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http://purl.uniprot.org/citations/23564083http://www.w3.org/1999/02/22-rdf-syntax-ns#typehttp://purl.uniprot.org/core/Journal_Citation
http://purl.uniprot.org/citations/23564083http://www.w3.org/2000/01/rdf-schema#comment"

Aim

To investigate the reverse mode function of Na(+)/Ca(2+) exchangers NCX1.1 and NCX1.5 expressed in CHO cells as well as their modulations by PKC and PKA.

Methods

CHO-K1 cells were transfected with pcDNA3.1 (+) plasmid carrying cDNA of rat cardiac NCX1.1 and brain NCX1.5. The expression of NCX1.1 and NCX1.5 was examined using Western blot analysis. The intracellular Ca(2+) level ([Ca(2+)]i) was measured using Ca(2+) imaging. Whole-cell NCX currents were recorded using patch-clamp technique. Reverse mode NCX activity was elicited by perfusion with Na(+)-free medium. Ca(2+) paradox was induced by Ca(2+)-free EBSS medium, followed by Ca(2+)-containing solution (1.8 or 3.8 mmol/L CaCl2).

Results

The protein levels of NCX1.1 and NCX1.5 expressed in CHO cells had no significant difference. The reverse modes of NCX1.1 and NCX1.5 in CHO cells exhibited a transient increase of [Ca(2+)]i, which was followed by a Ca(2+) level plateau at higher external Ca(2+) concentrations. In contrast, the wild type CHO cells showed a steady increase of [Ca(2+)]i at higher external Ca(2+) concentrations. The PKC activator PMA (0.3-10 μmol/L) and PKA activator 8-Br-cAMP (10-100 μmol/L) significantly enhanced the reverse mode activity of NCX1.1 and NCX1.5 in CHO cells. NCX1.1 was 2.4-fold more sensitive to PKC activation than NCX1.5, whereas the sensitivity of the two NCX isoforms to PKA activation had no difference. Both PKC- and PKA-enhanced NCX reverse mode activities in CHO cells were suppressed by NCX inhibitor KB-R7943 (30 μmol/L).

Conclusion

Both NCX1.1 and NCX1.5 are functional in regulating and maintaining stable [Ca(2+)]i in CHO cells and differentially regulated by PKA and PKC. The two NCX isoforms might be useful drug targets for heart and brain protection."xsd:string
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http://purl.uniprot.org/citations/23564083http://purl.uniprot.org/core/author"Wang L."xsd:string
http://purl.uniprot.org/citations/23564083http://purl.uniprot.org/core/author"Long Y."xsd:string
http://purl.uniprot.org/citations/23564083http://purl.uniprot.org/core/author"Yuan H."xsd:string
http://purl.uniprot.org/citations/23564083http://purl.uniprot.org/core/author"Feng N."xsd:string
http://purl.uniprot.org/citations/23564083http://purl.uniprot.org/core/author"Wang W.P."xsd:string
http://purl.uniprot.org/citations/23564083http://purl.uniprot.org/core/author"Wang X.L."xsd:string
http://purl.uniprot.org/citations/23564083http://purl.uniprot.org/core/author"Ma S.P."xsd:string
http://purl.uniprot.org/citations/23564083http://purl.uniprot.org/core/date"2013"xsd:gYear
http://purl.uniprot.org/citations/23564083http://purl.uniprot.org/core/name"Acta Pharmacol Sin"xsd:string
http://purl.uniprot.org/citations/23564083http://purl.uniprot.org/core/pages"691-698"xsd:string
http://purl.uniprot.org/citations/23564083http://purl.uniprot.org/core/title"Functional comparison of the reverse mode of Na+/Ca2+ exchangers NCX1.1 and NCX1.5 expressed in CHO cells."xsd:string
http://purl.uniprot.org/citations/23564083http://purl.uniprot.org/core/volume"34"xsd:string
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