Neurons were manually traced and reconstructed in 3-dimensions with a 63×/1.4 N.A., Plan-Apochromat oil immersion objective on a Zeiss Axio Imager Vario microscope equipped with a motorized stage, video camera system, and Neurolucida morphometry software (MBF Bioscience, Williston, VT, USA). To be included in the analysis, a loaded neuron had to satisfy the following criteria: (1) reside within the pyramidal layer of the CA1 as defined by cytoarchitectural characteristics; (2) demonstrate complete filling of dendritic tree, as evidenced by well-defined endings; and (3) demonstrate intact tertiary branches, with the exception of branches that extended beyond 50 μm in radial distance from the cell soma [33 (link),34 (link),75 (link)].Using NeuroExplorer software (MBF Bioscience) total dendritic length, number of intersections, and the amount of dendritic material per radial distance from the soma, in 30-μm increments [34 (link)]. were analyzed in order to assess morphological cellular diversity and potential differences between the animal groups. A total of 50 cells were reconstructed for controls (~8 cells per animal) and a total of 49 cells were reconstructed for neutron irradiated mice (~8 cells per animal).
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