Visualizing secretion and synaptic transmission with pH-sensitive green fluorescent proteins Journal Article
| Authors: | Miesenböck, G.; De Angelis, D. A.; Rothman, J. E. |
| Article Title: | Visualizing secretion and synaptic transmission with pH-sensitive green fluorescent proteins |
| Abstract: | In neural systems, information is often carried by ensembles of cells rather than by individual units. Optical indicators provide a powerful means to reveal such distributed activity, particularly when protein-based and encodable in DNA: 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 vesicles 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. |
| Keywords: | human cell; nonhuman; mutant protein; animals; animal tissue; cells, cultured; green fluorescent protein; fluorescent dyes; ph; hela cell; hela cells; luminescent proteins; molecular sequence data; recombinant fusion proteins; membrane protein; rat; green fluorescent proteins; rats; mutagenesis; hydrogen-ion concentration; exocytosis; indicators and reagents; hippocampus; synaptic transmission; cytoplasmic granules; synaptic vesicles; biosensor; secretory granule; humans; human; priority journal; article |
| Journal Title: | Nature |
| Volume: | 394 |
| Issue: | 6689 |
| ISSN: | 0028-0836 |
| Publisher: | Nature Publishing Group |
| Date Published: | 1998-07-09 |
| Start Page: | 192 |
| End Page: | 195 |
| Language: | English |
| DOI: | 10.1038/28190 |
| PUBMED: | 9671304 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Article -- Export Date: 12 December 2016 -- Source: Scopus |
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120Rothman -
17Miesenboeck -
10
De Angelis
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