L-Type Ca2+ Channels and SK Channels in Mouse Embryonic Stem Cells and Their Contribution to Cell Proliferation

Abstract

Mouse embryonic stem cells (mESCs) are capable of both self-renewal and multilineage differentiation; thus, they can be expanded in vivo or in vitro and differentiated to produce different cell types. Despite their biological and medical interest, many physiological properties of undifferentiated mESCs, such as ion channel function, are not fully understood. Ion channels are thought to be involved in cell proliferation and differentiation. The aim of this study was to characterize functional ion channels in cultured undifferentiated mESCs and their role in cell proliferation. L-type voltage-activated Ca(2+) channels sensitive to nifedipine and small-conductance Ca(2+)-activated K(+) (SK) channels sensitive to apamin were identified. Ca(2+)-activated K(+) currents were blocked by millimolar concentrations of tetraethylammonium. The effects of Ca(2+) channel and Ca(2+)-activated K(+) channel blockers on the proliferation of undifferentiated mESCs were investigated by bromodeoxyuridine (BrdU) incorporation. Dihydropyridine derivatives, such as nifedipine, inhibited cell growth and BrdU incorporation into the cells, whereas apamin, which selectively blocks SK channels, had no effect on cell growth. These results demonstrate that functional voltage-operated Ca(2+) channels and Ca(2+)-activated K(+) channels are present in undifferentiated mESCs. Moreover, voltage-gated L-type Ca(2+) channels, but not SK channels, might be necessary for proliferation of undifferentiated mESCs.

Patient-Specific iPSC-Based Models of Huntington’s Disease as a Tool to Study Store-Operated Calcium Entry Drug Targeting

“…The corresponding SOCE channels are characterized by a nearly linear I / V curve and relatively low selectivity for calcium. It has been shown that drug targeting to calcium channels results in significant alterations of electrophysiological signals ( Vegara-Meseguer et al, 2015 ; Ryazantseva et al, 2017 ). To gain a deeper insight into the mechanism of drug targeting, it is important to discriminate between currents maintained by CRAC and TRPC channels.…”

L‐type voltage‐gated calcium channels in stem cells and tissue engineering

“…54 L-VGCCs have been demonstrated to regulate the proliferation of mouse ESCs (mESCs). 55 In addition, neural differentiation of mESCs is also strongly enhanced by L-VGCCs. 56,57 Experiments performed by Lepski indicates that L-VGCCs-mediated maturation of NSCs depends on enhanced intracellular cAMP levels.…”

“…Huang et al carried out experiments on human ESC lines H7 and H9, indicating that these might be voltage‐insensitive cells, as depolarization‐induced Ca 2+ entry by a high K + solution did not change the intracellular free Ca 2+ concentration . L‐VGCCs have been demonstrated to regulate the proliferation of mouse ESCs (mESCs) . In addition, neural differentiation of mESCs is also strongly enhanced by L‐VGCCs .…”

Surface markers of human embryonic stem cells: a meta analysis of membrane proteomics reports

“…Acquisition of electrophysiological features is a prominent manifestation of the differentiation of cells into neural and muscular lineage. However, pluripotent human stem cells (hESCs and induced pluripotent stem cells) functionally express specialized ion channels [33,34] which play important roles in the regulation of cell proliferation [as reviewed in [35,36]]. The expression analysis of ion channels in hESCs at the transcriptomics level has revealed approximately 100 voltage-gated ion channel genes, ten transcripts of which were significantly expressed [33].…”