Keywords :
Automatic Data Processing/methods; Cells, Cultured; Dendrites/metabolism; Diagnostic Imaging; Hippocampus/cytology; Intracellular Signaling Peptides and Proteins/metabolism; Lysosome-Associated Membrane Glycoproteins/metabolism; Membrane Proteins/metabolism; Mice; Mice, Mutant Strains; Microtubule-Associated Proteins/metabolism; Munc18 Proteins/genetics; Neurites/metabolism; Neurons/cytology/physiology; Neuropeptide Y/metabolism; Receptors, Transferrin/metabolism; Software; Synapses/physiology; Synaptic Vesicles/metabolism; Time Factors; Vesicle-Associated Membrane Protein 2/metabolism
Abstract :
[en] The shape, structure and connectivity of nerve cells are important aspects of neuronal function. Genetic and epigenetic factors that alter neuronal morphology or synaptic localization of pre- and post-synaptic proteins contribute significantly to neuronal output and may underlie clinical states. To assess the impact of individual genes and disease-causing mutations on neuronal morphology, reliable methods are needed. Unfortunately, manual analysis of immuno-fluorescence images of neurons to quantify neuronal shape and synapse number, size and distribution is labor-intensive, time-consuming and subject to human bias and error. We have developed an automated image analysis routine using steerable filters and deconvolutions to automatically analyze dendrite and synapse characteristics in immuno-fluorescence images. Our approach reports dendrite morphology, synapse size and number but also synaptic vesicle density and synaptic accumulation of proteins as a function of distance from the soma as consistent as expert observers while reducing analysis time considerably. In addition, the routine can be used to detect and quantify a wide range of neuronal organelles and is capable of batch analysis of a large number of images enabling high-throughput analysis.
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