Endogenous Gastric Mediators: Patho‐Physiological Role and Measurements
The protocols described in this unit are designed to present the major endogenous gastric mediators involved in the control of gastric acid secretion, namely gastrin and histamine, and in the regulation of gastric motility, which include motilin and ghrelin, under physiological and pathological conditions. The measurement of these mediators in plasma or serum of humans and animals by radioimmunoassay are described and their pathophysiological role is discussed. Curr. Protoc. Toxicol. 45:21.6.1-21.6.24. © 2010 by John Wiley & Sons, Inc.
Keywords: gastrin; histamine; motilin; ghrelin; stomach
Basic Protocol 1: Measurement of Gastrin
Basic Protocol 2: Measurement of Histamine
NOTE: All above reagents are ready to use except the acylation reagent, which is supplied as a concentrated solution and equalizing reagent is supplied lyophilized, to be reconstituted with distilled water. The histamine RIA kit contains materials for 100 quantitative determinations (including the standards) of histamine.
NOTE: For safety measures and basic radioprotection rules, see
CAUTION: The reagents should be stored at 2° to 8°C. Avoid microbiological contamination of the reagents. Do not use components beyond the expiration date shown on the kit labels. Do not mix various lots of any kit component within an individual assay. Inappropriate handling of the test samples or deviations from the test instructions can affect the result.
Basic Protocol 3: Measurement of Motilin
NOTE: The standard size of the RIA kit is sufficient for 125 RIA tubes. The
Basic Protocol 4: Measurement of Ghrelin
NOTE: The standard size of the kit is sufficient for 125 RIA tubes. The
CAUTION: Due to the short half-life of iodine-125 (the tracer used with this kit), it is highly recommended to use the kit as soon as possible after receiving. All solutions should be used on the same day of rehydration. Never store reconstituted kit components as they are unstable. Use of stored reconstituted kit components to perform RIA may yield poor and unacceptable results. When using the ghrelin (human) RIA kit (Phoenix Pharmaceuticals), carefully follow the procedure for utilization of the RIA as described in General protocol for radio immunoassay kit of peptides.
Figure 21.6.2 Neuro-hormonal mechanisms involved in gastric acid secretion during cephalic and gastric phase of gastric acid secretion.
Figure 21.6.4 Hypergastrinemia, accompanying infection with H. pylori or treatment with potent gastric acid inhibitors, results in growth promotion of the oxyntic mucosa and gastric timorous tissue.
Figure 21.6.5 Gastric infection with H. pylori and accompanying changes in the release of gastrin, somatostatin, and histamine with subsequent alteration in gastric acid secretion.
Figure 21.6.7 Similarities between the amino acid sequence of ghrelin and motilin.
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