ULTRASTRUCTURAL CORRELATES OF FUNCTIONAL PROPERTIES OF INDIVIDUAL CNS SYNAPSES

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。使用共聚焦和多光子显微镜以及荧光Ca 2+指示剂，我们能够在脑切片制备物中可视化单个突触终扣和树突棘的激活（Emptage等，1999，2001）。这样，在有利的兴奋性突触，我们可以评估突触功能的量子参数，如递质释放概率和量子幅度。通过经由细胞内记录微电极注射可固定的标记物，并在生理成像实验结束时固定制备物，我们能够在电子显微镜水平上鉴定成像的突触（Reid等，2001）。目前，这是通过从连续切片（75 nm）进行三维重建来完成的。 已完成一个数据集的一张断层图像。我们正在收集倾斜系列与第二组数据。

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.