Hu Yanqiu, Lu Ying, Zhou Zuomin, Du Yong, Xing Jun, Wang Lei, Lin Min, Sha Jiahao
Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, 140 Hanzhong Road, Nanjing, Jiangsu Province, People's Republic of China.
J Mol Med (Berl). 2006 May;84(5):416-24. doi: 10.1007/s00109-005-0028-y. Epub 2006 Apr 13.
Antibody to the Galpha12-subunit of guanine nucleotide regulatory proteins was used to determine whether the Galpha12 is present in adult human spermatogenic cells and to determine its role in dyszoospermia. Immunoblots from testes and spermatozoa demonstrated the presence of Galpha12 in the samples. Immunohistochemical analyses of testes found that Galpha12 was expressed in the cytoplasm of Leydig cells and was expressed in spermatids from the elongating Sb phase to mature sperm. Indirect immunofluorescence of human spermatozoa revealed the presence of Galpha12 in the neck region and the midpiece of the sperm. Galpha12 in spermatids and spermatozoa partially co-localized with F-actin and alpha-tubulin. Immunohistochemical analyses of tissues from three patients with non-obstructive azoospermia showed abnormal expression of Galpha12 in more than 45% of spermatids. Furthermore, Western blots and indirect immunofluorescence found defective expression of Galpha12 in low-motility spermatozoa with midpieces that were bent on themselves. Therefore, it suggests that Galpha12 plays a role in polarity and tail formation as spermatids mature. Furthermore, Galpha12 may be a candidate protein responsible for azoospermia caused by spermatogenic disturbance or midpiece deformities.
使用针对鸟嘌呤核苷酸调节蛋白Gα12亚基的抗体来确定Gα12是否存在于成年人类生精细胞中，并确定其在精子生成障碍中的作用。睾丸和精子的免疫印迹显示样品中存在Gα12。睾丸的免疫组织化学分析发现，Gα12在Leydig细胞的细胞质中表达，并在从伸长的Sb期到成熟精子的精子细胞中表达。人类精子的间接免疫荧光显示Gα12存在于精子的颈部区域和中段。精子细胞和精子中的Gα12与F-肌动蛋白和α-微管蛋白部分共定位。对三名非梗阻性无精子症患者的组织进行免疫组织化学分析，结果显示超过45%的精子细胞中Gα12表达异常。此外，蛋白质免疫印迹和间接免疫荧光发现，中段自身弯曲的低活力精子中Gα12表达缺陷。因此，这表明Gα12在精子细胞成熟过程中的极性和尾部形成中发挥作用。此外，Gα12可能是导致生精障碍或中段畸形引起的无精子症的候选蛋白。
