热休克蛋白在食蟹猴出生后睾丸发育过程中的表达研究

发布时间：2018-05-02 23:20

本文选题：热休克蛋白 + 免疫组织化学　；参考：《广西医科大学》2007年硕士论文

【摘要】： 目的观察热休克蛋白60、70、90α(HSP60、70、90α)在出生后食蟹猴(Macaca Fascicularis)睾丸发育成熟过程中的表达特征,探讨三者在睾丸发育和精子发生中的作用。材料与方法30只雄性食蟹猴根据睾丸曲细精管发育程度分为5个发育阶段: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。获取的睾丸组织均用Bouin's溶液固定4～12小时,常规石蜡包埋,切片厚度6μM,贴附在多聚赖氨酸处理过的载玻片上。切片脱蜡后常规HE染色。用Eli Vision~(TM)免疫组织化学技术检测食蟹猴从新生猴期至成年猴期不同发育阶段睾丸HSP60、HSP70和HSP 90α的表达。用病理图像分析系统计算各发育阶段睾丸的阳性细胞率,用SPSS13.0统计软件包对睾丸各发育阶段的阳性细胞率进行统计分析。结果30只食蟹猴睾丸均表达HSP60、HSP70和HSP 90α。HSP60在各个发育阶段精原细胞及支持细胞的胞浆中均有表达;在青春猴期和成年猴期的初级精母细胞胞浆中也有表达。各发育阶段,HSP60在间质细胞胞浆中偶有表达。各发育阶段睾丸细胞HSP60阳性率比较在统计学上有显著性差异(P＜0.01),其阳性细胞率在童猴后期最高,成年猴期最低,呈先升后降趋势。HSP70在新生猴期、童猴前期、童猴后期表达于精原细胞胞浆;在青春猴期主要表达于初级精母细胞胞浆,而在成年猴期主要表达于精母细胞和精子细胞胞浆。前三个发育阶段睾丸精原细胞HSP70阳性率比较在统计学上有显著性差异(P＜0.01),其阳性细胞率在新生猴期最低,童猴后期最高。HSP90α在各发育阶段均表达于精原细胞胞浆,其余各级生精细胞、支持细胞和间质细胞均未见表达。各发育阶段睾丸精原细胞HSP90α阳性率比较在统计学上有显著性差异(P＜0.01),其阳性细胞率在新生猴期最低,童猴后期最高,随后呈下降趋势。结论生后的不同发育阶段,食蟹猴睾丸中HSP60、HSP70和HSP90α的表达发生动态性变化,具有显著差异,提示三者对睾丸发育及精子发生有重要调节作用。
[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.

【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R33

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