title: Conditional pulmonary over-expression of Claudin 6 during embryogenesis delays lung morphogenesis.
abstract: Claudin 6 (Cldn6) is a tetraspanin protein expressed by barrier epithelial cells. In order to assess the effects of persistent tight junctions involving Cldn6 during lung development, a doxycycline (dox)-inducible conditional transgenic mouse was generated that up-regulates Cldn6 in the distal lung. Pups had unlimited access to dox from conception until sacrifice date at embryonic day (E) 18.5. Quantitative PCR, immunoblotting, and immunohistochemistry revealed significantly elevated Cldn6 expression in transgenic mice compared to non-transgenic controls. There were no differences in terms of lung size, lung weight, or whole body weight at the time of necropsy. Histological evaluations led to the discovery that E18.5 Cldn6 transgenic pups appeared to be in the early canalicular stage of development coincident with fewer, thickened respiratory airspaces. In contrast, controls appeared to have entered the saccular stage characterized by thin airspace walls and spherical architecture. Immunostaining for transcriptional regulators including TTF-1 and FoxA2 was conducted to assess cell differentiation and specific cell types were identified via staining for pro-surfactant protein C (alveolar type II epithelial cells) or Clara Cell Secretory Protein (cub or Clara cells). Lastly, cell turnover was qualitatively measured via staining for cell proliferation or apoptosis. These data suggest that Cldn6 is an important junctional protein potentially involved in the programming of epithelial cells during lung development. Furthermore, genetic down-regulation of Cldn6 as development proceeds may influence differentiation observed in the transition from the canalicular to the saccular lung.
authors.affiliation: Lung and Placenta Research Laboratory, Brigham Young University, Physiology and Developmental Biology, Provo, Utah, USA.
authors.forename: Felix R
authors.initials: FR
authors.lastname: Jimenez
authors.forename: Samuel T
authors.initials: ST
authors.lastname: Belgique
authors.forename: Joshua B
authors.initials: JB
authors.lastname: Lewis
authors.forename: Scott A
authors.initials: SA
authors.lastname: Albright
authors.forename: Cameron M
authors.initials: CM
authors.lastname: Jones
authors.forename: Brian M
authors.initials: BM
authors.lastname: Howell
authors.forename: Aleksander P
authors.initials: AP
authors.lastname: Mika
authors.forename: Tyson R
authors.initials: TR
authors.lastname: Jergensen
authors.forename: Jason R
authors.initials: JR
authors.lastname: Gassman
authors.forename: Ryan J
authors.initials: RJ
authors.lastname: Morris
authors.forename: Juan A
authors.initials: JA
authors.lastname: Arroyo
authors.forename: Paul R
authors.initials: PR
authors.lastname: Reynolds
mesh_terms: D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D049109:Cell Proliferation; D002478:Cells, Cultured; D057167:Claudins / Q000502:physiology*; D004622:Embryo, Mammalian / Q000378:metabolism / Q000473:pathology*; D005260:Female; D005455:Fluorescent Antibody Technique; D007124:Immunoenzyme Techniques; D047468:Immunoprecipitation; D019426:Integrases / Q000378:metabolism; D008168:Lung / Q000196:embryology* / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D057167:Claudins; D012333:RNA, Messenger; C045073:Cre recombinase; D019426:Integrases; C122101:claudin 6