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Investigation of effects of Giardia duodenalis on transcellular and paracellular transport in enterocytes using in vitro Ussing chamber experiments

Published online by Cambridge University Press:  14 November 2014

KRISTOFFER R. TYSNES  and
LUCY J. ROBERTSON
KRISTOFFER R. TYSNES
Affiliation:
Department of Food Safety and Infection Biology, Norwegian University of Life Sciences, PO Box 8146, N-0033 Oslo, Norway
LUCY J. ROBERTSON*
Affiliation:
Department of Food Safety and Infection Biology, Norwegian University of Life Sciences, PO Box 8146, N-0033 Oslo, Norway
*
* Corresponding author. Department of Food Safety and Infection Biology, Norwegian University of Life Sciences, PO Box 8146, N-0033 Oslo, Norway. E-mail: lucy.robertson@nmbu.no
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Summary

The mechanisms by which different genotypes of Giardia duodenalis result in different symptoms remain unresolved. In particular, we lack detailed knowledge on which transport mechanisms (transcellular or paracellular) are affected by different Giardia isolates. Using horse radish peroxidase (HRP) and creatinine as transcellular and paracellular probes, respectively, we developed a robust assay that can be used with an Ussing chamber to investigate epithelial transport, as well as short-circuit current as an indicator of net ion transport. We investigated 2 Giardia isolates, both Assemblage A, one a lab-adapted strain and the other a field isolate. Results indicate that products from sonicated Giardia trophozoites increase both transcellular and paracellular transport. A non-significant increase in transepithelial electrical resistance (TEER) and short-circuit current were also noted. The paracellular transport was increased significantly more in the field isolate than in the lab-adapted strain. Our results indicate that while both transcellular and paracellular transport mechanisms may be increased following exposure of cells to Giardia trophozoite sonicate, perhaps by inducing non-specific increases in cellular traffic, it is important that in vitro studies of Giardia pathophysiology are conducted with different Giardia isolates, not just lab-attenuated strains.

Keywords

GiardiaUssing chamberCaco-2HRPcreatininetranscellularparacellularTEERELISA
Type
Research Article
Information
Parasitology , Volume 142 , Issue 5 , April 2015 , pp. 691 - 697
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Copyright
Copyright © Cambridge University Press 2014 

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