{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T05:44:38Z","timestamp":1792820678611,"version":"4.5.0"},"reference-count":43,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2020,7,20]],"date-time":"2020-07-20T00:00:00Z","timestamp":1595203200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Dr.-August-und-Erika-Appenrodt-Foundation","award":["not applicable"],"award-info":[{"award-number":["not applicable"]}]},{"DOI":"10.13039\/501100002347","name":"Bundesministerium f\u00fcr Bildung und Forschung","doi-asserted-by":"publisher","award":["01GM1515F"],"award-info":[{"award-number":["01GM1515F"]}],"id":[{"id":"10.13039\/501100002347","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Mutations of the Pkhd1 gene cause autosomal recessive polycystic kidney disease (ARPKD). Pkhd1 encodes fibrocystin\/polyductin (FPC), a ciliary type I membrane protein of largely unknown function, suggested to affect adhesion signaling of cells. Contributions of epithelial cell adhesion and contractility to the disease process are elusive. Here, we link loss of FPC to defective epithelial morphogenesis in 3D cell culture and altered cell contact formation. We study Pkhd1-silenced Madin-Darby Canine Kidney II (MDCKII) cells using an epithelial morphogenesis assay based on micropatterned glass coverslips. The assay allows analysis of cell adhesion, polarity and lumen formation of epithelial spheroids. Pkhd1 silencing critically affects the initial phase of the morphogenesis assay, leading to a reduction of correctly polarized spheroids by two thirds. Defects are characterized by altered cell adhesion and centrosome positioning of FPC-deficient cells in their 1-\/2-cell stages. When myosin II inhibitor is applied to reduce cellular tension during the critical early phase of the assay, Pkhd1 silencing no longer inhibits formation of correctly polarized epithelia. We propose that altered sensing and cell interaction of FPC-deficient epithelial cells promote progressive epithelial defects in ARPKD.<\/jats:p>","DOI":"10.3390\/ijms21145140","type":"journal-article","created":{"date-parts":[[2020,7,20]],"date-time":"2020-07-20T10:59:38Z","timestamp":1595242778000},"page":"5140","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Fibrocystin Is Essential to Cellular Control of Adhesion and Epithelial Morphogenesis"],"prefix":"10.3390","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4529-1916","authenticated-orcid":false,"given":"Wolfgang H.","family":"Ziegler","sequence":"first","affiliation":[{"name":"Department of Pediatric Kidney, Liver and Metabolic Diseases, Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Birga","family":"Soetje","sequence":"additional","affiliation":[{"name":"Department of Pediatric Kidney, Liver and Metabolic Diseases, Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa P.","family":"Marten","sequence":"additional","affiliation":[{"name":"Department of Pediatric Kidney, Liver and Metabolic Diseases, Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jana","family":"Wiese","sequence":"additional","affiliation":[{"name":"Department of Pediatric Kidney, Liver and Metabolic Diseases, Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mithila","family":"Burute","sequence":"additional","affiliation":[{"name":"Universit\u00e9 Paris Diderot, CEA, INSERM, H\u00f4pital Saint Louis, Institut Universitaire d\u2019Hematologie, UMRS1160, CytoMorpho Lab, 75010 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dieter","family":"Haffner","sequence":"additional","affiliation":[{"name":"Department of Pediatric Kidney, Liver and Metabolic Diseases, Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,7,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1533","DOI":"10.1056\/NEJMra1010172","article-title":"Ciliopathies","volume":"364","author":"Hildebrandt","year":"2011","journal-title":"N. Engl. J. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1056\/NEJMra022161","article-title":"Polycystic kidney disease","volume":"350","author":"Wilson","year":"2004","journal-title":"N. Engl. J. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"50:1","DOI":"10.1038\/s41572-018-0047-y","article-title":"Polycystic kidney disease","volume":"4","author":"Bergmann","year":"2018","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1915","DOI":"10.1007\/s00467-013-2634-1","article-title":"Clinical manifestations of autosomal recessive polycystic kidney disease (ARPKD): Kidney-related and non-kidney-related phenotypes","volume":"29","author":"Buscher","year":"2014","journal-title":"Pediatr. Nephrol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1002\/humu.20029","article-title":"PKHD1 mutations in autosomal recessive polycystic kidney disease (ARPKD)","volume":"23","author":"Bergmann","year":"2004","journal-title":"Hum. Mutat."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2266","DOI":"10.1681\/ASN.2010111173","article-title":"Epitope-tagged Pkhd1 tracks the processing, secretion, and localization of fibrocystin","volume":"22","author":"Bakeberg","year":"2011","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"3306","DOI":"10.1093\/hmg\/ddq233","article-title":"Polycystic kidney disease protein fibrocystin localizes to the mitotic spindle and regulates spindle bipolarity","volume":"19","author":"Zhang","year":"2010","journal-title":"Hum. Mol. Genet."