Anurans (frogs and toads) generate lymphatic fluid at 10 times the rate in mammals, largely as a consequence of their very 'leaky' vasculature and high interstitial compliance. Lymph is ultimately pumped into the venous system by paired, dorsally located lymph hearts. At present, it is unclear how lymphatic fluid that accumulates in central body subcutaneous lymph sacs is moved to the anterior and posterior lymph hearts in the axillary regions and how lymph is moved, against gravity, to the dorsally located lymph hearts. In this study, we tested the hypothesis that lung ventilation, through its consequent effects on lymph sac pressure, contributes to the vertical movement of lymphatic fluid in the cane toad (Chaunus marinus) and the North American bullfrog (Lithobates catesbeiana). We measured pressure in the dorsal, lateral and subvertebral lymph sacs of anesthetized cane toads and bullfrogs during artificial lung inflation and deflation. We also measured pressure in the subvertebral lymph sac, which adheres to the dorsal surface of the lungs, simultaneously with brachial (forelimb) and pubic (posterior) sac pressure during ventilation in freely behaving animals. There were highly significant (P<0.001) relationships between lung pressure and lymph sac pressures (r(2)=0.19-0.72), indicating that pulmonary pressure is transmitted to the highly compliant lymph sacs that surround the lungs. Subvertebral sac pressure of resting animals was not significantly different between L. catesbeiana (518+/-282 Pa) and C. marinus (459+/-111 Pa). Brachial sac compliance (ml kPa(-1) kg(-1)) also did not differ between the two species (33.6+/-5.0 in L. catesbeiana and 37.0+/-9.4 in C. marinus). During expiration (lung deflation), reductions in expanding subvertebral sac pressure are communicated to the brachial lymph sac. Changes in brachial and pubic lymph sac pressures were correlated almost entirely during expiration rather than inspiration. The change in brachial sac pressure during expiration was 235+/-43 Pa for C. marinus and 215+/-50 Pa for L. catesbeiana, which is of sufficient magnitude to move lymph the estimated 0.5-1.0 cm vertical distance from the forelimb to the vicinity of the anterior lymph hearts. We suggest that lymph is moved during expiration to the subvertebral sac from anterior and posterior lymph sacs. During lung inflation, increased lymph sac pressure moves lymph to axillary regions, where lymph hearts can return lymph to the vascular space. Consequently, pulmonary ventilation has an important role for lymph movement and, hence, blood volume regulation in anurans.
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