Choongkittaworn N M, Kim K H, Danner D B, Griswold M D
Department of Biochemistry and Biophysics, Washington State University, Pullman 99164-4660.
Biol Reprod. 1993 Aug;49(2):300-10. doi: 10.1095/biolreprod49.2.300.
Subtractive hybridization was used to isolate cDNAs highly expressed in stages IX-XI of the cycle of the seminiferous epithelium in the rat. One of the cloned cDNAs was sequenced and shown to be homologous to a previously described cDNA encoding rat prohibitin. Northern blot analyses showed that 1.9- and 1.2-kb transcripts were present in Sertoli cells whereas 1.5-, 1.2-, and 0.7-kb transcripts were expressed in germ cells. Western blot analyses with anti-peptide antibody to prohibitin revealed only a single 30-kDa protein in testis. Immunocytochemistry demonstrated that prohibitin protein was expressed constitutively in adult Leydig cells and Sertoli cells at all stages. Immunoreactivity of prohibition was very low in preleptotene spermatocytes, very high in leptotene spermatocytes, and very low in zygotene spermatocytes. In pachytene spermatocytes, immunoreactivity was very high in stages VII-XI and was minimal during stages XII and XIV. No protein was detected in spermatogonia and spermatocytes undergoing mitotic and meiotic divisions, respectively. These studies show that the prohibitin gene is expressed differentially in testis. The expression pattern of the prohibitin gene in rat testis appears to correlate with a proposed antiproliferative role of prohibitin.
采用消减杂交技术分离在大鼠生精上皮周期IX-XI阶段高表达的cDNA。对其中一个克隆的cDNA进行测序，结果显示它与先前描述的编码大鼠抑制素的cDNA具有同源性。Northern印迹分析表明，支持细胞中存在1.9 kb和1.2 kb的转录本，而生殖细胞中表达1.5 kb、1.2 kb和0.7 kb的转录本。用抗抑制素肽抗体进行的Western印迹分析显示，睾丸中仅有一种30 kDa的单一蛋白。免疫细胞化学表明，抑制素蛋白在成年睾丸间质细胞和支持细胞的所有阶段均持续表达。抑制素在细线前期精母细胞中的免疫反应性很低，在偶线期精母细胞中非常高，而在粗线期精母细胞中则很低。在粗线期精母细胞中，免疫反应性在VII-XI阶段非常高，而在XII和XIV阶段则最低。在精原细胞以及分别进行有丝分裂和减数分裂的精母细胞中均未检测到蛋白。这些研究表明，抑制素基因在睾丸中呈差异表达。大鼠睾丸中抑制素基因的表达模式似乎与抑制素所推测的抗增殖作用相关。
