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Article: Functional ion channels and cell proliferation in 3T3-L1 preadipocytes

TitleFunctional ion channels and cell proliferation in 3T3-L1 preadipocytes
Authors
Issue Date2012
PublisherJohn Wiley & Sons, Inc. The Journal's web site is located at http://www3.interscience.wiley.com/cgi-bin/jhome/31010
Citation
Journal Of Cellular Physiology, 2012, v. 227 n. 5, p. 1972-1979 How to Cite?
AbstractMouse 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'-disulfonic 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. © 2011 Wiley Periodicals, Inc.
Persistent Identifierhttp://hdl.handle.net/10722/145564
ISSN
2015 Impact Factor: 4.155
2015 SCImago Journal Rankings: 1.842
ISI Accession Number ID
Funding AgencyGrant Number
Research Grant Council of Hong KongHKU 770108M
Committee on Research and Conference Grant, University of Hong Kong200811159173
Funding Information:

Contract grant sponsor: Research Grant Council of Hong Kong;

References
Grants

 

DC FieldValueLanguage
dc.contributor.authorZhang, XHen_HK
dc.contributor.authorZhang, YYen_HK
dc.contributor.authorSun, HYen_HK
dc.contributor.authorJin, MWen_HK
dc.contributor.authorLi, GRen_HK
dc.date.accessioned2012-02-28T01:54:53Z-
dc.date.available2012-02-28T01:54:53Z-
dc.date.issued2012en_HK
dc.identifier.citationJournal Of Cellular Physiology, 2012, v. 227 n. 5, p. 1972-1979en_HK
dc.identifier.issn0021-9541en_HK
dc.identifier.urihttp://hdl.handle.net/10722/145564-
dc.description.abstractMouse 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'-disulfonic 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. © 2011 Wiley Periodicals, Inc.en_HK
dc.languageengen_US
dc.publisherJohn Wiley & Sons, Inc. The Journal's web site is located at http://www3.interscience.wiley.com/cgi-bin/jhome/31010en_HK
dc.relation.ispartofJournal of Cellular Physiologyen_HK
dc.rightsJournal of Cellular Physiology. Copyright © John Wiley & Sons, Inc.-
dc.subject.mesh3T3-L1 Cells - cytology - physiology-
dc.subject.meshCell Proliferation-
dc.subject.meshChloride Channels - genetics - metabolism-
dc.subject.meshIntermediate-Conductance Calcium-Activated Potassium Channels - genetics - metabolism-
dc.subject.meshPotassium Channels, Calcium-Activated - genetics - metabolism-
dc.titleFunctional ion channels and cell proliferation in 3T3-L1 preadipocytesen_HK
dc.typeArticleen_HK
dc.identifier.emailLi, GR:grli@hkucc.hku.hken_HK
dc.identifier.authorityLi, GR=rp00476en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1002/jcp.22925en_HK
dc.identifier.pmid21732368-
dc.identifier.scopuseid_2-s2.0-84862934285en_HK
dc.identifier.hkuros198635en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-84862934285&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume227en_HK
dc.identifier.issue5en_HK
dc.identifier.spage1972en_HK
dc.identifier.epage1979en_HK
dc.identifier.isiWOS:000299373900021-
dc.publisher.placeUnited Statesen_HK
dc.relation.projectCalcium Signals and Cell Proliferation in Human Cardiac Fibroblasts-
dc.identifier.scopusauthoridZhang, XH=55265324500en_HK
dc.identifier.scopusauthoridZhang, YY=55265645700en_HK
dc.identifier.scopusauthoridSun, HY=35723049200en_HK
dc.identifier.scopusauthoridJin, MW=35932258500en_HK
dc.identifier.scopusauthoridLi, GR=7408462932en_HK
dc.identifier.citeulike10523715-

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