Abstract
Early gestation lung development is characterized by branching morphogenesis of the airways and basic lung structure formation. Androgens delay late-gestation lung development if the androgen exposure begins in early gestation. We hypothesized that there would be effects of early gestation androgens on lung development. Embryonic mouse lungs (d 11.5) were cultured with dihydrotestosterone (DHT), DHT plus flutamide, or with nothing as controls. Branching morphogenesis was significantly increased after 24, 48, and 72 h of culture. This effect was blocked by simultaneous flutamide treatment. Fetal sex did not influence the DHT response. DHT increased cell proliferation as measured by [3H]thymidine incorporation into DNA. Autoradiography showed prominent [3H]thymidine labeling of epithelia and mesenchyme in regions of new bud formation. DHT treatment significantly increased the thymidine-labeling index of fibroblasts and airway epithelial cells. Programmed cell death, which is found in developing organs in association with cell proliferation during structure formation and tissue remodeling, was studied using terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling assay. In control lungs, programmed cell death occurred in the peripheral mesenchyme surrounding newly forming buds and underlying airway branch points. DHT treatment increased programmed cell death in association with increased branching morphogenesis. Evaluation of near-adjacent sections (control and DHT-treated lungs) showed that apoptotic mesenchymal cells were flanked by [3H]thymidine-labeled fibroblasts and epithelial cells, suggesting a coordination of these processes in the progression of branching morphogenesis. We conclude that androgen enhances the process of early lung morphogenesis by increasing cell proliferation and programmed cell death and by promoting the structural progression of branching morphogenesis.
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Abbreviations
- DHT:
-
dihydrotestosterone
- PCD:
-
programmed cell death
- TUNEL:
-
terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling
- SP-C:
-
surfactant protein C
- E9.5, E11.5, E13.5, E18.5:
-
embryonic day 9.5, 11.5, 13.5, 18.5
- EGF:
-
epidermal growth factor
- KGF:
-
keratinocyte growth factor
References
Warburton D, Seth R, Shum L, Horcher PG, Hall FL, Werb Z, Slavkin HC 1992 Epigenetic role of epidermal growth factor expression and signalling in embryonic mouse lung morphogenesis. Dev Biol 149: 123–133.
Nielsen HC, Martin A, Volpe MV, Hatzis D, Vosatka RJ 1997 Growth factor control of growth and epithelial differentiation in embryonic lungs. Biochem Mol Med 60: 38–48.
Serra R, Pelton RW, Moses HL 1994 TGFβ1 inhibits branching morphogenesis and N-myc expression in lung bud organ cultures. Development 120: 2153–2161.
Minoo P, Hamdan H, Bu D, Warburton D, Stepanik P 1995 TTF-1 regulates lung epithelial morphogenesis. Dev Biol 172: 694–698.
Oshika E, Liu S, Singh G, Michalopoulos GK, Shinozuka H, Katyal SL 1998 Antagonistic effects of dexamethasone and retinoic acid on rat lung morphogenesis. Pediatr Res 43: 315–324.
Goldin GV, Wessells NK 1979 Mammalian lung development: the possible role of cell proliferation in the formation of supernumerary tracheal buds and in branching morphogenesis. J Exp Zool 208: 337–346.
Hjalmarson O 1981 Epidemiology and classification of acute, neonatal respiratory disorders. Acta Paediar Scand 70: 773–783.
Reed DM, Bakketeig LS, Nugent RP 1978 The epidemiology of respiratory distress syndrome in Norway. Am J Epidemiol 107: 299–310.
Nielsen HC, Kirk WO, Sweezey N, Torday JS 1990 Coordination of growth and differentiation in the fetal lung. Exp Cell Res 188: 89–96.
Glucksmann A 1974 Sexual dimorphism in mammals. Biol Rev 49: 423–475.
Adamson IYR, King GM 1984 Sex differences in development of fetal rat lung. Lab Invest 50: 456–468.
Adamson IYR, King GM 1984 Sex differences in development of fetal rat lung. Lab Invest 40: 461–468.
