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Visualizing secretion and synaptic transmission with pH-sensitive green fluorescent proteins

Author

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  • Gero Miesenböck

    (Cellular Biochemistry and Biophysics Program, Memorial Sloan-Kettering Cancer Center)

  • Dino A. De Angelis

    (Cellular Biochemistry and Biophysics Program, Memorial Sloan-Kettering Cancer Center)

  • James E. Rothman

    (Cellular Biochemistry and Biophysics Program, Memorial Sloan-Kettering Cancer Center)

Abstract

In neural systems, information is often carried by ensembles of cells rather than by individual units. Optical indicators1 provide a powerful means to reveal such distributed activity, particularly when protein-based and encodable in DNA2,3,4: encodable probes can be introduced into cells, tissues, or transgenic organisms by genetic manipulation, selectively expressed in anatomically or functionally defined groups of cells, and, ideally, recorded in situ, without a requirement for exogenous cofactors. Here we describe sensors for secretion and neurotransmission that fulfil these criteria. We have developed pH-sensitive mutants of green fluorescent protein (‘pHluorins’) by structure-directed combinatorial mutagenesis, with the aim of exploiting the acidic pH inside secretory vesicles5,6 to monitor vesicle exocytosis and recycling. When linked to a vesicle membrane protein, pHluorins were sorted to secretory and synaptic vesicles and reported transmission at individual synaptic boutons, as well as secretion and fusion pore ‘flicker’ of single secretory granules.

Suggested Citation

  • Gero Miesenböck & Dino A. De Angelis & James E. Rothman, 1998. "Visualizing secretion and synaptic transmission with pH-sensitive green fluorescent proteins," Nature, Nature, vol. 394(6689), pages 192-195, July.
    • Handle: RePEc:nat:nature:v:394:y:1998:i:6689:d:10.1038_28190
    DOI: 10.1038/28190
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    Cited by:

    1. Marley D Kass & Andrew H Moberly & John P McGann, 2013. "Spatiotemporal Alterations in Primary Odorant Representations in Olfactory Marker Protein Knockout Mice," PLOS ONE, Public Library of Science, vol. 8(4), pages 1-10, April.
    2. Albert Pineda Rodó & Libuše Váchová & Zdena Palková, 2012. "In Vivo Determination of Organellar pH Using a Universal Wavelength-Based Confocal Microscopy Approach," PLOS ONE, Public Library of Science, vol. 7(3), pages 1-12, March.
    3. Jung-Hwan Choi & Lauren Bayer Horowitz & Niels Ringstad, 2021. "Opponent vesicular transporters regulate the strength of glutamatergic neurotransmission in a C. elegans sensory circuit," Nature Communications, Nature, vol. 12(1), pages 1-15, December.
    4. Mable Lam & Koji Takeo & Rafael G. Almeida & Madeline H. Cooper & Kathryn Wu & Manasi Iyer & Husniye Kantarci & J. Bradley Zuchero, 2022. "CNS myelination requires VAMP2/3-mediated membrane expansion in oligodendrocytes," Nature Communications, Nature, vol. 13(1), pages 1-21, December.
    5. Joshua J. Rennick & Cameron J. Nowell & Colin W. Pouton & Angus P. R. Johnston, 2022. "Resolving subcellular pH with a quantitative fluorescent lifetime biosensor," Nature Communications, Nature, vol. 13(1), pages 1-13, December.
    6. Shanley F. Longfield & Mahdie Mollazade & Tristan P. Wallis & Rachel S. Gormal & Merja Joensuu & Jesse R. Wark & Ashley J. Waardenberg & Christopher Small & Mark E. Graham & Frédéric A. Meunier & Ramó, 2023. "Tau forms synaptic nano-biomolecular condensates controlling the dynamic clustering of recycling synaptic vesicles," Nature Communications, Nature, vol. 14(1), pages 1-20, December.
    7. Christian Hoffmann & Jakob Rentsch & Taka A. Tsunoyama & Akshita Chhabra & Gerard Aguilar Perez & Rajdeep Chowdhury & Franziska Trnka & Aleksandr A. Korobeinikov & Ali H. Shaib & Marcelo Ganzella & Gr, 2023. "Synapsin condensation controls synaptic vesicle sequestering and dynamics," Nature Communications, Nature, vol. 14(1), pages 1-13, December.

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