PMID:9671304
| Citation |
Miesenböck, G, De Angelis, DA and Rothman, JE (1998) Visualizing secretion and synaptic transmission with pH-sensitive green fluorescent proteins. Nature 394:192-5 |
|---|---|
| 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. |
| Links |
PubMed Online version:10.1038/28190 |
| Keywords |
Animals; Cells, Cultured; Cytoplasmic Granules/physiology; Exocytosis; Fluorescent Dyes; Green Fluorescent Proteins; HeLa Cells; Hippocampus/cytology; Humans; Hydrogen-Ion Concentration; Indicators and Reagents; Luminescent Proteins/genetics; Molecular Sequence Data; Mutagenesis; Rats; Recombinant Fusion Proteins; Synaptic Transmission; Synaptic Vesicles/physiology |
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