INSULIN-RECEPTORS AND INSULIN-LIKE GROWTH FACTOR-I RECEPTORS IN EMBRYOS FROM GASTRULA UNTIL ORGANOGENESIS

The demonstration of growth factor receptors in very young embryos is limited by the difficulty in obtaining sufficient tissue to yield adequate membrane preparations. We have developed an in situ binding technique that allowed quantitation of [I-125]insulin and [I-125]insulin-like growth factor-I (IGF-I) binding to individual chick embryos. Specific binding per embryo increased from the youngest stage studied (Hamburger and Hamilton (HH) stages 3-4, gastrulating embryo of almost-equal-to 18-20 h) until the third day of development. At all ages, the binding of [I-125]IGF-I was several fold higher than the binding of [I-125]insulin. Autophosphorylation of the beta-subunit of the receptors was stimulated by insulin and IGF-I in a stage- and dose-dependent manner. The two peptides did not have an additive effect. The present studies further support our previous data showing the early developmental appearance of insulin and IGF-I receptors, which very likely are essential for normal embryo development. In additon, this in situ method for demonstration of receptors can be applied to other types of receptors present in isolated organs and young embryos.