Document Detail


Reversible translocation of EYFP-tagged STIM1 is coupled to calcium influx in insulin secreting beta-cells.
MedLine Citation:
PMID:  18452988     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Natalia A Tamarina; Andrey Kuznetsov; Louis H Philipson
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2008-05-02
Journal Detail:
Title:  Cell calcium     Volume:  44     ISSN:  1532-1991     ISO Abbreviation:  Cell Calcium     Publication Date:  2008 Dec 
Date Detail:
Created Date:  2008-11-10     Completed Date:  2009-01-30     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8006226     Medline TA:  Cell Calcium     Country:  Scotland    
Other Details:
Languages:  eng     Pagination:  533-44     Citation Subset:  IM    
Affiliation:
Department of Medicine, University of Chicago, 5841 S. Maryland Avenue, Chicago, IL 60637, USA.
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MeSH Terms
Descriptor/Qualifier:
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:
DK-20595/DK/NIDDK NIH HHS; DK-48494/DK/NIDDK NIH HHS; DK-63493/DK/NIDDK NIH HHS
Chemical
Reg. No./Substance:
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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