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Epidermal growth factor signaling induces behavioral quiescence in Caenorhabditis elegans

Nature Neuroscience volume 10, pages 1300–1307 (2007)Cite this article

Abstract

The epidermal growth factor receptor (EGFR)/ErbB receptor tyrosine kinases regulate several aspects of development, including the development of the mammalian nervous system. ErbB signaling also has physiological effects on neuronal function, with influences on synaptic plasticity and daily cycles of activity. However, little is known about the effectors of EGFR activation in neurons. Here we show that EGF signaling has a nondevelopmental effect on behavior in Caenorhabditis elegans. Ectopic expression of the EGF-like ligand LIN-3 at any stage induces a reversible cessation of feeding and locomotion. These effects are mediated by neuronal EGFR (also called LET-23) and phospholipase C–γ (PLC-γ), diacylglycerol-binding proteins, and regulators of synaptic vesicle release. Activation of EGFR within a single neuron, ALA, is sufficient to induce a quiescent state. This pathway modulates the cessation of pharyngeal pumping and locomotion that normally occurs during the lethargus period that precedes larval molting. Our results reveal an evolutionarily conserved role for EGF signaling in the regulation of behavioral quiescence.

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Figure 1: The hs:LIN-3 movement defect is mediated by LET-23, PLC-γ and EGL-4.
Figure 2: LET-23 and PLC-γ act in neurons to mediate the hs:LIN-3 movement defect.
Figure 3: LET-23 activity in the ALA neuron mediates the behavioral effects of LIN-3.
Figure 4: let-23 and plc-3 mutants show increased activity during lethargus.

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References

  1. Buonanno, A. & Fischbach, G.D. Neuregulin and ErbB receptor signaling pathways in the nervous system. Curr. Opin. Neurobiol. 11, 287–296 (2001).

    Article  CAS  Google Scholar 

  2. Huang, Y.Z. et al. Regulation of neuregulin signaling by PSD-95 interacting with ErbB4 at CNS synapses. Neuron 26, 443–455 (2000).

    Article  CAS  Google Scholar 

  3. Kwon, O-B., Longart, M., Vullhorst, D., Hoffman, D.A. & Buonanno, A. Neuregulin-1 reverses long-term potentiation at CA1 hippocampal synapses. J. Neurosci. 25, 9378–9383 (2005).

    Article  CAS  Google Scholar 

  4. Kramer, A. et al. Regulation of daily locomotor activity and sleep by hypothalamic EGF receptor signaling. Science 294, 2511–2515 (2001).

    Article  CAS  Google Scholar 

  5. Snodgrass-Belt, P., Gilbert, J.L. & Davis, F.C. Central administration of transforming growth factor–alpha and neuregulin-1 suppress active behaviors and cause weight loss in hamsters. Brain Res. 1038, 171–182 (2005).

    Article  CAS  Google Scholar 

  6. Jorissen, R.N. et al. Epidermal growth factor receptor: mechanisms of activation and signalling. Exp. Cell Res. 284, 31–53 (2003).

    Article  CAS  Google Scholar 

  7. Hill, R.J. & Sternberg, P.W. The gene lin-3 encodes an inductive signal for vulval development in C. elegans. Nature 358, 470–476 (1992).

    Article  CAS  Google Scholar 

  8. Aroian, R.V., Koga, M., Mendel, J.E., Ohshima, Y. & Sternberg, P.W. The let-23 gene necessary for Caenorhabditis elegans vulval induction encodes a tyrosine kinase of the EGF receptor subfamily. Nature 348, 693–699 (1990).

    Article  CAS  Google Scholar 

  9. Moghal, N. & Sternberg, P.W. The epidermal growth factor system in Caenorhabditis elegans. Exp. Cell Res. 284, 150–159 (2003).

    Article  CAS  Google Scholar 

  10. Clandinin, T.R., DeModena, J.A. & Sternberg, P.W. Inositol trisphosphate mediates a RAS-independent response to LET-23 receptor tyrosine kinase activation in C. elegans. Cell 92, 523–533 (1998).

