Reconstruction of Hippocampal Pyramidal Neurons

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 soma42 (link)43 (link)44 (link). Neurons meeting these criteria were reconstructed in three-dimensions (3D) with a 40×/1.4 N.A., Plan-Apochromat oil immersion objective on a Zeiss Axiophot 2 microscope equipped with a motorized stage, video camera system, and Neurolucida morphometry software (MBF Bioscience). 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 were analyzed in order to assess morphological cellular diversity and potential differences among animals45 (link).

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Lazarczyk M.J., Kemmler J.E., Eyford B.A., Short J.A., Varghese M., Sowa A., Dickstein D.R., Yuk F.J., Puri R., Biron K.E., Leist M., Jefferies W.A, & Dickstein D.L. (2016). Major Histocompatibility Complex class I proteins are critical for maintaining neuronal structural complexity in the aging brain. Scientific Reports, 6, 26199.

Publication 2016

Axiophot 2 microscope Neurolucida morphometry software NeuroExplorer software

Cellular Dendritic Immersion Intersections Microscope Neurons Radial Tree

Corresponding Organization :

Other organizations : Icahn School of Medicine at Mount Sinai, University of Konstanz, Canada's Michael Smith Genome Sciences Centre, University of British Columbia, Allen Institute for Brain Science

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Variable analysis

independent variables

Neuron location within the pyramidal layer of the CA1 as defined by cytoarchitectural characteristics
Completeness of dendritic tree filling, as evidenced by well-defined endings
Intactness of tertiary branches, with the exception of branches that extended beyond 50 μm in radial distance from the cell soma

dependent variables

Total dendritic length
Number of intersections
Amount of dendritic material per radial distance from the soma, in 30 μm increments

control variables

Neurons residing within the pyramidal layer of the CA1 as defined by cytoarchitectural characteristics
Neurons with complete filling of dendritic tree, as evidenced by well-defined endings
Neurons with intact tertiary branches, with the exception of branches that extended beyond 50 μm in radial distance from the cell soma

positive controls

None specified

negative controls

None specified

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