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

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.

[1]  Shreedhar Gadge,et al.  THE MOLECULAR STRUCTURE OF GREEN FLUORESCENT PROTEIN , 2022 .

[2]  L M Loew,et al.  Probing membrane potential with nonlinear optics. , 1993, Biophysical journal.

[3]  Roger Y. Tsien,et al.  Improved green fluorescence , 1995, Nature.

[4]  M. W. Hardisty.  The nervous system , 1979 .

[5]  J. Berg.  Genome sequence of the nematode C. elegans: a platform for investigating biology. , 1998, Science.

[6]  M Linial,et al.  Nonlinear optical measurement of membrane potential around single molecules at selected cellular sites. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[7]  S. Lowen.  The Biophysical Journal , 1960, Nature.

[8]  L. Loew,et al.  Infrared nonlinear optical measurements of membrane potential in photoreceptor cells. , 1996, Biophysical journal.

[9]  S. Brenner,et al.  The structure of the nervous system of the nematode Caenorhabditis elegans. , 1986, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[10]  M. Chalfie,et al.  A stomatin-like protein necessary for mechanosensation in C. elegans , 1995, Nature.

[11]  W. Wood.  The Nematode Caenorhabditis elegans , 1988 .

[12]  Leslie M. Loew,et al.  Second-harmonic generation of biological interfaces: probing the membrane protein bacteriorhodopsin and imaging membrane potential around GFP molecules at specific sites in neuronal cells of C. elegans , 1999 .

[13]  M. Chalfie,et al.  Green fluorescent protein as a marker for gene expression. , 1994, Science.

[14]  M. Chalfie,et al.  A mutated acetylcholine receptor subunit causes neuronal degeneration in C. elegans , 1995, Neuron.

[15]  Roger Y. Tsien,et al.  Crystal Structure of the Aequorea victoria Green Fluorescent Protein , 1996, Science.

[16]  Klony Lieberman,et al.  A micropipette force probe suitable for near‐field scanning optical microscopy , 1992 .

[17]  Klony Lieberman,et al.  A fully integrated near‐field optical, far‐field optical, and normal‐force scanned probe microscope , 1996 .

[18]  M. Chalfie,et al.  Two functionally dependent acetylcholine subunits are encoded in a single Caenorhabditis elegans operon. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[19]  S. Lockery,et al.  Active Currents Regulate Sensitivity and Dynamic Range in C. elegans Neurons , 1998, Neuron.

[20]  Jung Y. Huang,et al.  Second-harmonic generation in purple membrane−poly(vinyl alcohol) films: probing the dipolar characteristics of the bacteriorhodopsin chromophore in bR570 and M412 , 1989 .

[21]  N. Munakata.  [Genetics of Caenorhabditis elegans]. , 1989, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme.

[22]  R. Tsien,et al.  green fluorescent protein , 2020, Catalysis from A to Z.

[23]  R. Tsien,et al.  Fluorescent indicators for Ca2+based on green fluorescent proteins and calmodulin , 1997, Nature.

[24]  M. Taussig.  The Nervous System , 1991 .