Cookies on this website

We use cookies to ensure that we give you the best experience on our website. If you click 'Accept all cookies' we'll assume that you are happy to receive all cookies and you won't see this message again. If you click 'Reject all non-essential cookies' only necessary cookies providing core functionality such as security, network management, and accessibility will be enabled. Click 'Find out more' for information on how to change your cookie settings.

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.

Original publication

DOI

10.1038/28190

Type

Journal article

Journal

Nature

Publication Date

09/07/1998

Volume

394

Pages

192 - 195

Keywords

Animals, Cells, Cultured, Cytoplasmic Granules, Exocytosis, Fluorescent Dyes, Green Fluorescent Proteins, HeLa Cells, Hippocampus, Humans, Hydrogen-Ion Concentration, Indicators and Reagents, Luminescent Proteins, Molecular Sequence Data, Mutagenesis, Rats, Recombinant Fusion Proteins, Synaptic Transmission, Synaptic Vesicles