Lymphatic muscle contraction is critical for the centripetal movement of lymph that regulates fluid balance, protein homeostasis, lipid absorption, and immune function. We have demonstrated that lymphatic muscle has both smooth and striated muscle contractile elements; however, the basic contractile properties of this tissue remain poorly defined. We hypothesized that contractile characteristics of lymphatic myofilaments would be different from vascular smooth muscle myofilaments. To test this hypothesis, -log[Ca(2+)] (pCa)-tension relationship was determined for alpha-toxin permeabilized mesenteric lymphatics, arteries, and veins. The Ca(2+) sensitivity (pCa(50)) of mesenteric lymphatics was significantly lower compared with arteries (6.16 +/- 0.05 vs. 6.44 +/- 0.02; P < 0.05), whereas there was no difference in pCa(50) between lymphatics and veins (6.16 +/- 0.05 vs. 6.00 +/- 0.10; not significant). The Hill coefficient for alpha-toxin-permeabilized lymphatics was not significantly different from arteries but was significantly greater than that of the veins (1.98 +/- 0.19 vs. 1.21 +/- 0.18; P < 0.05). In addition, the maximal tension and pCa(50) values were significantly greater in alpha-toxin-permeabilized lymphatics compared with beta-escin-permeabilized lymphatics (0.27 +/- 0.03 vs. 0.15 +/- 0.01 and 6.16 +/- 0.05 vs. 5.86 +/- 0.06 mN/mm, respectively; P < 0.05), whereas the Hill coefficient was significantly greater in beta-escin-permeabilized lymphatics. Western blot analyses revealed that CPI-17 levels were significantly decreased by about 50% in beta-escin-permeabilized lymphatics, compared with controls, whereas no change in the level of calmodulin was detected. Our data constitute the first description of the pCa-tension relationship in permeabilized lymphatic muscle. It suggests that differences in myofilament Ca(2+) sensitivity and cooperativity among lymphatic muscle and vascular smooth muscles contribute to the functional differences that exist between these tissues.