L-type voltage-gated Ca 2+ channel Ca V 1.2 regulates chondrogenesis during limb development

Proc Natl Acad Sci U S A. 2019 Oct 22;116(43):21592-21601. doi: 10.1073/pnas.1908981116. Epub 2019 Oct 7.

Abstract

All cells, including nonexcitable cells, maintain a discrete transmembrane potential (V mem), and have the capacity to modulate V mem and respond to their own and neighbors' changes in V mem Spatiotemporal variations have been described in developing embryonic tissues and in some cases have been implicated in influencing developmental processes. Yet, how such changes in V mem are converted into intracellular inputs that in turn regulate developmental gene expression and coordinate patterned tissue formation, has remained elusive. Here we document that the V mem of limb mesenchyme switches from a hyperpolarized to depolarized state during early chondrocyte differentiation. This change in V mem increases intracellular Ca2+ signaling through Ca2+ influx, via CaV1.2, 1 of L-type voltage-gated Ca2+ channels (VGCCs). We find that CaV1.2 activity is essential for chondrogenesis in the developing limbs. Pharmacological inhibition by an L-type VGCC specific blocker, or limb-specific deletion of CaV1.2, down-regulates expression of genes essential for chondrocyte differentiation, including Sox9, Col2a1, and Agc1, and thus disturbs proper cartilage formation. The Ca2+-dependent transcription factor NFATc1, which is a known major transducer of intracellular Ca2+ signaling, partly rescues Sox9 expression. These data reveal instructive roles of CaV1.2 in limb development, and more generally expand our understanding of how modulation of membrane potential is used as a mechanism of developmental regulation.

Keywords: calcium channel; chondrogenesis; limb development; membrane potential.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aggrecans / metabolism
  • Animals
  • Calcium Channels, L-Type / metabolism*
  • Cartilage / embryology*
  • Chick Embryo
  • Chickens
  • Chondrogenesis / physiology*
  • Collagen Type II / metabolism
  • Extremities / embryology*
  • Gene Expression Regulation, Developmental / genetics
  • Membrane Potentials / physiology*
  • Mice
  • Mice, Transgenic
  • NFATC Transcription Factors / metabolism
  • SOX9 Transcription Factor / metabolism

Substances

  • Acan protein, mouse
  • Aggrecans
  • CACNA1C protein, mouse
  • Calcium Channels, L-Type
  • Col2a1 protein, mouse
  • Collagen Type II
  • NFATC Transcription Factors
  • Nfatc2 protein, mouse
  • SOX9 Transcription Factor
  • Sox9 protein, mouse