DFG Research Training Group "TJ-Train" (GRK 2318/2)
Tight junctions and their proteins
Molecular features and actions in health and disease

       Project A1     1st period

Dorothee Günzel1 , Jörg Scheffel2  
1Clinical Physiology / Nutritional Medicine, Campus Benjamin Franklin, Charité - Universitätsmedizin Berlin
2Department of Dermatology, Venerology and Allergology, Charité - Universitätsmedizin Berlin

Impedance analysis of epithelial barrier properties


Background: Electrophysiological methods have a long tradition in epithelial physiology. However, new developments both in the molecular understanding of the tight junction (TJ) and in the measuring technology make new approaches possible.

Hypotheses: Combining impedance spectroscopy with specific alterations of partial epithelial resistances will allow us to quantify the contribution of different epithelial components towards general barrier function of simple and stratified epithelia. Refinement by combining brief, reversible alteration with fast multisinus impedance spectroscopy will generate a method for a detailed analysis of the mechanisms underlying the action of toxins and drugs.

Aims: This project aims to resolve the relative contributions of trans- and paracellular pathways as well as stratum corneum components towards barrier function in simple and stratified epithelia, respectively. Healthy and diseased tissues will be compared and effects of pharmacological treatment on different barrier components will be evaluated.

Methods: Standard molecular biological methods will be used for the knockout/knockdown of specific claudins and for the over-expression of ion channels or TJ components. Resulting cell clones will be cultivated (standard and 3D cell culture) and used for Ussing-chamber-based impedance spectroscopy. Multisinus technique for fast impedance spectroscopy has recently been implemented and will be rigorously tested in simple as well as stratified epithelial cell culture. Data will be evaluated by fitting to various equivalent circuit models and by using COMSOL Multiphysics® Modeling Software. Morphological conclusions from impedance measurements will be cross-validated with optical methods (confocal laserscanning microscopy; transmission electron microscopy).

Thesis project: As part of this PhD project we will adapt equivalent electric circuits by employing established cell culture models for simple and stratified epithelia. Expression of TJ proteins will be perturbed (e.g. by specific knock-down of TJ proteins or by application of cytokines). The transcellular pathway will be affected by application of ionophores, pore-forming bacterial toxins or by overexpression/activation of ion channels). Trans- and paracellular barrier components will be quantified by modelling impedance spectroscopic data, and visualized by various optical techniques.

Suggested reading:  See Project-related publications, especially Refs. 2, 3, and 8

2nd cohort PhD doctoral student

2nd cohort MD doctoral student

  • Selma Dib

  • Denisa Daci

1st cohort PhD doctoral student

  • Roman Mannweiler
    19.08.21: Doctoral examination passed, Dr. ing., Technische Universität Berlin, magna cum laude

Mannweiler R, Bergmann S, Vidal-Y-Sy S, Brandner JM, Günzel D (2021) Direct assessment of individual skin barrier components by electrical impedance spectroscopy. Allergy 76(10): 3094-3106 (13 pages). doi: 10.1111/all.14851, Supplement PDF (°IF 13.1)

1st cohort MD doctoral student, 1st cohort

  • Ahmad Mouna  

Alzahrani AS, Hussein M, Alswailem M, Mouna A, Albalawi L, Moria Y, Jabbar MA, Shi Y, Günzel D*, Dasouki M* (*equally contributing) (2021) A novel claudin-10 mutation with a unique mechanism in two unrelated families with HELIX syndrome. Kidney Int. 100(2): 415-429, doi: 10.1016/j.kint.2021.02.023, Supplement (°IF 10.6)
 

Project-related publications

If a paper is not accessible, please mail to

  1. Milatz S*, Himmerkus N* (*shared first authorship), Wulfmeyer VC, Drewell H, Mutig K, Hou J, Breiderhoff T, Müller D, Fromm M, Bleich M°, Günzel D° (°shared last authorship) (2017) Mosaic expression of claudins in thick ascending limbs of Henle results in spatial separation of paracellular Na+ and Mg2+ transport. Proc. Natl. Acad. Sci. USA 114(2): E219-E227 [PubMed] [WebPage] [PDF+Supplement]. "Paper of the month" 03/2017 of the German Physiological Society

  2. Schmid T, Bogdan M, Günzel D (2013) Discerning apical and basolateral properties of HT-29/B6 and IPEC-J2 cell layers by impedance spectroscopy, mathematical modeling and machine learning. PLOS One 8(7): e62913 (printed pages 1-12) [PubMed] [WebPage] [PDF] [Supplements, PDF]

  3. Günzel D, Zakrzewski S, Schmid T, Pangalos M, Wiedenhoeft J, Blasse C, Ozboda C, Krug SM (2012) From TER to trans- and paracellular resistance: Lessons from impedance spectroscopy. Ann. N.Y. Acad. Sci. 1257: 142-151 [PubMed] [WebPage] [PDF]

  4. Krug SM, Günzel D, Conrad MP, Rosenthal R, Fromm A, Amasheh S, Schulzke JD, Fromm M (2012) Claudin-17 forms tight junction channels with distinct anion selectivity. Cell. Mol. Life Sci. 69(16): 2765-2778 [PubMed] [WebPage] [PDF] [Supplement]

  5. Milatz S, Krug SM, Rosenthal R, Günzel D, Müller D, Schulzke JD, Amasheh S*, Fromm M* (*shared last authorship) (2010) Claudin-3 acts as a sealing component of the tight junction for ions of either charge and uncharged solutes. Biochim. Biophys. Acta Biomembr. 1798: 2048-2057 [PubMed] [WebPage] [PDF]]

  6. Krug SM, Amasheh S, Richter JF, Milatz S, Günzel D, Westphal JK, Huber O, Schulzke JD, Fromm M (2009) Tricellulin forms a barrier to macromolecules in tricellular tight junctions without affecting ion permeability. Mol. Biol. Cell 20: 3713-3724 [PubMed] [WebPage] [PDF] [Supplement text] [Supplement video]

  7. Günzel D, Stuiver M, Kausalya PJ, Haisch L, Krug SM, Rosenthal R, Meij IC, Hunziker W, Fromm M, Müller D (2009a) Claudin-10 exists in six alternatively spliced isoforms which exhibit distinct localization and function. J. Cell Sci. 122: 1507-1517 [PubMed] [WebPage] [PDF]

  8. Krug SM, Fromm M, Günzel D (2009) Two-path impedance spectroscopy for measuring paracellular and transcellular epithelial resistance. Biophys. J. 97(8): 2202-2211 [PubMed] [WebPage] [PDF] [Supplement]

  9. Zeissig S, Bürgel N, Günzel D, Richter JF, Mankertz J, Wahnschaffe U, Kroesen AJ, Zeitz M, Fromm M, Schulzke JD (2007) Changes in expression and distribution of claudin-2, -5 and -8 lead to discontinuous tight junctions and barrier dysfunction in active Crohn's disease. Gut 56(1): 61-72 [PubMed] [WebPage] [PDF]

  10. Reiter B, Kraft R, Günzel D, Zeissig S, Schulzke JD, Fromm M, Harteneck C (2006) TRPV4-mediated regulation of epithelial permeability. FASEB J. 20: 1802-1812 [PubMed] [WebPage] [PDF]

  11. Tunggal JA*, Helfrich I* (*shared first authorship), Schmitz A, Schwarz H, Günzel D, Fromm M, Kemler R, Krieg T, Niessen CM (2005) E-cadherin is essential for in vivo epidermal barrier function by regulating tight junctions. EMBO J. 24: 1146-1156 [PubMed] [WebPage] [PDF]