Research Resource: A Chromogranin A Reporter for Serotonin and Histamine Secreting Enteroendocrine Cells

• Published in: Molecular endocrinology (Baltimore, Md.) Md.), 2015-11, Vol.29 (11), p.1658-1671
• Main Authors: Engelstoft, Maja S, Lund, Mari L, Grunddal, Kaare V, Egerod, Kristoffer L, Osborne-Lawrence, Sherri, Poulsen, Steen Seier, Zigman, Jeffrey M, Schwartz, Thue W
• Format: Article
• Language: English
• Subjects: Animals; Chromogranin A - genetics; Colon - metabolism; Enterochromaffin Cells - metabolism; Enterochromaffin Cells - secretion; Enteroendocrine Cells - metabolism; Enteroendocrine Cells - secretion; Genes, Reporter - genetics; Green Fluorescent Proteins - genetics; Histamine - metabolism; Histamine Release - physiology; Insulin-Secreting Cells - metabolism; Insulin-Secreting Cells - secretion; Intestinal Mucosa - metabolism; Intestinal Mucosa - secretion; Intestine, Small - metabolism; Mice; Mice, Transgenic; Protein Binding; Research Resource; Serotonin - secretion; Stomach - metabolism
• ISSN: 0888-8809; 1944-9917
• DOI: 10.1210/me.2015-1106

Chromogranin A (ChgA) is an acidic protein found in large dense-core secretory vesicles and generally considered to be expressed in all enteroendocrine cells of the gastrointestinal (GI) tract. Here, we characterize a novel reporter mouse for ChgA, ChgA-humanized Renilla reniformis (hr)GFP. The hrGFP reporter was found in the monoamine-storing chromaffin cells of the adrenal medulla, where ChgA was originally discovered. hrGFP also was expressed in enteroendocrine cells throughout the GI tract, faithfully after the expression of ChgA, as characterized by immunohistochemistry and quantitative PCR analysis of fluorescence-activated cell sorting-purified cells, although the expression in the small intestine was weak compared with that of the stomach and colon. In the stomach, hrGFP was highly expressed in almost all histamine-storing enterochromaffin (EC)-like cells, at a lower level in the majority of serotonin-storing EC cells and ghrelin cells, in a small fraction of somatostatin cells, but was absent from gastrin cells. In the small intestine, the hrGFP reporter was selectively, but weakly expressed in EC cells, although not in any peptide-storing enteroendocrine cells. In the colon, hrGFP was exclusively expressed in EC cells but absent from the peptide-storing enteroendocrine cells. In contrast, in the pancreas, hrGFP was expressed in β-cells, α-cells, and a fraction of pancreatic polypeptide cells. It is concluded that ChgA-hrGFP in the GI tract functions as an effective reporter, particularly for the large populations of still poorly characterized monoamine-storing enteroendocrine cells. Furthermore, our findings substantiate the potential function of ChgA as a monoamine-binding protein that facilitates the regulated endocrine secretion of large amounts of monoamines from enteroendocrine cells.