Membrane potential drives the exit from pluripotency and cell fate commitment via calcium and mTOR

Abstract

Transitioning from pluripotency to differentiated cell fates is fundamental to both embryonic development and adult tissue homeostasis. Improving our understanding of this transition would facilitate our ability to manipulate pluripotent cells into tissues for therapeutic use. Here, we show that membrane voltage (Vm) regulates the exit from pluripotency and the onset of germ layer differentiation in the embryo, a process that affects both gastrulation and left-right patterning. By examining candidate genes of congenital heart disease and heterotaxy, we identify KCNH6, a member of the ether-a-go-go class of potassium channels that hyperpolarizes the Vm and thus limits the activation of voltage gated calcium channels, lowering intracellular calcium. In pluripotent embryonic cells, depletion of kcnh6 leads to membrane depolarization, elevation of intracellular calcium levels, and the maintenance of a pluripotent state at the expense of differentiation into ectodermal and myogenic lineages. Using high-resolution temporal transcriptome analysis, we identify the gene regulatory networks downstream of membrane depolarization and calcium signaling and discover that inhibition of the mTOR pathway transitions the pluripotent cell to a differentiated fate. By manipulating Vm using a suite of tools, we establish a bioelectric pathway that regulates pluripotency in vertebrates, including human embryonic stem cells.

A controllable perfusion microfluidic chip for facilitating the development of retinal ganglion cells in human retinal organoids

“…64 Besides, KCNH6 and Ca 2+ channels affect the transition from pluripotency to differentiated cell fates during embryonic development. 65 Consistent with previous studies, our results show that the activity of voltage-gated potassium and calcium ion channels regulates neuronal differentiation, cell fate commitment, and central nervous system (CNS) development. However, other techniques, such as patch-clamp electrophysiological and ion channel detection techniques, will be needed in future research to test the function of ion channels in ROs under perfused conditions.…”

Multi‐Catcher Polymers Regulate the Nucleolin Cluster on the Cell Surface for Cancer Therapy

“…Indeed, the rapid calcium influx caused the loss of mitochondrial membrane potential. [ 42–45 ] Subsequently, the depolarization of mitochondrial membrane potential (ΔΨ m ) caused by the calcium influx was further confirmed by Rhodamine 123 staining (red) (Figure S12, Supporting Information). The effect of polyvalent aptamers in GC‐chol‐apt‐cDNA in the production of a similar physiological stimulation was further evaluated.…”

A red-emitting carborhodamine for monitoring and measuring membrane potential

“…Further, values of membrane potentials in nonelectrically excitable cells may play important roles in signaling, differentiation, and cell cycle progression. [5][6][7] Reflecting the importance of membrane potential, some 10 to 50% of oxygen consumption is directed towards setting the "resting" membrane potential through the action of the ATP-driven Na-K exchanger. 8 The primary mode of monitoring changes in and measuring values of membrane potential is patch-clamp electrophysiology.…”