Wu T C, Jih M H, Wang L, Wan Y J
Department of Obstetrics and Gynecology, UCLA School of Medicine 90024.
Mol Reprod Dev. 1994 May;38(1):9-15. doi: 10.1002/mrd.1080380103.
Activins, the dimeric polypeptides of inhibin beta-subunits, exhibit paracrine effects on cell proliferation, differentiation, and various other cell functions. The complex biological response to activin appears to involve multiple receptors. In the present study, we examined the isoform mRNA expression of both activin receptor type II (ActR-II) and type IIB (ActR-IIB) genes in mouse reproductive organs, cumulus-oocyte complexes (COCs), and ovulated oocytes. Northern blot analyses of female and male reproductive organs with single-stranded ActR-II cDNA probes revealed that mouse ovaries expressed high levels of the 6.0 kilobase (kb) mRNA, whereas the 3.0 kb transcript was the major mRNA species found in the testis. Reverse transcriptase-polymerase chain reaction (RT-PCR) showed that both COCs and oocytes contained ActR-II mRNA. To examine the expression of ActR-IIB gene, primer selection was made outside the two alternative splicing sites in order to amplify the cDNAs of all four distinct receptor isoforms. The results of RT-PCR demonstrated that isoforms IIB2 and IIB4 were the major mRNA species expressed in both female and male gonads and extragonal reproductive tissues. The ovary expressed all four mRNA isoforms, whereas the testes expressed only three isoforms. whereas the testes expressed only three isoforms. Furthermore, COCs and oocytes contained only the ActR-IIB2 isoform. The differential expression of both activin receptor mRNA isoforms in the reproductive organs suggests that distinct alternative splicing mechanisms are involved in activin receptor gene expression in male and female gonads, and that each of the activin receptors may have its own biological function in reproduction.(ABSTRACT TRUNCATED AT 250 WORDS)
激活素是抑制素β亚基的二聚体多肽,对细胞增殖、分化及其他多种细胞功能具有旁分泌作用。激活素引发的复杂生物学反应似乎涉及多种受体。在本研究中,我们检测了激活素II型受体(ActR-II)和IIB型受体(ActR-IIB)基因的亚型mRNA在小鼠生殖器官、卵丘-卵母细胞复合体(COC)及排卵后的卵母细胞中的表达情况。用单链ActR-II cDNA探针进行的Northern印迹分析显示,雌性和雄性生殖器官中,小鼠卵巢表达高水平的6.0千碱基(kb)mRNA,而3.0 kb转录本是睾丸中主要的mRNA种类。逆转录聚合酶链反应(RT-PCR)表明,COC和卵母细胞均含有ActR-II mRNA。为检测ActR-IIB基因的表达,在两个可变剪接位点之外选择引物,以扩增所有四种不同受体亚型的cDNA。RT-PCR结果表明,IIB2和IIB4亚型是在雌性和雄性性腺及性腺外生殖组织中表达的主要mRNA种类。卵巢表达所有四种mRNA亚型,而睾丸仅表达三种亚型。此外,COC和卵母细胞仅含有ActR-IIB2亚型。激活素受体mRNA亚型在生殖器官中的差异表达表明,雄性和雌性性腺中激活素受体基因表达涉及不同的可变剪接机制,且每种激活素受体在生殖过程中可能具有其自身的生物学功能。(摘要截短于250词)