Developmental lung expression and transcriptional regulation of Claudin-6 by TTF-1, Gata-6, and FoxA2

Respiratory Research

Paul R Reynolds¹  Tyler T Wood¹  Samuel T Belgique¹  Joshua B Lewis¹  Felix R Jimenez¹ 

[1] Department of Physiology and Developmental Biology, Brigham Young University, 3054 Life Sciences Building, Provo, UT 84602, USA

关键词: Immunofluorescence;    Transcription;    Claudin-6;    Lung;   

Others  :  809175
DOI  :  10.1186/1465-9921-15-70

 received in 2014-03-11, accepted in 2014-06-23,  发布年份 2014

【 摘 要 】

Background

Claudins are transmembrane proteins expressed in tight junctions that prevent paracellular transport of extracellular fluid and a variety of other substances. However, the expression profile of Claudin-6 (Cldn6) in the developing lung has not been characterized.

Methods and results

Cldn6 expression was determined during important periods of lung organogenesis by microarray analysis, qPCR and immunofluorescence. Expression patterns were confirmed to peak at E12.5 and diminish as lung development progressed. Immunofluorescence revealed that Cldn6 was detected in cells that also express TTF-1 and FoxA2, two critical transcriptional regulators of pulmonary branching morphogenesis. Cldn6 was also observed in cells that express Sox2 and Sox9, factors that influence cell differentiation in the proximal and distal lung, respectively. In order to assess transcriptional control of Cldn6, 0.5, 1.0, and 2.0-kb of the proximal murine Cldn6 promoter was ligated into a luciferase reporter and co-transfected with expression vectors for TTF-1 or two of its important transcriptional co-regulators, FoxA2 and Gata-6. In almost every instance, TTF-1, FoxA2, and Gata-6 activated gene transcription in cell lines characteristic of proximal airway epithelium (Beas2B) and distal alveolar epithelium (A-549).

Conclusions

These data revealed for the first time that Cldn6 might be an important tight junctional component expressed by pulmonary epithelium during lung organogenesis. Furthermore, Cldn6-mediated aspects of cell differentiation may describe mechanisms of lung perturbation coincident with impaired cell junctions and abnormal membrane permeability.

