太行山羊附睾头细胞Lcn5免疫荧光定位

Localization of Lcn5 in Epididymal Caput Cells in vitro of Taihang Goat

试验旨在研究太行山羊附睾头细胞Lcn5免疫荧光定位,采用细胞免疫荧光技术通过荧光显微镜和激光共聚焦显微镜观察太行山羊附睾头细胞Lcn5免疫荧光情况。结果表明:①不同一抗浓度对Lcn5呈现荧光图像影响较大,浓度稀释比例越大,浓度越小,荧光阳性信号越弱,适宜的一抗浓度稀释比为1∶50;不同二抗浓度对Lcn5阳性信号无显著影响,荧光二抗稀释比例为1∶100时荧光更清晰。②激光共聚焦显微镜扫描结果显示,Lcn5在细胞核和细胞质中均有表达;细胞培养到4 h时Lcn5在细胞核中大量表达,出现较强的阳性信号,并且在视野下发现培养液中有颗粒状的阳性信号。综上所述,推测Lcn5可能在附睾头细胞培养的过程中在细胞内合成并分泌到细胞外,对附睾微环境的构建起到了作用,同时可能参与精子的成熟过程。

The study aims to investigate the localization of Lcn5 in the epididymal caput cells of Taihang goats.The goats were selected and surgeried for epididymal caput cells and immunofluorescence of epididymal caput cells were carried out.The results were as follows:①the concentration of different primary antibodies had a greater influence on the image presented for Lcn5 in epididymal caput cells.The larger the dilution ratio and the smaller the concentration,the weaker the positive signals were in the graph;the different fluorescent secondary antibody concentrations had no significant influence on Lcn5 positive signal;the optimal concentration of primary antibody immunofluorescence was 1∶50,the dilution ratio of fluorescent secondary antibody was 1∶100.②Compared with the control group,Lcn5 was expressed both in the cytoplast and cytoplasm.When the cells were cultured for 4 h,Lcn5 was abundantly expressed in the nucleus with a strong positive signal appeared.The positive signal of granularity was also found in the fluid with the scanning field of laser confocal microscope.In summary,Lcn5 may be regulated by intranuclear cells and intracellularly synthesized during cell culture,secreted into the fluid.It maybe play a key role in the construction of the epididymal microenvironment,and involve in the sperm maturation process.