The objective was to evaluate the toxicity effect of gossypol on ultrastructure of mouse testis and the expression of Bax mRNA and Bcl-2 mRNA of sperm cells in mice. Forty-eight male mice were randomly divided into fo...The objective was to evaluate the toxicity effect of gossypol on ultrastructure of mouse testis and the expression of Bax mRNA and Bcl-2 mRNA of sperm cells in mice. Forty-eight male mice were randomly divided into four groups： control group, L-group （30 mg-kgt. d）, M-group （60 mg·kg-1 ·d） and H-group （120 mg·kg-1· d） and were orally administrated with gossypol diluted by sodium carboxymethyl cellulose （SCC） or SCC （control group） for 20 days. On the 21st day, all the mice were killed and ultrastructure changes of testis were observed by TEM. mRNA expression of Bax and Bcl-2 in testis was measured by semiquantitative RT-PCR. The results showed that the testicular ultrastructure in three treated groups was gradually damaged, according to the dosage of gossypol and cellular structure disordered and organdie degenerated, manifesting vacuolation of mitochondria, expansion of endoplasmie reticulum, mRNA expression of Bcl-2 in testis significantly increased （p〈0.05） in L-group and then significantly decreased （p〈0.05, p〈0.01） in M-group and H-group compared with that in the control group; mRNA expression of Bcl-2 in M-group and H-group significantly decreased （p〈0.05, p〈0.01） than that in L-group and Bcl-2 mRNA expression in H-group showed a significant decrease （p〈0.05） compared with that in M-group. On the other hand, mRNA expression of Bax significant increased （p〈0.05,p〈0.01） in M-group and H-group than that in the control group. The ratio of Bcl-2/Bax significantly reduced 07〈0.05, p〈0.01） in the treated group than that in the control group and was found to be an obvious dose-dependent. It demonstrated that the gnssypol could induce the changes on ultrastructure of mice testis, down-regulate mRNA expression of Bcl-2 and up-regnlate mRNA expression of Bax, which indicated that sperm ceils were induced apoptosis.展开更多
【目的】分析谷胱甘肽过氧化物酶-5(glutathione peroxidase-5,GPx5)在正常及隐睾症双峰驼睾丸及附睾中的分布与表达,探索其在双峰驼隐睾时精子的抗氧化对生殖的调控作用。【方法】选取成年(5岁)双峰驼正常睾丸附睾10对及隐睾睾丸附睾6...【目的】分析谷胱甘肽过氧化物酶-5(glutathione peroxidase-5,GPx5)在正常及隐睾症双峰驼睾丸及附睾中的分布与表达,探索其在双峰驼隐睾时精子的抗氧化对生殖的调控作用。【方法】选取成年(5岁)双峰驼正常睾丸附睾10对及隐睾睾丸附睾6对,利用HE染色、胶原染色、网状染色观察其组织学结构特征,通过免疫组织化学染色、Western blotting、免疫荧光染色结合Image Pro Plus 13.0图像分析研究GPx5的表达及分布。【结果】与正常组相比,隐睾组双峰驼生精小管发育不良且管腔直径极显著缩小(P<0.01),间质细胞减少,附睾管腔缩小且上皮形成空泡,睾丸及附睾间质网状纤维和胶原纤维较丰富。免疫组织化学结果显示,GPx5在正常组双峰驼睾丸中未见明显表达,在隐睾组支持细胞中可见明显表达。Western blotting结果显示,与正常组相比,GPx5在隐睾组双峰驼睾丸和附睾体中表达量极显著升高(P<0.01),在附睾头中表达量极显著降低(P<0.01),在附睾尾中表达量无显著差异(P>0.05)。免疫荧光定位显示,与正常组相比,GPx5在隐睾组双峰驼睾丸和附睾尾中表达量极显著升高(P<0.01),在附睾头中表达量极显著降低(P<0.01),在附睾体中表达量无显著差异(P>0.05)。【结论】双峰驼隐睾的睾丸及附睾趋向纤维化;GPx5蛋白表达量在双峰驼隐睾睾丸中显著增加,且在支持细胞中表达增强,生精小管中的氧化应激影响了精子的正常生成;在附睾头的主细胞表达微弱,提示附睾微环境的抗氧化异常,精子损伤明显。展开更多
