Scribble is required for normal epithelial cell–cell contacts and lumen morphogenesis in the mammalian lung
Authors: Laura L Yates
Helen Hilton
Lauren Chessum
Carsten Schnatwinkel
Andy Greenfield
Caroline Formstone
Jeremy Sanderson
Jennifer N Murdoch
Lee A Niswander
Lee Hazelwood
M Rosario Romero
Charlotte H Dean
Anju Paudyal
Jonathan Wilde
Debora Bogani
Journal: Developmental Biology (2013)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: lung development; epithelial cell arrangements; arborized network; lumen diameter; adherens junctions
Affiliations: Mammalian Genetics Unit, Medical Research Council, Harwell, UK.

Abstract:  During lung development, proper epithelial cell arrangements are critical for the formation of an arborized network of tubes. Each tube requires a lumen, the diameter of which must be tightly regulated to enable optimal lung function. Lung branching and lumen morphogenesis require close epithelial cell-cell contacts that are maintained as a result of adherens junctions, tight junctions and by intact apical-basal (A/B) polarity. However, the molecular mechanisms that maintain epithelial cohesion and lumen diameter in the mammalian lung are unknown. Here we show that Scribble, a protein implicated in planar cell polarity (PCP) signalling, is necessary for normal lung morphogenesis. Lungs of the Scrib mouse mutant Circletail (Crc) are abnormally shaped with fewer airways, and these airways often lack a visible, 'open' lumen. Mechanistically we show that Scrib genetically interacts with the core PCP gene Vangl2 in the developing lung and that the distribution of PCP pathway proteins and Rho mediated cytoskeletal modification is perturbed in Scrib(Crc/Crc) lungs. However A/B polarity, which is disrupted in Drosophila Scrib mutants, is largely unaffected. Notably, we find that Scrib mediates functions not attributed to other PCP proteins in the lung. Specifically, Scrib localises to both adherens and tight junctions of lung epithelia and knockdown of Scrib in lung explants and organotypic cultures leads to reduced cohesion of lung epithelial cells. Live imaging of Scrib knockdown lungs shows that Scrib does not affect bud bifurcation, as previously shown for the PCP protein Celsr1, but is required to maintain epithelial cohesion. To understand the mechanism leading to reduced cell-cell association, we show that Scrib associates with β-catenin in embryonic lung and the sub-cellular distribution of adherens and tight junction proteins is perturbed in mutant lung epithelia. Our data reveal that Scrib is required for normal lung epithelial organisation and lumen morphogenesis by maintaining cell-cell contacts. Thus we reveal novel and important roles for Scrib in lung development operating via the PCP pathway, and in regulating junctional complexes and cell cohesion. 

Retracted: No

PubPeer comments:
 This article was selected for a replication project that replicated social science studies published in Science and Nature. The main finding did not replicate in a much larger sample. Thus, this article should not be cited as evidence for the claims made in this article. Camerer CF, Dreber A, Holzmeister F, Ho T-H, Huber J, Johannesson M, et al. Evaluating the replicability of social science experiments in Nature and Science between 2010 and 2015. Nat Hum Behav. 2018; 2: 637u2013644. https: doi.org 10.1038 s41562-018-0399-z Peers also had low confidence in the replicability of this study (33%). Gordon M, Viganola D, Dreber A, Johannesson M, Pfeiffer T (2021) Predicting replicabilityu2014Analysis of survey and prediction market data from large-scale forecasting projects. PLoS ONE 16(4): e0248780. https: doi.org 10.1371 journal.pone.0248780 ,