    Article  CAS  Google Scholar 

  11. Yin, X., Gower, N.J.D., Baylis, H.A. & Strange, K. Inositol 1,4,5-trisphosphate signaling regulates rhythmic contractile activity of myoepithelial sheath cells in Caenorhabditis elegans. Mol. Biol. Cell 15, 3938–3949 (2004).

    Article  CAS  Google Scholar 

  12. Katz, W.S., Hill, R.J., Clandinin, T.R. & Sternberg, P.W. Different levels of the C. elegans growth factor LIN-3 promote distinct vulval precursor fates. Cell 82, 297–307 (1995).

    Article  CAS  Google Scholar 

  13. Johnson, T.E., Mitchell, D.H., Kline, S., Kemal, R. & Foy, J. Arresting development arrests aging in the nematode Caenorhabditis elegans. Mech. Ageing Dev. 28, 23–40 (1984).

    Article  CAS  Google Scholar 

  14. Timmons, L., Court, D.L. & Fire, A. Ingestion of bacterially expressed dsRNAs can produce specific and potent genetic interference in Caenorhabditis elegans. Gene 263, 103–112 (2001).

    Article  CAS  Google Scholar 

  15. Yang, C. & Kazanietz, M.G. Divergence and complexities in DAG signaling: looking beyond PKC. Trends Pharmacol. Sci. 24, 602–608 (2003).

    Article  CAS  Google Scholar 

  16. Tabuse, Y. Protein kinase C isotypes in C. elegans. J. Biochem. 132, 519–522 (2002).

    Article  CAS  Google Scholar 

  17. van der Linden, A.M., Moorman, C., Cuppen, E., Korswagen, H.C. & Plasterk, R. Hyperactivation of the G12-mediated signaling pathway in Caenorhabditis elegans induces a developmental growth arrest via protein kinase C. Curr. Biol. 13, 516–521 (2003).

    Article  CAS  Google Scholar 

  18. Richmond, J.E., Weimer, R.M. & Jorgensen, E.M. An open form of syntaxin bypasses the requirement for UNC-13 in vesicle priming. Nature 412, 338–341 (2001).

    Article  CAS  Google Scholar 

  19. Richmond, J.E., Davis, W.S. & Jorgensen, E.M. UNC-13 is required for synaptic vesicle fusion in C. elegans. Nat. Neurosci. 2, 959–964 (1999).

    Article  CAS  Google Scholar 

  20. Avery, L. & Horvitz, H.R. Pharyngeal pumping continues after laser killing of the pharyngeal nervous system of C. elegans. Neuron 3, 473–485 (1989).

    Article  CAS  Google Scholar 

  21. Saifee, O., Wei, L.P. & Nonet, M.L. The Caenorhabditis elegans unc-64 locus encodes a syntaxin that interacts genetically with synaptobrevin. Mol. Biol. Cell 9, 1235–1252 (1998).

    Article  CAS  Google Scholar 

  22. Weimer, R.M. et al. Defects in synaptic vesicle docking in unc-18 mutants. Nat. Neurosci. 6, 1023–1030 (2003).

    Article  CAS  Google Scholar 

  23. Avery, L., Bargmann, C.I. & Horvitz, H.R. The Caenorhabditis elegans unc-31 gene affects multiple nervous system-controlled functions. Genetics 134, 455–464 (1993).

    CAS  PubMed  PubMed Central  Google Scholar 

  24. Ann, K., Kowalchyk, J.A., Loyet, K.M. & Martin, T.F.J. Novel Ca2+-binding protein (CAPS) related to UNC-31 required for Ca2+-activated exocytosis. J. Biol. Chem. 272, 19637–19640 (1997).

    Article  CAS  Google Scholar 

  25. Treinin, M. & Chalfie, M. A mutated acetylcholine receptor subunit causes neuronal degeneration in C. elegans. Neuron 14, 871–877 (1995).

