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219 Use of a genetically-engineered swine line to elucidate the role of GnRH-II and its receptor in gilts

机译:219使用遗传工程猪线以阐明GnRH-II及其受体在吉尔茨的作用

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摘要

The second form of GnRH (GnRH-II) and its receptor (GnRHR-II) are produced in only one livestock species, the pig. Paradoxically, their interaction does not stimulate gonadotropin secretion. Instead, both have been implicated in autocrine/paracrine regulation of steroidogenesis. To elucidate their role in ovarian function, our laboratory generated transgenic swine with ubiquitous knockdown (KD) of GnRHR-II. Blood samples were collected from GnRHR-II KD (n = 8) and littermate control (n = 7) gilts at the onset of estrus (follicular) and 10 d later (luteal). Serum samples were subjected to HPLC-MS/MS to quantify concentrations of 16 steroid hormones. At euthanasia, ovarian weight, ovulation rate and weight of each excised corpus luteum (CL) were recorded; HPLC-MS/MS was also performed on CL tissue. A line (GnRHR-II KD versus control) x phase (follicular versus luteal) interaction was detected for serum progesterone concentrations; levels were reduced in transgenic compared with control gilts during the luteal phase (P = 0.0329). A tendency for a line effect was observed for 11-deoxycorticosterone and 11-deoxycortisol; transgenic females tended to produce less of these corticosteroids (P < 0.10). A phase effect was detected for cortisone, 11-deoxycortisol, cortisol, corticosterone, androstenedione, androsterone, testosterone, estrone and 17β-estradiol (P < 0.05); concentrations were greater in follicular versus luteal samples (P < 0.05). Conversely, 17α-hydroxyprogesterone concentrations were elevated in luteal samples (P < 0.05). Ovarian weight did not differ between lines, although ovulation rate was reduced in GnRHR-II KD versus control gilts (P = 0.0123). However, average CL weight was greater in GnRHR-II KD compared with control females (P < 0.0001); therefore, total CL weight tended to be reduced in transgenic gilts (P = 0.0958). In tissue samples, concentrations of progesterone and estrone tended to be reduced in transgenics females (P ≤ 0.10). Ultimately, these data suggest that GnRH-II and its receptor may help regulate ovulation rate, CL development and progesterone production in gilts. Supported by USDA/NIFA AFRI-ELI predoctoral fellowship (2017-67011-26036; ATD) and AFRI (2017-67015-26508; BRW) funds.
机译:第二种形式的GnRH(GNRH-II)及其受体(GNRHR-II)仅在猪牲畜中生产。矛盾的是,它们的相互作用不刺激促性腺激素分泌。相反,两者都涉及对类固化性的自分泌/旁静脉调节。为了阐明它们在卵巢功能中的作用,我们的实验室产生的转基因猪,GNRHR-II的无处不在的敲低(KD)。从GNRHR-II KD(n = 8)和凋落物(n = 7)吉尔收集血液样品,在雌性(卵泡)和10d以后(肺炎)的发作。对血清样品进行HPLC-MS / MS以量化16种类固醇激素的浓度。在安乐死,记录卵巢体重,排卵率和每次切除的菌毛酸盐(CL)的重量; HPLC-MS / MS也对CL组织进行。检测到血清孕酮浓度的线(GNRHR-II KD与对照)X相(滤泡与肺部)相互作用;转基因中的水平与肺癌期间的对照吉尔相比降低(P = 0.0329)。对于11-脱氧曲线酮和11-脱氧曲面醇,观察到线效应的趋势;转基因雌性倾向于产生少量这些皮质类固醇(P <0.10)。检测到基质,11-脱氧酸甲溶胶,皮质醇,皮质酮,雄甾醇,雄甾酮,睾酮,雌激素和17β-雌二醇(P <0.05)中检测到相效应;卵泡与氯化尿样品的浓度更大(P <0.05)。相反,17α-羟丙酮浓度在耐力样品中升高(P <0.05)。卵巢重量在线之间没有区别,尽管GNRHR-II KD与对照吉尔(P = 0.0123)中排卵率降低了排卵率。然而,与对照雌性相比,GNRHR-II Kd的平均Cl重量更大(P <0.0001);因此,在转基因胃肠中倾向于减少总Cl重量(P = 0.0958)。在组织样品中,孕酮的浓度和雌激素倾向于转基因雌性(P≤0.10)。最终,这些数据表明GNRH-II及其受体可以帮助调节胃肠中的排卵率,CL开发和孕酮产生。由USDA / NIFA AFRI-ELI PERVOCTORAL FORDOWSHIP(2017-67011-26036; ATD)和AFRI(2017-67015-26508; BRW)资金支持。

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