Lamsam-Casalotti S, Onoda M, Papadopoulos V, Dym M
Department of Cell Biology, Georgetown University Medical Center, Washington, DC 20007.
J Reprod Fertil. 1993 Nov;99(2):487-95. doi: 10.1530/jrf.0.0990487.
The expression of guanine nucleotide-binding proteins (G proteins) during the development of rat testes was investigated. Immunohistochemical studies on frozen sections and isolated testicular cells demonstrated that the expression of the GTP-binding proteins was developmentally regulated and specific for different cell types. The alpha subunit of the cholera toxin-sensitive stimulatory G protein (Gs alpha) was first detected in testes from 7-day-old rats; its value reached a maximum at 23 days and then decreased to very low or undetectable amounts in testes of 45-day-old and adult rats (60-90 days of age). The Gs alpha subunit appears to be expressed by Sertoli, peritubular myoid and interstitial cells. The common beta subunit (G beta) was present at all ages during development and was more prominent around the periphery of the tubules in younger animals but then became more evident in the cytoplasm of germ cells with increasing age. The pertussis toxin-sensitive inhibitory G proteins, Gi1/2 and Gi3, showed a similar pattern of expression. Sertoli cells and peritubular cells expressed Gi1/2 and Gi3 at very low levels at all ages, whereas pachytene spermatocytes and round spermatids expressed the inhibitory binding proteins only at later ages of development (45-day-old and adult testis). Northern blot analysis showed that with increasing age the Gs alpha mRNA in the testis decreased and this was confirmed by in situ hybridization. These latter studies showed localization of the transcripts to somatic cells but not to germ cells.(ABSTRACT TRUNCATED AT 250 WORDS)
研究了大鼠睾丸发育过程中鸟嘌呤核苷酸结合蛋白(G蛋白)的表达。对冰冻切片和分离的睾丸细胞进行的免疫组织化学研究表明,GTP结合蛋白的表达受发育调控,且对不同细胞类型具有特异性。霍乱毒素敏感的刺激性G蛋白(Gsα)的α亚基最早在7日龄大鼠的睾丸中检测到;其含量在23日龄时达到最高,然后在45日龄和成年大鼠(60 - 90日龄)的睾丸中降至非常低或无法检测到的水平。Gsα亚基似乎由支持细胞、睾丸肌样细胞和间质细胞表达。常见的β亚基(Gβ)在发育的各个年龄段均存在,在较年轻动物的小管周边更为突出,但随着年龄增长,在生殖细胞的细胞质中变得更加明显。百日咳毒素敏感的抑制性G蛋白Gi1/2和Gi3表现出类似的表达模式。支持细胞和睾丸肌样细胞在所有年龄段均以非常低的水平表达Gi1/2和Gi3,而粗线期精母细胞和圆形精子细胞仅在发育后期(45日龄和成年睾丸)表达抑制性结合蛋白。Northern印迹分析表明,随着年龄增长,睾丸中的Gsα mRNA减少,原位杂交证实了这一点。这些后续研究表明转录本定位于体细胞而非生殖细胞。(摘要截短于250字)