Download PDF
Eur J Pediatr Surg 2012; 22(02): 148-156
DOI: 10.1055/s-0032-1308700
Original Article
Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

Compensatory Lung Growth in NOS3 Knockout Mice Suggests Synthase Isoform Redundancy

Stefan Pokall
1   Division of Pediatric General, Thoracic, and Fetal Surgery, Center for Molecular Fetal Therapy, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States
2   Department of Cardiovascular Surgery, German Heart Center Munich, Technical University Munich, Munich, Bavaria, Germany
,
Arturo R. Maldonado
1   Division of Pediatric General, Thoracic, and Fetal Surgery, Center for Molecular Fetal Therapy, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States
3   Division of Biomedical Informatics, Center for Computational Medicine, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States
,
Charles A. Klanke
1   Division of Pediatric General, Thoracic, and Fetal Surgery, Center for Molecular Fetal Therapy, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States
,
Shuichi Katayama
1   Division of Pediatric General, Thoracic, and Fetal Surgery, Center for Molecular Fetal Therapy, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States
,
Lee M. Morris
1   Division of Pediatric General, Thoracic, and Fetal Surgery, Center for Molecular Fetal Therapy, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States
,
Jose F. Vuletin
1   Division of Pediatric General, Thoracic, and Fetal Surgery, Center for Molecular Fetal Therapy, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States
,
Foong-Yen Lim
1   Division of Pediatric General, Thoracic, and Fetal Surgery, Center for Molecular Fetal Therapy, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States
,
Timothy M. Crombleholme
1   Division of Pediatric General, Thoracic, and Fetal Surgery, Center for Molecular Fetal Therapy, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States
› Author Affiliations
Further Information

Publication History

10 October 2011

07 November 2011

Publication Date:
19 April 2012 (online)

    Permissions and Reprints

Abstract

Nitric oxide synthase 3 (NOS3) produces nitric oxide (NO) in endothelial cells, which stimulates cyclic guanosine monophosphate (cGMP) production and thereby mediates pulmonary vasodilation. Inhibition of cGMP enzymatic cleavage by sildenafil might be involved in lung growth stimulating processes in pulmonary hypoplasia. The aim of this study was to discover insights into the transcriptional regulation of NOS3 in a mouse model of compensatory lung growth (CLG). CLG was studied in wild type animals (WT) and NOS3 knockout mice (NOS3−/−) by dry weight, DNA, and protein quantification as well as relative quantification of NOS mRNA. All assessments were done on adult female mice, 10 days after left pneumonectomy (PNX) or sham thoracotomy. Weight ratios of right NOS3−/− lungs were no different than controls. There was a compensatory increase in DNA and a noncompensating increase in protein ratios in NOS3−/− mice compared with controls. Pharmacological knockdown with the pan-NOS inhibitor l-NAME (nitro-arginine methyl ester) reduced CLG by only 8% compared with the d-NAME treated control mice. Relative quantification of lung mRNA revealed no up-regulation of NOS3 expression in WT lungs after PNX, but NOS3−/− lungs showed a 2.6-fold higher inducible NOS2 expression compared with shams. These data suggest that NOS3 loss of function alone does not impair CLG in mice, possibly because of redundancy mechanisms involving NOS2.

