本文選題：Mahoganoid　切入點：生精細胞　出處：《華中科技大學》2006年碩士論文　論文類型：學位論文

【摘要】： 第一部分Mahoganoid蛋白及其mRNA在成年大鼠睪丸和附睪中的表達 目的探討Md蛋白及其mRNA在雄性成鼠睪丸和附睪組織中的分布。 方法健康成年SD大鼠20只,取睪丸和附睪組織4%多聚甲醛固定,石蠟切片。用免疫酶組織化學S-P法檢測Md蛋白、原位雜交方法檢測Mahoganoid mRNA在成年大鼠睪丸和附睪組織中的表達及定位。 結(jié)果1.免疫組織化學SP法顯示:睪丸組織間質(zhì)細胞、睪丸精曲小管管周肌樣細胞和各級生精細胞(精原細胞、初級精母細胞和精子細胞)、支持細胞呈陽性反應,在附睪組織,棕色免疫陽性反應物質(zhì)可見于輸出管的高柱狀細胞、低柱狀細胞的胞膜及胞質(zhì),以及附睪管的高柱狀細胞、主細胞、基細胞的胞膜及胞質(zhì)。其中,在附睪尾部的上皮細胞中僅見微弱的棕色免疫陽性反應物質(zhì)。Mahoganoid蛋白主要表達于胞膜及胞質(zhì)。 2.原位雜交方法顯示:睪丸組織間質(zhì)細胞、睪丸精曲小管管周肌樣細胞和各級生精細胞(精原細胞、初級精母細胞和精子細胞)、支持細胞上也均有Mahoganoid mRNA陽性棕色顆粒雜交信號。在附睪組織,Mahoganoid mRNA陽性棕色顆粒雜交信號也均可見于高柱狀細胞、主細胞、基細胞。Mahoganoid mRNA主要表達于胞漿。 結(jié)論Mahoganoid蛋白及其mRNA在成熟大鼠睪丸和附睪組織中均有表達,二者分布基本一致,提示Mahoganoid在精子發(fā)生中可能具有一定的作用,并且睪丸和附睪具有自主合成Mahoganoid蛋白的能力。為進一步闡明Mahoganoid在精子發(fā)生中的作用奠定基礎(chǔ)。 第二部分Mahoganoid蛋白及其mRNA在不同天齡大鼠睪丸和附睪中的表達 目的已知成熟大鼠睪丸及附睪組織中有Mahoganoid蛋白及其mRNA的廣泛表達,本文旨在進一步研究Mahoganoid蛋白及其mRNA在各年齡段大鼠睪丸組織中的分布情況。 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)大鼠睪丸和附睪組織固定,采用酶免疫組織化學和原位雜交方法檢測Mahoganoid蛋白及其mRNA在不同天齡大鼠睪丸和附睪組織中的表達。每張切片光鏡下拍照并行圖像分析,分析數(shù)據(jù)結(jié)果由SPSS11.0軟件包處理,以單因素方差分析進行統(tǒng)計學分析。 結(jié)果1.免疫組化法檢測Mahoganoid蛋白的實驗結(jié)果顯示的棕色陽性信號可見于各天齡段大鼠睪丸的生精細胞、支持細胞、間質(zhì)細胞和管周肌樣細胞以及附睪輸出管的高柱狀、低柱狀上皮細胞和附睪管的上皮細胞、主細胞、基細胞。其中Mahoganoid蛋白主要表達于胞膜和胞漿。并且Mahoganoid蛋白在50天齡和70天齡組的表達顯著強于20天齡組(P〈0.05),而在20天齡組的表達顯著強于0天齡組和5天齡組(P〈0.05)。 2.原位雜交檢測Mahoganoid mRNA的實驗結(jié)果顯示的棕色陽性信號可見于各天齡段大鼠睪丸的生精細胞、支持細胞、間質(zhì)細胞和管周肌樣細胞以及附睪輸出管的高柱狀、低柱狀上皮細胞和附睪管的上皮細胞、主細胞、基細胞。Mahoganoid mRNA主要表達于胞漿。其中Mahoganoid mRNA在50天齡和70天齡組的表達顯著強于20天齡組(P〈0.05),在20天齡組的表達顯著強于0天齡組和5天齡組(P〈0.05)。 結(jié)論Mahoganoid蛋白及其mRNA在各天齡大鼠睪丸和附睪組織中均有廣泛的分布,分布基本一致。且成年期(70天,50天)的表達顯著強于青春中期(20天齡)和青春前期(0天,5天),青春中期(20天齡)的表達又顯著強于青春前期(0天,5天),提示Mahoganoid基因的功能與性成熟及精子發(fā)生密切相關(guān)。其具體生理功能和機制尚有待進一步探討。
[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.

【學位授予單位】：華中科技大學
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R742.9

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本文編號：1563173