Filopodyan: An open-source pipeline for the analysis of filopodia
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Filopodia have important sensory and mechanical roles in motile cells. The recruitment of actin regulators, such as ENA/ VASP proteins, to sites of protrusion underlies diverse molecular mechanisms of filopodia formation and extension. We developed Filopodyan (filopodia dynamics analysis) in Fiji and R to measure fluorescence in filopodia and at their tips and bases concurrently with their morphological and dynamic properties. Filopodyan supports high-throughput phenotype characterization as well as detailed interactive editing of filopodia reconstructions through an intuitive graphical user interface. Our highly customizable pipeline is widely applicable, capable of detecting filopodia in four different cell types in vitro and in vivo. We use Filopodyan to quantify the recruitment of ENA and VASP preceding filopodia formation in neuronal growth cones, and uncover a molecular heterogeneity whereby different filopodia display markedly different responses to changes in the accumulation of ENA and VASP fluorescence in their tips over time.
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1540-8140
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Wellcome Trust (085314/Z/08/Z)
Wellcome Trust (095829/Z/11/Z)
Isaac Newton Trust (Minute 1106(v))
European Research Council (281971)
European Commission and European Federation of Pharmaceutical Industries and Associations (EFPIA) FP7 Innovative Medicines Initiative (IMI) (115439)
European Research Council (322817)
Wellcome Trust (203144/Z/16/Z)
Cancer Research UK (C6946/A24843)
Medical Research Council (MR/L023784/2)
Medical Research Council (MR/L023784/1)