Skip to main content
PMC home page
Biophysical Journal logoLink to Biophysical Journal
. 2000 Oct;79(4):2199–2208. doi: 10.1016/S0006-3495(00)76468-X

The use of pHluorins for optical measurements of presynaptic activity.

S Sankaranarayanan 1, D De Angelis 1, J E Rothman 1, T A Ryan 1
PMCID: PMC1301110  PMID: 11023924

Abstract

Genetically encoded reporters for optical measurements of presynaptic activity hold significant promise for measurements of neurotransmission within intact or semi-intact neuronal networks. We have characterized pH-sensitive green fluorescent protein-based sensors (pHluorins) of synaptic vesicle cycling at nerve terminals. pHluorins have a pK approximately 7.1, which make them ideal for tracking synaptic vesicle lumen pH upon cycling through the plasma membrane during action potentials. A theoretical analysis of the expected signals using this approach and guidelines for future reporter development are provided.

Full Text

The Full Text of this article is available as a PDF (468.6 KB).

Selected References

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

  1. Denk W., Svoboda K. Photon upmanship: why multiphoton imaging is more than a gimmick. Neuron. 1997 Mar;18(3):351–357. doi: 10.1016/s0896-6273(00)81237-4. [DOI] [PubMed] [Google Scholar]
  2. Haupts U., Maiti S., Schwille P., Webb W. W. Dynamics of fluorescence fluctuations in green fluorescent protein observed by fluorescence correlation spectroscopy. Proc Natl Acad Sci U S A. 1998 Nov 10;95(23):13573–13578. doi: 10.1073/pnas.95.23.13573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Helmchen F., Svoboda K., Denk W., Tank D. W. In vivo dendritic calcium dynamics in deep-layer cortical pyramidal neurons. Nat Neurosci. 1999 Nov;2(11):989–996. doi: 10.1038/14788. [DOI] [PubMed] [Google Scholar]
  4. Kneen M., Farinas J., Li Y., Verkman A. S. Green fluorescent protein as a noninvasive intracellular pH indicator. Biophys J. 1998 Mar;74(3):1591–1599. doi: 10.1016/S0006-3495(98)77870-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Liu Y., Krantz D. E., Waites C., Edwards R. H. Membrane trafficking of neurotransmitter transporters in the regulation of synaptic transmission. Trends Cell Biol. 1999 Sep;9(9):356–363. doi: 10.1016/s0962-8924(99)01605-0. [DOI] [PubMed] [Google Scholar]
  6. Matz M. V., Fradkov A. F., Labas Y. A., Savitsky A. P., Zaraisky A. G., Markelov M. L., Lukyanov S. A. Fluorescent proteins from nonbioluminescent Anthozoa species. Nat Biotechnol. 1999 Oct;17(10):969–973. doi: 10.1038/13657. [DOI] [PubMed] [Google Scholar]
  7. Miesenböck G., De Angelis D. A., Rothman J. E. Visualizing secretion and synaptic transmission with pH-sensitive green fluorescent proteins. Nature. 1998 Jul 9;394(6689):192–195. doi: 10.1038/28190. [DOI] [PubMed] [Google Scholar]
  8. Nelson N. Structure and function of V-ATPases in endocytic and secretory organelles. J Exp Biol. 1992 Nov;172:149–153. doi: 10.1242/jeb.172.1.149. [DOI] [PubMed] [Google Scholar]
  9. Roos A., Boron W. F. Intracellular pH. Physiol Rev. 1981 Apr;61(2):296–434. doi: 10.1152/physrev.1981.61.2.296. [DOI] [PubMed] [Google Scholar]
  10. Ryan T. A. Inhibitors of myosin light chain kinase block synaptic vesicle pool mobilization during action potential firing. J Neurosci. 1999 Feb 15;19(4):1317–1323. doi: 10.1523/JNEUROSCI.19-04-01317.1999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Sankaranarayanan S., Ryan T. A. Real-time measurements of vesicle-SNARE recycling in synapses of the central nervous system. Nat Cell Biol. 2000 Apr;2(4):197–204. doi: 10.1038/35008615. [DOI] [PubMed] [Google Scholar]
  12. Schikorski T., Stevens C. F. Quantitative ultrastructural analysis of hippocampal excitatory synapses. J Neurosci. 1997 Aug 1;17(15):5858–5867. doi: 10.1523/JNEUROSCI.17-15-05858.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Threadgill R., Bobb K., Ghosh A. Regulation of dendritic growth and remodeling by Rho, Rac, and Cdc42. Neuron. 1997 Sep;19(3):625–634. doi: 10.1016/s0896-6273(00)80376-1. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Walch-Solimena C., Blasi J., Edelmann L., Chapman E. R., von Mollard G. F., Jahn R. The t-SNAREs syntaxin 1 and SNAP-25 are present on organelles that participate in synaptic vesicle recycling. J Cell Biol. 1995 Feb;128(4):637–645. doi: 10.1083/jcb.128.4.637. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES