Effects of biochemical inhibitors on positional signalling in the chick limb bud

In 3- to 4-day embryonic chick limb buds, a region at the posterior margin of the limb, the zone of polarizing activity, appears to be responsible for signalling positional information along the anteroposterior axis. Our experiments were designed to test which biosynthetic processes are required for polarizing activity. We have treated polarizing regions with biochemical inhibitors, and then assayed their abilities to induce limb reduplications when grafted into anterior sites on host limb buds and also measured their capacities for protein, RNA, and DNA synthesis. DNA synthesis, and possibly oxidative phosphorylation, do not seem to be required for polarizing activity. But, glycolysis and protein and RNA synthesis are necessary, although not sufficient, for polarizing region activity. Activity seems particularly sensitive to inhibitors (actinomycin D and alpha-amanitin) of RNA synthesis.