Abstract
The role of nitric oxide (NO) in the intestinal basal ion transport and under conditions of enterotoxin-induced ion secretion is controversial. Namely it is not clear whether NO enhances or counteracts intestinal ion secretion and whether the effects on transport result from a direct interaction with the enterocyte. The cell origin of NO is also unclear. We have tested the hypothesis that NO produced by the enterocyte directly regulates ion transport processes either in basal condition or in response to cholera toxin-induced secretion. Electrical variables reflecting transepithelial ion transport were measured in Caco-2 cell monolayers mounted in Ussing chambers exposed to the NO synthase inhibitor Nω-nitro-L-arginine methyl ester, in the presence or absence of cholera toxin. cAMP concentrations were also measured. NO release was determined by nitrite-nitrate concentration. NO synthase activities were assayed by Western blot analysis. Nω-nitro-L-arginine methyl ester had a secretory effect, as judged by increased basal short-circuit current and cAMP concentration. It also increased cholera toxin-induced electrical response and cAMP production. Either cholera toxin or the cAMP analog 8-bromo-cAMP induced a rapidly progressive and Ca2+-dependent increase in NO concentration, suggesting a homeostatic up-regulation of the constitutive form of NO synthase. Western blot analysis showed an increase in constitutive NO synthase enzyme isoform. These results indicate that the enterocyte regulates its own ion transport processes, either in basal condition or in the presence of active secretion, through the activation of a constitutive NO synthase-NO pathway, functioning as a braking force of cAMP-induced ion secretion.
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Abbreviations
- NO:
-
nitric oxide
- NOS:
-
nitric oxide synthase
- cNOS:
-
constitutive nitric oxide synthase
- iNOS:
-
inducible nitric oxide synthase
- NO2−/NO3−:
-
nitrite/nitrate
- L-NAME:
-
Nω-nitro-L-arginine methyl ester
- L-NIL:
-
N-iminoethyl-L-lysine
- CT:
-
cholera toxin
- G :
-
conductance
- I sc :
-
short circuit current
- PD:
-
transepithelial potential difference
- 8Br-cAMP:
-
8-bromo-cAMP
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Supported in part by a grant from the Ministero della Sanità, AIDS research project 1999 program 50.c.28, and by a grant from CNR n.94.02505 CT04.
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Canani, R., Cirillo, P., Buccigrossi, V. et al. Nitric Oxide Produced by the Enterocyte Is Involved in the Cellular Regulation of Ion Transport. Pediatr Res 54, 64–68 (2003). https://doi.org/10.1203/01.PDR.0000069841.52414.A5
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DOI: https://doi.org/10.1203/01.PDR.0000069841.52414.A5
