Prolactin enables normal development of ACh-stimulated current in cultured larval bullfrog skin

The response to acetylcholine (ACh) can be used as a marker for larval-type bullfrog skin because apically applied ACh induces an increase in short-circuit current (SCC) in larval-type but not adult-type skin. EDTA-treated larval skin, which contains only basal cells and does not respond to ACh, was used as the starting material for our culture. ACh, carbamylcholine, and choline stimulated SCC in skin that had been cultured with aldosterone (5 x 10(-7) M) supplemented with prolactin (PRL; 2 micrograms/ml). Atropine and d-tubocurarine each inhibited the ACh-induced stimulation of SCC in skin so cultured. Eserine, an inhibitor of acetylcholinesterase, also inhibited the ACh response. Amiloride stimulated SCC itself, but it reduced the ACh response. All of these results are quite similar to those seen in intact larval skin, suggesting that a larval-skin had differentiated from the basal cells used as the starting point for our culture. This is the first physiological report that PRL induces differentiation in vitro into a true larval-type bullfrog skin.