Keywords

compensatory lung growth - nitric oxide synthase - lung growth - nitric oxide
 

  • References

  • 1 Laros CD, Westermann CJ. Dilatation, compensatory growth, or both after pneumonectomy during childhood and adolescence. A thirty-year follow-up study. J Thorac Cardiovasc Surg 1987; 93 (4) 570-576
    CrossrefPubMedGoogle Scholar
  • 2 Fehrenbach H, Voswinckel R, Michl V , et al. Neoalveolarisation contributes to compensatory lung growth following pneumonectomy in mice. Eur Respir J 2008; 31 (3) 515-522
    CrossrefPubMedGoogle Scholar
  • 3 Macchiarini P, Jungebluth P, Go T , et al. Clinical transplantation of a tissue-engineered airway. Lancet 2008; 372 (9655) 2023-2030
    CrossrefPubMedGoogle Scholar
  • 4 Price AP, England KA, Matson AM, Blazar BR, Panoskaltsis-Mortari A. Development of a decellularized lung bioreactor system for bioengineering the lung: the matrix reloaded. Tissue Eng Part A 2010; 16 (8) 2581-2591
    CrossrefPubMedGoogle Scholar
  • 5 Voswinckel R, Motejl V, Fehrenbach A , et al. Characterisation of post-pneumonectomy lung growth in adult mice. Eur Respir J 2004; 24 (4) 524-532
    CrossrefPubMedGoogle Scholar
  • 6 Brody JS. Time course of and stimuli to compensatory growth of the lung after pneumonectomy. J Clin Invest 1975; 56 (4) 897-904
    CrossrefPubMedGoogle Scholar
  • 7 Cagle PT, Langston C, Goodman JC, Thurlbeck WM. Autoradiographic assessment of the sequence of cellular proliferation in postpneumonectomy lung growth. Am J Respir Cell Mol Biol 1990; 3 (2) 153-158
    CrossrefPubMedGoogle Scholar
  • 8 Hoffman AM, Shifren A, Mazan MR , et al. Matrix modulation of compensatory lung regrowth and progenitor cell proliferation in mice. Am J Physiol Lung Cell Mol Physiol 2010; 298 (2) L158-L168
    CrossrefPubMedGoogle Scholar
  • 9 Hsia CC, Wu EY, Wagner E, Weibel ER. Preventing mediastinal shift after pneumonectomy impairs regenerative alveolar tissue growth. Am J Physiol Lung Cell Mol Physiol 2001; 281 (5) L1279-L1287
    PubMedGoogle Scholar
  • 10 Hsia CC, Zhou XS, Bellotto DJ, Hagler HK. Regenerative growth of respiratory bronchioles in dogs. Am J Physiol Lung Cell Mol Physiol 2000; 279 (1) L136-L142
    PubMedGoogle Scholar
  • 11 McBride JT. Lung volumes after an increase in lung distension in pneumonectomized ferrets. J Appl Physiol 1989; 67 (4) 1418-1421
    PubMedGoogle Scholar
  • 12 Olson LE, Hoffman EA. Lung volumes and distribution of regional air content determined by cine X-ray CT of pneumonectomized rabbits. J Appl Physiol 1994; 76 (4) 1774-1785
    PubMedGoogle Scholar
  • 13 Olson MF, Ashworth A, Hall A. An essential role for Rho, Rac, and Cdc42 GTPases in cell cycle progression through G1. Science 1995; 269 (5228) 1270-1272
    CrossrefPubMedGoogle Scholar
  • 14 Chaturvedi LS, Marsh HM, Basson MD. Src and focal adhesion kinase mediate mechanical strain-induced proliferation and ERK1/2 phosphorylation in human H441 pulmonary epithelial cells. Am J Physiol Cell Physiol 2007; 292 (5) C1701-C1713
    CrossrefPubMedGoogle Scholar
  • 15 Correa-Meyer E, Pesce L, Guerrero C, Sznajder JI. Cyclic stretch activates ERK1/2 via G proteins and EGFR in alveolar epithelial cells. Am J Physiol Lung Cell Mol Physiol 2002; 282 (5) L883-L891
    PubMedGoogle Scholar
  • 16 Chatterjee A, Black SM, Catravas JD. Endothelial nitric oxide (NO) and its pathophysiologic regulation. Vascul Pharmacol 2008; 49 (4-6) 134-140
    CrossrefPubMedGoogle Scholar
  • 17 Geller DA, Billiar TR. Molecular biology of nitric oxide synthases. Cancer Metastasis Rev 1998; 17 (1) 7-23
    CrossrefPubMedGoogle Scholar