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"942","DOI":"10.1093\/hmg\/ddm039","article-title":"Polyductin undergoes notch-like processing and regulated release from primary cilia","volume":"16","author":"Kaimori","year":"2007","journal-title":"Hum. Mol. Genet."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"887","DOI":"10.1038\/nrm2523","article-title":"From cells to organs: Building polarized tissue","volume":"9","author":"Bryant","year":"2008","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1035","DOI":"10.1038\/ncb2106","article-title":"A molecular network for de novo generation of the apical surface and lumen","volume":"12","author":"Bryant","year":"2010","journal-title":"Nat. Cell Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1161","DOI":"10.1007\/s00467-012-2337-z","article-title":"Cell polarity and cystic kidney disease","volume":"28","author":"Fedeles","year":"2013","journal-title":"Pediatr. Nephrol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1016\/j.cub.2008.02.076","article-title":"Cell-polarity dynamics controls the mechanism of lumen formation in epithelial morphogenesis","volume":"18","author":"Yu","year":"2008","journal-title":"Curr. Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1011","DOI":"10.1083\/jcb.201203075","article-title":"Cell confinement controls centrosome positioning and lumen initiation during epithelial morphogenesis","volume":"198","author":"Auzan","year":"2012","journal-title":"J. Cell Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"C831","DOI":"10.1152\/ajpcell.00032.2009","article-title":"Abnormalities in focal adhesion complex formation, regulation, and function in human autosomal recessive polycystic kidney disease epithelial cells","volume":"298","author":"Israeli","year":"2010","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1083\/jcb.200407072","article-title":"Gp135\/podocalyxin and NHERF-2 participate in the formation of a preapical domain during polarization of MDCK cells","volume":"168","author":"Meder","year":"2005","journal-title":"J. Cell Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"8:1","DOI":"10.1186\/1471-2091-7-8","article-title":"Identification of glycosylated marker proteins of epithelial polarity in MDCK cells by homology driven proteomics","volume":"7","author":"Fullekrug","year":"2006","journal-title":"BMC Biochem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"8128:1","DOI":"10.1038\/ncomms9128","article-title":"JAM-A regulates cortical dynein localization through Cdc42 to control planar spindle orientation during mitosis","volume":"6","author":"Tuncay","year":"2015","journal-title":"Nat. Commun."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1681\/ASN.2007070770","article-title":"Fibrocystin\/polyductin modulates renal tubular formation by regulating polycystin-2 expression and function","volume":"19","author":"Kim","year":"2008","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e11530:1","DOI":"10.1371\/journal.pone.0011530","article-title":"The vinculin-DeltaIn20\/21 mouse: Characteristics of a constitutive, actin-binding deficient splice variant of vinculin","volume":"5","author":"Marg","year":"2010","journal-title":"PLoS ONE"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"746","DOI":"10.1002\/jcp.22084","article-title":"Contractility modulates cell adhesion strengthening through focal adhesion kinase and assembly of vinculin-containing focal adhesions","volume":"223","author":"Dumbauld","year":"2010","journal-title":"J. Cell Physiol."},{"key":"ref_21","first-page":"7524:1","article-title":"A plastic relationship between vinculin-mediated tension and adhesion complex area defines adhesion size and lifetime","volume":"6","author":"Berge","year":"2015","journal-title":"Nat. Commun."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"19771","DOI":"10.1073\/pnas.0609267103","article-title":"Anisotropy of cell adhesive microenvironment governs cell internal organization and orientation of polarity","volume":"103","author":"Thery","year":"2006","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1","DOI":"10.18637\/jss.v031.i10","article-title":"CircStat: A MATLAB Toolbox for Circular Statistics","volume":"31","author":"Berens","year":"2009","journal-title":"J. Stat. Softw."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"136","DOI":"10.4161\/sgtp.24303","article-title":"Mechanical control of epithelial lumen formation","volume":"4","year":"2013","journal-title":"Small GTPases"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1425","DOI":"10.1242\/jcs.077388","article-title":"Actomyosin-generated tension controls the molecular kinetics of focal adhesions","volume":"124","author":"Wolfenson","year":"2011","journal-title":"J. Cell Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"4652","DOI":"10.1091\/mbc.e09-02-0137","article-title":"Trafficking of Crumbs3 during cytokinesis is crucial for lumen formation","volume":"20","author":"Schluter","year":"2009","journal-title":"Mol. Biol. Cell"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"4398","DOI":"10.1091\/mbc.e04-11-1019","article-title":"Inhibition of Pkhd1 impairs tubulomorphogenesis of cultured IMCD cells","volume":"16","author":"Mai","year":"2005","journal-title":"Mol. Biol. Cell"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"066006:1","DOI":"10.1088\/1478-3975\/ab39fa","article-title":"The transmembrane protein fibrocystin\/polyductin regulates cell mechanics and cell motility","volume":"16","author":"Puder","year":"2019","journal-title":"Phys. