| Reversible translocation of EYFP-tagged STIM1 is coupled to calcium influx in insulin secreting beta-cells. | |
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MedLine Citation:
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PMID: 18452988 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Calcium (Ca(2+)) signaling regulates insulin secretion in pancreatic beta-cells. STIM1 has been proposed to function as an endoplasmic reticulum (ER) Ca(2+) sensor regulating store-operated Ca(2+) entry (SOCE). Here we studied the translocation of EYFP-STIM1 in response to ER calcium depletion in mouse insulinoma MIN6 cells by fluorescent microscopy. While in resting cells EYFP-STIM1 is co-localized with an ER marker, in thapsigargin (Tg)-stimulated cells it occupied highly defined areas of the peri-PM space in punctae adjacent to, but not entirely coincident with the ER. Co-staining with fluorescent phalloidin revealed that EYFP-STIM1 punctae was located in actin-poor areas. Use of the SOCE blocker in MIN6 cells, 2-aminoethoxy diphenylborate (2-APB), prevented store depletion-dependent translocation of EYFP-STIM1 to the PM in a concentration-dependent (3.75-100muM) and reversible manner. TIRF microscopy revealed that 2-APB treatment led to the reversible disappearance of peri-PM EYFP-STIM1 punctae, while the ER structure in this compartment remained grossly unaffected. We conclude from this data that in these cells EYFP-STIM1 is delivered to a peri-PM location from the ER upon store depletion and this trafficking is reversibly blocked by 2-APB. |
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Authors:
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Natalia A Tamarina; Andrey Kuznetsov; Louis H Philipson |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural Date: 2008-05-02 |
Journal Detail:
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Title: Cell calcium Volume: 44 ISSN: 1532-1991 ISO Abbreviation: Cell Calcium Publication Date: 2008 Dec |
Date Detail:
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Created Date: 2008-11-10 Completed Date: 2009-01-30 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 8006226 Medline TA: Cell Calcium Country: Scotland |
Other Details:
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Languages: eng Pagination: 533-44 Citation Subset: IM |
Affiliation:
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Department of Medicine, University of Chicago, 5841 S. Maryland Avenue, Chicago, IL 60637, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Bacterial Proteins / metabolism* Boron Compounds / pharmacology Calcium / pharmacology Calcium Signaling* / drug effects Cell Membrane / drug effects, metabolism Endoplasmic Reticulum / drug effects, metabolism Insulin-Secreting Cells / cytology, drug effects, metabolism* Luminescent Proteins / metabolism* Membrane Glycoproteins / metabolism* Mice Protein Transport / drug effects Recombinant Fusion Proteins / metabolism* |
| Grant Support | |
ID/Acronym/Agency:
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DK-20595/DK/NIDDK NIH HHS; DK-48494/DK/NIDDK NIH HHS; DK-63493/DK/NIDDK NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Bacterial Proteins; 0/Boron Compounds; 0/Luminescent Proteins; 0/Membrane Glycoproteins; 0/Recombinant Fusion Proteins; 0/Stim1 protein, mouse; 0/yellow fluorescent protein, Bacteria; 7440-70-2/Calcium; 83075-94-9/diphenylborate |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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