Smooth muscle strips from the distal rectum of 11 patients who underwent surgery for imperforate anus and cloacal malformations, were studied in vitro to assess the motility response to electrical field stimulations (EFS) and to pharmacological stimulation with adrenergic and cholinergic agonists. EFS induced a nonadrenergic, noncholinergic inhibition in most strips. Acetylcholine caused either a modest contraction, no response, or a relaxation. Following atropine administration, acetylcholine caused a nonadrenergic and tetrodotoxin-resistant relaxation. The alpha-adrenergic agonist phenylephrine induced contractions in all strips. The response was abolished by alpha-adrenoceptor blockade with phentolamine, but was resistant to atropine and tetrodotoxin. beta-Adrenergic stimulation caused a relaxation that was abolished by propranolol. Function of the distal rectal smooth muscle, resected during correction of anorectal malformations, shows similarities to the function reported previously on normal anal smooth muscle evaluated in vitro.