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经阴道途径给予重组兔β神经生长因子对兔精子的生理影响及兔母兔生殖反应

Physiological effects on rabbit sperm and reproductive response to recombinant rabbit beta nerve growth factor administered by intravaginal route in rabbit does.

作者信息

Sanchez-Rodriguez Ana, Arias-Álvarez María, Millán Pilar, Lorenzo Pedro L, García-García Rosa M, Rebollar Pilar G

机构信息

Department of Physiology, School of Veterinary Science, Complutense University of Madrid, Madrid, Spain.

Department of Animal Production, School of Veterinary Science, Complutense University of Madrid, Madrid, Spain.

出版信息

Theriogenology. 2020 Nov;157:327-334. doi: 10.1016/j.theriogenology.2020.08.003. Epub 2020 Aug 8.

Abstract

Beta nerve growth factor (β-NGF) is present in the seminal plasma of some species, including rabbits, acting as an ovulation-inducing factor in camelids. Traditionally, GnRH analogues are used to induce ovulation by intramuscular route when artificial insemination (AI) is performed in rabbit does. A specific rabbit recombinant β-NGF (rrβ-NGF) produced in our laboratory was tested as an alternative method to conventional treatment with GnRH analogues to induce ovulation. In the present work, different concentrations (0, 20, 100 ng/mL and 1, 20 and 100 μg/mL) of rrβ-NGF were added to diluted semen to assess its effect on sperm traits (viability and motility parameters). rrβ-NG was used also, incorporated to the AI dose, to evaluate ovulation response (LH and progesterone plasma concentrations, ovulation rate (OR) and embryo implantation at Day 7) after intravaginal administration. A negative control group stimulated with an empty catheter, and a positive control group inseminated and intramuscularly treated as usual with GnRH were also set up. Results showed that seminal quality was influenced by rrβ-NGF depending on the concentration added, being the highest concentrations tested deleterious for semen. Whereas the highest OR was found in the positive control group (100%), concentrations of 20 ng/mL, 1 μg/mL and 20 μg/mL of rrβ-NGF triggered intermediate OR (30, 60 and 42.9%, respectively), and 100 ng/mL and 100 μg/mL had the lowest OR (20 and 14.3%, respectively). Although LH peak was not observed in the first 2 h after AI in the ovulated females from rrβ-NGF groups, plasma progesterone significantly increased at Day 7, except in those females treated with 20 and 100 μg/mL. Also, 98.4% of ovulated females were pregnant on Day 7. In conclusion, rrβ-NGF added to diluted semen affects seminal quality and provokes ovulation, the development of functional CL and conception by intravaginal route in rabbit does, depending on the concentration added.

摘要

β神经生长因子(β-NGF)存在于包括兔子在内的一些物种的精浆中,在骆驼科动物中作为排卵诱导因子。传统上,在对兔母兔进行人工授精(AI)时,促性腺激素释放激素(GnRH)类似物通过肌肉注射途径用于诱导排卵。我们实验室生产的一种特异性兔重组β-NGF(rrβ-NGF)作为GnRH类似物传统治疗诱导排卵的替代方法进行了测试。在本研究中,将不同浓度(0、20、100 ng/mL和1、20和100 μg/mL)的rrβ-NGF添加到稀释的精液中,以评估其对精子特性(活力和运动参数)的影响。rrβ-NG也被用于加入AI剂量中,以评估经阴道给药后排卵反应(血浆促黄体生成素(LH)和孕酮浓度、排卵率(OR)以及第7天的胚胎着床情况)。还设立了用空导管刺激的阴性对照组和像往常一样进行授精并肌肉注射GnRH治疗的阳性对照组。结果表明,rrβ-NGF对精液质量的影响取决于添加的浓度,所测试的最高浓度对精液有害。阳性对照组的排卵率最高(100%),而20 ng/mL、1 μg/mL和20 μg/mL的rrβ-NGF浓度引发的排卵率处于中等水平(分别为30%、60%和42.9%),100 ng/mL和100 μg/mL的排卵率最低(分别为20%和14.3%)。尽管在rrβ-NGF组排卵的母兔人工授精后的前2小时内未观察到LH峰值,但除了用20和100 μg/mL处理的母兔外,第7天血浆孕酮显著升高。此外,98.4%的排卵母兔在第7天怀孕。总之,添加到稀释精液中的rrβ-NGF会影响精液质量,并通过经阴道途径在兔母兔中引发排卵、功能性黄体的发育和受孕,这取决于添加的浓度。

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