Reconstruction and morphometric analysis of hippocampal neurons from mice expressing fluorescent proteins

Abstract

Dendritic morphology and its alterations determine the strength of the integrated signal of individual neurons. Changes in the dendritic arborization accompany neuronal development, memory formation, and neurodegenerative processes. Region-specific dendritic simplification is a key feature in Alzheimer's disease and other neurodegenerative disorders. Here we describe a method to analyze whole-neuron morphology in brain slices prepared from transgenic mice expressing enhanced green fluorescent protein (EGFP) in different subpopulations of neurons. Complete pyramidal neurons are imaged using high-resolution confocal microscopy and analyzed after semi-automated 3D reconstruction. The approach allows evaluation of different morphological features, such as total extent of dendrites and number of branching points, as well as 3D Sholl analysis, e.g., of apical and basal dendritic subtrees of neurons from different hippocampal or cortical subregions. © 2014 Springer Science+Business Media New York.