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Cellular and subcellular localization of cholecystokinin (CCK)-1 receptors in the pancreas, gallbladder, and stomach of mice

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Abstract

Information concerning the cellular localization of cholecystokinin (CCK)-1 receptors has been discrepant and remained scanty at ultrastructural levels. The present immunohistochemical study at light and electron microscopic levels revealed the distinct localization of CCK1 receptors in visceral organs. Immunohistochemistry by use of a purified antibody against mouse CCK1 receptor was applied to fixed tissue sections of the pancreas, gallbladder, stomach, and intestine of mice. A silver-intensified immunogold method revealed the subcellular localization under electron microscope. The immunoreactivity for CCK1 receptors was selectively found in the basolateral membrane of pancreatic acinar cells and gastric chief cells but was absent in pancreatic islets and gastric D cells. Another intense expression in the gut was seen in the myenteric nerve plexus of the antro-duodenal region and some populations of c-Kit-expressing pacemaker cells in the duodenal musculature. The gallbladder contained smooth muscle fibers with an intense immunoreactivity of CCK1 receptors on cell surfaces. The restricted localization of CCK1 receptors on the basolateral membrane of pancreatic acinar cells and gastric chief cells, along with their absence in the islets of Langerhans and gastric D cells, provides definitive information concerning the regulatory mechanism by circulating CCK. Especially, the subcellular localization in the acinar cells completes the investigation for the detection of circulating CCK by the basolateral membrane.

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Acknowledgments

This study is supported by Grants-in-Aid for Scientific Research 19100005 (to M. W.) and Core Research for Evolutional Science and Technology (CREST) from Japan Science and Technology Cooperation (to M.W.).

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Authors and Affiliations

  1. Department of Anatomy, Hokkaido University Graduate School of Medicine, N15W7, Kita-ku, Sapporo, 060-8638, Japan

    Kohtarou Konno, Hiromi Takahashi-Iwanaga, Motokazu Uchigashima, Masahiko Watanabe & Toshihiko Iwanaga

  2. Department of Physiology and Nutrition, Tokyo Kasei University, Tokyo, 173-8602, Japan

    Kyoko Miyasaka

  3. Division of Pancreatology, Department of Internal Medicine, Fukuoka Sanno Hospital, 3-6-45 Momochihama, Sawara-ku, Fukuoka, 814-0001, Japan

    Akihiro Funakoshi

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  1. Kohtarou Konno
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  2. Hiromi Takahashi-Iwanaga
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Correspondence to Toshihiko Iwanaga.

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418_2014_1281_MOESM1_ESM.jpg

Immunostaining of the CCK1 receptor in wild-type (A, C, and E) and CCK1 receptor-deficient mice (B, D, and F). The immunoreactivities in the muscle layer of the gallbladder (A), the chief cells of the stomach (C), and ICC cells of the duodenum disappear in CCK1 receptor-deficient mice (B, D, and F). Nuclei are stained green in A, B, E, and F. Bar, 20 μm (A, B, E, and F), 50 μm (C and D) (JPEG 786 kb)

418_2014_1281_MOESM2_ESM.jpg

Immunostaining of Gαq11 in the pancreas (A), stomach (B), and gallbladder (C). The basolateral membrane of pancreatic acinar cells and gastric chief cells is positive in reaction. In figure B, the positive immunoreactivities are also found in endocrine cells (arrow) and lamina muscularis mucosae. The gallbladder (C) displays the smooth muscle layer with a Gαq11 immunoreactivity. Nuclei are counterstained green with SYTOX green. Bar, 20 μm (JPEG 479 kb)

418_2014_1281_MOESM3_ESM.jpg

Immunostaining of the CCK1 receptor in the nodose ganglion (A and B) and the myenteric nerve plexus of the duodenum (C). Several neurons in the nodose ganglion show a positive immunoreactivity for the CCK1 receptor. Nuclei are stained green in figure B. In figure C, double staining with PGP9.5, a neuronal marker, of a whole mount preparation demonstrates a dot-like distribution of the CCK1 receptor immunoreactivity on the myenteric nerve plexus. Arrows indicate immunoreactive cell bodies of neurons. In figure D, in situ hybridization analysis for the CCK1 receptor and neuron-specific enolase (NSE) on a tissue section detects a CCK1 receptor mRNA-expressing neuron in the myenteric nerve plexus of the duodenum. Bar, 50 μm (A), 20 μm (B–D) (JPEG 553 kb)

418_2014_1281_MOESM4_ESM.jpg

CCK1 receptor immunoreactivities in the pancreas of the guinea pig (A) and rat (B). The CCK1 receptor is localized in pancreatic acinar cells but not islets (I). In figure A, nuclei are stained green, while in figure B insulin-containing cells are stained green. Bar, 20 μm (JPEG 519 kb)

418_2014_1281_MOESM5_ESM.jpg

Surface view of pancreatic acinar cells by scanning electron microscope (SEM). After removal of the basement membrane and connective tissue by the NaOH maceration method, the basal (B) and lateral (L) surfaces of acinar cells were observed by SEM. The basal and lateral surfaces are equipped with small domains composed of microfolds (arrows). An asterisk indicates the lumen of an acinus. Bar, 1 μm (JPEG 463 kb)

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Konno, K., Takahashi-Iwanaga, H., Uchigashima, M. et al. Cellular and subcellular localization of cholecystokinin (CCK)-1 receptors in the pancreas, gallbladder, and stomach of mice. Histochem Cell Biol 143, 301–312 (2015). https://doi.org/10.1007/s00418-014-1281-3

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