Immunohistochemical detection and regulation of α5 nicotinic acetylcholine receptor (nAChR) subunits by FoxA2 during mouse lung organogenesis

Respiratory Research

Paul R Reynolds¹  Charles P Willnauer¹  Alex J Geyer¹  Benjamin R Bukey¹  Jason L Porter¹ 

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

关键词: nAChR;    lung;    epithelium;    development;    alpha 5;   

Others  :  796840
DOI  :  10.1186/1465-9921-12-82

 received in 2011-05-13, accepted in 2011-06-17,  发布年份 2011

【 摘 要 】

Background

α₅ nicotinic acetylcholine receptor (nAChR) subunits structurally stabilize functional nAChRs in many non-neuronal tissue types. The expression of α₅ nAChR subunits and cell-specific markers were assessed during lung morphogenesis by co-localizing immunohistochemistry from embryonic day (E) 13.5 to post natal day (PN) 20. Transcriptional control of α₅ nAChR expression by FoxA2 and GATA-6 was determined by reporter gene assays.

Results

Steady expression of α₅ nAChR subunits was observed in distal lung epithelial cells during development while proximal lung expression significantly alternates between abundant prenatal expression, absence at PN4 and PN10, and a return to intense expression at PN20. α₅ expression was most abundant on luminal edges of alveolar type (AT) I and ATII cells, non-ciliated Clara cells, and ciliated cells in the proximal lung at various periods of lung formation. Expression of α₅ nAChR subunits correlated with cell differentiation and reporter gene assays suggest expression of α₅ is regulated in part by FoxA2, with possible cooperation by GATA-6.

Conclusions

Our data reveal a highly regulated temporal-spatial pattern of α₅ nAChR subunit expression during important periods of lung morphogenesis. Due to specific regulation by FoxA2 and distinct identification of α₅ in alveolar epithelium and Clara cells, future studies may identify possible mechanisms of cell differentiation and lung homeostasis mediated at least in part by α₅-containing nAChRs.

