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  • 1.
    Durrieu, Lucía
    et al.
    IFIByNE, DFBMC, FCEN, UBA, Buenos Aires, Argentine.
    Johansson, Rikard
    Linköping University, Department of Biomedical Engineering, Division of Biomedical Engineering. Linköping University, Faculty of Science & Engineering.
    Bush, Alan
    IFIByNE, DFBMC, FCEN, UBA, Buenos Aires, Argentine.
    Janzén, David
    Linköping University, Department of Clinical and Experimental Medicine, Division of Cell Biology. Linköping University, Faculty of Medicine and Health Sciences.
    Gollvik, Martin
    Linköping University, Faculty of Medicine and Health Sciences. Linköping University, Department of Biomedical Engineering, Division of Biomedical Engineering.
    Cedersund, Gunnar
    Linköping University, Faculty of Science & Engineering. Linköping University, Department of Biomedical Engineering, Division of Biomedical Engineering.
    Colman-Lerner, Alejandro
    IFIByNE, DFBMC, FCEN, UBA, Buenos Aires, Argentine.
    Quantification of nuclear transport in single cells2014Other (Other academic)
    Abstract [en]

    Regulation of nuclear transport is a key cellular function involved in many central processes, such as gene expression regulation and signal transduction. Rates of protein movement between cellular compartments can be measured by FRAP. However, no standard and reliable methods to calculate transport rates exist. Here we introduce a method to extract import and export rates, suitable for noisy single cell data. This method consists of microscope procedures, routines for data processing, an ODE model to fit to the data, and algorithms for parameter optimization and error estimation. Using this method, we successfully measured import and export rates in individual yeast. For YFP, average transport rates were 0.15 sec-1. We estimated confidence intervals for these parameters through likelihood profile analysis. We found large cell-to-cell variation (CV = 0.79) in these rates, suggesting a hitherto unknown source of cellular heterogeneity. Given the passive nature of YFP diffusion, we attribute this variation to large differences among cells in the number or quality of nuclear pores. Owing to its broad applicability and sensitivity, this method will allow deeper mechanistic insight into nuclear transport processes and into the largely unstudied cell-to-cell variation in kinetic rates.

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