|
DFG Research Unit FOR 721/1 Molecular Structure and Function of the Tight Junction |
|
|
Project 4 |
Dominik Müller, MD
Pediatric Nephrology,
Campus Virchow Klinikum, Charité, Freie Universität and Humboldt-Universität, Berlin
Thomas Willnow, Prof. PhD
Max Delbrueck Center and
Inst. of Clinical Pharmacology,
Campus Benjamin Franklin, Charité, Freie Universität and Humboldt-Universität, Berlin
Tight junction proteins: role in ion homeostasis and blood pressure regulation
Paracellular transport mediated by the intercellular tight junction play a major role in water and ion homeostasis. Thus it is plausible that individual renal tight junction proteins are essential for blood pressure regulation.
A goal of the project is it to examine in vivo the meaning of the tight junction proteins claudin-3 and claudin-10 for ion and water homeostasis and thus blood pressure regularization. In a first subproject different stress models (salts, water, drug components) will be applied in rats and mice. Kidneys will be investigated by means of real-time PCR, immune histochemistry, and Western blot for the regulation of claudines under these conditions.
In a second subproject mice with claudin-10 and claudin-3 gene defects will be generated by means of Cre lox technology and then functionally characterised. In a multi-step process the genes are to be inactivated ubiquitously, in order to get first insight about the function of claudin-3 and claudin-10 in the intact organism. In the following, conditional inactivation in the kidney will be performed by means of the ApoE Cre line already established as well as in the PT via a Cre line (sglt2Cre).
Apart from the morhpologic processing (immune histochemistry, freeze fracture) animals will be functionally characterized regarding blood pressure by means of telemetry, glomerular filtration rate, and behavior under salt resp. volume load.
Additionally, we will investigate the effect of furosemide in order to further characterise the function of claudin-3 and -10 and to form the basis for future pharmacologic interventions.
Granted by project 1, FOR 721/1
Dipl. Biol. Kathrin Kopplin
Dipl. Biochem. Dr. rer. nat. Constanze Will
Publications 2007 - 2012
Breiderhoff T, Himmerkus N, Stuiver M, Mutig K, Will C, Meij IC, Bachmann S, Bleich M, Willnow TE, Müller D (2012) Deletion of Cldn10 in the thick ascending limb impairs paracellular sodium permeability and leads to hypermagnesemia and nephrocalcinosis. Proc. Natl. Acad. Sci. USA 109(35): 14241-14246 (IF 9.7) [PubMed] [WebPage] [PDF w. Supplements]
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]
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. 298: F1152-F1161 [PubMed] [WebPage] [PDF]
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]
Will C, Fromm M, Müller D (2008) Claudin tight junction proteins: novel aspects in paracellular transport. Periton. Dialysis Int. 28(6): 577-584 [PubMed] [PDF] (Review)
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]
Zhang J, Piontek J, Wolburg H, Piehl C, Liss M, Otten C, Christ A, Willnow TE, Blasig IE, Abdelilah-Seyfried S (2010) Establishment of a neuroepithelial barrier by claudin-5a is essential for zebrafish brain ventricular lumen expansion. Proc. Natl. Acad. Sci. 107(4): 1425-1430 [PubMed] [WebPage] [PDF]
Doctoral thesis
Will C (2011) [Generation and characterisation of claudin-16- and claudin-10-deficient mice] Doctoral thesis, Dr. rer. nat., Humboldt-Universität zu Berlin, Supervision: Müller D, 18.02.11: Magna cum laude
Haisch L (2010) [Barrier function of claudins of the thick ascensing loop of Henles loop] Doctoral thesis, Dr. med., Charité Berlin. Supervision: Müller D and Günzel D. , Summa cum laude
Previous work
Hammes A, Andreassen T, Spoelgen R, Raila J, Hübner N, Schulz H, Metzger J, Schweigert FJ, Luppa PB, Nykjaer A, Willnow TE (2005) Role of endocytosis in cellular uptake of sex steroids. Cell 122: 751-762.
Hoenderop JGJ, Müller D, Suzuki M, Ishibashi K, Imai M, Hartog A, van der Kemp AWCM, Sweep F, Willems PGHM, Bindels RJM (2001) Calcitriol controls the epithelial calcium channel in the kidney. J. Am. Soc. Nephrol. 12: 1342-1349.
Kausalya JP*, Amasheh S* (*shared first authorship), Günzel D, Wurps H, Müller D, Fromm M, Hunziker W (2006) Disease-associated mutations affect intracellular traffic and paracellular Mg2+ transport function of Claudin-16. J. Clin. Invest. 116(4): 878-891.
Leheste JR, Melsen F, Wellner M, Jansen P, Schlichting U, Renner-Muller I, Andreassen T, Wolf E, Bachmann S, Nykjaer A, Willnow TE (2003) Hypocalcemia and osteopathy in mice with kidney-specific megalin gene defect. FASEB J. 17: 247-249.
Marr N, Bichet DG, Hoefs S, Savelkoul PJM, Koenings IBM, de Mattia F, Graat MPJ, Arthus MF, Lonergan M, Fujiwara M, Knoers NVAM, Landau D, Balfe J, Oksche A, Rosenthal W, Müller D, van Os CH, Deen PMT (2002) Cell-biological and functional analysis of five new aquaporin-2 missense mutations that cause recessive nephrogenic diabetes insipidus. J. Am. Soc. Nephrol. 13: 2267-2277.
Müller D, Hoenderop JGJ, Claverie-Martin F, Garcia-Nieto V, Mèhes K, Karoly F, Eggert P, van Os CH, Vennekes R, Nilius B, Bindels RJM (2002a) Epithelial Ca2+ channel (ECAC1) in autosomal dominant hypercalciuria. Nephrol. Dial. Transplant. 17: 1614-1620.
Müller D, Hoenderop JGJ, Meij IC, Knoers NVAM, van den Heuvel B, Den Hollander A, Eggert P, Garcia-Nieto V, Claverie-Martin F, Bindels RJM (2000) Molecular cloning, tissue distribution and chromosomal mapping of the human epithelial calcium channel, ECaC1. Genomics 67: 48-53.
Müller D, Kausalya PJ, Bockenhauer D, Thumfart J, Meij IC, Dillon MJ, van't Hoff W, Hunziker W (2006) Unusual clinical presentation and possible rescue of a novel claudin-16 mutation. J. Clin. Endocrinol. Metab. 91(8): 3076-3079 [Abstract] [PDF]
Müller D, Kausalya PJ, Claverie-Martin F, Meij I, Eggert P, Garcia-Nieto V, Hunziker W (2003) A novel mutation in claudin-16 associated with childhood hypercalciuria abolishes binding to ZO-1 and leads to lysosomal mistargeting. Am. J. Hum. Genet. 73: 1293-1301.
Müller D, Kausalya JP, Meij IC, Hunziker W (2006) Familial hypomagnesemia with hypercalciuria and nephrocalcinosis: blocking endocytosis restores surface expression of a novel Claudin-16 mutant that lacks the entire C-terminal cytosolic tail. Hum. Mol. Genet. 15(7): 1049-1058.
Müller D, Marr N, Ankermann T, Eggert P, Deen PMT (2002b) Desmopressin for nocturnal enuresis in nephrogenic diabetes insipidus. Lancet 359: 495-497.
Nykjaer A, Dragun D, Walther D, Vorum H, Jacobsen C, Herz J, Melsen F, Christensen EI, Willnow TE (1999) An endocytic pathway essential for renal uptake and activation of the steroid 25-(OH) vitamin D3. Cell 96: 507-515.