Neuronal regeneration in C. elegans requires subcellular calcium release by ryanodine receptor channels and can be enhanced by optogenetic stimulation
Department of Neurobiology; Alkema Lab; Graduate School of Biomedical Sciences, Neuroscience Program
Medical Subject Headings
Animals; Animals, Genetically Modified; Caenorhabditis elegans; Calcium; Mechanotransduction, Cellular; Nerve Regeneration; Neurons; Optogenetics; Rhodopsin; Ryanodine Receptor Calcium Release Channel; Subcellular Fractions
Neuroscience and Neurobiology
Regulated calcium signals play conserved instructive roles in neuronal repair, but how localized calcium stores are differentially mobilized, or might be directly manipulated, to stimulate regeneration within native contexts is poorly understood. We find here that localized calcium release from the endoplasmic reticulum via ryanodine receptor (RyR) channels is critical in stimulating initial regeneration following traumatic cellular damage in vivo. Using laser axotomy of single neurons in Caenorhabditis elegans, we find that mutation of unc-68/RyR greatly impedes both outgrowth and guidance of the regenerating neuron. Performing extended in vivo calcium imaging, we measure subcellular calcium signals within the immediate vicinity of the regenerating axon end that are sustained for hours following axotomy and completely eliminated within unc-68/RyR mutants. Finally, using a novel optogenetic approach to periodically photo-stimulate the axotomized neuron, we can enhance its regeneration. The enhanced outgrowth depends on both amplitude and temporal pattern of excitation and can be blocked by disruption of UNC-68/RyR. This demonstrates the exciting potential of emerging optogenetic technology to beneficially manipulate cell physiology in the context of neuronal regeneration and indicates a link to the underlying cellular calcium signal. Taken as a whole, our findings define a specific localized calcium signal mediated by RyR channel activity that stimulates regenerative outgrowth, which may be dynamically manipulated for beneficial neurotherapeutic effects.
Rights and Permissions
Citation: J Neurosci. 2014 Nov 26;34(48):15947-56. doi: 10.1523/JNEUROSCI.4238-13.2014. Link to article on publisher's site
DOI of Published Version
The Journal of neuroscience : the official journal of the Society for Neuroscience
Sun, Lin; Shay, James; McLoed, Melissa; Roodhouse, Kevin; Chung, Samuel H.; Clark, Christopher M.; Pirri, Jennifer K.; Alkema, Mark J.; and Gabel, Christopher V., "Neuronal regeneration in C. elegans requires subcellular calcium release by ryanodine receptor channels and can be enhanced by optogenetic stimulation" (2014). GSBS Student Publications. 1876.