DFG-FOR 721

DFG-Forschergruppe FOR 721/2
Molekulare Struktur und Funktion der Tight Junction

Teilprojekt 7

Institut für Klinische Physiologie, Campus Benjamin Franklin, Charité Berlin

Renale Claudine und ihre Interaktionen

Ziel des Teilprojektes ist zum einen die Analyse der cis- und trans-Interaktionen nierentypischer Claudine, zum anderen die Aufklärung der Interaktion von Claudins-16 mit dem apikalen Cl^--Kanal Bestrophin. Die Experimente werden an einfach- und doppelttransfizierten MDCK C7-Zellen durchgeführt und beinhalten die Herstellung von Claudin-Chimären durch Austausch des 2. extrazellulären Loops, fluoreszenzoptische Untersuchungen von YFP- bzw. CFP-markierten Claudinen und Gefrierbruch-Elektronenmikroskopische Analyse der gebildeten TJ.

Die Untersuchungen der Wechselwirkung zwischen Claudin-16 und den beiden Bestrophin-Isoformen 1 und 4 werden mittels Überexpression bzw. knock-down (siRNA) der Bestrophine in MDCK-C7-Zellen mit oder ohne Claudin-16-Coexpression durchgeführt. Die funktionellen Auswirkungen aller untersuchten Interaktionen werden elektrophysiologisch charakterisiert und quantifiziert.

DFG-Mitarbeiter des Teilprojekts

Dr. rer. nat. Susanne Milatz

Publikationen 2010 - present (FOR 721/2)

2015

Conrad MP*, Piontek J* (*shared first authorship), Günzel D, Fromm M, Krug SM (2016, online 2015) Molecular basis of claudin-17 anion selectivity. Cell. Mol. Life Sci. 73(1): 185-200 (°IF 5.8) [PubMed] [WebPage] [PDF] [Supplement] TP1+TP6+TP7+TPZ{FFEM}

2014

Bücker R, Schulz E, Günzel D, Bojarski C, Lee IM, John LJ, Wiegand S, Janßen T, Wieler LH, Dobrindt U, Beutin L, Ewers C, Fromm M, Siegmund B, Troeger H, Schulzke JD (2014) α-Haemolysin of Escherichia coli: a potentiator of inflammatory activity in the colon. Gut 63(12): 1893-1901 (IF 14.7) [PubMed] [WebPage] [PDF] [Supplements] TP2+TP7+TP1

Hering NA*, Richter JF* (*shared first authorship), Fromm A, Wieser A, Hartmann S, Günzel D, Bücker R, Fromm M, Schulzke JD° (°corresponding author), Troeger H (2014) TcpC protein from E. coli Nissle improves epithelial barrier function involving PKCζ and ERK1/2 signalling in HT-29/B6 cells. Mucosal Immunol. 7(2): 369-378 (IF 7.0) [PubMed] [WebPage] [PDF] TP2+TP1+TP7+TPZ{2PI}

Richter JF, Pieper R, Zakrzewski SS, Günzel D, Schulzke JD, Van Kessel AG (2014) Diets high in fermentable protein and fiber alter tight junction protein composition with minor effect on barrier function in piglet colon. Br. J. Nutr. 111(6): 1040-1049 (IF 3.3) [PubMed] [WebPage] [PDF] [Supplements] TP7+TP2

Rossa J, Plöger C, Vorreiter F, Saleh T, Protze J, Günzel D, Wolburg H, Krause G, Piontek J (2014) Claudin-3 and claudin-5 protein folding and assembly into the tight junction are controlled by non-conserved residues in transmembrane2 (TM3) and extracellular loop 2 (ECL2) segments. J. Biol. Chem. 289(11): 7641-7653 (IF 4.7) [29.01.14 Accepted] [PubMed] [WebPage] [PDF] [Supplements] TP6+TP7

