Mahoganoid在大鼠睾丸和附睾中的表达

发布时间：2018-03-03 23:26

本文选题：Mahoganoid　切入点：生精细胞　出处：《华中科技大学》2006年硕士论文　论文类型：学位论文

【摘要】： 第一部分Mahoganoid蛋白及其mRNA在成年大鼠睾丸和附睾中的表达目的探讨Md蛋白及其mRNA在雄性成鼠睾丸和附睾组织中的分布。方法健康成年SD大鼠20只,取睾丸和附睾组织4%多聚甲醛固定,石蜡切片。用免疫酶组织化学S-P法检测Md蛋白、原位杂交方法检测Mahoganoid mRNA在成年大鼠睾丸和附睾组织中的表达及定位。结果1.免疫组织化学SP法显示:睾丸组织间质细胞、睾丸精曲小管管周肌样细胞和各级生精细胞(精原细胞、初级精母细胞和精子细胞)、支持细胞呈阳性反应,在附睾组织,棕色免疫阳性反应物质可见于输出管的高柱状细胞、低柱状细胞的胞膜及胞质,以及附睾管的高柱状细胞、主细胞、基细胞的胞膜及胞质。其中,在附睾尾部的上皮细胞中仅见微弱的棕色免疫阳性反应物质。Mahoganoid蛋白主要表达于胞膜及胞质。 2.原位杂交方法显示:睾丸组织间质细胞、睾丸精曲小管管周肌样细胞和各级生精细胞(精原细胞、初级精母细胞和精子细胞)、支持细胞上也均有Mahoganoid mRNA阳性棕色颗粒杂交信号。在附睾组织,Mahoganoid mRNA阳性棕色颗粒杂交信号也均可见于高柱状细胞、主细胞、基细胞。Mahoganoid mRNA主要表达于胞浆。结论Mahoganoid蛋白及其mRNA在成熟大鼠睾丸和附睾组织中均有表达,二者分布基本一致,提示Mahoganoid在精子发生中可能具有一定的作用,并且睾丸和附睾具有自主合成Mahoganoid蛋白的能力。为进一步阐明Mahoganoid在精子发生中的作用奠定基础。第二部分Mahoganoid蛋白及其mRNA在不同天龄大鼠睾丸和附睾中的表达目的已知成熟大鼠睾丸及附睾组织中有Mahoganoid蛋白及其mRNA的广泛表达,本文旨在进一步研究Mahoganoid蛋白及其mRNA在各年龄段大鼠睾丸组织中的分布情况。方法取出生第1天(新生鼠)、第5天(青春前期)、第20天(青春中期)、第50天(青春后期)及第70天(成年期)大鼠睾丸和附睾组织固定,采用酶免疫组织化学和原位杂交方法检测Mahoganoid蛋白及其mRNA在不同天龄大鼠睾丸和附睾组织中的表达。每张切片光镜下拍照并行图像分析,分析数据结果由SPSS11.0软件包处理,以单因素方差分析进行统计学分析。结果1.免疫组化法检测Mahoganoid蛋白的实验结果显示的棕色阳性信号可见于各天龄段大鼠睾丸的生精细胞、支持细胞、间质细胞和管周肌样细胞以及附睾输出管的高柱状、低柱状上皮细胞和附睾管的上皮细胞、主细胞、基细胞。其中Mahoganoid蛋白主要表达于胞膜和胞浆。并且Mahoganoid蛋白在50天龄和70天龄组的表达显著强于20天龄组(P〈0.05),而在20天龄组的表达显著强于0天龄组和5天龄组(P〈0.05)。 2.原位杂交检测Mahoganoid mRNA的实验结果显示的棕色阳性信号可见于各天龄段大鼠睾丸的生精细胞、支持细胞、间质细胞和管周肌样细胞以及附睾输出管的高柱状、低柱状上皮细胞和附睾管的上皮细胞、主细胞、基细胞。Mahoganoid mRNA主要表达于胞浆。其中Mahoganoid mRNA在50天龄和70天龄组的表达显著强于20天龄组(P〈0.05),在20天龄组的表达显著强于0天龄组和5天龄组(P〈0.05)。结论Mahoganoid蛋白及其mRNA在各天龄大鼠睾丸和附睾组织中均有广泛的分布,分布基本一致。且成年期(70天,50天)的表达显著强于青春中期(20天龄)和青春前期(0天,5天),青春中期(20天龄)的表达又显著强于青春前期(0天,5天),提示Mahoganoid基因的功能与性成熟及精子发生密切相关。其具体生理功能和机制尚有待进一步探讨。
[Abstract]:Part 1 expression of Mahoganoid protein and its mRNA in the testis and epididymis of adult rats
Objective to investigate the distribution of Md protein and its mRNA in the testis and epididymis of male adult rats.
Methods 20 healthy adult SD rats were fixed by paraformaldehyde and paraffin section. The Md protein was detected by immuno enzyme histochemical S-P method. The expression and location of Mahoganoid mRNA in the testis and epididymis tissues of adult rats were detected by in situ hybridization method. Results the expression of Mahoganoid mRNA in the testis and epididymis tissues of adult rats was detected.
Results 1. SP immunohistochemical method showed that testicular interstitial cells, seminiferous tubule and peritubular myoid cells and spermatogenic cells (spermatogonia, primary spermatocytes and sperm cells), Sertoli cells were positive in the epididymis, brown immunoreactive substances can be found in high columnar cells output tube, cell membrane and cytoplasmic low columnar cells, and epididymal principal cells, Gao Zhu cells, cell membrane and cytoplasm of basal cells. Among them, the main expression of positive Brown immune substances.Mahoganoid protein in epididymal epithelial cells were faint in cell membrane and cytoplasm.
Methods: in situ hybridization showed that 2. testicular interstitial cells, seminiferous tubule and peritubular myoid cells and spermatogenic cells (spermatogonia, primary spermatocytes and sperm cells), Sertoli cells were also Mahoganoid mRNA positive hybridization signal. Brown granules in the epididymis tissue, brown granules of Mahoganoid mRNA positive hybridization signal it can be found in the high columnar cells, primary cells, basal cells.Mahoganoid mRNA was mainly expressed in the cytoplasm.
