TY - JOUR T1 - Filopodyan: an open-source pipeline for the analysis of filopodia JF - bioRxiv DO - 10.1101/138610 SP - 138610 AU - Vasja Urbančič AU - Richard Butler AU - Benjamin Richier AU - Manuel Peter AU - Julia Mason AU - Frederick J. Livesey AU - Christine E. Holt AU - Jennifer L. Gallop Y1 - 2017/01/01 UR - http://biorxiv.org/content/early/2017/07/19/138610.abstract N2 - 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.eTOC summary: Urbančič et al. developed an open-source platform called Filopodyan (filopodia dynamics analysis) in Fiji and R to measure fluorescence in filopodia, at their tips, and bases, concurrently with their morphological and dynamic properties. This customizable tool therefore enables researchers to determine the relationship between protein localization and filopodium behavior. ER -