Developmental Changes of Ni2+ Sensitivity and Automaticity in Nkx2.5-Positive Cardiac Precursor Cells From Murine Embryonic Stem Cell

Manabe, Kasumi; Miake, Junichiro; Sasaki, Norihito; Furuichi, Hitomi; Yano, Shuichi; Mizuta, Einosuke; Yamamoto, Yorihiro; Hoshikawa, Yasushi; Yamazaki, Hidetoshi; Tajima, Fumihito; Shiota, Goshi; Nanba, Eiji; Ohgi, Shigetsugu; Hidaka, Kumi; Morisaki, Takayuki; Kurata, Yasutaka; Lee, Jong-Kook; Igawa, Osamu; Shigemasa, Chiaki; Hisatome, Ichiro

doi:10.1253/circj.68.724

Cited by 8 publications

(6 citation statements)

References 11 publications

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“…Furthermore, our previous results suggested subtype switching of the T-type Ca 2+ channel from Cav3.2 to Cav3.1 as another mechanism for the change in Ni 2+ -sensitivity to automaticity during cellular differentiation. 15 Both the contribution of Cav3.1, but not Cav3.2, to chamber-specified Mlc2v-positive cells derived from ES cells 12 and the subtype switching of T-type Ca 2+ channel during differentiation have been reported, 6,10 supporting the subtype switching idenitified in the present study. On the other hand, the expression of the L-type Ca 2+ channel, another depolarizing current responsible for automaticity, may explain the change in Ni 2+ -sensitivity to automaticity during differentiation.…”

Section: Discussionsupporting

confidence: 82%

“…They differentiate into sinus nodeor conduction-system-type cells, as well as myocytes. 14 We previously reported that the Nkx2.5/GFP(+) cells showed Ni 2+ -sensitive automaticity in the early and intermediate stages of their differentiation, but not in the late stage, 15 a result that suggested the subtypes of cardiac T-type Ca 2+ channels changed during their differentiation. The purpose of the current study was to study the subtype switch of the T-type Ca 2+ channels in beating Nkx2.5/GFP(+) cells.…”

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confidence: 98%

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Subtype Switching of T-Type Ca 2+ Channels From Cav3.2 to Cav3.1 During Differentiation of Embryonic Stem Cells to Cardiac Cell Lineage

Mizuta

Miake

Yano

et al. 2005

Circ J

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Background The developmental changes of Ni 2+ -sensitivity to automaticity of Nkx2.5-positive cells derived from mouse embryonic stem cell have been identified, suggesting developmental regulation of expressing Ni 2+ -sensitive T-type Ca 2+ 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 Ni 2+ -sensitive T-type Ca 2+ channel, Ni 2+ -insensitive T-type Ca 2+ channel, and L-type Ca 2+ 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. ConclusionThe results indicate that the mechanism of change of Ni 2+ -sensitivity is partly, if not completely, the subtype switch of T-type Ca 2+ channel from Cav3.2 to Cav3.1 at the transcriptional level, and that the expression of the L-type Ca 2+ channel might have an attenuating effect on Ni 2+ -sensitivity to automaticity in the late stage of differentiation. (Circ J 2005; 69: 1284 -1289

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Section: Discussionsupporting

confidence: 82%

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confidence: 98%

Subtype Switching of T-Type Ca 2+ Channels From Cav3.2 to Cav3.1 During Differentiation of Embryonic Stem Cells to Cardiac Cell Lineage

Mizuta

Miake

Yano

et al. 2005

Circ J

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“…Nkx2.5 is a common marker of cardiac progenitor cells whereas Tbx5 and Isl-1 are markers of the first and secondary heart field, respectively. All these markers genes are expressed at this stage, (9,11). Thus, we examined the distribution of Cav3.1 and Cav3.2 mRNAs and the marker gene transcripts in E10.5 hearts.…”

Section: Resultsmentioning

confidence: 99%

Different distribution of Cav3.2 and Cav3.1 transcripts encoding T-type Ca2+ channels in the embryonic heart of mice

Mizuta¹,

Shirai

Arakawa³

et al. 2010

Biomed. Res.

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We investigated the distribution of T-type Ca 2+ channel mRNAs in the mouse embryonic heart. Cav3.2, but not Cav3.1, was expressed in the E8.5 embryonic heart along with cardiac progenitor markers (Nkx2.5, Tbx5, Isl-1) and contractile proteins (alpha and beta MHC). In the E10.5 heart, the distribution of Cav3.1 mRNA was confirmed in the AV-canal and overlapped with that of MinK or Tbx2. Cav3.2 mRNA was observed not only in the AV-canal but also in the outflow tract, along with MinK and Isl-1, indicating the expression of Cav3.2 in the secondary heart field. Thus, Cav3.2 may contribute to the development of the outflow tract from the secondary heart field in the embryonic heart, whereas Cav3.1 may be involved in the development of the cardiac conduction-system together with Cav3.2.

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“…For inactivation-curve assessment, 200 mol/L NiCl2 was added to external solution to inhibit the T-type Ca 2+ channel. [20][21][22] Test pulses to +10 mV were preceded by a conditioning pulse ranging from -100 mV to 10 mV for 1 s from the holding potential of -50 mV. For activation-curve assessment, conductance (g) was obtained by dividing peak Ca 2+ channel current at the test potential by the difference between test and reverse potential.…”

Section: Electrophysiological Experimentsmentioning

confidence: 99%

Subtype Switching of L-Type Ca 2+ Channel From Cav1.3 to Cav1.2 in Embryonic Murine Ventricle

Takemura

Yasui²,

Opthof

et al. 2005

Circ J

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Developmental Changes of Ni2+ Sensitivity and Automaticity in Nkx2.5-Positive Cardiac Precursor Cells From Murine Embryonic Stem Cell

Cited by 8 publications

References 11 publications

Subtype Switching of T-Type Ca 2+ Channels From Cav3.2 to Cav3.1 During Differentiation of Embryonic Stem Cells to Cardiac Cell Lineage

Subtype Switching of T-Type Ca 2+ Channels From Cav3.2 to Cav3.1 During Differentiation of Embryonic Stem Cells to Cardiac Cell Lineage

Different distribution of Cav3.2 and Cav3.1 transcripts encoding T-type Ca2+ channels in the embryonic heart of mice

Subtype Switching of L-Type Ca 2+ Channel From Cav1.3 to Cav1.2 in Embryonic Murine Ventricle

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