Sex differences in the incidence of varicose veins have been suggested; however, the venous mechanisms involved are unclear. We hypothesized sex-related differences in venous function and underlying distinctions in intracellular free calcium, [Ca(2+)](i), signaling and Ca(2+)-dependent mechanisms of venous contraction. Circular segments of inferior vena cava (IVC) from male and female Sprague-Dawley rats were suspended between two hooks, labeled with fura-2, and placed in a cuvet inside a spectrofluorometer for simultaneous measurement of isometric contraction and the 340/380 fluorescence ratio (indicative of [Ca(2+)](i)). In male IVC, phenylephrine (PHE; 10(-5) M) caused significant increase in contraction and [Ca(2+)](i). In female IVC, PHE-induced contraction was significantly reduced, but [Ca(2+)](i) did not differ significantly from males. Membrane depolarization by KCl (96 mM), which stimulates Ca(2+) influx, caused parallel increases in contraction and [Ca(2+)](i) in male IVC, and the KCl-induced contraction was significantly reduced in parallel with [Ca(2+)](i) in female IVC. In male IVC stimulated with 0 Ca(2+) KCl solution, the addition of increasing concentrations of extracellular Ca(2+) ([Ca(2+)](e)) (0.1, 0.3, 0.6, 1, and 2.5 mM) caused stepwise increases in contraction and [Ca(2+)](i), and both the KCl-induced [Ca(2+)](e)-contraction curve and the [Ca(2+)](e)-[Ca(2+)](i) curve were reduced in female IVC, suggesting reduced Ca(2+) entry via voltage-gated channels. The PHE-induced [Ca(2+)](e)-contraction curve was significantly reduced in females, but the [Ca(2+)](e)-[Ca(2+)](i) curve was similar in female and male IVC, suggesting the involvement of other mechanisms in addition to Ca(2+) entry. The [Ca(2+)](e)-contraction and [Ca(2+)](e)-[Ca(2+)](i) curves were used to construct the [Ca(2+)](i)-contraction relationship. The KCl-induced [Ca(2+)](i)-contraction relationship was superimposed in male and female IVC. In contrast, the PHE-induced [Ca(2+)](i)-contraction relationship was reduced and located to the right in female compared with male IVC, suggesting reduced [Ca(2+)](i) sensitivity of the venous contractile myofilaments. The reduced contraction, [Ca(2+)](i), and [Ca(2+)](i) sensitivity in female veins render them more prone to dilation. These sex-specific reductions in venous function, if they also occur in human veins, may play a role in the greater incidence of varicose veins in females.