2013

Cording J, Berg J, Käding N, Bellmann C, Tscheik C, Westphal JK, Milatz S, Günzel D, Wolburg H, Piontek J, Huber O, Blasig IE (2013) Tight junctions: Claudins regulate the interactions between occludin, tricellulin and marvelD3, which, inversely, modulate the claudin oligomerization. J. Cell Sci. 126: 554-564 (IF 6.1) [PubMed] [WebPage] [PDF] [Supplement] TP5+TP7+TP6+TP3

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) (IF 3.5) [PubMed] [WebPage] [PDF] [Supplements, PDF] TP7

Schulzke JD, Siegmund B, Günzel D (2013) New insights in intestinal secretion. Gut ###: ###-### (IF 10.7) (Commentary) [PubMed] [WebPage] [PDF] TP2+TP7

Zakrzewski SS, Richter JF, Krug SM, Jebautzke B, Lee IM, Rieger J, Sachtleben M, Bondzio A, Schulzke JD, Fromm M, Günzel D (2013) Improved cell line IPEC-J2, characterized as a model for porcine jejunal epithelium. PLOS One 8(11): e79643 (print pages 1-12) (IF 3.5) [PubMed] [WebPage] [PDF] [Supplements] TP7+TP1+TP2+TPZ{2PI, FFEM}

Reviews

Günzel D, Yu AS (2013) Claudins and the modulation of tight junction permeability. Physiol. Rev. 93(2): 525-569 (IF 26.9) [PubMed] [WebPage] [PDF] TP7

2012

Kirschner N, Rosenthal R, Günzel D, Moll I, Brandner JM (2012) Tight junctions and differentiation – a chicken or the egg question? Exp. Dermatol. 21(3): 171-175 (IF 4.2) [PubMed] [WebPage] [PDF] TP9+TP1+TP7

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 (IF 6.6) [PubMed] [WebPage] [PDF] [Supplement] TP1+TP7+TP2

Rosenthal R*, Günzel D* (*shared first authorship), Finger C, Krug SM, Richter JF, Schulzke JD, Fromm M, Amasheh S (2012) The effect of chitosan on transcellular and paracellular mechanisms in the intestinal epithelial barrier. Biomaterials 33(9): 2791-2800 (IF 7.9) [PubMed] [WebPage] [PDF] [Supplement] TP1+TP7+TP2+TPZ

Schumann M, Günzel D, Buergel N, Richter JF, Troeger H, May C, Fromm A, Sorgenfrei D, Daum S, Bojarski C, Heyman M, Zeitz M, Fromm M, Schulzke JD (2012) Cell polarity-determining proteins Par-3 and PP-1 are involved in epithelial tight junction defects in celiac disease. Gut 61(2): 220-228 (IF 10.1) [PubMed] [WebPage] [PDF] [Supplements] TP2+TP7+TP1

Reviews

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 (IF 3.2) [PubMed] [WebPage] [PDF] (Review) TP7+TP1

Krug SM, Günzel D, Conrad MP, Lee IM, Amasheh S, Fromm M, Yu ASL (2012) Charge-selective claudin channels. Ann. N.Y. Acad. Sci. 1257: 20-28 (IF 3.2) [PubMed] [WebPage] [PDF] (Review) TP1+TP7

Plöger S, Stumpff F, Penner GB, Schulzke JD, Gäbel G, Martens H, Shen Z, Günzel D, Aschenbach JR (2012) Microbial butyrate and its role for barrier function in the gastrointestinal tract. Ann. N.Y. Acad. Sci. 1258: 52-59 (IF 3.2) [PubMed] [WebPage] [PDF] (Review) TP2+TP7

Schulzke JD, Günzel D, John LJ, Fromm M. Perspectives in tight junction research. Ann. N.Y. Acad. Sci. 1257: 1-19 (IF 3.2) [PubMed] [WebPage] [PDF] (Superinducing review) TP2+TP7+TP1

Handbook article

Günzel D, Fromm M (2012) Claudins and other tight junction proteins. Compreh. Physiol. (former Handbook of Physiology) 2(3): 1819-1852 [PubMed] [WebPage] [PDF] TP7+TP1

