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Conference Paper: Ion channels and their role in cell proliferration of 3T3-L1 preadipocytes

TitleIon channels and their role in cell proliferration of 3T3-L1 preadipocytes
Authors
KeywordsCardiovascular disease
Issue Date2009
PublisherHong Kong College of Cardiology. The Journal's web site is located at http://www.hkcchk.com/journals.php#3
Citation
The 13th Annual Scientific Meeting of the Institute of Cardiovascular Science and Medicine (ICSM), Hong Kong, 12 December 2009. In Journal of the Hong Kong College of Cardiology, 2009, v. 17 n. 2, p. 68, abstract no. P29 How to Cite?
AbstractBACKGROUND: Mouse 3T3-L1 peradipocytes are widely used for metabolic study; however, cellular physiology (e.g. functional ion channel expression) is not fully understood. The present study was to investigate ion channel expression and functional role of them in regulating cell proliferation using whole cell patch voltage clamp technique, RT-PCR, Western blot, and cell proliferation assay in undifferentiated 3T3-L1 preadipocytes. RESULTS: We found that three types of ionic currents were present in 3T3-L1 preadipocytes, including a Ca2+-activated K+ current (IKCa) in 39% cells, an inwardly-rectifying K+ current (IKir) in 15% cells, and a chloride current (ICl) only in 8% cells under isotonic conditions. Interestingly, ICl was observed in all cells with hypotonic (0.8T) insult, suggesting that it is a volumesensitive ICl (ICl.vol). IKir was inhibited by Ba2+, and IKCa was inhibited by the intermediate conductance IKCa channel blocker clotrimazole. ICl was reduced by the chloride channel blockers DIDS. RT-PCR revealed significant expression of mRNAs: KCa3.1 for IKCa, Kir2.1 for IKir, and Clcn3 for ICl.vol. Proteins of these channels were detected using Western blot analysis. Proliferation assay demonstrated that blockade of IKCa with clotrimazole or ICl.vol with DIDS inhibited cell proliferation in a concentration-dependent manner. Flowcytometry analysis showed that clotrimazole (3 μM) and DIDS (200 μM) accumulated the cells at G0/G1 phase (from control 49.91±2.8% to 57.05±3.6% for clotrimazole, P<0.05; to 61.08±4.3% for DIDS, P<0.05). CONCLUSIONS: These results demonstrate the first information that three types of functional ion channel currents, including intermediate-conductance IKCa, ICl.vol, and IKir, are heterogeneously present in 3T3-L1 preadipocytes. IKCa and ICl.vol participate in the regulation of cell proliferation.
DescriptionPoster Presentation: abstract no. P29
Persistent Identifierhttp://hdl.handle.net/10722/129845
ISSN
2015 SCImago Journal Rankings: 0.102

 

DC FieldValueLanguage
dc.contributor.authorZhang, XHen_US
dc.contributor.authorJin, MWen_US
dc.contributor.authorLi, GRen_US
dc.date.accessioned2010-12-23T08:42:51Z-
dc.date.available2010-12-23T08:42:51Z-
dc.date.issued2009en_US
dc.identifier.citationThe 13th Annual Scientific Meeting of the Institute of Cardiovascular Science and Medicine (ICSM), Hong Kong, 12 December 2009. In Journal of the Hong Kong College of Cardiology, 2009, v. 17 n. 2, p. 68, abstract no. P29en_US
dc.identifier.issn1027-7811-
dc.identifier.urihttp://hdl.handle.net/10722/129845-
dc.descriptionPoster Presentation: abstract no. P29-
dc.description.abstractBACKGROUND: Mouse 3T3-L1 peradipocytes are widely used for metabolic study; however, cellular physiology (e.g. functional ion channel expression) is not fully understood. The present study was to investigate ion channel expression and functional role of them in regulating cell proliferation using whole cell patch voltage clamp technique, RT-PCR, Western blot, and cell proliferation assay in undifferentiated 3T3-L1 preadipocytes. RESULTS: We found that three types of ionic currents were present in 3T3-L1 preadipocytes, including a Ca2+-activated K+ current (IKCa) in 39% cells, an inwardly-rectifying K+ current (IKir) in 15% cells, and a chloride current (ICl) only in 8% cells under isotonic conditions. Interestingly, ICl was observed in all cells with hypotonic (0.8T) insult, suggesting that it is a volumesensitive ICl (ICl.vol). IKir was inhibited by Ba2+, and IKCa was inhibited by the intermediate conductance IKCa channel blocker clotrimazole. ICl was reduced by the chloride channel blockers DIDS. RT-PCR revealed significant expression of mRNAs: KCa3.1 for IKCa, Kir2.1 for IKir, and Clcn3 for ICl.vol. Proteins of these channels were detected using Western blot analysis. Proliferation assay demonstrated that blockade of IKCa with clotrimazole or ICl.vol with DIDS inhibited cell proliferation in a concentration-dependent manner. Flowcytometry analysis showed that clotrimazole (3 μM) and DIDS (200 μM) accumulated the cells at G0/G1 phase (from control 49.91±2.8% to 57.05±3.6% for clotrimazole, P<0.05; to 61.08±4.3% for DIDS, P<0.05). CONCLUSIONS: These results demonstrate the first information that three types of functional ion channel currents, including intermediate-conductance IKCa, ICl.vol, and IKir, are heterogeneously present in 3T3-L1 preadipocytes. IKCa and ICl.vol participate in the regulation of cell proliferation.-
dc.languageengen_US
dc.publisherHong Kong College of Cardiology. The Journal's web site is located at http://www.hkcchk.com/journals.php#3-
dc.relation.ispartofJournal of the Hong Kong College of Cardiologyen_US
dc.rightsOpen Access journal-
dc.subjectCardiovascular disease-
dc.titleIon channels and their role in cell proliferration of 3T3-L1 preadipocytesen_US
dc.typeConference_Paperen_US
dc.identifier.emailZhang, XH: zhangxiaohua1981@126.comen_US
dc.identifier.emailJin, MW: mwjin@mails.tjmu.edu.cnen_US
dc.identifier.emailLi, GR: grli@hkucc.hku.hk-
dc.identifier.authorityLi, GR=rp00476en_US
dc.description.naturelink_to_OA_fulltext-
dc.identifier.hkuros177395en_US
dc.identifier.volume17en_US
dc.identifier.issue2en_US
dc.identifier.spage68en_US
dc.identifier.epage68-
dc.publisher.placeHong Kong-
dc.description.otherThe 13th Annual Scientific Meeting of the Institute of Cardiovascular Science and Medicine (ICSM), Hong Kong, 12 December 2009. In Journal of the Hong Kong College of Cardiology, 2009, v. 17 n. 2, p. 68, abstract no. P29-

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