application) In secretory granules of sympathoadrenal chromaffin cells, co-stored catecholamines and peptides are cosecreted by exocytosis in response to acetylcholine or to the novel non-cholinergic neurotransmitter pituitary adenylyl cyclase-activating polypeptide (PACAP). The chromogranins/ secretogranins (Cg/Sg), a family of acidic, soluble proteins found in the cores of amine and peptide hormone and neurotransmitter secretory vesicles, have a virtually ubiquitous occurrence in such vesicles throughout the neuroendocrine system. Their proteolytic fragments exert effects on hormone or neurotransmitter release as well as target cells at a variety of sites. For example, chromogranin A (CgA) fragments vasostatin and catestatin control vasoreactivity and catecholamine release, and CgA fragment pancreastatin elevates blood glucose. This proposal develops 3 specific Aims in seeking to understand secretory granule events which occur in response to secretory stimuli in neuroendocrine cells such as the chromaffin cell. The responses to be investigated are secretion and gene expression (transcription).
In Aim 1, we will investigate how catecholamine secretion and CgA gene transactivation responses desensitize following pre-ganglionic stimulation of chromaffin cells by PACAP and whether desensitization of these responses involves G protein-coupled receptor kinases (GRK), beta-arrestins and/or second messenger-activated kinases.
In Aim 2, we will use the fluorescence of a novel CgA-EGFP chimera (Enhanced Green Fluorescent Protein) to determine in situ whether discrete CgA primary structural domains are responsible for the trafficking of CgA into the regulated pathway of secretion, and how CgA interacts with other secreted or membrane proteins of the secretory granule. Finally, we will use this chimeric CgA-EGFP protein as a calibrated H+ (pH) sensor (Aim 3) to investigate in situ, the dynamics of intravesicular pH and its role in the secretory process triggered by the physiologic secretagogues, such as PACAP or acetylcholine, or by sympathomimetic amines.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Scientist Development Award - Research & Training (K01)
Project #
1K01DK059628-01
Application #
6322497
Study Section
Diabetes, Endocrinology and Metabolic Diseases B Subcommittee (DDK)
Program Officer
Hyde, James F
Project Start
2001-06-01
Project End
2004-05-31
Budget Start
2001-06-01
Budget End
2002-05-31
Support Year
1
Fiscal Year
2001
Total Cost
$84,797
Indirect Cost
Name
University of California San Diego
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
077758407
City
La Jolla
State
CA
Country
United States
Zip Code
92093
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Stettler, Hansruedi; Beuret, Nicole; Prescianotto-Baschong, Cristina et al. (2009) Determinants for chromogranin A sorting into the regulated secretory pathway are also sufficient to generate granule-like structures in non-endocrine cells. Biochem J 418:81-91
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Taupenot, Laurent; Harper, Kimberly L; O'Connor, Daniel T (2005) Role of H+-ATPase-mediated acidification in sorting and release of the regulated secretory protein chromogranin A: evidence for a vesiculogenic function. J Biol Chem 280:3885-97
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