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牛精液中胰岛素样生长因子I及其在精子上的受体的鉴定:对精子活力的影响。

Identification of insulin-like growth factor I in bovine seminal plasma and its receptor on spermatozoa: influence on sperm motility.

作者信息

Henricks D M, Kouba A J, Lackey B R, Boone W R, Gray S L

机构信息

Endocrine Physiology Laboratory. Animal and Veterinary Sciences Department, Clemson University, Clemson,South Carolina 29634, USA.

出版信息

Biol Reprod. 1998 Aug;59(2):330-7. doi: 10.1095/biolreprod59.2.330.

Abstract

Insulin-like growth factor I (IGF-I) has been identified in human seminal plasma. This study was conducted to determine whether IGF-I is present in bovine seminal plasma, whether sperm cells express the IGF-I receptor (IGF-IR), and whether IGF-I affects sperm motility. Semen samples were collected from bulls by electroejaculation and maintained at 37 degrees C, and motility of sperm was assessed. After centrifugation to separate sperm cells from seminal plasma, the seminal plasma was submitted to a validated heterologous RIA for IGF-I. Significant concentrations of IGF-I (116.29 +/- 40.83 ng/ml expressed as mean +/- SD) were measured in bovine seminal plasma. Sperm cells were washed with buffer and subjected to either radioreceptor assay (RRA) or immunocytochemistry (IC). RRA revealed a single high affinity for the IGF-IR with a Kd of 0.83 nM as determined by the computer program LIGAND. IC, using three monoclonal antibodies, localized the IGF-IR to the acrosomal region of the sperm. Computer-assisted sperm-motion analysis was used to determine the effects of IGF-I and IGF-II on bovine sperm motility parameters. Both IGF-I and IGF-II increased sperm motility and straight-line velocity (p < 0.05) relative to the control. The presence of IGF-IR on sperm, the presence of IGF-I in semen, and the ability of IGF-I to stimulate sperm motility provide evidence that the IGF system may be involved in the fertilization process in the bovine species.

摘要

胰岛素样生长因子I(IGF-I)已在人类精浆中被鉴定出来。本研究旨在确定IGF-I是否存在于牛精浆中,精子细胞是否表达IGF-I受体(IGF-IR),以及IGF-I是否影响精子活力。通过电刺激法从公牛采集精液样本,并维持在37摄氏度,评估精子活力。离心以从精浆中分离出精子细胞后,将精浆提交给经过验证的用于IGF-I的异源放射免疫分析。在牛精浆中检测到显著浓度的IGF-I(以平均值±标准差表示为116.29±40.83 ng/ml)。用缓冲液洗涤精子细胞,然后进行放射受体分析(RRA)或免疫细胞化学(IC)。RRA显示对IGF-IR有单一高亲和力,通过计算机程序LIGAND测定其解离常数(Kd)为0.83 nM。IC使用三种单克隆抗体将IGF-IR定位到精子的顶体区域。使用计算机辅助精子运动分析来确定IGF-I和IGF-II对牛精子活力参数的影响。相对于对照组,IGF-I和IGF-II均增加了精子活力和直线速度(p<0.05)。精子上存在IGF-IR、精液中存在IGF-I以及IGF-I刺激精子活力的能力提供了证据,表明IGF系统可能参与牛的受精过程。

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