【 授权许可】

   
2011 Porter et al; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Hogan BL, Kolodziej PA: Organogenesis: Molecular Mechanisms of Tubulogenesis. Nat Rev Genet 2002, 3.7:513-23.
• [2]Zhou L, Dey CR, Wert SE, Yan C, Costa RH, Whitsett JA: Hepatocyte Nuclear Factor-3 beta Limits Cellular Diversity in the Developing Respiratory Epithelium and Alters Lung Morphogenesis in Vivo. Dev Dyn 1997, 210.3:305-14.
• [3]Liu C, Morrisey EE, Whitsett JA: GATA-6 is required for maturation of the lung in late gestation. Am J Physiol-Lung C 2002, 283:L468-L475.
• [4]Costa RH, Kalinichenko V, Lim L: Transcription Factors in Mouse Lung Development and Function. Am J Physiol-Lung C 2001, 280.5:L823-38.
• [5]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, 14.9:5671-81.
• [6]Ikeda K, Shaw-White JR, Wert SE, Whitsett JA: Hepatocyte Nuclear Factor 3 Activates Transcription of Thyroid Transcription Factor 1 in Respiratory Epithelial Cells. Mol Cell Biol 1996, 16.7:3626-36.
• [7]Wan H, Dingle S, Xu Y, Besnard V, Kaestner KH, Ang SL, Wert S, Stahlman MT, Whitsett JA: Compensatory roles of Foxa1 and Foxa2 during lung morphogenesis. J Biol Chem 2005, 280:13809-13816.
• [8]Wan H, Kaestner KH, Ang SL, Ikegami M, Finkelman FD, Stahlman MT, Fulkerson PC, Rothenberg ME, Whitsett JA: Foxa2 regulates alveolarization and goblet cell hyperplasia. Development 2004, 131:953-964.
• [9]Wan H, Xu Y, Ikegami M, Stahlman MT, Kaestner KH, Ang SL, Whitsett JA: Foxa2 is required for transition to air breathing at birth. Proc Natl Acad Sci USA 2004, 101:14449-14454.
• [10]Morrisey EE, Tang Z, Sigrist K, Lu MM, Jiang F, Ip HS, Parmacek MS: GATA-6 regulates HNF4 and is required for differentiation of visceral endoderm in the mouse embryo. Genes Dev 1998, 12:3579-3590.
• [11]Yang H, Lu MM, Zhang L, Whitsett JA, Morrisey EE: GATA-6 regulates differentiation of distal lung epithelium. Development 2002, 129:2233-2246.
• [12]Koutsourakis M, Keijzer R, Visser P, Post M, Tibboel D, Grosveld F: Branching and differentiation defects in pulmonary epithelium with elevated Gata6 expression. Mech Dev 2001, 105:105-114.
• [13]Lindstrom J: Nicotinic Acetylcholine Receptors in Health and Disease. Mol Neurobiol 1997, 15.2:193-222.
• [14]Maouche K, Polette M, Jolly T, Medjber K, Cloëz-Tayarani I, Changeux JP, Burlet H, Terryn C, Coraux C, Zahm JM, Birembaut P, Tournier JM: {Alpha}7 Nicotinic Acetylcholine Receptor Regulates Airway Epithelium Differentiation by Controlling Basal Cell Proliferation. Am J Pathol 2009, 175.5:1868-82.
• [15]Wessler I, Kirkpatrick CJ: Acetylcholine Beyond Neurons: The Non-Neuronal Cholinergic System in Humans. Brit J Pharmacol 2008, 154.8:1558-71.
• [16]Bertrand D, Changeux J: Orthodontic correction of maxillary flaring using provisional restorations. Neuroscience 1995, 7:75-90.
• [17]Paleari L, Grozio A, Cesario A, Russo P: The Cholinergic System and Cancer. Semin Cancer Biol 2008, 18.3:211-7.
• [18]Rogers SW, Mandelzys A, Deneris ES, Cooper E, Heinemann S: The expression of nicotinic acetylcholine receptors by PC12 cells treated with NGF. J Neurosci 1992, 12(12):4611-4623.
• [19]Reynolds PR, Mucenski ML, Whitsett JA: Thyroid Transcription Factor (TTF) -1 Regulates the Expression of Midkine (MK) during Lung Morphogenesis. Dev Dyn 2003, 227.2:227-37.
• [20]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, 280.37:32548-54.
• [21]Reynolds PR, Allisson CH, Willnauer CP: TTF-1 regulates alpha5 nicotinic acetylcholine receptor (nAChR) subunits in proximal and distal lung epithelium. Respir Res 2010, 11:175. BioMed Central Full Text
• [22]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(4):333-45.
• [23]Warburton D, Schwarz , Tefft D, Flores-Delgado G, Anderson KD, Cardoso WV: The molecular basis of lung morphogenesis. Mech Develop 2000, 92:55-81.
• [24]Maus AD, Pereira EF, Karachunski PI, Horton RM, Navaneetham D, Macklin K, Cortes WS, Albuquerque EX, Conti-Fine BM: Human and Rodent Bronchial Epithelial Cells Express Functional Nicotinic Acetylcholine Receptors. Mol Pharm 1998, 54.5:779-88.
• [25]Zia S, Ndoye A, Nguyen VT, Grando SA: Nicotine Enhances Expression of the Alpha 3, Alpha 4, Alpha 5, and Alpha 7 Nicotinic Receptors Modulating Calcium Metabolism and Regulating Adhesion and Motility of Respiratory Epithelial Cells. Res Commun Mol Path 1997, 97.3:243-62.
• [26]Wang Y, Pereira EF, Maus AD, Ostlie NS, Navaneetham D, Lei S, Albuquerque EX, Conti-Fine BM: Human Bronchial Epithelial and Endothelial Cells Express alpha7 Nicotinic Acetylcholine Receptors. Mol Pharacol 2001, 60.6:1201-9.
• [27]Lazzaro D, Price M, de Felice M, Di Lauro R: The Transcription Factor TTF-1 is Expressed at the Onset of Thyroid and Lung Morphogenesis and in Restricted Regions of the Foetal Brain. Development 1991, 113.4:1093-104.
• [28]Zhou L, Lim L, Costa RH, Whitsett JA: Thyroid Transcription Factor-1, Hepatocyte Nuclear Factor-3 beta, Surfactant Protein B, C, and Clara Cell Secretory Protein in Developing Mouse Lung. J Histochem Cytochem 1996, 44.10:1183-93.
• [29]Kimura S, Hara Y, Pineau T, Fernandez-Salguero P, Fox CH, Ward JM, Gonzalez 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, 10.1:60-9.
• [30]Besnard V, Wert SE, Hull WM, Whitsett JA: Immunohistochemical Localization of Foxa1 and Foxa2 in Mouse Embryos and Adult Tissues. Gene Exp Patterns 2004, 193-208.
• [31]Maeda Y, Vrushank D, Whitsett JA: Transcriptional Control of Lung Morphogenesis. Physiol Rev 2007, 87.1:219-44.
• [32]Gomperts BN, Xiulan G, Hackett BP: Foxj1 Regulates Basal Body Anchoring to the Cytoskeleton of Ciliated Pulmonary Epithelial Cells. J Cell Sci 2004, 117:1329-37.
• [33]Huang T, You Y, Spoor MS, Richer EJ, Kudva VV, Paige RC, Seiler MP, Liebler JM, Zabner J, Plopper CG, Brody SL: Foxj1 is Required for Apical Localization of Ezrin in Airway Epithelial Cells. J Cell Sci 2003, 116:4935-45.
• [34]Reynolds SD, Malkinson AM: Clara Cell: Progenitor for the Bronchiolar Epithelium. Int J Biochem Cell B 2010, 42.1:1-4.
• [35]Hsia SH, Schulman SR, Meliones JN, Canada AT, Chen SC: Effects of Maternal Nicotine Exposure on Branching Morphogenesis of Mouse Fetal Lung: In Vivo and in Vitro Studies. Acta Paediatr Tai 2003, 44.3:150-4.
• [36]Taylor B, Wadsworth J: Maternal Smoking during Pregnancy and Lower Respiratory Tract Illness in Early Life. Arch Dis Child 1987, 62.8:786-91.
• [37]Tager IB, Hanrahan JP, Tosteson TD, Castile RG, Brown RW, Weiss ST, Speizer FE: Lung Function, Pre- and Post-Natal Smoke Exposure, and Wheezing in the First Year of Life. Am Rev Respir Dis 1993, 147.4:811-7.
• [38]Sandberg K, Poole SD, Hamdan A, Arbogast P, Sundell HW: Altered Lung Development After Prenatal Nicotine Exposure in Young Lambs. Pediatr Res 2004, 56.3:432-9.
• [39]Carlisle DL, Hopkins TM, Gaither-Davis A, Silhanek MJ, Luketich JD, Christie NA, Siegfried JM: Nicotine signals through muscle-type and neuronal nicotinic acetylcholine receptors in both human bronchial epithelial cells and airway fibroblasts. Respir Res 2004, 5:27. BioMed Central Full Text