Measuring exocytosis rate in arabidopsis pollen tubes using corrected fluorescence recovery after photoconversion (cfrapc) technique

Abstract

Exocytosis is a fundamental process essential for many cellular functions by targeting signal peptides, proteins, and cell wall components to the plasma membrane (PM) or extracellular matrix. Conventional methods, such as FRAP, often underestimate the exocytosis rate of a specific molecule, because retrieval of the molecules from the PM by endocytosis can impact the measurement. To overcome this issue, we have previously established a novel method, corrected fluorescence recovery after photoconversion (cFRAPc), which allows us to accurately measure the exocytosis rate by monitoring both exocytosis-dependent and exocytosis-independent events. In this chapter, we provide detailed procedures for the cFRAPc method to measure the exocytosis rate of Arabidopsis receptor-like kinase PRK1 in pollen tubes. This method should be widely applicable to various cell types.