Expression of cholecystokinin a receptors in neurons innervating the rat stomach and intestine

C. Sternini, H. Wong, T. Pham, R. De Giorgio, L. J. Miller, S. M. Kuntz, Jr Reeve J.R., J. H. Walsh, Helen E Raybould

Research output: Contribution to journalArticle

80 Citations (Scopus)

Abstract

Background and Aims: Two distinct receptors, cholecystokinin (CCK)-A and CCK-B, mediate CCK effects in the digestive system. The aim of this study was to elucidate the cellular sites of expression of CCK-A receptor in the rat stomach and small intestine. Methods: We developed and characterized an antibody to the N-terminal region (LDQPQPSKEWQSA) of rat CCK-A receptor and used it for localization studies with immunohistochemistry. Results: Specificity of the antiserum was demonstrated by (1) detection of a broad band at 85-95 kilodaltons in Western blots of membranes from CCK-A receptor CHO-transfected cells; (2) cell surface staining of CCK-A receptor- transfected cells, (3) translocation of CCK-A receptor immunostaining in CCK- A receptor-transfected cells after exposure to CCK; and (4) abolition of tissue immunostaining by preadsorbtion of the antibody with the peptide used for immunization. CCK-A receptor immunoreactivity was localized to myenteric neurons and to fibers in the muscle and mucosa. In the stomach, myenteric neurons and mucosal fibers were abundant. Many CCK-A receptor myenteric neurons contained the inhibitory transmitter vasoactive intestinal polypeptide, and some were immunoreactive for the excitatory transmitter substance P. Subdiaphragmatic vagotomy reduced the density of CCK-A receptor fibers in the gastric mucosa by approximately 50%, whereas celiac/superior mesenteric ganglionectomy had no detectable effect on fiber density. Conclusions: CCK-A receptor is expressed in functionally distinct neurons of the gastrointestinal tract. CCK-A receptor may mediate reflexes stimulated by CCK through the release of other transmitters from neurons bearing the receptor.

Original languageEnglish (US)
Pages (from-to)1136-1146
Number of pages11
JournalGastroenterology
Volume117
Issue number5
DOIs
StatePublished - 1999
Externally publishedYes

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Cholecystokinin A Receptor
Cholecystokinin Receptors
Intestines
Stomach
Neurons
Cholecystokinin
Tetragastrin
Ganglionectomy
Digestive System
CHO Cells
Antibodies
Vagotomy
Vasoactive Intestinal Peptide
Substance P
Gastric Mucosa
Abdomen
Small Intestine
Reflex

ASJC Scopus subject areas

  • Gastroenterology

Cite this

Expression of cholecystokinin a receptors in neurons innervating the rat stomach and intestine. / Sternini, C.; Wong, H.; Pham, T.; De Giorgio, R.; Miller, L. J.; Kuntz, S. M.; Reeve J.R., Jr; Walsh, J. H.; Raybould, Helen E.

In: Gastroenterology, Vol. 117, No. 5, 1999, p. 1136-1146.

Research output: Contribution to journalArticle

Sternini, C, Wong, H, Pham, T, De Giorgio, R, Miller, LJ, Kuntz, SM, Reeve J.R., J, Walsh, JH & Raybould, HE 1999, 'Expression of cholecystokinin a receptors in neurons innervating the rat stomach and intestine', Gastroenterology, vol. 117, no. 5, pp. 1136-1146. https://doi.org/10.1016/S0016-5085(99)70399-9
Sternini, C. ; Wong, H. ; Pham, T. ; De Giorgio, R. ; Miller, L. J. ; Kuntz, S. M. ; Reeve J.R., Jr ; Walsh, J. H. ; Raybould, Helen E. / Expression of cholecystokinin a receptors in neurons innervating the rat stomach and intestine. In: Gastroenterology. 1999 ; Vol. 117, No. 5. pp. 1136-1146.
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abstract = "Background and Aims: Two distinct receptors, cholecystokinin (CCK)-A and CCK-B, mediate CCK effects in the digestive system. The aim of this study was to elucidate the cellular sites of expression of CCK-A receptor in the rat stomach and small intestine. Methods: We developed and characterized an antibody to the N-terminal region (LDQPQPSKEWQSA) of rat CCK-A receptor and used it for localization studies with immunohistochemistry. Results: Specificity of the antiserum was demonstrated by (1) detection of a broad band at 85-95 kilodaltons in Western blots of membranes from CCK-A receptor CHO-transfected cells; (2) cell surface staining of CCK-A receptor- transfected cells, (3) translocation of CCK-A receptor immunostaining in CCK- A receptor-transfected cells after exposure to CCK; and (4) abolition of tissue immunostaining by preadsorbtion of the antibody with the peptide used for immunization. CCK-A receptor immunoreactivity was localized to myenteric neurons and to fibers in the muscle and mucosa. In the stomach, myenteric neurons and mucosal fibers were abundant. Many CCK-A receptor myenteric neurons contained the inhibitory transmitter vasoactive intestinal polypeptide, and some were immunoreactive for the excitatory transmitter substance P. Subdiaphragmatic vagotomy reduced the density of CCK-A receptor fibers in the gastric mucosa by approximately 50{\%}, whereas celiac/superior mesenteric ganglionectomy had no detectable effect on fiber density. Conclusions: CCK-A receptor is expressed in functionally distinct neurons of the gastrointestinal tract. CCK-A receptor may mediate reflexes stimulated by CCK through the release of other transmitters from neurons bearing the receptor.",
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T1 - Expression of cholecystokinin a receptors in neurons innervating the rat stomach and intestine