【 授权许可】

   
2014 Jimenez et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Burri PH: Structural aspects of postnatal lung development – alveolar formation and growth. Neonatol 2006, 89:313-322.
• [2]Schneeberger EE, Lynch RD: The tight junction: a multifunctional complex. Am J Physiol Cell Physiol 2004, 286:C1213-C1228.
• [3]Balkovetz DF: Claudins at the gate: determinants of renal epithelial tight junction paracellular permeability. Am J Physiol Renal Physiol 2006, 290:F572-F579.
• [4]Harhaj NS, Antonetti DA: Regulation of tight junctions and loss of barrier function in pathophysiology. Int J Biochem Cell Biol 2004, 36:1206-1237.
• [5]Aijaz S, Balda MS, Matter K: Tight junctions: molecular architecture and function. Int Rev Cytol 2006, 248:261-298.
• [6]Chiba H, Osanai M, Murata M, Kojima T, Sawada N: Transmembrane proteins of tight junctions. Biochim Biophys Acta 2008, 1778:588-600.
• [7]Will C, Fromm M, Muller D: Claudin tight junction proteins: novel aspects in paracellular transport. Perit Dial Int 2008, 28:577-584.
• [8]Findley MK, Koval M: Regulation and roles for claudin-family tight junction proteins. IUBMB Life 2009, 61:431-437.
• [9]Turksen K, Troy TC: Barriers built on claudins. J Cell Sci 2004, 117:2435-2447.
• [10]Lal-Nag M, Morin PJ: The claudins. Genome Biol 2009, 10:235.
• [11]Van Itallie CM, Anderson JM: Claudins and epithelial paracellular transport. Annu Rev Physiol 2006, 68:403-429.
• [12]Turksen K, Troy TC: Claudin-6: a novel tight junction molecule is developmentally regulated in mouse embryonic epithelium. Dev Dyn 2001, 222:292-300.
• [13]Arabzadeh A, Troy T-C, Turksen K: Role of the Cldn6 cytoplasmic tail domain in membrane targeting and epidermal differentiation in vivo. Mol Cell Biol 2006, 26:5876-5887.
• [14]Turksen K, Troy TC: Permeability barrier dysfunction in transgenic mice overexpressing claudin 6. Development 2002, 129:1775-1784.
• [15]Quan C, Lu SJ: Identification of genes preferentially expressed in mammary epithelial cells of Copenhagen rat using subtractive hybridization and microarrays. Carcinogenesis 2003, 24:1593-1599.
• [16]Morin PJ: Claudin proteins in human cancer: promising new targets for diagnosis and therapy. Cancer Res 2005, 65:9603-9606.
• [17]Kominsky SL: Claudins: emerging targets for cancer therapy. Exp Rev Mol Med 2006, 8:1-11.
• [18]Wu Q, Liu Y, Ren Y, Xu X, Yu L, Li Y, Quan C: Tight junction protein, claudin-6, downregulates the malignant phenotype of breast carcinoma. Eur J Cancer Prev 2010, 19:186-194.
• [19]Reynolds P, Mucenski M, Whitestt JA: Thyroid transcription factor (TTF)-1 regulates the expression of Midkine (MK) during lung morphogenesis. Dev Dyn 2003, 227:227-237.
• [20]Reynolds PR, Mucenski ML, LeCras TD, Nichols WC, Whitsett JA: Midkine (MK) induces myocardin during hypoxia and causes pulmonary vascular remodeling. J Biol Chem 2004, 279(35):37124-37132.
• [21]Xu Y, Wang Y, Besnard V, Ikegami M, Wert SE, Heffner C, Murray SA, Donahue LR, Whitsett JA: Transcriptional programs controlling perinatal lung maturation. PLoS One 2012, 7(8):e37046.
• [22]Reynolds PR, Allison C, Willnauer C: TTF-1 regulates alpha5 nicotinic acetylcholine receptor (nAChR) subunits in proximal and distal lung epithelium. Respir Res 2010, 11:175.
• [23]Porter JL, Bukey BR, Geyer AJ, Willnauer CP, Reynolds PR: Immunohistochemical detection and regulation of α5 nicotinic acetylcholine receptor (nAChR) subunits by FoxA2 during mouse lung organogenesis. Respir Res 2011, 12:82.
• [24]Sporty JL, Horalkova L, Ehrhardt C: In vitro cell culture models for the assessment of pulmonary drug disposition. Expert Opin Drug Metab Toxicol 2008, 4:333-345.
• [25]Reynolds PR, Hoidal JR: Temporal-spatial expression and transcriptional regulation of alpha7 nicotinic acetylcholine receptor by thyroid transcription factor-1 and early growth response factor-1 during murine lung development. J Biol Chem 2005, 280:32548-32554.
• [26]Guazzi S, Prince M, De Felice M, Damante G, Mattei MG, Di Lauro R: Thyroid nuclear factor 1 (TTF-1) contains a homeodomain and displays a novel DNA binding specificity. EMBO J 1990, 9(11):3631-3639.
• [27]Bohinski RJ, Di Lauro R, Whitsett JA: The lung-specific surfactant protein B gene promoter is a target for thyroid transcription factor 1 and hepatocyte nuclear factor 3, indicating common factors for organ-specific gene expression along the foregut axis. Mol Cell Biol 1994, 14(9):5671-5681.
