Infrared laser-induced gene expression for tracking development and function of single C. elegans embryonic neurons

Anupriya Singhal and Shai Shaham ()
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Anupriya Singhal: Laboratory of Developmental Genetics, The Rockefeller University
Shai Shaham: Laboratory of Developmental Genetics, The Rockefeller University

Nature Communications, 2017, vol. 8, issue 1, 1-13

Abstract: Abstract Visualizing neural-circuit assembly in vivo requires tracking growth of optically resolvable neurites. The Caenorhabditis elegans embryonic nervous system, comprising 222 neurons and 56 glia, is attractive for comprehensive studies of development; however, embryonic reporters are broadly expressed, making single-neurite tracking/manipulation challenging. We present a method, using an infrared laser, for reproducible heat-dependent gene expression in small sublineages (one to four cells) without radiation damage. We go beyond proof-of-principle, and use our system to label and track single neurons during early nervous-system assembly. We uncover a retrograde extension mechanism for axon growth, and reveal the aetiology of axon-guidance defects in sax-3/Robo and vab-1/EphR mutants. We also perform cell-specific rescues, determining DAF-6/patched-related site of action during sensory-organ development. Simultaneous ablation and labelling of cells using our system reveals roles for glia in dendrite extension. Our method can be applied to other optically/IR-transparent organisms, and opens the door to high-resolution systematic analyses of C. elegans morphogenesis.

Date: 2017
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DOI: 10.1038/ncomms14100

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