Caldés T, Alemany J, Robcis H L, de Pablo F
Diabetes Branch, National Institutes of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892.
J Biol Chem. 1991 Nov 5;266(31):20786-90.
We, and others, have recently reported that insulin-like growth factor I (IGF-I) mRNA is expressed in multiple tissues during embryogenesis and in whole embryos during early organogenesis. Therefore, it is likely that, in addition to any effect on embryo growth, IGF-I plays a paracrine/autocrine role in development. The embryonic chicken lens, an avascular organ composed by a single type of cell that undergoes differentiation in vivo and in vitro, is an ideal model to characterize the paracrine/autocrine action of IGF-I. The lens cells express IGF-I receptors, and respond to exogenous IGF-I with induction of fiber cells differentiation and stimulation of delta-crystallin gene transcription. Whether embryonic lens cells express IGF-I was uncertain. In the present study, we used a sensitive semiquantitative method (reverse transcription of RNA followed by amplification with the polymerase chain reaction) to analyze IGF-I gene expression. An amplified product of the expected length (209 base pairs) was found in days 8, 12, 15, and 19 lenses. At all embryo ages studied, the product was more readily detected in the lens than in the liver, while in eye tissues (excluding lens), IGF-I expression was relatively high. After microdissection of the epithelial cells from the fully differentiated fiber cells, IGF-I expression was detected exclusively in the epithelial cells. IGF-I immunoactivity was found using high performance liquid chromatography followed by radioimmunoassay in the days 8-19 lens extracts, and in primary cultures of isolated epithelial cells. Our previous and present findings show that the lens has all the elements for IGF-I autocrine/paracrine action in development.
我们及其他研究人员最近报道,胰岛素样生长因子I(IGF-I)mRNA在胚胎发生期间的多种组织以及器官发生早期的整个胚胎中均有表达。因此,IGF-I除了对胚胎生长有任何影响外,很可能在发育过程中发挥旁分泌/自分泌作用。鸡胚晶状体是一种无血管器官,由单一类型的细胞组成,该细胞在体内和体外都会发生分化,是表征IGF-I旁分泌/自分泌作用的理想模型。晶状体细胞表达IGF-I受体,并对外源IGF-I作出反应,诱导纤维细胞分化并刺激δ-晶状体蛋白基因转录。胚胎晶状体细胞是否表达IGF-I尚不确定。在本研究中,我们使用一种灵敏的半定量方法(RNA逆转录后用聚合酶链反应进行扩增)来分析IGF-I基因表达。在第8、12、15和19天的晶状体中发现了预期长度(209个碱基对)的扩增产物。在所研究的所有胚胎发育阶段,该产物在晶状体中比在肝脏中更容易检测到,而在眼组织(不包括晶状体)中,IGF-I表达相对较高。从完全分化的纤维细胞中显微分离出上皮细胞后,仅在上皮细胞中检测到IGF-I表达。使用高效液相色谱法,随后进行放射免疫测定,在第8 - 19天的晶状体提取物以及分离的上皮细胞原代培养物中发现了IGF-I免疫活性。我们之前和现在的研究结果表明,晶状体具备IGF-I在发育过程中自分泌/旁分泌作用的所有要素。