Rat vasa deferentia were cultured for 3 days in Dulbecco's modified Eagle's medium in the absence or presence of 1 microM noradrenaline (NA) to investigate if the lack of NA release is the key factor to explain the selective reduction of the Na(+)/K(+)-ATPase alpha(2) isoform previously observed after in vivo denervation of this organ (Quintas et al., Biochem Pharmacol 2000;60:741-7). The lack of effects of the indirect sympathomimetic tyramine and the neuronal amine uptake blocker cocaine on NA curves indicated that cultured organs were denervated completely. Organ culture induced supersensitivity, expressed as a 6.3-fold increase of pD(2) and a 42% elevation of maximal contraction for NA but not for Ba(2+). Western blotting indicated that the level of the alpha(1) isoform of Na(+)/K(+)-ATPase was unchanged after organ culture, but the alpha(2) isoform was down-regulated drastically to levels that were barely detectable. The addition of NA to the culture medium did not prevent the reduction of alpha(2) expression although it did impede NA supersensitivity (in fact a 4-fold decrease of pD(2) and a 32% reduction of maximal response were observed after incubation in the presence of NA). A striking reduction of L-type Ca(2+) channel expression also was observed, indicated by an 85% decrease of [3H]isradipine binding sites. These data suggest that NA is a trophic factor relevant to the control of muscle contraction, mediated by alpha(1)-adrenoceptors, but not to the expression of either Na(+)/K(+)-ATPase or the L-type Ca(2+) channel.