试验旨在探讨成年宁都黄鸡睾丸的蛋白质表达谱,筛选大小睾丸个体差异表达蛋白和功能通路,为繁殖性能选育提供基础数据。以6只22周龄宁都黄鸡的睾丸组织为研究对象,分为大睾丸组(H-TES)和小睾丸组(L-TES),采用串联质量标签(tandem mass t...试验旨在探讨成年宁都黄鸡睾丸的蛋白质表达谱,筛选大小睾丸个体差异表达蛋白和功能通路,为繁殖性能选育提供基础数据。以6只22周龄宁都黄鸡的睾丸组织为研究对象,分为大睾丸组(H-TES)和小睾丸组(L-TES),采用串联质量标签(tandem mass tags, TMT)技术对2组个体睾丸蛋白质进行差异表达、功能富集和基因互作分析,初步筛选候选蛋白质,分析这些蛋白质在6个个体的表达量与6个睾丸性状指标(左侧睾丸重、右侧睾丸重、总睾丸重、左侧睾丸指数、右侧睾丸指数、总睾丸指数)之间的相关性,分析候选蛋白质的功能。结果表明,宁都黄鸡睾丸中共鉴定到4 683个蛋白质,筛选到144个差异表达蛋白质。GO分析结果表明,差异蛋白质主要参与精子结构组成,具有尿蛋白-谷氨酸连接酶活性、胸腺嘧啶结合、脱氢抗坏血酸跨膜转运体活性等功能。差异蛋白质显著富集的KEGG通路为精氨酸和脯氨酸代谢、泛酸和辅酶A的生物合成、β-丙氨酸代谢等。蛋白质互作网络中有34个差异蛋白质,初步筛选出30个候选蛋白质。结合相关性分析和文献查阅,筛选出16个关键候选蛋白质,即SPAG6、TPPP2、ENKUR、ODF2、SAXO1、TCP11、DNALI1、CCDC63、TSSK3、TEKT2、TEKT3、ALDH2、DNAH5、CCDC40、CCDC39、DNAH10。结合关键候选蛋白质的功能,初步确定精氨酸和脯氨酸代谢、泛酸和辅酶A生物合成、β-丙氨酸代谢、紧密连接、间隙连接是睾丸性状调控关键KEGG通路。研究结果为地方鸡种睾丸质量性状选育提供了基础依据。展开更多
文摘The objective was to evaluate the toxicity effect of gossypol on ultrastructure of mouse testis and the expression of Bax mRNA and Bcl-2 mRNA of sperm cells in mice. Forty-eight male mice were randomly divided into four groups： control group, L-group （30 mg-kgt. d）, M-group （60 mg·kg-1 ·d） and H-group （120 mg·kg-1· d） and were orally administrated with gossypol diluted by sodium carboxymethyl cellulose （SCC） or SCC （control group） for 20 days. On the 21st day, all the mice were killed and ultrastructure changes of testis were observed by TEM. mRNA expression of Bax and Bcl-2 in testis was measured by semiquantitative RT-PCR. The results showed that the testicular ultrastructure in three treated groups was gradually damaged, according to the dosage of gossypol and cellular structure disordered and organdie degenerated, manifesting vacuolation of mitochondria, expansion of endoplasmie reticulum, mRNA expression of Bcl-2 in testis significantly increased （p〈0.05） in L-group and then significantly decreased （p〈0.05, p〈0.01） in M-group and H-group compared with that in the control group; mRNA expression of Bcl-2 in M-group and H-group significantly decreased （p〈0.05, p〈0.01） than that in L-group and Bcl-2 mRNA expression in H-group showed a significant decrease （p〈0.05） compared with that in M-group. On the other hand, mRNA expression of Bax significant increased （p〈0.05,p〈0.01） in M-group and H-group than that in the control group. The ratio of Bcl-2/Bax significantly reduced 07〈0.05, p〈0.01） in the treated group than that in the control group and was found to be an obvious dose-dependent. It demonstrated that the gnssypol could induce the changes on ultrastructure of mice testis, down-regulate mRNA expression of Bcl-2 and up-regnlate mRNA expression of Bax, which indicated that sperm ceils were induced apoptosis.