• [28]Costa RH, Kalinichenko VV, Lim L: Transcription factors in mouse lung development and function. Am J Physiol Lung Cell Mol Physiol 2001, 280:L823-L838.
• [29]Ang SL, Rossant J: HNF-3 beta is essential for node and notochord formation in mouse development. Cell 1994, 78:561-574.
• [30]Besnard V, Wert SE, Hull WM, Whitsett JA: Immunohistochemical localization of Foxa1 and Foxa2 in mouse embryos and adult tissues. Gene Expr Patterns 2004, 5:193-208.
• [31]Hashimoto S, Chen H, Que J, Brockway BL, Drake JA, Snyder JC, Randell SH, Stripp BR: B-catenin-SOX2 signaling regulates the fate of developing airway epithelium. J Cell Sci 2011, 125:932-942.
• [32]Bowles J, Schepers G, Koopman P: Phylogeny of the SOX family of developmental transcription factors based on sequence and structural indicators. Dev Biol 2000, 227:239-255.
• [33]Okubo T, Knoepfler PS, Eisenman RN, Hogan BL: Nmyc plays an essential role during lung development as a dosage-sensitive regulator of progenitor cell proliferation and differentiation. Development 2005, 132:1363-1374.
• [34]Perl AK, Kist R, Shan Z, Scherer G, Whitsett JA: Normal lung development and function after Sox9 inactivation in the respiratory epithelium. Genesis 2005, 41:23-32.
• [35]Morrisey EE, Tang Z, Sigrist K, Lu MM, Jiang F, Ip HS, Parmacek MS: GATA6 regulates HNF4 and is required for differentiation of visceral endoderm in the mouse embryo. Genes Dev 1998, 12:3579-3590.
• [36]Liu C, Morrisey EE, Whitsett JA: GATA-6 is required for maturation of the lung in late gestation. Am J Physiol Lung Cell Mol Physiol 2002, 283:L468-L475.
• [37]Moriwaki K, Tsukita S, Feruse M: Tight junctions containing claudins 4 and 6 are essential for blastocyst formation in preimplantation mouse embryos. Dev Biol 2007, 312:509-522.
• [38]Zhang J, Liss M, Wolburg H, Blasig IE, Abdelilah-Seyfried S: Involvement of claudins in zebrafish brain ventricle morphogenesis. Ann NY Acad Sci 2012, 1257:193-198.
• [39]Lei Z, Maeda T, Tamura A, Nakahama T, Yamazaki Y, Shiratori H, Yashiro K, Tsukita A, Hamada H: EpCAM contributes to formation of functional tight junction in the intestinal epithelium by recruiting claudin proteins. Dev Biol 2012, 371(2):136-145.
• [40]Westmoreland JJ, Drosos Y, Kelly J, Ye J, Means AL, Washington MK, Sosa-Pineda B: Dynamic distribution of claudin proteins in pancreatic epithelia undergoing morphogenesis or neoplastic transformation. Dev Dyn 2012, 241(3):583-594.
• [41]Haddad N, El Andalousi J, Khairaallah H, Yu M, Ryan AK, Gupta IR: The tight junction protein claudin-3 shows conserved expression in the nephric duct and ureteric bud and promotes tubulogenesis in vitro. Am J Physiol Renal Physiol 2011, 310(5):F1057-F1065.
• [42]Volckaert T, Campbell A, Dill E, Li C, Minoo P, De Langhe S: Localized Fgf10 expression is not required for lung branching morphogenesis but prevents differentiation of epithelial progenitors. Development 2013, 140(18):3731-3742.
• [43]Lu J, Zhang S, Nakano H, Simmons DG, Wang S, Kong S, Wang Q, Shen L, Tu Z, Wang W, Wang B, Wang H, Wang Y, van Es JH, Clevers H, Leone G, Cross JC, Wang H: A positive feedback loop involving Gcm1 and Fzd5 directs chorionic branching morphogenesis in the placenta. PLoS Biol 2013, 11(4):e1001536.
• [44]Maeda Y, Dave V, Whitsett JA: Transcriptional control of lung morphogenesis. Physiol Rev 2007, 87:219-244.
• [45]Perl AKT, Whitsett JA: Molecular mechanisms controlling lung morphogenesis. Clin Genet 1999, 56:14-27.
• [46]Kimura S, Hara Y, Pineau T, Fernandez-Salguero P, Fox CH, Ward JM, Gonzales FJ: The T/EBP null mouse: thyroid specific enhancer-binding protein is essential for the organogenesis of the thyroid, lung, ventral forebrain and pituitary. Genes Dev 1996, 10:60-69.
• [47]Zhou L, Lim L, Costa RH, Whitsett JA: Thyroid transcription factor-1 hepatocyte nuclear factor-3beta, surfactant protein B, C, and Clara cell secretory protein in developing mouse lung. J Histochem Cytochem 1996, 44:1183-1193.
• [48]Keijzer R, van Tuyl M, Meijers C, Post M, Tibboel D, Grosveld F, Koutsourakis M: The transcription factor Gata-6 is essential for branching morphogenesis and epithelial cell differentiation during fetal pulmonary development. Development 2001, 128:503-511.