贵州香猪睾丸发育中支持细胞和生精细胞数量变化观察

Observation on the testis sertoli and germ cells in the Guizhou Xiang pig

为了解香猪睾丸发育过程中生殖细胞和支持细胞数变化规律,用手术取出30,40,50,70,90和110日龄(每个年龄组n=3～4)香猪右侧睾丸,经中性多聚甲醛固定,石蜡包埋,组织切片采用免疫组化SP法,用单克隆抗体GATA-4检测睾丸支持细胞的特异生长转录因子-4,经DAB显色、苏木素复染。光镜下核呈棕色者为支持细胞,核呈蓝色者则为生殖细胞;经显微照相并用Scion image软件测量生精小管及管壁面积。结果:30～110日龄睾丸支持细胞数维持在稳定水平(P>0.05),而生殖细胞数随日龄增加而增多,70日龄生殖细胞数快速增多(P<0.05),持续到110日龄。同样,从70日龄开始睾丸生精小管和管壁面积显著性增大(P<0.05)。香猪睾丸支持细胞快速增殖发生在30日龄前,而生殖细胞数随着日龄的增长而增多。

To investigate the sertoli and germ ceils in the testis of xiang pig. Right testis from 30, 40, 50, 70, 90 and 110-day-old pig （ number= 3 - 4） were collected by surgical operation. The collected tissue was fixed in 4% （w/v） paraformaldehyde for 24 h and processed to paraffin wax. Five-micron-thick sections were cut for Sertoli cell detection by immunohistochemistry using a specific monoclonal antibody to the GATA-4 protein, and the peroxidase activity detected using DAB and stained with Hematoxylin. In addition, the number of sertoli and germ cells were counted under microscopy. The area of seminiferous tubule was measured with Scion image software after micrography. The results showed that the Sertoli ceils maintained at a stable level （P〉0. 05） from 30 to 110 days old, while the number of germ cells increased with increasing age. The number of germ cells increased rapidly （P〈0. 05） from 70 days to 110 days of age. Similarly, the testis seminiferous tubules and the tubules wall area were significantly increased （P〈0. 05 ）. In conclusion, the rapid proliferation of testis Sertoli cells oc- curred before the 30 days old and the number of germ cells increased with the increasing of age in the Xiang pig.