文摘【目的】分析谷胱甘肽过氧化物酶-5(glutathione peroxidase-5,GPx5)在正常及隐睾症双峰驼睾丸及附睾中的分布与表达,探索其在双峰驼隐睾时精子的抗氧化对生殖的调控作用。【方法】选取成年(5岁)双峰驼正常睾丸附睾10对及隐睾睾丸附睾6对,利用HE染色、胶原染色、网状染色观察其组织学结构特征,通过免疫组织化学染色、Western blotting、免疫荧光染色结合Image Pro Plus 13.0图像分析研究GPx5的表达及分布。【结果】与正常组相比,隐睾组双峰驼生精小管发育不良且管腔直径极显著缩小(P<0.01),间质细胞减少,附睾管腔缩小且上皮形成空泡,睾丸及附睾间质网状纤维和胶原纤维较丰富。免疫组织化学结果显示,GPx5在正常组双峰驼睾丸中未见明显表达,在隐睾组支持细胞中可见明显表达。Western blotting结果显示,与正常组相比,GPx5在隐睾组双峰驼睾丸和附睾体中表达量极显著升高(P<0.01),在附睾头中表达量极显著降低(P<0.01),在附睾尾中表达量无显著差异(P>0.05)。免疫荧光定位显示,与正常组相比,GPx5在隐睾组双峰驼睾丸和附睾尾中表达量极显著升高(P<0.01),在附睾头中表达量极显著降低(P<0.01),在附睾体中表达量无显著差异(P>0.05)。【结论】双峰驼隐睾的睾丸及附睾趋向纤维化;GPx5蛋白表达量在双峰驼隐睾睾丸中显著增加,且在支持细胞中表达增强,生精小管中的氧化应激影响了精子的正常生成;在附睾头的主细胞表达微弱,提示附睾微环境的抗氧化异常,精子损伤明显。
文摘试验旨在探讨成年宁都黄鸡睾丸的蛋白质表达谱,筛选大小睾丸个体差异表达蛋白和功能通路,为繁殖性能选育提供基础数据。以6只22周龄宁都黄鸡的睾丸组织为研究对象,分为大睾丸组(H-TES)和小睾丸组(L-TES),采用串联质量标签(tandem mass tags, TMT)技术对2组个体睾丸蛋白质进行差异表达、功能富集和基因互作分析,初步筛选候选蛋白质,分析这些蛋白质在6个个体的表达量与6个睾丸性状指标(左侧睾丸重、右侧睾丸重、总睾丸重、左侧睾丸指数、右侧睾丸指数、总睾丸指数)之间的相关性,分析候选蛋白质的功能。结果表明,宁都黄鸡睾丸中共鉴定到4 683个蛋白质,筛选到144个差异表达蛋白质。GO分析结果表明,差异蛋白质主要参与精子结构组成,具有尿蛋白-谷氨酸连接酶活性、胸腺嘧啶结合、脱氢抗坏血酸跨膜转运体活性等功能。差异蛋白质显著富集的KEGG通路为精氨酸和脯氨酸代谢、泛酸和辅酶A的生物合成、β-丙氨酸代谢等。蛋白质互作网络中有34个差异蛋白质,初步筛选出30个候选蛋白质。结合相关性分析和文献查阅,筛选出16个关键候选蛋白质,即SPAG6、TPPP2、ENKUR、ODF2、SAXO1、TCP11、DNALI1、CCDC63、TSSK3、TEKT2、TEKT3、ALDH2、DNAH5、CCDC40、CCDC39、DNAH10。结合关键候选蛋白质的功能,初步确定精氨酸和脯氨酸代谢、泛酸和辅酶A生物合成、β-丙氨酸代谢、紧密连接、间隙连接是睾丸性状调控关键KEGG通路。研究结果为地方鸡种睾丸质量性状选育提供了基础依据。
