Vagus nerve participates in CCK-induced Fos expression in hindbrain but not myenteric plexus.

CCK activates neurons in rat hindbrain and small intestinal myenteric ganglia. Activation of neurons at both sites is mediated through type A CCK receptors. CCK-induced activation of hindbrain neurons is mediated by capsaicin-sensitive vagal fibers. Therefore, it is possible that CCK-induced activation of myenteric neurons also depends upon vagal activation. To test this hypothesis, we examined hindbrain and myenteric neuronal expression of Fos immunoreactivity following CCK injection in rats that had undergone bilateral subdiaphragmatic vagotomy or systemic treatment with capsaicin, a neurotoxin that destroys small unmyelinated primary sensory neurons in the vagus, as well as in other peripheral nerves. We found that CCK (2 or 10 microg/kg) significantly increased Fos expression in both the brains and small intestinal myenteric plexuses of control rats. CCK-induced Fos expression was abolished or attenuated in the brains of vagotomized or capsaicin-treated animals. However, vagotomy or capsaicin treatment did not diminish CCK-induced Fos expression in the small intestinal myenteric plexus. We conclude that CCK-induced activation of intestinal myenteric neurons does not depend on activation of vagal sensory or motor neurons, while activation of neurons in the dorsal hindbrain is mediated, at least in part by CCK-induced activity of small unmyelinated vagal sensory neurons.

CCK-A receptor activation induces fos expression in myenteric neurons of rat small intestine.

Cholecystokinin (CCK), a hormone secreted from endocrine cells of the small intestine, participates in the control of motility and secretion in the gastrointestinal tract, and in the control of food intake. At least some of the effects of CCK on intestinal function appear to be mediated via activation of intrinsic neurons in the myenteric plexus. However, the distribution of CCK-responsive enteric neurons within the rat small intestine is not known. Neither has the role of CCK-A receptors in the activation of rat myenteric neurons been investigated. Therefore, to determine the distribution of CCK-responsive neurons in the small intestinal myenteric plexus we utilized immunohistochemical detection of Fos, the protein product of the immediate early gene c-fos, to identify neurons that were activated by exogenous CCK. We also monitored Fos expression in the dorsal hindbrain, and examined CCK-induced Fos expression in the presence or absence of a receptor antagonist for the type-A CCK receptor. We found that CCK significantly increased Fos expression in the hindbrain and in myenteric neurons of the duodenum and jejunum, but not the ileum. Neuronal Fos responsiveness in both brain and myenteric neurons was mediated by CCK-A receptors, as CCK-induced Fos expression was eliminated in rats pretreated with a CCK-A receptor antagonist. We conclude that CCK activates small intestinal myenteric neurons, via CCK-A receptors. Activation of these intrinsic intestinal neurons may participate in reflexes and behaviors that are mediated by CCK.

Purification of two equine pepsinogens by use of high-performance liquid chromatography.

OBJECTIVES: To purify and characterize pepsinogens in equine gastric mucosa. SAMPLE POPULATION: Stomachs collected from 2 healthy horses at necropsy. PROCEDURE: After collection, stomachs were placed immediately in ice before storage at -48 C. After slow thawing, the mucosa was scraped off while the tissue was immersed in 0.1M potassium phosphate (pH 7.4) at 4 C, then was homogenized. The filtered extract was subjected to anion-exchange chromatography. Fractions that were found to contain pepsin or pepsinogen were further chromatographed. Individual fractions were tested for pepsinogen or pepsin content by monitoring proteolytic activity at pH 2 and 3, respectively. Fractions from all columns were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to confirm molecular weight of pepsinogens and pepsin. RESULTS: Two pepsinogens and at least 1 pepsin were purified from equine gastric mucosa. CONCLUSIONS: On the basis of molecular mass, equine gastric mucosa contains 2 pepsinogens. CLINICAL RELEVANCE: Results of this study will enable future development of an ELISA or radioimmunoassay for use in the diagnosis of equine gastric ulceration.