2011

Bücker R, Krug SM, Rosenthal R, Günzel D, Fromm A, Zeitz M, Chakraborty T, Fromm M, Epple HJ, Schulzke JD (2011) Aerolysin from Aeromonas hydrophila perturbs tight junction integrity and cell lesion repair in intestinal epithelial HT-29/B6 cells. J. Infect. Dis. 204 (8): 1283-1292 [PubMed] [WebPage] [PDF] TP2+TP1+TP7+TPZ

Hering NA, Richter JF, Krug SM, Günzel D, Fromm A, Bohn E, Rosenthal R, Bücker R, Fromm M, Troeger H, Schulzke JD (2011) Yersinia enterocolitica induces epithelial barrier dysfunction through regional tight junction changes in colonic HT-29/B6 cell monolayers. Lab. Invest. 91: 310-324 [PubMed] [WebPage] [PDF] [Supplement DOC]

Nielsen HL, Nielsen H, Ejlertsen T, Engberg J, Günzel D, Zeitz M, Hering NA, Fromm M, Schulzke JD, Bücker R (2011) Oral and fecal Campylobacter concisus strains perturb barrier function by apoptosis induction in HT-29/B6 intestinal epithelial cells. PlosOne 6(8): e23858 (print pages 1-8) [PubMed] [WebPage] [PDF] TP2+TP7+TP1

Schulzke JD*, Andres S*, Amasheh M* (*shared first authorship), Fromm A, Günzel D (2011) Anti-diarrheal mechanism of the traditional remedy Uzara via reduction of active chloride secretion. PLoS One 6(3): e18107 (print 1-9) [PubMed] [WebPage] [PDF]

Stumpff F, Georgi MI, Mundhenk L, Rabbani I, Fromm M, Martens H, Günzel D (2011) Sheep rumen and omasum primary cultures and source epithelia: Barrier function aligns with expression of tight junction proteins. J. Exp. Biol. 214: 2871-2882 [PubMed] [WebPage] [PDF] [Supplement] TP7+TP1

2010

Amasheh S, Fromm M, Günzel D (2010) Claudins of intestine and nephron - a correlation of molecular tight junction structure and barrier function. Acta Physiol. 201(1): 133-140 [PubMed] [WebPage] [PDF] (Review) TP1+TP7

Günzel D, Krug SM, Rosenthal R, Fromm M (2010) Biophysical methods to study tight junction permeability. Curr. Top. Membr. 65: 39-78 [Directory] [WebPage] [PDF] (Review / book chapter)

Hillenbrand B, Günzel D, Richter JF, Höhne, Schreier E, Schulzke JD, Mankertz J (2010) Norovirus non-structural protein p20 leads to impaired restitution of epithelial defects by inhibition of actin cytoskeleton remodelling. Scand. J. Gastroent. 45(11):1307-1319 [PubMed] [WebPage] [PDF]

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]

Rosenthal R, Milatz S, Krug SM, Oelrich B, Schulzke JD, Amasheh S, Günzel D, Fromm M (2010) Claudin-2, a component of the tight junction, forms a paracellular water channel. J. Cell Sci. 123(11): 1913-1921 [PubMed] [WebPage] [PDF] [Supplement]

Schmid T, Günzel D, Bogdan M (2010) Using an artificial neural network to determine electrical properties of epithelia. In: Diamantaras K, Duch W, Iliadis LS Eds., ICANN 2010, Part I, Lect. Notes Comput. Sci. 6352: 211-216 [26.07.10 In press]

Will C, Breiderhoff T, Thumfart J, Stuiver M, Kopplin K, Sommer K, Günzel D, Querfeld U, Meij IC, Shan Q, Bleich M, Willnow TE, Müller D (2010) Targeted deletion of murine Cldn16 identifies extra- and intrarenal compensatory mechanisms of Ca²⁺ and Mg²⁺ wasting. Am. J. Physiol. Renal Physiol. [PubMed] [WebPage] [Full text preprint PDF]

