Permeability analyses and three dimensional imaging of interferon gamma-induced barrier disintegration in intestinal organoids
Bardenbacher M, Ruder B, Britzen-Laurent N, Schmid B, Waldner M, Naschberger E, Scharl M, Mueller W, Günther C, Becker C, Stürzl M, Tripal P (2019)
Publication Type: Journal article
Publication year: 2019
Journal
Book Volume: 35
DOI: 10.1016/j.scr.2019.101383
Abstract
The aberrant regulation of the epithelial barrier integrity is involved in many diseases of the digestive tract, including inflammatory bowel diseases and colorectal cancer. Intestinal epithelial cell organoid cultures provide new perspectives for analyses of the intestinal barrier in vitro. However, established methods of barrier function analyses from two dimensional cultures have to be adjusted to the analysis of three dimensional organoid structures. Here we describe the methodology for analysis of epithelial barrier function and molecular regulation in intestinal organoids. Barrier responses to interferon-y of intestinal organoids with and without epithelial cellspecific deletion of the interferon-y-receptor 2 gene were used as a model system. The established method allowed monitoring of the kinetics of interferon-y-induced permeability changes in living organoids. Proteolytic degradation and altered localization of the tight junction proteins claudin-2, -7, and - 15 was detected using confocal spinning disc microscopy with 3D reconstruction. Hessian analysis was used for quantification of relocalization of claudins. In summary, we provide a novel methodologic approach for quantitative analyses of intestinal epithelial barrier functions in the 3D organoid model.
Authors with CRIS profile
Nathalie Britzen-Laurent
Professur für Molekulare und Experimentelle Chirurgie
Benjamin Schmid
Optical Imaging Center Erlangen (OICE)
Maximilian Waldner
Lehrstuhl für Innere Medizin I
Elisabeth Naschberger
Medizinische Fakultät
Claudia Günther
Department of Medicine 1 – Gastroenterology, Pneumology and Endocrinology
Christoph Becker
Professur für Molekulare Gastroenterologie
Michael Stürzl
Professur für Molekulare und Experimentelle Chirurgie
Philipp Tripal
Department of Surgery
Additional Organisation(s)
Involved external institutions
Switzerland (CH)
United Kingdom (GB)How to cite
APA:
Bardenbacher, M., Ruder, B., Britzen-Laurent, N., Schmid, B., Waldner, M., Naschberger, E.,... Tripal, P. (2019). Permeability analyses and three dimensional imaging of interferon gamma-induced barrier disintegration in intestinal organoids. Stem Cell Research, 35. https://dx.doi.org/10.1016/j.scr.2019.101383
MLA:
Bardenbacher, Marco, et al. "Permeability analyses and three dimensional imaging of interferon gamma-induced barrier disintegration in intestinal organoids." Stem Cell Research 35 (2019).
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