Document Detail

Functional ion channels and cell proliferation in 3T3-L1 preadipocytes.
MedLine Citation:
PMID:  21732368     Owner:  NLM     Status:  Publisher    
Mouse 3T3-L1 preadipocytes are widely used for metabolic study of obesity; however, their cellular physiology is not fully understood. The present study investigates functional ion channels and their role in the regulation of cell proliferation using whole-cell patch voltage-clamp, RT-PCR, Western blot, and cell proliferation assay in undifferentiated 3T3-L1 preadipocytes. We found three types of ionic currents present in 3T3-L1 preadipocytes, including an inwardly-rectifying K(+) current (I(Kir) , recorded in 15% of cells) inhibited by Ba(2+) , a Ca(2+) -activated intermediate K(+) current (IK(Ca) , recorded in 44% of cells) inhibited by clotrimazole (or TRAM-34) as well as a chloride current (I(Cl) ) inhibited by 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS) in 12% of cells, which can be activated in all cells with hypotonic (0.8T) insult, implicating a volume-sensitive I(Cl) (I(Cl.vol) ). RT-PCR and Western blot analysis revealed the expression of KCa3.1 (for IK(Ca) ), Kir2.1 (for I(Kir) ), and Clcn3 (for I(Cl.vol) ). Blockade of IK(Ca) with TRAM-34 or I(Cl.vol) with DIDS inhibited cell proliferation in a concentration-dependent manner. Knockdown of KCa3.1 or Clcn3 with specific siRNAs also suppressed cell proliferation. Flow cytometry analysis showed that blockade or silencing of KCa3.1 or Clcn3 channels with corresponding blockers or siRNAs caused an accumulation of cells at the G0/G1 phase. These results demonstrate that three functional ion channel currents, I(KCa) , I(Cl.vol) , and I(Kir) , are heterogeneously present in 3T3-L1 preadipocytes. I(KCa) and I(Cl.vol) participate in the regulation of cell proliferation. J. Cell. Physiol. © 2011 Wiley-Liss, Inc.
Xiao-Hua Zhang; Ying-Ying Zhang; Hai-Ying Sun; Man-Wen Jin; Gui-Rong Li
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-7-5
Journal Detail:
Title:  Journal of cellular physiology     Volume:  -     ISSN:  1097-4652     ISO Abbreviation:  -     Publication Date:  2011 Jul 
Date Detail:
Created Date:  2011-7-6     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0050222     Medline TA:  J Cell Physiol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2011 Wiley-Liss, Inc.
Department of Pharmacology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China; Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.
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