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microPublication / Biology / daf-2 modulates regeneration of mechanosensory...
daf-2 modulates regeneration of mechanosensory neurons II
Zehra C. Abay1, Michelle Yu-Ying Wong1 and Brent Neumann1
1Neuroscience Program, Monash Biomedicine Discovery Institute and Department of Anatomy and Developmental Biology, Monash University, Melbourne VIC 3800, Australia
Correspondence to: Brent Neumann (brent.neumann@monash.edu)

Description

​The daf-2 gene encodes an insulin-like growth factor/IGF-1 receptor that regulates C. elegans embryonic and larval development. It has previously been shown that DAF-2 inhibits neurite regeneration of the GABAergic motor neurons and PVD sensory neurons in an age-dependent fashion (Bryne et al., 2014; Kravtsov et al., 2017). Following injury, the posterior lateral microtubule (PLM) neurons are capable of regenerating through axonal fusion, a highly efficient regrowth mechanism in which separated fragments fuse back together (Ghosh-Roy et al., 2010; Neumann et al., 2011; Neumann et al. 2015; Abay et al., 2017). We previously established that a critical event for axonal fusion to occur is the exposure of injury-induced phosphatidylserine (PS) ‘save-me’ signals (Neumann et al., 2015). The level of PS exposed increases with advancing age (Abay et al., 2017). To determine if daf-2 is involved in this age-dependent modulation of PS exposure, we visualised and quantified the level of PS exposed after PLM axotomy using a secreted, tagged version of Annexin V (Neumann et al., 2011; Mapes et al., 2012; Neumann et al. 2015). Mutation of daf-2 had no effect on PS exposure 1 h post-axotomy, with no significant differences observed on either the distal or proximal axon segments (Table 1).​

Table 1. Quantification of the relative level of PS exposed 1 h post-axotomy.

Genotype PS exposed on distal axon
(relative to pre-axotomy)
n PS exposed on proximal axon
(relative to pre-axotomy)
n
wild-type 1.53 ± 0.105 28 1.44 ± 0.0855 28
daf-2(e1370) 1.51 ± 0.167 26 1.57 ± 0.166 26

Reagents

One-day-old adult hermaphrodites were used for all experiments, and were grown under standard conditions at 20°C. The BXN301 [daf-2(e1370); smIs95(Phsp-16.2::sAnxV::mRFP); zdIs5(Pmec-4::GFP)] strain was used along with the CU4204 [smIs95(Phsp-16.2::sAnxV::mRFP); zdIs5(Pmec-4::GFP)] control strain. The daf-2(e1370) allele has been considered temperature sensitive for the dauer phenotype, but not for the long-lived phenotype. At 20°C, daf-2(e1370) animals display a greater than 2-fold increase in lifespan compared to the wild-type (Kenyon et al., 1993). Laser axotomy, microscopy and quantification of data was performed as previously described (Abay et al 2017).

Acknowledgments

We thank Ding Xue for sharing strains.

References

Abay, Z.C., M.Y. Wong, J.S. Teoh, T. Vijayaraghavan, M.A. Hilliard, and B. Neumann, Phosphatidylserine save-me signals drive functional recovery of severed axons in Caenorhabditis elegans. Proc Natl Acad Sci U S A, 2017. 114(47): p. E10196-E10205.
PubMed
Byrne, A.B., T. Walradt, K.E. Gardner, A. Hubbert, V. Reinke, and M. Hammarlund, Insulin/IGF1 signaling inhibits age-dependent axon regeneration. Neuron, 2014. 81(3): p. 561-73.
PubMed
Ghosh-Roy, A., Z. Wu, A. Goncharov, Y. Jin, and A.D. Chisholm, Calcium and cyclic AMP promote axonal regeneration in Caenorhabditis elegans and require DLK-1 kinase. J Neurosci, 2010. 30(9): p. 3175-83.
PubMed
Kenyon, C., J. Chang, E. Gensch, A. Rudner, and R. Tabtiang, A C. elegans mutant that lives twice as long as wild type. Nature, 1993. 366(6454): p. 461-4.
PubMed
Kravtsov, V., M. Oren-Suissa, and B. Podbilewicz, The fusogen AFF-1 can rejuvenate the regenerative potential of adult dendritic trees by self-fusion. Development, 2017. 144(13): p. 2364-2374.
PubMed
Mapes, J., Y.Z. Chen, A. Kim, S. Mitani, B.H. Kang, and D. Xue, CED-1, CED-7, and TTR-52 regulate surface phosphatidylserine expression on apoptotic and phagocytic cells. Curr Biol, 2012. 22(14): p. 1267-75.
PubMed
Neumann, B., K.C. Nguyen, D.H. Hall, A. Ben-Yakar, and M.A. Hilliard, Axonal regeneration proceeds through specific axonal fusion in transected C. elegans neurons. Dev Dyn, 2011. 240(6): p. 1365-72.
PubMed
Neumann, B., S. Coakley, R. Giordano-Santini, C. Linton, E.S. Lee, A. Nakagawa, D. Xue, and M.A. Hilliard, EFF-1-mediated regenerative axonal fusion requires components of the apoptotic pathway. Nature, 2015. 517(7533): p. 219-22.
PubMed

Funding

This work was supported by National Health and Medical Research Council (NHMRC) Project Grant 1101974.

Reviewed By

Rachid El Bejjani

History

Received: November 10, 2017
Accepted: November 26, 2017
Published: December 1, 2017

Copyright

© 2017 by the authors. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Citation

Abay, ZC; Wong, MYY; Neumann, B (2017). daf-2 modulates regeneration of mechanosensory neurons II. microPublication Biology. 10.17912/W2SM1T.
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