The invention relates to methods and compositions which utilize the
emission of light to monitor changes in microenvironments involving
cells. The invention is especially useful for monitoring exocytotic
activity such as detecting quantal release of synaptic vesicles. Fusion
proteins of Cypridina luciferase and synaptotagmin-I or
VAMP/synaptobrevin-2 were targeted to synaptic vesicles and, upon
exocytosis, formed light-emitting complexes with luciferin present in the
extracellular medium. Photon emissions in the presence of a depolarizing
stimulus can be observed with these systems. pH-sensitive mutants of
green fluorescent protein are also provided, which are useful for
visualizing exocytosis and for imaging and measuring the pH of
intracellular compartments.