Beta-adrenoreceptor stimulation with isoproterenol (IP) rapidly and reversibly rounded and arborized smooth muscle cells (SMC) cultured from rat aorta. The arborized SMC remained firmly attached to the substratum via a network of long, dendritelike processes. Arborization of the SMC correlated closely with increases in cellular cAMP produced by IP and a variety of other compounds. The intracellular calcium antagonist 8-(N,N-diethylamino)-octyl-3,4,5-trimethoxybenzoate (TMB-8) also rounded and arborized the SMC. Antitubulin compounds potently blocked arborized by IP, dibutyryl cAMP, and TMB-8. The release of cell-bound 45Ca2+ was followed in the presence and absence of IP. IP strikingly increased the amount of 45Ca2+ that remained cell bound between 20 and 120 min. Propranolol and colchicine prevented IP from inhibiting the release of cell-bound 45Ca2+. These results suggest that the modulation of Ca2+ transport is involved in the arborization of cultured SMC by cAMP.