Groigno L, Bonnec G, Wolff J, Joly J, Boujard D
URA 256 CNRS, Department of Cell Biology and Reproduction, University of Rennes, France.
Endocrinology. 1996 Sep;137(9):3856-63. doi: 10.1210/endo.137.9.8756557.
To study Xenopus insulin-like growth factor I (IGF-I) receptor messenger expression during oogenesis, we isolated a complementary DNA (cDNA) corresponding to the beta-subunit of the receptor. There is a high degree of conservation between the deduced polypeptide and the three mammalian sequences previously described for the IGF-I receptor (75% homology) even though it is lower than the homology among mammals themselves (95% homology). IGF-I receptor messenger RNAs were specifically detected by reverse transcription-PCR in oocytes, embryos, and adult liver and muscle. By in situ hybridization, these messenger RNAs could be visualized only in oocytes. Quantification showed that they accumulated from the previtellogenic stage until early vitellogenesis. No specific labeling could be detected in oocytes after stage IV of vitellogenesis. Thus, the IGF-I receptor messenger stock does not seem to increase further beyond this point or may even decrease. The long 3'-untranslated sequence (1.8 kilobases) included in the cDNA presents no homology with those of mammalian receptor cDNAs and could be longer, as no polyadenylated tail is observed. Some motifs corresponding to sequence described as cytoplasmic polyadenylation element or that have been described in unstable messengers could be observed. Moreover, a deadenylation of this RNA occurs in the postvitellogenic stage. These results suggest that translation occurred very early during oogenesis. Therefore, IGF-I receptor could play a role early on, during oocyte growth, in addition to its involvement in the maturation process.
为了研究非洲爪蟾胰岛素样生长因子I(IGF-I)受体信使在卵子发生过程中的表达,我们分离出了一段与该受体β亚基对应的互补DNA(cDNA)。尽管推导的多肽与先前描述的三种哺乳动物IGF-I受体序列之间的同源性程度很高(75%同源性),但低于哺乳动物自身之间的同源性(95%同源性)。通过逆转录-聚合酶链反应(RT-PCR)在卵母细胞、胚胎以及成年肝脏和肌肉中特异性检测到了IGF-I受体信使核糖核酸(mRNA)。通过原位杂交,这些mRNA仅能在卵母细胞中可视化。定量分析表明,它们从前卵黄生成阶段一直积累到早期卵黄生成阶段。在卵黄生成IV期之后的卵母细胞中未检测到特异性标记。因此,IGF-I受体信使储备似乎在此之后不会进一步增加,甚至可能减少。cDNA中包含的长3'非翻译序列(1.8千碱基)与哺乳动物受体cDNA的序列没有同源性,并且可能更长,因为未观察到多聚腺苷酸尾巴。可以观察到一些与被描述为细胞质聚腺苷酸化元件的序列相对应的基序,或者在不稳定信使中描述过的基序。此外,这种RNA在卵黄生成后期会发生去腺苷酸化。这些结果表明翻译在卵子发生过程中很早就发生了。因此,IGF-I受体除了参与成熟过程外,可能在卵母细胞生长早期就发挥作用。