Sonic hedgehog signaling is decoded by calcium spike activity in the developing spinal cord.
Evolutionarily conserved hedgehog proteins orchestrate the patterning of embryonic tissues, and dysfunctions in their signaling can lead to tumorigenesis. In vertebrates, Sonic hedgehog (Shh) is essential for nervous system development, but the mechanisms underlying its action remain unclear. Early electrical activity is another developmental cue important for proliferation, migration, and differentiation of neurons. Here we demonstrate the interplay between Shh signaling and Ca(2+) dynamics in the developing spinal cord. Ca(2+) imaging of embryonic spinal cells shows that Shh acutely increases Ca(2+) spike activity through activation of the Shh coreceptor Smoothened (Smo) in neurons. Smo recruits a heterotrimeric GTP-binding protein-dependent pathway and engages both intracellular Ca(2+) stores and Ca(2+) influx. The dynamics of this signaling are manifested in synchronous Ca(2+) spikes and inositol triphosphate transients apparent at the neuronal primary cilium. Interaction of Shh and electrical activity modulates neurotransmitter phenotype expression in spinal neurons. These results indicate that electrical activity and second-messenger signaling mediate Shh action in embryonic spinal neurons.
PubMed ID: 21368195
PMC ID: PMC3060219
Article link: Proc Natl Acad Sci U S A.
Genes referenced: itpr1 mnx1 ptch1 sag shh smo socs1 sox2 trpc1
Morpholinos referenced: trpc1 MO2
Article Images: [+] show captions
|Fig. 1. Spiking cells in the developing neural tube are postmitotic neurons. (A) Ca2+ imaging of the ventral spinal cord of a stage-24 (26-h-postfertilization) embryo for 20 min. Circles identify cells spiking during 20-min recording. Inset shows Ca2+ spike activity for the cell outlined in yellow. (B) (Left) After imaging, the same preparation was whole-mount immunostained for Sox2 and N-β-tubulin. (Right) Immunostaining of a transverse section of the spinal cord from a stage-24 embryo. (C) Ca2+ imaging of an open-book spinal cord preparation. (D) Whole-mount immunostaining of the same preparation for Sox2 and N-β-tubulin. (E) Diagram of the open-book spinal cord preparation shown in C and D. D, dorsal; V, ventral. (Scale bars, 20 μm.)|