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Three‐dimensional quantification of mossy‐fiber. Presynaptic boutons in living hippocampal slices using confocal microscopy
Author(s) -
Yu TzuPing,
Brown Thomas H.
Publication year - 1994
Publication title -
synapse
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.809
H-Index - 106
eISSN - 1098-2396
pISSN - 0887-4476
DOI - 10.1002/syn.890180304
Subject(s) - hippocampal formation , mossy fiber (hippocampus) , stratum , confocal , stratum granulosum , confocal microscopy , dentate gyrus , neuroscience , confocal laser scanning microscopy , chemistry , biophysics , anatomy , biology , stratum corneum , physics , optics , paleontology , genetics
Confocal laser scanning microscopy (CLSM) was used to visualize presynaptic elements of mossy‐fiber synapses in living rat hippocampal slices. Mossy fiber (mf) axons and their boutons were labeled in transverse hippocampal slices by injecting one of three fluorescent dyes (diI, diA, or fast diI) into stratum granulosum of the dentate gyrus and/or stratum lucidum of CA3. Three‐dimensional (3D) images of the mf boutons were obtained from serial optical sections in stratum lucidum. The 3D data were used to quantify and characterize the size and shape of the boutons based on their volumes (V) and surface areas (A), and the latter were compared with conventional 2D analyses. Various geometric models were fitted to the V‐A relationship, one of which provided a reasonable approximation to the data. The results demonstrate that this approach is useful for quantifying and characterizing the size and shape of mf expansions and raise the possibility of detecting and analyzing in real time activity‐dependent modifications in presynaptic structure. © 1994 Wiley‐Liss, Inc.