Journal
CIRCULATION JOURNAL
Volume 69, Issue 10, Pages 1284-1289Publisher
JAPANESE CIRCULATION SOC
DOI: 10.1253/circj.69.1284
Keywords
cardiac cell lineage; differentiation; Nkx2.5; subtype switch; T-type Ca2+ channels
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Background The developmental changes of Ni2+-sensitivity to automaticity of Nkx2.5-positive cells derived from mouse embryonic stem cell have been identified, suggesting developmental regulation of expressing Ni2+-sensitive T-type Ca2+ channel, although the mechanism of the change has not been fully studied. Methods and Results Transcripts of Cav3.2, Cav3.1 and Cav1.2 genes of beating Nkx2.5-positive cells, which encode the Ni2+-sensitive T-type Ca2+ channel, Ni2+-insensitive T-type Ca2+ channel, and L-type Ca2+ channel, respectively, were investigated by real-time reverse-transcriptase-polymerase chain reaction, and the current density of each channel was measured by patch-clamp techniques at the early and late stages of differentiation. The expression of the Cav3.2 transcript predominated in the early stage whereas those of Cav3.1 and Cav1.2 transcripts were upregulated in the late stage, which was consistent with the change in each current density, suggesting the expression of channel proteins is largely determined at the transcriptional level. Conclusion The results indicate that the mechanism of change of Ni2+-sensitivity is partly, if not completely, the subtype switch of T-type Ca2+ channel from Cav3.2 to Cav3.1 at the transcriptional level, and that the expression of the L-type Ca2+ channel might have an attenuating effect on Ni2+-sensitivity to automaticity in the late stage of differentiation.
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