發(fā)布時間：2018-05-02 23:20

  本文選題：熱休克蛋白 + 免疫組織化學(xué)�。� 參考：《廣西醫(yī)科大學(xué)》2007年碩士論文

【摘要】： 目的觀察熱休克蛋白60、70、90α(HSP60、70、90α)在出生后食蟹猴(Macaca Fascicularis)睪丸發(fā)育成熟過程中的表達(dá)特征,探討三者在睪丸發(fā)育和精子發(fā)生中的作用。 材料與方法30只雄性食蟹猴根據(jù)睪丸曲細(xì)精管發(fā)育程度分為5個發(fā)育階段:新生猴期5只、童猴前期5只、童猴后期6只、青春猴期6只、成年猴期8只。獲取的睪丸組織均用Bouin's溶液固定4～12小時,常規(guī)石蠟包埋,切片厚度6μM,貼附在多聚賴氨酸處理過的載玻片上。切片脫蠟后常規(guī)HE染色。用Eli Vision~(TM)免疫組織化學(xué)技術(shù)檢測食蟹猴從新生猴期至成年猴期不同發(fā)育階段睪丸HSP60、HSP70和HSP 90α的表達(dá)。用病理圖像分析系統(tǒng)計算各發(fā)育階段睪丸的陽性細(xì)胞率,用SPSS13.0統(tǒng)計軟件包對睪丸各發(fā)育階段的陽性細(xì)胞率進(jìn)行統(tǒng)計分析。 結(jié)果30只食蟹猴睪丸均表達(dá)HSP60、HSP70和HSP 90α。HSP60在各個發(fā)育階段精原細(xì)胞及支持細(xì)胞的胞漿中均有表達(dá);在青春猴期和成年猴期的初級精母細(xì)胞胞漿中也有表達(dá)。各發(fā)育階段,HSP60在間質(zhì)細(xì)胞胞漿中偶有表達(dá)。各發(fā)育階段睪丸細(xì)胞HSP60陽性率比較在統(tǒng)計學(xué)上有顯著性差異(P＜0.01),其陽性細(xì)胞率在童猴后期最高,成年猴期最低,呈先升后降趨勢。HSP70在新生猴期、童猴前期、童猴后期表達(dá)于精原細(xì)胞胞漿;在青春猴期主要表達(dá)于初級精母細(xì)胞胞漿,而在成年猴期主要表達(dá)于精母細(xì)胞和精子細(xì)胞胞漿。前三個發(fā)育階段睪丸精原細(xì)胞HSP70陽性率比較在統(tǒng)計學(xué)上有顯著性差異(P＜0.01),其陽性細(xì)胞率在新生猴期最低,童猴后期最高。HSP90α在各發(fā)育階段均表達(dá)于精原細(xì)胞胞漿,其余各級生精細(xì)胞、支持細(xì)胞和間質(zhì)細(xì)胞均未見表達(dá)。各發(fā)育階段睪丸精原細(xì)胞HSP90α陽性率比較在統(tǒng)計學(xué)上有顯著性差異(P＜0.01),其陽性細(xì)胞率在新生猴期最低,童猴后期最高,隨后呈下降趨勢。 結(jié)論生后的不同發(fā)育階段,食蟹猴睪丸中HSP60、HSP70和HSP90α的表達(dá)發(fā)生動態(tài)性變化,具有顯著差異,提示三者對睪丸發(fā)育及精子發(fā)生有重要調(diào)節(jié)作用。
[Abstract]:Objective To observe the expression of heat shock protein 60,70,90 alpha (HSP60,70,90 alpha) in the development of testicular maturation in postnatal crab macaque (Macaca Fascicularis), and to explore the role of the three in the development of testis and spermatogenesis.
Materials and methods 30 male cynomolgus monkeys were divided into 5 developmental stages according to the development degree of testicular seminiferous tubules: 5 in the new monkey period, 5 in the early monkey, 6 in the late monkey, 6 in the young monkey, 8 in the adult monkey period. The obtained testis tissues were fixed for 4~12 hours with Bouin's solution, and the thickness of the slice was 6 M, attached to polylysine. HE staining was performed on the treated slides. Eli Vision~ (TM) immunohistochemical technique was used to detect the expression of HSP60, HSP70 and HSP 90 alpha in the testis of cynomolgus monkey from the new monkey period to the adult monkey stage. The positive cell rate of each developmental stage of the testis was calculated by the pathological image analysis system, and the SPSS13.0 statistical package was used. The percentage of positive cells in each stage of testicular development was analyzed statistically.
Results the testis of 30 cynomolgus monkeys expressed HSP60, HSP70 and HSP 90 alpha.HSP60 expressed in the cytoplasm of spermatogonial cells and support cells at various developmental stages, and expressed in the cytoplasm of primary spermatocyte in young monkey and adult monkey stage. HSP60 was occasionally expressed in the cytoplasm of interstitial cells. HSP60 of each developmental stage testis cells The positive rate was significantly different (P < 0.01). The positive cell rate was highest in the late stage of the monkey and the lowest in the adult monkey period. The positive cell rate was first rising and then descending in the new monkey period, the early stage of the monkey and the late monkey monkey expression in the spermatogonial cytoplasm, and the main expression in the primary spermatocyte in the young monkey period, and the main expression in the adult monkey period. The HSP70 positive rate of spermatogonial cells in the first three stages was significantly different (P < 0.01). The positive cell rate was the lowest in the newborn monkey period. The highest.HSP90 alpha at the late stage of the monkey was expressed in the spermatogonio cytoplasm, the other spermatogenic cells, the supporting cells and the interspermatogonial cells. The positive rate of HSP90 alpha in spermatogonial cells at various developmental stages was significantly different (P < 0.01), and the positive cell rate was the lowest in the newborn monkey period, the highest in the late monkey and then the descending trend.
Conclusion the expression of HSP60, HSP70 and HSP90 alpha in the testicles of cynomolgus monkeys has a significant difference in different developmental stages, suggesting that the three are important for the regulation of testicular development and spermatogenesis.

【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2007
【分類號】：R33

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本文編號：1835916