AU - Sternini, C.

AU - Wong, H.

AU - Pham, T.

AU - De Giorgio, R.

AU - Miller, L. J.

AU - Kuntz, S. M.

AU - Reeve J.R., Jr

AU - Walsh, J. H.

AU - Raybould, Helen E

PY - 1999

Y1 - 1999

N2 - Background and Aims: Two distinct receptors, cholecystokinin (CCK)-A and CCK-B, mediate CCK effects in the digestive system. The aim of this study was to elucidate the cellular sites of expression of CCK-A receptor in the rat stomach and small intestine. Methods: We developed and characterized an antibody to the N-terminal region (LDQPQPSKEWQSA) of rat CCK-A receptor and used it for localization studies with immunohistochemistry. Results: Specificity of the antiserum was demonstrated by (1) detection of a broad band at 85-95 kilodaltons in Western blots of membranes from CCK-A receptor CHO-transfected cells; (2) cell surface staining of CCK-A receptor- transfected cells, (3) translocation of CCK-A receptor immunostaining in CCK- A receptor-transfected cells after exposure to CCK; and (4) abolition of tissue immunostaining by preadsorbtion of the antibody with the peptide used for immunization. CCK-A receptor immunoreactivity was localized to myenteric neurons and to fibers in the muscle and mucosa. In the stomach, myenteric neurons and mucosal fibers were abundant. Many CCK-A receptor myenteric neurons contained the inhibitory transmitter vasoactive intestinal polypeptide, and some were immunoreactive for the excitatory transmitter substance P. Subdiaphragmatic vagotomy reduced the density of CCK-A receptor fibers in the gastric mucosa by approximately 50%, whereas celiac/superior mesenteric ganglionectomy had no detectable effect on fiber density. Conclusions: CCK-A receptor is expressed in functionally distinct neurons of the gastrointestinal tract. CCK-A receptor may mediate reflexes stimulated by CCK through the release of other transmitters from neurons bearing the receptor.

AB - Background and Aims: Two distinct receptors, cholecystokinin (CCK)-A and CCK-B, mediate CCK effects in the digestive system. The aim of this study was to elucidate the cellular sites of expression of CCK-A receptor in the rat stomach and small intestine. Methods: We developed and characterized an antibody to the N-terminal region (LDQPQPSKEWQSA) of rat CCK-A receptor and used it for localization studies with immunohistochemistry. Results: Specificity of the antiserum was demonstrated by (1) detection of a broad band at 85-95 kilodaltons in Western blots of membranes from CCK-A receptor CHO-transfected cells; (2) cell surface staining of CCK-A receptor- transfected cells, (3) translocation of CCK-A receptor immunostaining in CCK- A receptor-transfected cells after exposure to CCK; and (4) abolition of tissue immunostaining by preadsorbtion of the antibody with the peptide used for immunization. CCK-A receptor immunoreactivity was localized to myenteric neurons and to fibers in the muscle and mucosa. In the stomach, myenteric neurons and mucosal fibers were abundant. Many CCK-A receptor myenteric neurons contained the inhibitory transmitter vasoactive intestinal polypeptide, and some were immunoreactive for the excitatory transmitter substance P. Subdiaphragmatic vagotomy reduced the density of CCK-A receptor fibers in the gastric mucosa by approximately 50%, whereas celiac/superior mesenteric ganglionectomy had no detectable effect on fiber density. Conclusions: CCK-A receptor is expressed in functionally distinct neurons of the gastrointestinal tract. CCK-A receptor may mediate reflexes stimulated by CCK through the release of other transmitters from neurons bearing the receptor.

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