Adamson IYR, King GM 1984 Sex-related differences in cellular composition and surfactant synthesis of developing fetal rat lungs. Am Rev Respir Dis 129: 130–134.
Kotas RV, Avery ME 1980 The influence of sex on fetal rabbit lung maturation and on the response to glucocorticoids. Am Rev Respir Dis 121: 377–380.
Nielsen HC, Torday JS 1981 Sex differences in fetal rabbit pulmonary surfactant production. Pediatr Res 15: 1245–1247.
Torday JS 1984 The sex difference in type II cell surfactant synthesis originates in the fibroblast in vitro. Exp Lung Res 7: 187–194.
Nielsen HC, Zinman HM, Torday JS 1982 Dihydrotestosterone inhibits fetal rabbit pulmonary surfactant production. J Clin Invest 69: 611–616.
Nielsen HC 1986 The development of surfactant synthesis in fetal rabbit lung organ culture exhibits a sex dimorphism. Biochim Biophys Acta 883: 373–379.
Torday JS, Nielsen HC 1987 The sex difference in fetal lung surfactant production. Exp Lung Res 12: 1–19.
Nielsen HC, Torday JS 1983 Anatomy of fetal rabbit gonads and the sexing of fetal rabbits. Lab Anim 17: 148–150.
Kunieda T, Xian M, Kobayashi E, Imamichi T, Moriwaki K, Toyoda Y 1992 Sexing of mouse preimplantation embryos by detection of Y chromosome-specific sequences using polymerase chain reaction. Biol Reprod 46: 692–697.
Burton K 1956 A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochemistry 62: 315–323.
Pointis G, Latreille T, Cedard L 1980 Gonado-pituitary relationships in the fetal mouse at various times during sexual differentiation. J Endocrinol 86: 483–488.
Wasner G, Hennermann I, Kratochwil K 1983 Ontogeny of mesenchymal androgen receptors in the embryonic mouse mammary gland. Endocrinology 113: 1771–1780.
Yokouchi Y, Sakiyama J, Kameda T, Iba H, Suzuki A, Ueno N, Kuroiwa A 1996 BMP-2/-4 mediate programmed cell death in chicken limb buds. Development 122: 3725–3734.
Alarid ET, Rubin JS, Young P, Chedid M, Ron D, Aaronson SA, Cunha GR 1994 Keratinocyte growth factor functions in epithelial induction during seminal vesicle development. Proc Natl Acad Sci USA 91: 1074–1078.
Shannon JM 1994 Induction of alveolar type II cell differentiation in fetal tracheal epithelium by grafted distal lung mesenchyme. Dev Biol 166: 600–614.
Vosatka RJ, Volpe MV, Nielsen HC 1995 Alterations of branching morphogenesis in embryonic lung by EGF and TGFβ analyzed by interpolated cinephotomicrography. Pediatr Res 37: 72Aabstr
Sultan C, Migeon BR, Rothwell SW, Maes M, Zerhouni N, Migeon CJ 1980 Androgen receptors and metabolism in cultured human fetal fibroblasts. Pediatr Res 13: 67–69.
Spooner BS, Wessels NK 1970 Mammalian lung development interactions in primordium formation and branching morphogenesis. J Exp Zool 175: 445–454.
Wessels NK 1970 Mammalian lung development: interactions in formation and morphogenesis of tracheal buds. J Exp Zool 175: 455–466.
Hilfer SR, Rayner RM, Brown JW 1985 Mesenchymal control of branching pattern in the fetal mouse lung. Tissue Cell 17: 523–538.
Wilson JD 1978 Sexual differentiation. Ann Rev Physiol 40: 279–306.
Wilson JD, Griffin JE, Leshin M, George FW 1981 Role of gonadal hormones in development of the sexual phenotypes. Hum Genet 58: 78–84.
Torday JS, Zinman HM, Nielsen HC 1986 Glucocorticoid regulation of DNA, protein and surfactant phospholipid in developing lung: temporal relationship between growth and development. Dev Pharmacol Ther 9: 124–131.
McMillan EM, King GM, Adamson IYR 1989 Sex hormones influence growth and surfactant production in fetal lung explants. Exp Lung Res 15: 167–179.
