LH and E2 had significant effect on the secretion of Ovn in the oviduct epithelial cells of the yak. The optimal action concentration of LH was 10 ng/mL and the E2 was 25 ng/mL. This study provides basic information for the mechanism of LH and E2 on Ovn secretion of epithelial cells of oviduct, and also provides theoretical foundation to regulate embryonic development of yak with related hormones.
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Received: 03 May 2016
Published: 06 August 2016
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