Abstract
Abnormalities of mucus viscosity play a critical role in the pathogenesis of several respiratory diseases, including cystic fibrosis. Currently, there are no approaches to assess the rheological properties of mucin granule matrices in live cells. This is the first example of the use of a molecular rotor, a BODIPY dye, to quantitatively visualize the viscosity of intragranular mucin matrices in a large population of individual granules in differentiated primary bronchial epithelial cells using fluorescence lifetime imaging microscopy.
Highlights
Mucus forms a sticky, gel-like layer that covers wet surfaces of various organs and tissues, including airways
In the course of our studies on developing BODIPY-based viscometers[16,17], we discovered that a simple BODIPY dye (Fig. 1, where the rotation of the phenyl group around the BODIPY core is sensitive to the viscosity of the surrounding media, i.e., BODIPY rotor) showed appreciable membrane permeability and accumulation inside mucin granules
We present the results on the use of BODIPY rotor to probe the viscosity of the intragranular mucin matrix of differentiated primary cultures of human bronchial epithelial cells from non-cystic fibrosis (CF) and CF patients (carrying the dominant CF mutation (F508del/F508), which affects 70 % of all CF patients worldwide23), which to the best of our knowledge, is the first example of a small molecule viscometer that is capable of reporting on the viscosity of intact mucin granule matrices in live cells
Summary
Mucus forms a sticky, gel-like layer that covers wet surfaces of various organs and tissues, including airways. Mucus layer act as the first line of defense against toxins and pathogens inhaled into the lungs It is produced by the secretion and hydration of gel-forming mucins, which are large fibrous biopolymers that are synthetized and stored as a condensed matrix inside the secretory granules in mucus/goblet cells. It is possible that CF mucus defects may already be present prior to mucin secretion during the early stages of biogenesis, which could impact the packaging and rheological properties of the intragranular mucin matrix. Fluorescent probes that could permeate the cell membrane, accumulate inside mucin granules, and whose lifetimes are sensitive to intragranular viscosity fluctuations would be a significant addition to the diagnostic/analytical tools of chemical biology
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