Imaging viscosity of intragranular mucin matrix in cystic fibrosis cells

Sebastian Requena; Olga Ponomarchuk; Marlius Castillo; Jonathan Rebik; Emmanuelle Brochiero; Julian Borejdo; Ignacy Gryczynski; Sergei V Dzyuba; Zygmunt Gryczynski; Ryszard Grygorczyk; Rafal Fudala

doi:10.1038/s41598-017-17037-2

Imaging viscosity of intragranular mucin matrix in cystic fibrosis cells

Sci Rep. 2017 Dec 1;7(1):16761. doi: 10.1038/s41598-017-17037-2.

Authors

Sebastian Requena¹, Olga Ponomarchuk^{2

3}, Marlius Castillo⁴, Jonathan Rebik⁴, Emmanuelle Brochiero^{2

5}, Julian Borejdo¹, Ignacy Gryczynski¹, Sergei V Dzyuba⁶, Zygmunt Gryczynski^{1

7}, Ryszard Grygorczyk^{8

9}, Rafal Fudala¹⁰

Affiliations

¹ University of North Texas Health Science Center, Department of Microbiology, Immunology & Genetics, Fort Worth, TX, 76107, USA.
² Centre de Recherche du CHUM (CRCHUM), Université de Montréal, Montréal, Québec, H2×0A9, Canada.
³ visiting graduate student from Moscow State University, Moscow, Russia.
⁴ Texas Christian University, Department of Chemistry and Biochemistry, Fort Worth, TX, 76129, USA.
⁵ Département de Médecine, Université de Montréal, Montréal, Québec, H3T 1J4, Canada.
⁶ Texas Christian University, Department of Chemistry and Biochemistry, Fort Worth, TX, 76129, USA. s.dzyuba@tcu.edu.
⁷ Texas Christian University, Department of Physics and Astronomy, Fort Worth, TX, 76129, USA.
⁸ Centre de Recherche du CHUM (CRCHUM), Université de Montréal, Montréal, Québec, H2×0A9, Canada. ryszard.grygorczyk@umontreal.ca.
⁹ Département de Médecine, Université de Montréal, Montréal, Québec, H3T 1J4, Canada. ryszard.grygorczyk@umontreal.ca.
¹⁰ University of North Texas Health Science Center, Department of Microbiology, Immunology & Genetics, Fort Worth, TX, 76107, USA. Rafal.Fudala@unthsc.edu.

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.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Cells, Cultured
Cystic Fibrosis / etiology
Cystic Fibrosis / metabolism*
Cytoplasmic Granules / metabolism*
Epithelial Cells / metabolism
Fluorescent Dyes / chemistry
Humans
Microscopy, Fluorescence
Molecular Imaging* / methods
Molecular Structure
Mucins / metabolism*
Respiratory Mucosa / metabolism
Viscosity*

Substances

Fluorescent Dyes
Mucins

Grants and funding

PJT148593 /CIHR/Canada