title: Automated analysis of neuronal morphology, synapse number and synaptic recruitment.
abstract: 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.
authors.affiliation: Functional Genomics, Center for Neurogenomics and Cognitive Research (CNCR), VU University Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The Netherlands.
authors.forename: Sabine K
authors.initials: SK
authors.lastname: Schmitz
authors.forename: J J Johannes
authors.initials: JJ
authors.lastname: Hjorth
authors.forename: Raoul M S
authors.initials: RM
authors.lastname: Joemai
authors.forename: Rick
authors.initials: R
authors.lastname: Wijntjes
authors.forename: Susanne
authors.initials: S
authors.lastname: Eijgenraam
authors.forename: Petra
authors.initials: P
authors.lastname: de Bruijn
authors.forename: Christina
authors.initials: C
authors.lastname: Georgiou
authors.forename: Arthur P H
authors.initials: AP
authors.lastname: de Jong
authors.forename: Arjen
authors.initials: A
authors.lastname: van Ooyen
authors.forename: Matthijs
authors.initials: M
authors.lastname: Verhage
authors.forename: L Niels
authors.initials: LN
authors.lastname: Cornelisse
authors.forename: Ruud F
authors.initials: RF
authors.lastname: Toonen
authors.forename: Wouter J H
authors.initials: WJ
authors.lastname: Veldkamp
authors.forename: Wouter
authors.initials: W
authors.lastname: Veldkamp
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003712:Dendrites / Q000378:metabolism; D003952:Diagnostic Imaging; D000076122:Disks Large Homolog 4 Protein; D001330:Electronic Data Processing / Q000379:methods*; D051528:Guanylate Kinases; D006624:Hippocampus / Q000166:cytology; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D008239:Lysine / Q000031:analogs & derivatives / Q000378:metabolism; D051907:Lysosomal Membrane Proteins / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008817:Mice, Mutant Strains; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D051938:Munc18 Proteins / Q000235:genetics; D016501:Neurites / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000502:physiology*; D009478:Neuropeptide Y / Q000378:metabolism; D011990:Receptors, Transferrin / Q000378:metabolism; D012984:Software; D013569:Synapses / Q000502:physiology*; D013572:Synaptic Vesicles / Q000378:metabolism; D013997:Time Factors; D050988:Vesicle-Associated Membrane Protein 2 / Q000378:metabolism; D052118:Lysosomal-Associated Membrane Protein 1
chemical_list: D000076122:Disks Large Homolog 4 Protein; D051528:Guanylate Kinases; D047908:Intracellular Signaling Peptides and Proteins; D008239:Lysine; D051907:Lysosomal Membrane Proteins; D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins; D051938:Munc18 Proteins; D009478:Neuropeptide Y; D011990:Receptors, Transferrin; D050988:Vesicle-Associated Membrane Protein 2; C501447:Dlg4 protein, mouse; C052255:Lamp1 protein, mouse; D052118:Lysosomal-Associated Membrane Protein 1; C079240:Mtap2 protein, mouse; C013411:biocytin