Conclusion Mahoganoid and mRNA protein in mature rat testis and epididymis tissues. The expression of the two distribution is consistent, suggesting that Mahoganoid may play a role in spermatogenesis, and testis and epididymis with independent Mahoganoid protein synthesis. Lay the foundation for further elucidate the role of Mahoganoid in spermatogenesis.
The expression of the second part of Mahoganoid protein and its mRNA in the testis and epididymis of rats of different age
Objective to detect the widespread expression of Mahoganoid protein and mRNA in mature testis and epididymal tissues of adult rats. The purpose of this study is to further investigate the distribution of Mahoganoid protein and mRNA in testicular tissues of all age rats.
Methods first days after birth (neonatal rats), fifth days (prepubertal), Twentieth days, fiftieth days (Youth) (late youth) and 70 days (adult) of testis and epididymis of rats fixed expression by enzyme immunohistochemistry and in situ hybridization detection of Mahoganoid protein and mRNA in different days of age the rat testis and epididymis. Each slice under light microscope camera parallel image analysis, the results of data analysis by SPSS11.0 software package, using single factor analysis of variance was used for statistical analysis.
Spermatogenic cells, brown positive signal of 1. results of immunohistochemical detection of Mahoganoid protein showing results observed in the old rat testis sertoli cells, Leydig cells and peritubular myoid cells and epididymis tube output high columnar, low columnar epithelial cells and epididymal epithelial cells. The main cells, basal cells. The Mahoganoid protein was mainly expressed in the cytoplasm and membrane. And Mahoganoid protein expression in 50 day old and 70 day old group was significantly stronger than the 20 day old group (P < 0.05), while the expression in the 20 day old group was significantly stronger than the 0 day old group and 5 days of age group (P < 0.05).
Spermatogenic cells, brown positive signal of 2. in situ hybridization detection of Mahoganoid mRNA experiments showed that in the days of old rat testis sertoli cells, Leydig cells and peritubular myoid cells and epididymis tube output high columnar epithelial cells, epithelial cells and epididymal low column cells..Mahoganoid cells mRNA expressed mainly in the cytoplasm. The Mahoganoid mRNA in the 50 day old and 70 day old expression group was significantly stronger than the 20 day old group (P < 0.05), expressed in the 20 day old group was significantly stronger than the 0 day old group and 5 days group (P < 0.05).
Conclusion Mahoganoid protein and mRNA were widely distributed in different day old rat testis and epididymis, the distribution is basically the same. And the adult stage (70 days, 50 days) was significantly stronger than middle youth (20 day old) and adolescent (0 days, 5 days), youth (20 days of age) expression is significantly stronger than in prepubertal (0 days, 5 days), suggesting that the Mahoganoid gene function and sexual maturation and spermatogenesis are closely related to its specific physiological function and mechanism remains to be further discussed.

【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R742.9

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