Dissertationen

Milatz S (2010) Funktionelle Charakterisierung des Tight Junction-Proteins Claudin-3 in Epithel- und Endothelzellen. Biologie, Humboldt-Universität zu Berlin. Dr. rer. nat., Magna cum laude
Haisch L (2010) Barrierefunktion der Claudine des dicken aufsteigenden Astes der Henle-Schleife. Charité Berlin, Betreuung Müller D und Günzel D. Dr. med., Summa cum laude

Vorarbeiten

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]
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]
Günzel D, Amasheh S, Pfaffenbach S, Richter JF, Kausalya PJ, Hunziker W, Fromm M (2009) Claudin-16 affects transcellular Cl^- secretion in MDCK cells. J. Physiol. (Lond.) 587(15): 3777-3793 [PubMed] [WebPage] [PDF] [Supplement]
Günzel D, Stuiver M, Kausalya PJ, Haisch L, Krug SM, Rosenthal R, Meij IC, Hunziker W, Fromm M, Müller D (2009) Claudin-10 exists in six alternatively spliced isoforms which exhibit distinct localization and function. J. Cell Sci. 122: 1507-1517 [PubMed] [WebPage] [PDF]
Yu ASL, Cheng MH, Angelow S, Günzel D, Kanzawa SA, Schneeberger EE, Fromm M, Coalson RD (2009) Molecular basis for cation selectivity in claudin-2-based paracellular pores: Identification of an electrostatic interaction site. J. Gen. Physiol. 133(1):111-127 [PubMed] [WebPage] [PDF]
Amasheh S, Milatz S, Krug SM, Markov AG, Günzel D, Amasheh M, Fromm M (2009) Tight junction proteins as channel formers and barrier builders: claudin-2, -5, and -8. Ann. N.Y. Acad. Sci. 1165: 211-219 [PubMed] [WebPage] [PDF]
Günzel D, Haisch L, Pfaffenbach S, Krug SM, Milatz S, Amasheh S, Hunziker W, Müller D (2009) Claudin function in the thick ascending limb of Henle's loop. Ann. N.Y. Acad. Sci. 1165: 152-162 [PubMed] [WebPage] [PDF]
Fromm M, Krug SM, Zeissig S, Richter JF, Rosenthal R, Schulzke JD, Günzel D (2009) High resolution analysis of barrier function. Ann. N.Y. Acad. Sci. 1165: 74-81 [PubMed] [WebPage] [PDF]
Günzel D, Yu ASL (2009, Epub 16.09.2008) Function and regulation of claudins in the thick ascending limb of Henle. Pflügers Arch. 458(1): 77-88 [PubMed] [WebPage] [PDF]
Amasheh S*, Milatz S* (*shared first authorship), Krug SM, Bergs M, Amasheh M, Schulzke JD, Fromm M (2009, Epub 08.11.2008) Na⁺ absorption defends from paracellular back-leakage by claudin-8 upregulation. Biochem. Biophys. Res. Comm. 378(1): 45-50 [PubMed] [WebPage] [PDF]
Troeger H *, Richter JF* (*shared first authorship), Beutin L, Günzel D, Dobrindt U, Epple HJ, Gitter AH, Zeitz M, Fromm M, Schulzke JD (2007) E. coli alpha-hemolysin induces focal leaks in colonic epithelium – a novel mechanism of bacterial translocation. Cell. Microbiol. 9(10): 2530-2540 [PubMed] [WebPage] [PDF]
Gabriel TE, Günzel D (2007) Quantification of Mg²⁺ extrusion and cytosolic Mg²⁺-buffering in Xenopus oocytes. Arch. Biochem. Biophys. 458(1): 3-15 [PubMed] [WebPage] [PDF]
Markov AG, Veshnyakova A, Krug S, Milatz S (2007) EExpression of tight junction proteins in the epithelium of small intestine of the rat. Russ. J. Physiol. (Ross. Fiziol. Zh. Im. I. M. Sechenova) 93(9): 1043-1054 [PubMed]
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]