人精子蛋白ERp57的功能研究

发布时间：2018-01-10 18:25

本文关键词：人精子蛋白ERp57的功能研究　出处：《南京医科大学》2007年硕士论文　论文类型：学位论文

【摘要】： 男性不育是目前日趋严重的生殖问题，而且目前的一些辅助生殖技术存在着成功率低的问题，所以近年来，很多研究都着重于寻找受精相关的候选蛋白从而能预测这些辅助生殖技术的成功率。本实验室为了识别这些候选蛋白，在前期工作中，通过二维电泳和质谱的方法建立了人睾丸蛋白质谱系。在这个谱系中共鉴定出979个点对应于541种蛋白质。ERp57就是其中的一个蛋白质。ERp57属于蛋白质二硫键异构酶家族，在体细胞中主要表达于细胞质内质网中，，主要功能是对新合成的蛋白进行重折叠。在小鼠精子中，已证实ERp57定位于精子赤道部，在精卵融合过程中起着重要的作用。本研究着重探讨ERp57在人精子中的定位与功能，从而预测其在临床上的应用。首先western blot证实ERp57在人睾丸和人精子中确有表达，免疫组化显示ERp57定位于人睾丸的初级精母细胞以上的各级生精细胞胞质以及间质细胞中，但在Sertoli细胞中表达较弱。用二维电泳与质谱的方法鉴定出在人精子获能前后ERp57发生了修饰形式的变化。而通过凝集素荧光染色证实ERp57抗体对A23187诱导的精子顶体反应无作用。在体外，用人精子穿去透明带仓鼠卵实验证实抗ERp57的抗体能抑制人精子穿仓鼠卵，并且这种抑制效果呈现抗体浓度的依赖性。通过对临床收集的IVF患者的精子标本的分析，发现IVF成功(受精率＞60％)患者精子中ERp57含量与正常人精子标本中的含量之间没有统计学差异，而IVF失败(受精率＜60％)患者精子中ERp57量与前两者比较都明显降低并有统计学意义。总之，ERp57这个蛋白是人精子顶体蛋白的组分，它在精卵相互作用中起着重要的作用。通过临床研究提示ERp57可能成为男性不育的一个新的分子标记，并且有望成为筛选患者进行IVF治疗的指标。
[Abstract]:Male infertility is an increasingly serious reproductive problem, and some of the current assisted reproductive technology has a low success rate, so in recent years. Many studies have focused on finding fertilisation-related candidate proteins that can predict the success rate of these assisted reproductive techniques. The human testis protein lineage was established by two-dimensional electrophoresis and mass spectrometry. A total of 979 sites corresponding to 541 proteins. ERp57 is one of the proteins. ERp5. 7 belongs to the protein disulfide isomerase family. In somatic cells, mainly expressed in the cytoplasmic reticulum, the main function is to refolding the newly synthesized protein. In mouse spermatozoa, it has been confirmed that ERp57 is located in the equatorial part of sperm. ERp57 plays an important role in the process of spermatozoa fusion. This study focuses on the localization and function of ERp57 in human spermatozoa. In order to predict its clinical application, western blot confirmed the expression of ERp57 in human testis and human spermatozoa. Immunohistochemical staining showed that ERp57 was located in the cytoplasm and interstitial cells of spermatogenic cells above primary spermatocytes in human testis. However, the expression of ERp57 was weak in Sertoli cells. The modified form of ERp57 was identified by two-dimensional electrophoresis and mass spectrometry before and after human sperm capacitation. ERp was confirmed by agglutinin fluorescence staining. 57 antibody had no effect on sperm acrosome reaction induced by A23187 in vitro. The experiment of human spermatozoa piercing zona-free hamster egg confirmed that the antibody against ERp57 could inhibit human sperm from passing through hamster egg. And the inhibitory effect was concentration-dependent. The sperm samples collected from IVF patients were analyzed. It was found that there was no significant difference between the content of ERp57 in spermatozoa of patients with IVF success (fertilization rate > 60) and that of normal sperm samples. However, the amount of ERp57 in spermatozoa of patients with IVF failure (fertilization rate < 60) was significantly lower than that of the former. In short, the protein of ERp57 is a component of acrosome protein in human spermatozoa. It plays an important role in spermatozoa interaction. Clinical studies suggest that ERp57 may be a new molecular marker for male infertility and may be a potential marker for screening patients for IVF therapy.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R321

【相似文献】