The purpose of the present investigation was to examine the role of the internal and external anal sphincters in the maintenance of resting pressures in the anal canal. The studies were performed in opossums anesthetized with alpha-chloralose. The radial and axial pressures in the anal canal were monitored using a continuously perfused catheter assembly. Electromyography of the external anal sphincter was monitored using bipolar tungsten hook electrodes. To examine the contribution of the external anal sphincter and surrounding skeletal muscle to the resting tone in the anal canal, pancuronium bromide was administered in a dose that abolished the electromyographic activity of the external anal sphincter muscle. The abolition of external anal sphincter activity did not modify the peak anal canal pressures, suggesting that these pressures are due to the internal anal sphincter. The alpha-adrenergic antagonist, phentolamine, did not modify the anal canal pressure, suggesting that basal internal anal sphincter pressure is not due to tonic adrenergic activity. Tetrodotoxin in a dose that produced obliteration of the anorectal reflex causing anal sphincter relaxation did not produce any change in the peak anal canal pressures. These studies show that the resting pressures in the anal canal of opossums are due to myogenic properties of the internal anal sphincter.