Research Resource: A Chromogranin A Reporter for Serotonin and Histamine Secreting Enteroendocrine Cells.
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.
Novo Nordisk Foundation Center for Basic Metabolic Research (M.S.E., M.L.L., K.V.G., K.L.E., T.W.S.), Section for Metabolic Receptology, and Laboratory for Molecular Pharmacology (M.S.E., M.L.L., K.V.G., K.L.E., T.W.S.), Department of Neuroscience and Pharmacology, Faculty of Health and Medical Sciences, and Department of Biomedical Sciences (S.S.P.), Endocrinology Research Section, University of Copenhagen, Copenhagen DK-2200, Denmark; Danish Diabetes Academy (M.S.E.), Odense, Denmark; and Division of Hypothalamic Research (S.O.-L., J.M.Z.), Department of Internal Medicine, The University of Texas Southwestern Medical Center, Dallas, Texas 75390.
Maja S
MS
Engelstoft
Novo Nordisk Foundation Center for Basic Metabolic Research (M.S.E., M.L.L., K.V.G., K.L.E., T.W.S.), Section for Metabolic Receptology, and Laboratory for Molecular Pharmacology (M.S.E., M.L.L., K.V.G., K.L.E., T.W.S.), Department of Neuroscience and Pharmacology, Faculty of Health and Medical Sciences, and Department of Biomedical Sciences (S.S.P.), Endocrinology Research Section, University of Copenhagen, Copenhagen DK-2200, Denmark; Danish Diabetes Academy (M.S.E.), Odense, Denmark; and Division of Hypothalamic Research (S.O.-L., J.M.Z.), Department of Internal Medicine, The University of Texas Southwestern Medical Center, Dallas, Texas 75390.
Mari L
ML
Lund
Novo Nordisk Foundation Center for Basic Metabolic Research (M.S.E., M.L.L., K.V.G., K.L.E., T.W.S.), Section for Metabolic Receptology, and Laboratory for Molecular Pharmacology (M.S.E., M.L.L., K.V.G., K.L.E., T.W.S.), Department of Neuroscience and Pharmacology, Faculty of Health and Medical Sciences, and Department of Biomedical Sciences (S.S.P.), Endocrinology Research Section, University of Copenhagen, Copenhagen DK-2200, Denmark; Danish Diabetes Academy (M.S.E.), Odense, Denmark; and Division of Hypothalamic Research (S.O.-L., J.M.Z.), Department of Internal Medicine, The University of Texas Southwestern Medical Center, Dallas, Texas 75390.
Kaare V
KV
Grunddal
Novo Nordisk Foundation Center for Basic Metabolic Research (M.S.E., M.L.L., K.V.G., K.L.E., T.W.S.), Section for Metabolic Receptology, and Laboratory for Molecular Pharmacology (M.S.E., M.L.L., K.V.G., K.L.E., T.W.S.), Department of Neuroscience and Pharmacology, Faculty of Health and Medical Sciences, and Department of Biomedical Sciences (S.S.P.), Endocrinology Research Section, University of Copenhagen, Copenhagen DK-2200, Denmark; Danish Diabetes Academy (M.S.E.), Odense, Denmark; and Division of Hypothalamic Research (S.O.-L., J.M.Z.), Department of Internal Medicine, The University of Texas Southwestern Medical Center, Dallas, Texas 75390.
Kristoffer L
KL
Egerod
Novo Nordisk Foundation Center for Basic Metabolic Research (M.S.E., M.L.L., K.V.G., K.L.E., T.W.S.), Section for Metabolic Receptology, and Laboratory for Molecular Pharmacology (M.S.E., M.L.L., K.V.G., K.L.E., T.W.S.), Department of Neuroscience and Pharmacology, Faculty of Health and Medical Sciences, and Department of Biomedical Sciences (S.S.P.), Endocrinology Research Section, University of Copenhagen, Copenhagen DK-2200, Denmark; Danish Diabetes Academy (M.S.E.), Odense, Denmark; and Division of Hypothalamic Research (S.O.-L., J.M.Z.), Department of Internal Medicine, The University of Texas Southwestern Medical Center, Dallas, Texas 75390.
Sherri
S
Osborne-Lawrence
Novo Nordisk Foundation Center for Basic Metabolic Research (M.S.E., M.L.L., K.V.G., K.L.E., T.W.S.), Section for Metabolic Receptology, and Laboratory for Molecular Pharmacology (M.S.E., M.L.L., K.V.G., K.L.E., T.W.S.), Department of Neuroscience and Pharmacology, Faculty of Health and Medical Sciences, and Department of Biomedical Sciences (S.S.P.), Endocrinology Research Section, University of Copenhagen, Copenhagen DK-2200, Denmark; Danish Diabetes Academy (M.S.E.), Odense, Denmark; and Division of Hypothalamic Research (S.O.-L., J.M.Z.), Department of Internal Medicine, The University of Texas Southwestern Medical Center, Dallas, Texas 75390.
Steen Seier
SS
Poulsen
Novo Nordisk Foundation Center for Basic Metabolic Research (M.S.E., M.L.L., K.V.G., K.L.E., T.W.S.), Section for Metabolic Receptology, and Laboratory for Molecular Pharmacology (M.S.E., M.L.L., K.V.G., K.L.E., T.W.S.), Department of Neuroscience and Pharmacology, Faculty of Health and Medical Sciences, and Department of Biomedical Sciences (S.S.P.), Endocrinology Research Section, University of Copenhagen, Copenhagen DK-2200, Denmark; Danish Diabetes Academy (M.S.E.), Odense, Denmark; and Division of Hypothalamic Research (S.O.-L., J.M.Z.), Department of Internal Medicine, The University of Texas Southwestern Medical Center, Dallas, Texas 75390.
Jeffrey M
JM
Zigman
Novo Nordisk Foundation Center for Basic Metabolic Research (M.S.E., M.L.L., K.V.G., K.L.E., T.W.S.), Section for Metabolic Receptology, and Laboratory for Molecular Pharmacology (M.S.E., M.L.L., K.V.G., K.L.E., T.W.S.), Department of Neuroscience and Pharmacology, Faculty of Health and Medical Sciences, and Department of Biomedical Sciences (S.S.P.), Endocrinology Research Section, University of Copenhagen, Copenhagen DK-2200, Denmark; Danish Diabetes Academy (M.S.E.), Odense, Denmark; and Division of Hypothalamic Research (S.O.-L., J.M.Z.), Department of Internal Medicine, The University of Texas Southwestern Medical Center, Dallas, Texas 75390.
Thue W
TW
Schwartz
D000818:Animals; D053379:Chromogranin A / Q000235:genetics*; D003106:Colon / Q000378:metabolism; D004759:Enterochromaffin Cells / Q000378:metabolism; D019858:Enteroendocrine Cells / Q000378:metabolism*; D005753:Gastric Mucosa / Q000378:metabolism; D017930:Genes, Reporter / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics*; D006632:Histamine / Q000378:metabolism; D006636:Histamine Release / Q000502:physiology*; D050417:Insulin-Secreting Cells / Q000378:metabolism; D007413:Intestinal Mucosa / Q000378:metabolism; D007421:Intestine, Small / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D011485:Protein Binding; D012701:Serotonin / Q000378:metabolism*
D053379:Chromogranin A; D049452:Green Fluorescent Proteins; D012701:Serotonin; D006632:Histamine