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J Exp Biol. 2010 Sep 15;213(Pt 18):3161-6. doi: 10.1242/jeb.042044.

Lymph flux rates from various lymph sacs in the cane toad Rhinella marina: an experimental evaluation of the roles of compliance, skeletal muscles and the lungs in the movement of lymph.

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Department of Biology, Portland State University, Portland, OR 97207-0751, USA.


A new method for quantitatively determining lymph flux from various lymphatic sacs of an anuran, the cane toad, was developed. This method used the dye dilution principle of C(i)V(i)=C(f)V(f) following injection of Evans Blue into specific lymph sacs and measuring its appearance in the venous circulation. The apparent lymph volume was 57 ml kg(-1). The greatest rate of lymph return (0.5-0.8 ml kg(-1) min(-1)) and best linear fit of Evans Blue appearance in the circulation with time followed injections into the subvertebral lymph sac, which has direct connections to both the anterior and posterior pairs of lymphatic hearts. Rate of lymph flux from the pair of posterior lymph hearts was three times greater than the anterior pair. Rates of lymph flux were only influenced by injection volume in the crural lymph sacs, implicating lymph sac compliance as the source of the pressure for lymph movement from these sacs. Femoral lymph sac fluxes were decreased by 60% following ablation of the tendons of the sphincter ani cloacalis, abdominal crenators and piriformis. This supports a role for these muscles in generating the pressure for vertical lymph movement. Femoral lymph sac fluxes were also decreased by 70% by the insertion of a coil in the subvertebral lymph sac, preventing normal compression and expansion of this sac by the lungs. This supports a role for lung ventilation in generating the pressure for vertical movement of lymph. Contrary to previous hypotheses, fluxes from the brachial sac were not influenced by insertion of the coil into the subvertebral sac. A haemorrhage equivalent to 50% of the blood volume did not change lymph flux rates from the femoral lymph sacs. These data provide the first experimental evidence that actual lymph fluxes in the cane toad Rhinella marina depend on lymph sac compliance, contraction of specific skeletal muscles and lung ventilation to move lymph laterally and vertically to the dorsally located lymphatic hearts.

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