Differential actions of exogenous and intracellular spermine on contractile activity in smooth muscle of rat portal vein

Effects of the naturally occurring polyamine spermine on electrical and contractile properties of the rat portal vein were studied. 1 mM spermine nearly abolished spike activity and spontaneous contractions and decreased the intracellular Ca2+ concentration ([Ca2+]i). The phasic force responses to 0.1 and 1 microM phenylephrine were partially inhibited, but not the sustain plateau contraction caused by 5 microM phenylephrine. The Ca(2+)-force relation in high-K+ (128 mM)-depolarized veins was shifted to the right, EC50 for Ca2+ increasing from 0.50 +/- 0.03 mM (control, n = 8) to 0.65 +/- 0.06 and to 0.94 +/- 0.03 at 1 (n = 4) and 10 (n = 3) mM spermine, respectively. However, at a Ca2+ concentration of 2.5 mM, giving maximal force, there was no effect of spermine (1 mM) on either force or [Ca2+]i. Whereas extracellular spermine thus reduced contractile activity at moderate levels of stimulation, increased intracellular concentration of spermine potentiated the force response to Ca2+. Intracellular loading of spermine by reversible permeabilization increased its concentration by 2-3 times. The spontaneous activity and response to phenylephrine were unchanged. However, the Ca(2+)-force relation of depolarized veins was shifted to the left, EC50 decreasing from 0.51 +/- 0.04 mM in controls (n = 7) to 0.36 +/- 0.02 mM in the loaded veins (n = 9). Spermine increased Ca(2+)-activated force in portal veins permeabilized with beta-escin. The degree of potentiation was consistent with observed effects in spermine-loaded intact veins. The results suggest that spermine at physiological intracellular concentration may contribute to the determination of Ca2+ sensitivity in vascular smooth muscle cells.