    Article  CAS  Google Scholar 

  26. Albertson, D.G. & Thomson, J.N. The pharynx of Caenorhabditis elegans. Phil. Trans. R. Soc. Lond. B 275, 299–325 (1976).

    Article  CAS  Google Scholar 

  27. Keane, J. & Avery, L. Mechanosensory inputs influence Caenorhabditis elegans pharyngeal activity via ivermectin sensitivity genes. Genetics 164, 153–162 (2003).

    CAS  PubMed  PubMed Central  Google Scholar 

  28. Avery, L. The genetics of feeding in Caenorhabditis elegans. Genetics 133, 897–917 (1993).

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Raizen, D.M., Cullison, K.M., Pack, A.I. & Sundaram, M. A novel gain-of-function mutant of the cGMP-dependent protein kinase egl-4 affects multiple physiological processes in C. elegans. Genetics 173, 177–187 (2006).

    Article  CAS  Google Scholar 

  30. Nonet, M.L., Saifee, O., Zhao, H., Rand, J.B. & Wei, L. Synaptic transmission deficits in C. elegans synaptobrevin mutants. J. Neurosci. 18, 70–80 (1998).

    Article  CAS  Google Scholar 

  31. Okkema, P.G., Harrison, S.W., Plunger, V., Aryana, A. & Fire, A. Sequence requirements for myosin gene expression and regulation in Caenorhabditis elegans. Genetics 135, 385–404 (1993).

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Garcia, R.A., Vasudevan, K. & Buonanno, A. The neuregulin receptor ErbB-4 interacts with PDZ-containing proteins at neuronal synapses. Proc. Natl. Acad. Sci. USA 97, 3596–3601 (2000).

    Article  CAS  Google Scholar 

  33. Aroian, R.V., Lesa, G.M. & Sternberg, P.W. Mutations in the Caenorhabditis elegans let-23 EGFR-like gene define elements important for cell-type specificity and function. EMBO J. 13, 360–366 (1994).

    Article  CAS  Google Scholar 

  34. Kaech, S.M., Whitfield, C.W. & Kim, S.K. The LIN-2/LIN-7/LIN-10 complex mediates basolateral membrane localization of the C. elegans EGF receptor LET-23 in vulval epithelial cells. Cell 94, 761–771 (1998).

    Article  CAS  Google Scholar 

  35. Simske, J.S., Kaech, S.M., Harp, S.A. & Kim, S.K. LET-23 receptor localization by the cell junction protein LIN-7 during C. elegans vulval induction. Cell 85, 195–204 (1996).

    Article  CAS  Google Scholar 

  36. Chang, C., Newman, A.P. & Sternberg, P.W. Reciprocal EGF signaling back to the uterus from the induced C. elegans vulva coordinates morphogenesis of epithelia. Curr. Biol. 9, 237–246 (1999).

    Article  CAS  Google Scholar 

  37. Pujol, N., Torregrossa, P., Ewbank, J.J. & Brunet, J.-F. The homeodomain protein CePHOX2/CEH-17 controls antero-posterior axonal growth in C. elegans. Development 127, 3361–3371 (2000).

    CAS  PubMed  Google Scholar 

  38. Popovici, C., Isnardon, D., Birnbaum, D. & Roubin, R. Caenorhabditis elegans receptors related to mammalian vascular endothelial growth factor receptors are expressed in neural cells. Neurosci. Lett. 329, 116–120 (2002).

    Article  CAS  Google Scholar 

  39. Hwang, B.J. & Sternberg, P.W. A cell-specific enhancer that specifies lin-3 expression in the C. elegans anchor cell for vulval development. Development 131, 143–151 (2004).

    Article  CAS  Google Scholar 

  40. Cassada, R.C. & Russell, R.L. The dauerlarva, a post-embryonic developmental variant of the nematode C. elegans. Dev. Biol. 46, 326–342 (1975).