  • 18 Luong C, Rey-Perra J, Vadivel A , et al. Antenatal sildenafil treatment attenuates pulmonary hypertension in experimental congenital diaphragmatic hernia. Circulation 2011; 123 (19) 2120-2131
    CrossrefPubMedGoogle Scholar
  • 19 Gien J, Seedorf GJ, Balasubramaniam V, Markham N, Abman SH. Intrauterine pulmonary hypertension impairs angiogenesis in vitro: role of vascular endothelial growth factor nitric oxide signaling. Am J Respir Crit Care Med 2007; 176 (11) 1146-1153
    CrossrefPubMedGoogle Scholar
  • 20 Sinha S, Sridhara SR, Srinivasan S , et al. NO (nitric oxide): the ring master. Eur J Cell Biol 2011; 90 (1) 58-71
    CrossrefPubMedGoogle Scholar
  • 21 Cooke JP. NO and angiogenesis. Atheroscler Suppl 2003; 4 (4) 53-60
    PubMedGoogle Scholar
  • 22 Stahmann N, Woods A, Spengler K , et al. Activation of AMP-activated protein kinase by vascular endothelial growth factor mediates endothelial angiogenesis independently of nitric-oxide synthase. J Biol Chem 2010; 285 (14) 10638-10652
    CrossrefPubMedGoogle Scholar
  • 23 Leuwerke SM, Kaza AK, Tribble CG, Kron IL, Laubach VE. Inhibition of compensatory lung growth in endothelial nitric oxide synthase-deficient mice. Am J Physiol Lung Cell Mol Physiol 2002; 282 (6) L1272-L1278
    CrossrefPubMedGoogle Scholar
  • 24 Maxey TS, Fernandez LG, Reece TB, Keeling WB, Kron IL, Laubach VE. Endothelial nitric oxide synthase is essential for postpneumonectomy compensatory vasodilation. Ann Thorac Surg 2006; 81 (4) 1234-1238
    CrossrefPubMedGoogle Scholar
  • 25 Iwakiri Y, Cadelina G, Sessa WC, Groszmann RJ. Mice with targeted deletion of eNOS develop hyperdynamic circulation associated with portal hypertension. Am J Physiol Gastrointest Liver Physiol 2002; 283 (5) G1074-G1081
    PubMedGoogle Scholar
  • 26 Shesely EG, Maeda N, Kim HS , et al. Elevated blood pressures in mice lacking endothelial nitric oxide synthase. Proc Natl Acad Sci U S A 1996; 93 (23) 13176-13181
    Google Scholar
  • 27 Vaporidi K, Voloudakis G, Priniannakis G , et al. Effects of respiratory rate on ventilator-induced lung injury at a constant PaCO2 in a mouse model of normal lung. Crit Care Med 2008; 36 (4) 1277-1283
    CrossrefPubMedGoogle Scholar
  • 28 Daxhelet GA, Coene MM, Hoet PP, Cocito CG. Spectrofluorometry of dyes with DNAs of different base composition and conformation. Anal Biochem 1989; 179 (2) 401-403
    CrossrefPubMedGoogle Scholar
  • 29 Labarca C, Paigen K. A simple, rapid, and sensitive DNA assay procedure. Anal Biochem 1980; 102 (2) 344-352
    CrossrefPubMedGoogle Scholar
  • 30 Sekhon HS, Thurlbeck WM. A comparative study of postpneumonectomy compensatory lung response in growing male and female rats. J Appl Physiol 1992; 73 (2) 446-451
    PubMedGoogle Scholar
  • 31 Taft RA, Davisson M, Wiles MV. Know thy mouse. Trends Genet 2006; 22 (12) 649-653
    CrossrefPubMedGoogle Scholar
  • 32 Rannels DE, Rannels SR. Compensatory growth of the lung following partial pneumonectomy. Exp Lung Res 1988; 14 (2) 157-182
    CrossrefPubMedGoogle Scholar
  • 33 Kleinbongard P, Dejam A, Lauer T , et al. Plasma nitrite reflects constitutive nitric oxide synthase activity in mammals. Free Radic Biol Med 2003; 35 (7) 790-796
    CrossrefPubMedGoogle Scholar
  • 34 Tsutsui M, Shimokawa H, Morishita T, Nakashima Y, Yanagihara N. Development of genetically engineered mice lacking all three nitric oxide synthases. J Pharmacol Sci 2006; 102 (2) 147-154
    CrossrefPubMedGoogle Scholar
  • 35 Carlin JI, Hsia CC, Cassidy SS, Ramanathan M, Clifford PS, Johnson Jr RL. Recruitment of lung diffusing capacity with exercise before and after pneumonectomy in dogs. J Appl Physiol 1991; 70 (1) 135-142
    PubMedGoogle Scholar
