Michels K M, Saavedra J M
Section of Pharmacology, National Institute of Mental Health, Bethesda, Md.
Neuroendocrinology. 1991 Nov;54(5):504-14. doi: 10.1159/000125945.
We investigated the in vitro binding of [125I]insulin-like growth factor I ([125I]IGF-I) within the suprachiasmatic nucleus (SCN) and median eminence (ME) of the rat by quantitative autoradiography. Binding of [125I]IGF-I within the adult SCN was saturable, reversible and to a single class of sites with an estimated Kd of 3.01 x 10(-10) M and Bmax of 131 fmol/mg protein. Competition studies revealed [125I]IGF-I binding within the SCN and ME to have the pharmacological specificity characteristic of the brain IGF-I receptor. The most potent competitors were IGF-I and IGF-II, while insulin displaced binding with a much lower potency and nerve growth factor 7S had no effect. Binding within the SCN was evident by embryonic day 18, peaked perinatally, declined to adult levels by day 6 and remained constant through middle age. The potency of IGF-I to displace [125I]IGF-I binding from SCN was similar among all ages. Binding in the ME was not evident until postnatal day 2, peaked near the end of the 1st week, declined to adult levels by day 9 and remained constant thereafter. There was no difference in binding in SCN or ME between day and early night, in animals with acute or long-term eye enucleation, or in animals fed a high sucrose diet. These results demonstrate differential development of [125I]IGF-I binding within the SCN and ME and support a role for IGFs in both these nuclei during development and maturity.
我们通过定量放射自显影术研究了[125I]胰岛素样生长因子I([125I]IGF-I)在大鼠视交叉上核(SCN)和正中隆起(ME)内的体外结合情况。成年SCN内[125I]IGF-I的结合具有饱和性、可逆性,且针对单一类位点,估计解离常数(Kd)为3.01×10⁻¹⁰ M,最大结合容量(Bmax)为131 fmol/mg蛋白质。竞争研究表明,SCN和ME内[125I]IGF-I的结合具有脑IGF-I受体的药理学特异性特征。最有效的竞争者是IGF-I和IGF-II,而胰岛素以低得多的效力取代结合,神经生长因子7S则无作用。SCN内的结合在胚胎第18天就很明显,在围产期达到峰值,到第6天降至成年水平,并在中年期保持恒定。IGF-I从SCN取代[125I]IGF-I结合的效力在所有年龄段相似。ME内的结合直到出生后第2天才明显,在第1周快结束时达到峰值,到第9天降至成年水平,此后保持恒定。在白天和深夜、急性或长期摘除眼球的动物,或喂食高蔗糖饮食的动物中,SCN或ME内的结合没有差异。这些结果表明SCN和ME内[125I]IGF-I结合的发育存在差异,并支持IGF在这些核的发育和成熟过程中发挥作用。