    Article  CAS  Google Scholar 

  41. Lesa, G.M. & Sternberg, P.W. Positive and negative tissue-specific signaling by a nematode epidermal growth factor receptor. Mol. Biol. Cell 8, 779–793 (1997).

    Article  CAS  Google Scholar 

  42. Cronin, C.J. et al. An automated system for measuring parameters of nematode sinusoidal movement. BMC Genet. 6, 5 (2005).

    Article  Google Scholar 

  43. Majewski, H. & Iannazzo, L. Protein kinase C: A physiological mediator of enhanced transmitter output. Prog. Neurobiol. 55, 463–475 (1998).

    Article  CAS  Google Scholar 

  44. Rhee, J.-S. β-phorbol ester and diacylglycerol-induced augmentation of transmitter release is mediated by Munc13s and not by PKCs. Cell 108, 121–133 (2002).

    Article  CAS  Google Scholar 

  45. Kim, K. & Li, C. Expression and regulation of an FMRFamide-related neuropeptide gene family in Caenorhabditis elegans. J. Comp. Neurol. 475, 540–550 (2004).

    Article  CAS  Google Scholar 

  46. Dutt, A., Canevascini, S., Froehli-Hoier, E. & Hajnal, A. EGF signal propagation during C. elegans vulval development mediated by ROM-1 Rhomboid. PLoS Biol. 2, e334 (2004).

    Article  Google Scholar 

  47. Wong, R.W.C. & Guillaud, L. The role of epidermal growth factor and its receptors in mammalian CNS. Cytokine Growth Factor Rev. 15, 147–156 (2004).

    Article  CAS  Google Scholar 

  48. Brenner, S. The genetics of Caenorhabditis elegans. Genetics 77, 71–94 (1974).

    CAS  PubMed  PubMed Central  Google Scholar 

  49. Stringham, E.G., Dixon, D.K., Jones, D. & Candido, E.P. Temporal and spatial expression patterns of the small heat shock (hsp16) genes in transgenic Caenorhabditis elegans. Mol. Biol. Cell 3, 221–233 (1992).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank the Caenorhabditis Genetics Center for providing strains, S. Kim (Stanford University School of Medicine) for anti-LET-23 antibody, J. DeModena for antibody staining, S. Xu (Univ. Michigan) for the plc-3(sy698) allele, S. Mitani (Tokyo Women's Medical University School of Medicine) for the plc-3(tm1340) allele, N. Moghal, M. Kato, and S. Xu for critical reading of the manuscript, and C.J. Cronin for help with the automated tracking system and locomotion data analysis. This work was supported by a California Breast Cancer Research Program fellowship to C.V.B., by a US National Institute on Drug Abuse grant (DA018341) to P.W.S., and by the Howard Hughes Medical Institute, with which C.V.B. is an Associate and P.W.S. is an Investigator.

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Authors and Affiliations

  1. Division of Biology 156-29, Howard Hughes Medical Institute, California Institute of Technology, 1200 E. California Blvd., Pasadena, 91125, California, USA

    Cheryl Van Buskirk & Paul W Sternberg

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  1. Cheryl Van Buskirk
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Contributions

C.V.B. designed and conducted the experiments and wrote the manuscript; P.W.S. supervised the project.

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Correspondence to Paul W Sternberg.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–4, Tables 1–4, and Methods (PDF 2524 kb)

Supplementary Video 1

Wild-type pharyngeal pumping (MOV 651 kb)

Supplementary Video 2

hs:LIN-3 inhibits pharyngeal pumping (MOV 823 kb)

Supplementary Video 3

Wild-type locomotion (MOV 2327 kb)

Supplementary Video 4

hs:LIN-3 severely impairs locomotion (MOV 2357 kb)

Supplementary Video 5

LIN-3 animals move normally when prodded (MOV 2135 kb)

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Van Buskirk, C., Sternberg, P. Epidermal growth factor signaling induces behavioral quiescence in Caenorhabditis elegans. Nat Neurosci 10, 1300–1307 (2007). https://doi.org/10.1038/nn1981

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