Few studies have exploited the power of genetics and functional genomics to understand the mechanisms of regrowth of axons following injury. We have developed femtosecond laser axotomy to cut single axons in intact living C. elegans animals. Severed axons of several C. elegans cell types show robust regrowth and functional recovery. We have shown that several factors, including cell type, position of axotomy and life stage, can regulate whether axons regrow after injury. Conserved signaling pathways, including cyclic AMP signaling and ephrin signaling, regulate regenerative growth of axons. We also found an unexpected role for synaptic branches in regulating axon regrowth. The tractable genetic and genomic tools available in C. elegans facilitate large scale screens for new regeneration genes. A pilot screen has uncovered several new genes that promote or repress regenerative growth. Our three specific Aims build on these preliminary results: First, we will dissect the mechanism by which the synaptic branch regulates regeneration in mechanosensory neurons. We hypothesize that the synaptic branch point contains a sorting area that regulates membrane and organelle traffic after injury. We will analyze the transport of motors and cargoes required for regrowth and will specifically test the role of the Liprin pathway in promoting regrowth. Second we will define how cAMP signaling promotes C. elegans neuronal regeneration. We will test whether cAMP or its effectors are required for regrowth. We will examine the effects of axotomy on cAMP dynamics in vivo. We will test the role of a putative cAMP-regulated transcription factor that we have found is essential for regeneration. Third, we will perform a large scale functional genomic screen to identify new genes with roles in regenerative axon growth. The mechanisms of genes with strong pro- or anti-regeneration roles will be studied in detail. Relevance: This work will yield a systematic understanding of the pathways that regulate axon regeneration after injury in a simple model system. Knowledge of the conserved mechanisms controlling axon regeneration will allow their manipulation in therapies for nervous system disease and injury.

Public Health Relevance

We will analyze the molecular and genetic mechanisms underlying nerve regeneration in the nematode C. elegans. Specifically, we will define the roles of trafficking at axonal branch points and the function of cyclic AMP signaling. We will perform a large scale screen to discover novel genes involved in regenerative growth of neurons. The results will further our understanding of nerve regrowth and will aid therapies for trauma and disease of the nervous system.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS057317-04
Application #
8204622
Study Section
Neurodifferentiation, Plasticity, and Regeneration Study Section (NDPR)
Program Officer
Owens, David F
Project Start
2009-01-15
Project End
2013-12-31
Budget Start
2012-01-01
Budget End
2012-12-31
Support Year
4
Fiscal Year
2012
Total Cost
$298,089
Indirect Cost
$105,151
Name
University of California San Diego
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
804355790
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Chuang, Marian; Goncharov, Alexandr; Wang, Shaohe et al. (2014) The microtubule minus-end-binding protein patronin/PTRN-1 is required for axon regeneration in C. elegans. Cell Rep 9:874-83
Hubert, Thomas; Wu, Zilu; Chisholm, Andrew D et al. (2014) S6 kinase inhibits intrinsic axon regeneration capacity via AMP kinase in Caenorhabditis elegans. J Neurosci 34:758-63
Kurup, Naina; Sharifnia, Panid; Jin, Yishi (2013) Spatial and temporal dynamics of neurite regrowth. Curr Opin Neurobiol 23:1011-7
Wang, Zhiping; Hou, Yanli; Guo, Xing et al. (2013) The EBAX-type Cullin-RING E3 ligase and Hsp90 guard the protein quality of the SAX-3/Robo receptor in developing neurons. Neuron 79:903-16
Chisholm, Andrew D (2013) Cytoskeletal dynamics in Caenorhabditis elegans axon regeneration. Annu Rev Cell Dev Biol 29:271-97
Chen, Lizhen; Chisholm, Andrew D (2011) Axon regeneration mechanisms: insights from C. elegans. Trends Cell Biol 21:577-84
Chen, Lizhen; Wang, Zhiping; Ghosh-Roy, Anindya et al. (2011) Axon regeneration pathways identified by systematic genetic screening in C. elegans. Neuron 71:1043-57
Gotenstein, Jennifer R; Swale, Ryann E; Fukuda, Tetsuko et al. (2010) The C. elegans peroxidasin PXN-2 is essential for embryonic morphogenesis and inhibits adult axon regeneration. Development 137:3603-13
Ghosh-Roy, Anindya; Wu, Zilu; Goncharov, Alexandr et al. (2010) Calcium and cyclic AMP promote axonal regeneration in Caenorhabditis elegans and require DLK-1 kinase. J Neurosci 30:3175-83
Ghosh-Roy, Anindya; Chisholm, Andrew D (2010) Caenorhabditis elegans: a new model organism for studies of axon regeneration. Dev Dyn 239:1460-4

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