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Functional characterization of a putative disaccharide membrane transporter in crustacean intestine

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Abstract

Transepithelial absorption of dietary sucrose in the American lobster, Homarus americanus, was investigated by mounting an intestine in a perfusion chamber to characterize mucosal to serosal (MS) 14C-sucrose transport. These fluxes were measured by adding varying concentrations of 14C-sucrose to the perfusate and monitoring their appearance in the bathing solution. Transepithelial 14C-sucrose transport was the combination of a hyperbolic function of luminal concentration, following Michaelis–Menten kinetics, and apparent diffusion. The kinetic constants of the putative sucrose transporter were K M  = 20.50 ± 6.00 µM and J max = 1.81 ± 0.50 pmol/cm2 × min. Phloridzin, an inhibitor of Na+-dependent mucosal glucose transport, decreased MS 14C-sucrose transport. Decreased MS 14C-sucrose transport also occurred in the presence of luminal trehalose, a disaccharide containing d-glucose moieties. Thin-layer chromatography (TLC) identified the chemical nature of radioactively labeled sugars in the bath following transepithelial transport. TLC revealed 14C-sucrose was transported across the intestine largely intact with no 14C-glucose or 14C-fructose appearing in the serosal bath or luminal perfusate. Only 13 % of bath radioactivity was volatile metabolites. Results suggest that disaccharide sugars can be transported intact across crustacean intestine and support the occurrence of a functional disaccharide membrane transporter.

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Acknowledgments

This work has been supported by the USDA Grant No. 2010-65206-20617. The authors would like to express gratitude to Dr. Corey Causey and Dr. Julie Avery (University of North Florida) for providing materials and assistance during this study.

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Authors and Affiliations

  1. Department of Biology, University of North Florida, 1 UNF Drive, Jacksonville, FL, 32224, USA

    Rasheda Likely, Eric Johnson & Gregory A. Ahearn

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  1. Rasheda Likely
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  2. Eric Johnson
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  3. Gregory A. Ahearn
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Correspondence to Gregory A. Ahearn.

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Communicated by I.D. Hume.

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Likely, R., Johnson, E. & Ahearn, G.A. Functional characterization of a putative disaccharide membrane transporter in crustacean intestine. J Comp Physiol B 185, 173–183 (2015). https://doi.org/10.1007/s00360-014-0876-2

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  • DOI: https://doi.org/10.1007/s00360-014-0876-2

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