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1043","DOI":"10.1083\/jcb.200703036","article-title":"Vinculin controls focal adhesion formation by direct interactions with talin and actin","volume":"179","author":"Humphries","year":"2007","journal-title":"J. Cell Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"489","DOI":"10.1152\/physrev.00021.2007","article-title":"Regulation of actin assembly associated with protrusion and adhesion in cell migration","volume":"88","author":"Carlier","year":"2008","journal-title":"Physiol. Rev."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"3427","DOI":"10.1242\/jcs.206433","article-title":"Actin assembly mechanisms at a glance","volume":"130","author":"Rottner","year":"2017","journal-title":"J. Cell Sci."},{"key":"ref_32","first-page":"3217","article-title":"Integrins and epithelial cell polarity","volume":"127","author":"Lee","year":"2014","journal-title":"J. Cell Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1016\/j.devcel.2016.12.004","article-title":"Polarity Reversal by Centrosome Repositioning Primes Cell Scattering during Epithelial-to-Mesenchymal Transition","volume":"40","author":"Burute","year":"2017","journal-title":"Dev. Cell"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"4708","DOI":"10.1073\/pnas.1011123108","article-title":"Cell-ECM traction force modulates endogenous tension at cell-cell contacts","volume":"108","author":"Maruthamuthu","year":"2011","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"248:1","DOI":"10.3389\/fgene.2020.00248","article-title":"Application and Comparison of Supervised Learning Strategies to Classify Polarity of Epithelial Cell Spheroids in 3D Culture","volume":"11","author":"Soetje","year":"2020","journal-title":"Front. Genet."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1083\/jcb.149.2.317","article-title":"The respective contributions of the mother and daughter centrioles to centrosome activity and behavior in vertebrate cells","volume":"149","author":"Piel","year":"2000","journal-title":"J. Cell Biol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"RESEARCH0034:1-0034:12","DOI":"10.1186\/gb-2002-3-7-research0034","article-title":"Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes","volume":"3","author":"Vandesompele","year":"2002","journal-title":"Genome Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"934","DOI":"10.1354\/vp.43-6-934","article-title":"Selection of reference genes for quantitative real-time PCR analysis in canine mammary tumors using the GeNorm algorithm","volume":"43","author":"Etschmann","year":"2006","journal-title":"Vet. Pathol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1016\/j.ab.2006.06.001","article-title":"Development and evaluation of canine reference genes for accurate quantification of gene expression","volume":"356","author":"Brinkhof","year":"2006","journal-title":"Anal. Biochem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"C13","DOI":"10.1152\/ajpcell.00263.2008","article-title":"Cell confluency-induced Stat3 activation regulates NHE3 expression by recruiting Sp1 and Sp3 to the proximal NHE3 promoter region during epithelial dome formation","volume":"296","author":"Su","year":"2009","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2687","DOI":"10.1002\/j.1460-2075.1984.tb02194.x","article-title":"Development of cell surface polarity in the epithelial Madin-Darby canine kidney (MDCK) cell line","volume":"3","author":"Pfeiffer","year":"1984","journal-title":"EMBO J."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"2377","DOI":"10.1083\/jcb.107.6.2377","article-title":"The polarized distribution of an apical cell surface glycoprotein is maintained by interactions with the cytoskeleton of Madin-Darby canine kidney cells","volume":"107","author":"Ojakian","year":"1988","journal-title":"J. Cell Biol."},{"key":"ref_43","unstructured":"Meijering, E. (2018, February 13). FeatureJ: An ImageJ Plugin Suite for Image Feature Extraction. Available online: https:\/\/imagescience.org\/meijering\/software\/featurej\/."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/14\/5140\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:50:07Z","timestamp":1760176207000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/14\/5140"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,7,20]]},"references-count":43,"journal-issue":{"issue":"14","published-online":{"date-parts":[[2020,7]]}},"alternative-id":["ijms21145140"],"URL":"https:\/\/doi.org\/10.3390\/ijms21145140","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,7,20]]}}}