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First published online January 18, 2008
Journal of Experimental Biology 211, 327-336 (2008)
Published by The Company of Biologists 2008
doi: 10.1242/jeb.010132

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Fluid and osmolyte recovery in the common pond snail Lymnaea stagnalis following full-body withdrawal

Sue C. Ebanks^* and Martin Grosell

University of Miami, Rosenstiel School of Marine and Atmospheric Science, Division of Marine Biology and Fisheries, 4600 Rickenbacker Causeway, Miami, FL 33149, USA

View larger version (25K): [in this window] [in a new window]   Fig. 1. Fluid volume released during bleeding (A), osmotic pressure (B), and the concentrations of the main electrolytes (C) Na+, (D) Cl–, (E) Ca2+ and (F) total CO2; in hemolymph from L. stagnalis taken from all individuals at the initial time point (open circles) and again from subsets of the same initial snails in groups of eight individuals at any one time point between 2 and 72 h (closed circles) following initial sampling. Asterisks indicate values significantly different from initial time point. Values are means ± s.e.m. (N=114 at t0 and N=8 at later time points).

View larger version (9K): [in this window] [in a new window]   Fig. 2. (A) Copper and (B) total protein concentrations in hemolymph of L. stagnalis as a function of time. Individuals were bled at time 0 (open bars; N=54 for Cu, N=53 for proteins) and sub-groups (closed bars) were bled again at 6 h (N=8 for Cu and N=6 for proteins), 12 h (N=7 for Cu and N=8 for proteins), 1 week (N=10 for Cu and total proteins), 2 weeks (N=10 for Cu and total proteins), 5 weeks (N=10 for Cu and total proteins) and 10 weeks (N=9 for Cu and total proteins). Asterisks indicate values significantly different from initial time point values. Values are means ± s.e.m.

View larger version (9K): [in this window] [in a new window]   Fig. 3. Sodium uptake rates (nmol g–1 h–1) as a function of ambient Na+ ion concentration, by the pond snail L. stagnalis. Values are for bled (closed circles) and un-bled (open circles) snails. Regression lines are Michaelis–Menten curves (SigmaPlot 8.0 for Windows). Uptake values at 2 mmol l–1 [Na+] were excluded from the regression analysis because they did not conform to the saturation kinetics observed at and below the rearing concentration of 1 mmol l–1. N=10 for each data point. Values are means ± s.e.m.

View larger version (6K): [in this window] [in a new window]   Fig. 4. Assessment of Cl– dependency for Na+ uptake by un-bled and bled L. stagnalis as tested under control (with NaCl) and treatment (with sodium gluconate or sodium sulfate) conditions. Each treatment medium contained 500 µmol l–1 of Na+. Differences between un-bled and bled conditions within treatments were significantly different. There were no significant differences among Na+-treatment groups in un-bled or bled conditions. N=10 snails in all cases. Values are means ± s.e.m.

View larger version (8K): [in this window] [in a new window]   Fig. 5. Assessment of HCO3– dependency for Na+ uptake by bled and un-bled L. stagnalis as tested under control (sodium bicarbonate) and treatment (NaCl, sodium gluconate or sodium sulfate) conditions. Each treatment medium contained 500 µmol l–1 of Na+. Differences between un-bled and bled conditions within treatments were significantly different. There were no significant differences among sodium treatment groups in un-bled or bled conditions. N=10 in all cases except control (NaCl, N=8) and sodium gluconate treatment (N=9). Values are means ± s.e.m.

View larger version (8K): [in this window] [in a new window]   Fig. 6. Effect of 1% dimethyl sulfoxide (DMSO) and two amiloride concentrations on un-bled (closed circles) and bled (gray bars) snails. N=10 for each concentration/treatment. Both un-bled and bled snails were exposed to DMSO or amiloride. Asterisk indicates value significantly less than DMSO control value. Differences between un-bled and bled conditions within treatments were significantly different. There was no significant difference between uptake rates in regular control relative to rates in the DMSO carrier control medium. Values are means ± s.e.m.

View larger version (8K): [in this window] [in a new window]   Fig. 7. Effect of 1% dimethyl sulfoxide (DMSO), 100 µmol l–1 ethylisopropylamiloride (EIPA), 100 µmol l–1 ethoxzolamide (ETOX) and 1 µmol l–1 bafilomycin (Baf) on un-bled (closed circles) and bled (gray bars). N=10 snails for each except un-bled EIPA (N=5), un-bled bafilomycin (N=7), and bled non-DMSO control (N=8). Both un-bled and bled snails were treated as indicated. ABled snails in drug treatments had significantly lower Na+ uptake rates than those in DMSO carrier control. BUn-bled snails in experimental control and those treated with ETOX had significantly greater unidirectional influx rates relative to those in the DMSO carrier control medium. Values are means ± s.e.m.

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