Finch PW, Rubin JS, Miki T, Ron D, Aaronson SA 1989 Human KGF is FGF-related with properties of a paracrine effector of epithelial cell growth. Science 245: 752–755.
Peters K, Werner S, Liao X, Wert S, Whitsett J, Williams L 1994 Targeted expression of a dominant negative FGF receptor blocks branching morphogenesis and epithelial differentiation of the mouse lung. EMBO J 13: 3296–3301.
Shiratori M, Oshika E, Ung LP, Singh G, Shinozuka H, Warburton D, Michalopoulos G 1996 Keratinocyte growth factor and embryonic rat lung morphogenesis. Am J Respir Cell Mol Biol 15: 328–338.
Cardoso WV, Itoh A, Nogawa H, Mason I, Brody JS 1997 FGF-1 and FGF-7 induce distinct patterns of growth and differentiation in embryonic lung epithelium. Dev Dyn 208: 398–405.
Schuger L, Skubitz A, O'Shae KS, Chang JF, Varani J 1991 Identification of laminin domains involved in branching morphogenesis: effects of anti-laminin monoclonal antibodies on mouse embryonic lung development. Dev Biol 146: 531–541.
Sanders EJ, Wride MA 1995 Programmed cell death in development. Int Rev Cytol 163: 105–173.
Kresch MJ, Christian C, Wu F, Hussain N 1998 Ontogeny of apoptosis during lung development. Pediatr Res 43: 426–431.
Catlin EA, Tonnu VC, Ebb RG, Pacheco BA, Manganaro TV, Ezzell RM, Donahoe PK 1997 Müllerian inhibiting substance inhibits branching morphogenesis and induces apoptosis in fetal rat lung. Endocrinology 138: 790–796.
Jacobson MD, Weil M, Raff MC 1997 Programmed cell death in animal development. Cell 88: 347–354.
Furuya Y, Isaacs JT 1993 Differential gene regulation during programmed death (apoptosis) versus proliferation of prostatic glandular cells induced by androgen manipulation. Endocrinology 133: 2660–2666.
Meikrantz W, Schlegel R 1995 Apoptosis and the cell cycle. J Cell Biochem 58: 160–174.
King KL, Cidlowski JA 1995 Cell cycle and apoptosis: common pathways to life and death. J Cell Biochem 58: 175–180.
Kellogg CK, Nielsen HC, Cochran B 1991 C-myc expression in fetal lung development suggests a positive regulatory role in lung growth. Am Rev Respir Dis 143: 159Aabstr
Schmid P, Schulz WA, Hameister H 1989 Dynamic expression pattern of the myc proto-oncogene in midgestation mouse embryos. Science 243: 226–229.
Hirning U, Schmid P, Schulz WA, Rettenberger G, Hameister H 1991 A comparative analysis of N-myc and C-myc expression and cellular proliferation in mouse organogenesis. Mech Dev 33: 119–126.
Evan GI, Wyllie AH, Gilbert CS, Littlewood TD, Land H, Brooks M, Waters CM 1992 Induction of apoptosis in fibroblasts by c-myc protein. Cell 69: 119–128.
Hoang AT, Cohen KJ, Barrett JF, Bergstrom DA, Dang CV 1994 Participation of cyclin A in myc-induced apoptosis. Proc Natl Acad Sci USA 91: 6875–6879.
Hueber AO, Zornig M, Lyon D, Suda T, Nagata S, Evans GI 1997 Requirement for the CD95 receptor-ligand pathway in C-myc-induced apoptosis. Science 278: 1305–1309.
Acknowledgements
The authors thank Dr. MaryAnn V. Volpe and Cynthia A. Doyle for help and advice.
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Supported by National Institutes of Health grant HL37930.
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Levesque, B., Vosatka, R. & Nielsen, H. Dihydrotestosterone Stimulates Branching Morphogenesis, Cell Proliferation, and Programmed Cell Death in Mouse Embryonic Lung Explants. Pediatr Res 47, 481–491 (2000). https://doi.org/10.1203/00006450-200004000-00012
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DOI: https://doi.org/10.1203/00006450-200004000-00012
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