Skip to main content
PMC home page
Biophysical Journal logoLink to Biophysical Journal
. 2000 Nov;79(5):2345–2352. doi: 10.1016/S0006-3495(00)76480-0

GFP is a selective non-linear optical sensor of electrophysiological processes in Caenorhabditis elegans.

A Khatchatouriants 1, A Lewis 1, Z Rothman 1, L Loew 1, M Treinin 1
PMCID: PMC1301122  PMID: 11053114

Abstract

Electrophysiology of the nematode Caenorhabditis elegans has the potential to bridge the wealth of information on the molecular biology and anatomy of this organism with the responses of selected cells and cellular neural networks associated with a behavioral response. In this paper we report that the nonlinear optical phenomenon of second harmonic generation (SHG) can be detected using green fluorescent protein (GFP) chimeras expressed in selected cells of living animals. Alterations in the SHG signal as a result of receptor ligand interactions and mechanical stimulation of the mechanosensory cells indicate that this signal is very sensitive to membrane potential. The results suggest that this approach to membrane potential measurements in C. elegans and in other biological systems could effectively couple data on selective locations within specific cells with functional responses that are associated with behavioral and sensory processes.

Full Text

The Full Text of this article is available as a PDF (783.7 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ben-Oren I., Peleg G., Lewis A., Minke B., Loew L. Infrared nonlinear optical measurements of membrane potential in photoreceptor cells. Biophys J. 1996 Sep;71(3):1616–1620. doi: 10.1016/S0006-3495(96)79365-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bouevitch O., Lewis A., Pinevsky I., Wuskell J. P., Loew L. M. Probing membrane potential with nonlinear optics. Biophys J. 1993 Aug;65(2):672–679. doi: 10.1016/S0006-3495(93)81126-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brenner S. The genetics of Caenorhabditis elegans. Genetics. 1974 May;77(1):71–94. doi: 10.1093/genetics/77.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chalfie M., Tu Y., Euskirchen G., Ward W. W., Prasher D. C. Green fluorescent protein as a marker for gene expression. Science. 1994 Feb 11;263(5148):802–805. doi: 10.1126/science.8303295. [DOI] [PubMed] [Google Scholar]
  5. Goodman M. B., Hall D. H., Avery L., Lockery S. R. Active currents regulate sensitivity and dynamic range in C. elegans neurons. Neuron. 1998 Apr;20(4):763–772. doi: 10.1016/s0896-6273(00)81014-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Heim R., Cubitt A. B., Tsien R. Y. Improved green fluorescence. Nature. 1995 Feb 23;373(6516):663–664. doi: 10.1038/373663b0. [DOI] [PubMed] [Google Scholar]
  7. Huang M., Gu G., Ferguson E. L., Chalfie M. A stomatin-like protein necessary for mechanosensation in C. elegans. Nature. 1995 Nov 16;378(6554):292–295. doi: 10.1038/378292a0. [DOI] [PubMed] [Google Scholar]
  8. Miyawaki A., Llopis J., Heim R., McCaffery J. M., Adams J. A., Ikura M., Tsien R. Y. Fluorescent indicators for Ca2+ based on green fluorescent proteins and calmodulin. Nature. 1997 Aug 28;388(6645):882–887. doi: 10.1038/42264. [DOI] [PubMed] [Google Scholar]
  9. Ormö M., Cubitt A. B., Kallio K., Gross L. A., Tsien R. Y., Remington S. J. Crystal structure of the Aequorea victoria green fluorescent protein. Science. 1996 Sep 6;273(5280):1392–1395. doi: 10.1126/science.273.5280.1392. [DOI] [PubMed] [Google Scholar]
  10. Peleg G., Lewis A., Linial M., Loew L. M. Nonlinear optical measurement of membrane potential around single molecules at selected cellular sites. Proc Natl Acad Sci U S A. 1999 Jun 8;96(12):6700–6704. doi: 10.1073/pnas.96.12.6700. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Treinin M., Chalfie M. A mutated acetylcholine receptor subunit causes neuronal degeneration in C. elegans. Neuron. 1995 Apr;14(4):871–877. doi: 10.1016/0896-6273(95)90231-7. [DOI] [PubMed] [Google Scholar]
  12. Treinin M., Gillo B., Liebman L., Chalfie M. Two functionally dependent acetylcholine subunits are encoded in a single Caenorhabditis elegans operon. Proc Natl Acad Sci U S A. 1998 Dec 22;95(26):15492–15495. doi: 10.1073/pnas.95.26.15492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Tsien R. Y. The green fluorescent protein. Annu Rev Biochem. 1998;67:509–544. doi: 10.1146/annurev.biochem.67.1.509. [DOI] [PubMed] [Google Scholar]
  14. Yang F., Moss L. G., Phillips G. N., Jr The molecular structure of green fluorescent protein. Nat Biotechnol. 1996 Oct;14(10):1246–1251. doi: 10.1038/nbt1096-1246. [DOI] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES