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This Article Full Text Full Text (PDF) All Versions of this Article: 293/1/E347    most recent 00055.2007v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Web of Science (2) Citing Articles via Google Scholar Google Scholar Articles by Gartner, W. Articles by Wagner, L. Search for Related Content PubMed PubMed Citation Articles by Gartner, W. Articles by Wagner, L.

New functional aspects of the neuroendocrine marker secretagogin based on the characterization of its rat homolog

¹Department of Medicine III, ³Department of Immunology, and ⁴Division of Endocrinology and Metabolism, Medical University Vienna, Vienna, Austria; and ²Bulgarian Academy of Science, Sofia, Bulgaria

Submitted 23 January 2007 ; accepted in final form 4 April 2007

Secretagogin is a recently cloned human -cell-expressed EF-hand Ca²⁺-binding protein. Converging evidence indicates that it exerts Ca²⁺ sensor activity and is involved in regulation of insulin synthesis and secretion. To obtain a potent tool for the extension of its functional analysis in rat in vitro systems, we cloned the rat homolog of human secretagogin. Using comparative sequence analysis, immunostaining, and immunoblotting, we demonstrated a high degree of sequence homology and similar tissue expression patterns of human and rat secretagogin. Highest rat secretagogin expression levels were found in pancreatic -cells. On the basis of newly generated anti-rat secretagogin antibodies, we established a rat secretagogin-specific sandwich capture ELISA and demonstrated release of secretagogin from viable Rin-5F cells. Dexamethasone treatment of Rin-5F cells resulted in an increased secretagogin release rate, which was inversely correlated with insulin secretion. In contrast, the secretagogin transcription rate was markedly reduced. This resulted in a decreased intracellular secretagogin content under the influence of dexamethasone. Sucrose gradient cell fractionation analysis of Rin-5F cells confirmed the predominant cytosolic localization of secretagogin, with only limited association of secretagogin with insulin granules. The loss of intracellular secretagogin after dexamethasone treatment affected predominantly the insulin granule-associated secretagogin fractions. The sequence homology and the comparable tissue expression patterns of human and rat secretagogin indicate conserved intracellular functions. The effects of dexamethasone on the total secretagogin content in Rin-5F cells and on its intracellular distribution might result in an impaired Ca²⁺ sensitivity of dexamethasone-treated insulin-secreting cells.

EF-hand protein; insulin granules; dexamethasone; calcium sensor