ULTRASTRUCTURAL CORRELATES OF FUNCTIONAL PROPERTIES OF INDIVIDUAL CNS SYNAPSES

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Using confocal and multiphoton microscopy together with fluorescent Ca2+ indicators, we are able to visualise activation of individual synaptic boutons and dendritic spines in brain slice preparations (Emptage et al. 1999, 2001). In this way, at favourable excitatory synapses, we can assess quantal parameters of synaptic function, such as transmitter release probability and quantal amplitude. By injecting fixable markers via the intracellular recording microelectrode, and fixing the preparation at the end of the physiological imaging experiment, we are able to identify the imaged synapse at the electron microscopic level (Reid et al. 2001). This is currently done by 3-D reconstruction from serial ultrathin (75 nm) sections. One tomogram from one data set has been completed. We are proceeding in collecting a tilt series with a second set of data.