  • 36 McBride JT, Kirchner KK, Russ G, Finkelstein J. Role of pulmonary blood flow in postpneumonectomy lung growth. J Appl Physiol 1992; 73 (6) 2448-2451
    PubMedGoogle Scholar
  • 37 Le Cras TD, McMurtry IF. Nitric oxide production in the hypoxic lung. Am J Physiol Lung Cell Mol Physiol 2001; 280 (4) L575-L582
    PubMedGoogle Scholar
  • 38 Ballard PL, Merrill JD, Truog WE , et al. Surfactant function and composition in premature infants treated with inhaled nitric oxide. Pediatrics 2007; 120 (2) 346-353
    CrossrefPubMedGoogle Scholar
  • 39 Bland RD, Albertine KH, Carlton DP, MacRitchie AJ. Inhaled nitric oxide effects on lung structure and function in chronically ventilated preterm lambs. Am J Respir Crit Care Med 2005; 172 (7) 899-906
    CrossrefPubMedGoogle Scholar
  • 40 Lin YJ, Markham NE, Balasubramaniam V , et al. Inhaled nitric oxide enhances distal lung growth after exposure to hyperoxia in neonatal rats. Pediatr Res 2005; 58 (1) 22-29
    CrossrefPubMedGoogle Scholar
  • 41 Decker NK, Abdelmoneim SS, Yaqoob U , et al. Nitric oxide regulates tumor cell cross-talk with stromal cells in the tumor microenvironment of the liver. Am J Pathol 2008; 173 (4) 1002-1012
    CrossrefPubMedGoogle Scholar
  • 42 Mortensen KE, Conley LN, Nygaard I , et al. Increased sinusoidal flow is not the primary stimulus to liver regeneration. Comp Hepatol 2010; 9: 2
    CrossrefPubMedGoogle Scholar
  • 43 Fausto N, Campbell JS, Riehle KJ. Liver regeneration. Hepatology 2006; 43 (2, Suppl 1) S45-S53
    CrossrefPubMedGoogle Scholar
  • 44 Lim KH, Ancrile BB, Kashatus DF, Counter CM. Tumour maintenance is mediated by eNOS. Nature 2008; 452 (7187) 646-649
    CrossrefPubMedGoogle Scholar
  • 45 Le Cras TD, Fernandez LG, Pastura PA, Laubach VE. Vascular growth and remodeling in compensatory lung growth following right lobectomy. J Appl Physiol 2005; 98 (3) 1140-1148
    PubMedGoogle Scholar
  • 46 Hsia CC, Herazo LF, Fryder-Doffey F, Weibel ER. Compensatory lung growth occurs in adult dogs after right pneumonectomy. J Clin Invest 1994; 94 (1) 405-412
    CrossrefPubMedGoogle Scholar
  • 47 Konerding MA, Gibney BC, Houdek JP , et al. Spatial dependence of alveolar angiogenesis in post-pneumonectomy lung growth. Angiogenesis 2012; 15 (1) 23-32
    CrossrefPubMedGoogle Scholar
  • 48 Lan Y, Liu B, Yao H , et al. Essential role of endothelial Smad4 in vascular remodeling and integrity. Mol Cell Biol 2007; 27 (21) 7683-7692
    CrossrefPubMedGoogle Scholar
  • 49 Liu WF, Nelson CM, Tan JL, Chen CS. Cadherins, RhoA, and Rac1 are differentially required for stretch-mediated proliferation in endothelial versus smooth muscle cells. Circ Res 2007; 101 (5) e44-e52
    CrossrefPubMedGoogle Scholar
  • 50 Chamoto K, Gibney BC, Lee GS , et al. Cd34+ progenitor to endothelial cell transition in post-pneumonectomy angiogenesis. Am J Respir Cell Mol Biol 2012; 46 (3) 283-289
    CrossrefPubMedGoogle Scholar
  • 51 Takahashi Y, Izumi Y, Kohno M , et al. Thyroid transcription factor-1 influences the early phase of compensatory lung growth in adult mice. Am J Respir Crit Care Med 2010; 181 (12) 1397-1406
    CrossrefPubMedGoogle Scholar
  • 52 Wolff JC, Wilhelm J, Fink L, Seeger W, Voswinckel R. Comparative gene expression profiling of post-natal and post-pneumonectomy lung growth. Eur Respir J 2010; 35 (3) 655-666
    CrossrefPubMedGoogle Scholar
  • 53 Talukder MA, Fujiki T, Morikawa K , et al. Up-regulated neuronal nitric oxide synthase compensates coronary flow response to bradykinin in endothelial nitric oxide synthase-deficient mice. J Cardiovasc Pharmacol 2004; 44 (4) 437-445
    CrossrefPubMedGoogle Scholar