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Immunohistochemical approach reveals involvement of inducible nitric oxide synthase in rat late development

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Abstract

To better understand the role of nitric oxide (NO) in mammal development, specifically in the transition of the fetal stages at birth, we studied the timing of cell-specific expression of inducible NO synthase (iNOS) isoform during gestational periods of rats, mainly at the late stages of intra-uterine development. Before experimentation, the samples were collected (from 17th to 21st gestational days), fixed in 10% buffered formalin and embedded in paraffin for histological procedures. Hereafter, the sections (5 μm thickness) obtained from different embryos were immunostained by avidin–biotin–immunoperoxidase technique, by using antibody against iNOS isoform. The most of cell immunopositive was suggestive of granulocyte-like cells and those cells were resident close to the blood vessels in different organs, such as: lung, liver or bone marrow environment. Sometimes we noted immunopositive cells in the blood flow, as reported in the thymus. In agreement, iNOS expression, obtained by western blotting analysis, showed the same profile. Together, our data shows that iNOS expression increased gradually during the late stages of rat development (from E17 to E21) and it was executed by cells close to blood vessels. Thus, we can clearly to predict that this expression was finely modulated and it contributes for time-line dependent NO production during rat late development.

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Fig. 1
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Abbreviations

ROS:

Reactive oxygen species

RNS:

Reactive nitrogen species

PBS:

Phosphate-buffered saline

iNOS:

Inducible nitric oxide synthase

NO:

Nitric oxide

E:

Gestational period

RT:

Room temperature

References

  • Accorsi-Mendonça T, Zambuzzi WF, da Silva Paiva KB, Pereira Lauris JR, Cestari TM, Taga R, Granjeiro JM (2005) Expression of metalloproteinase 2 in the cell response to porous demineralized bovine bone matrix. J Mol Histol 36:311–316

    Article  PubMed  Google Scholar 

  • Accorsi-Mendonça T, Paiva KB, Zambuzzi WF, Cestari TM, Lara VS, Sogayar MC, Taga R, Granjeiro JM (2008) Expression of matrix metalloproteinases-2 and -9 and RECK during alveolar bone regeneration in rat. J Mol Histol 39:201–208

    Article  PubMed  Google Scholar 

  • Batista AC, Silva TA, Chun JH, Lara VS (2002) Nitric oxide synthesis and severity of human periodontal disease. Oral Dis 8:254–260

    Article  CAS  PubMed  Google Scholar 

  • Beauvais F, Michel L, Dubertret L (1995) The nitric oxide donors, azide and hydroxylamine, inhibit the programmed cell death of cytokine-deprived human eosinophils. FEBS Lett 361:229–232

    Article  CAS  PubMed  Google Scholar 

  • Bispo de Jesus M, Zambuzzi WF, Ruela de Sousa RR, Areche C, Santos de Souza AC, Aoyama H, Schmeda-Hirschmann G, Rodríguez JA, Monteiro de Souza Brito AR, Peppelenbosch MP, den Hertog J, de Paula E, Ferreira CV (2008) Ferruginol suppresses survival signaling pathways in androgen-independent human prostate cancer cells. Biochimie 90:843–854

    Article  CAS  PubMed  Google Scholar 

  • Chakraborti S, Mandal M, Das S, Mandal A, Chakraborti T (2003) Regulation of matrix metalloproteinases: an overview. Mol Cell Biochem 253:269–285

    Article  CAS  PubMed  Google Scholar 

  • de Fátima A, Zambuzzi WF, Modolo LV, Tarsitano CA, Gadelha FR, Hyslop S, de Carvalho JE, Salgado I, Ferreira CV, Pilli RA (2008) Cytotoxicity of goniothalamin enantiomers in renal cancer cells: involvement of nitric oxide, apoptosis and autophagy. Chem Biol Interact 176:143–150

    Article  PubMed  Google Scholar 

  • de Souza Queiroz KC, Zambuzzi WF, Santos de Souza AC, da Silva RA, Machado D, Justo GZ, Carvalho HF, Peppelenbosch MP, Ferreira CV (2007) A possible anti-proliferative and anti-metastatic effect of irradiated riboflavin in solid tumours. Cancer Lett 258:126–134

    Article  PubMed  Google Scholar 

  • Dimmeler S, Haendeler J, Nehls M, Zeiher AM (1997) Suppression of apoptosis by nitric oxide via inhibition of interleukin-1beta-converting enzyme (ICE)-like and cysteine protease protein (CPP)-32-like proteases. J Exp Med 185:601–607

    Article  CAS  PubMed  Google Scholar 

  • Garthwaite J, Charles SL, Chess-Williams R (1988) Endothelium-derived relaxing factor release on activation of NMDA receptors suggests role as intercellular messenger in the brain. Nature 336:385–388

    Article  CAS  PubMed  Google Scholar 

  • Geller DA, Billiar TR (1998) Molecular biology of nitric oxide synthases. Cancer Metastasis Rev 17:7–23

    Article  CAS  PubMed  Google Scholar 

  • Gess B, Sandner P, Kurtz A (1996) Differential effects of kinase inhibitors on erythropoietin and vascular endothelial growth factor gene expression in rat hepatocytes. Pflugers Arch 432:426–432

    Article  CAS  PubMed  Google Scholar 

  • Hartree EF (1972) Determination of proteins: a modification of Lowry method that give a linear photometric response. Anal Biochem 48:422–427

    Article  CAS  PubMed  Google Scholar 

  • Hortelano S, Dallaporta B, Zamzami N, Hirsch T, Susin SA, Marzo I, Boscá L, Kroemer G (1997) Nitric oxide induces apoptosis via triggering mitochondrial permeability transition. FEBS Lett 410:373–377

    Article  CAS  PubMed  Google Scholar 

  • Ignarro LJ, Byrns RE, Buga GM, Wood KS (1987) Endothelium-derived relaxing factor from pulmonary artery and vein possesses pharmacologic and chemical properties identical to those of nitric oxide radical. Circ Res 61:866–879

    CAS  PubMed  Google Scholar 

  • Kaneto H, Fujii J, Seo HG, Suzuki K, Matsuoka T, Nakamura M, Tatsumi H, Yamasaki Y, Kamada T, Taniguchi N (1995) Apoptotic cell death triggered by nitric oxide in pancreatic beta-cells. Diabetes 44:733–738

    Article  CAS  PubMed  Google Scholar 

  • Kim YM, de Vera ME, Watkins SC, Billiar TR (1997) Nitric oxide protects cultured rat hepatocytes from tumor necrosis factor-alpha-induced apoptosis by inducing heat shock protein 70 expression. J Biol Chem 272:1402–1411

    Article  CAS  PubMed  Google Scholar 

  • Lala PK, Orucevic A (1998) Role of nitric oxide in tumor progression: lessons from experimental tumors. Cancer Metastasis Rev 17:91–106

    Article  CAS  PubMed  Google Scholar 

  • Mannick JB, Asano K, Izumi K, Kieff E, Stamler JS (1994) Nitric oxide produced by human B lymphocytes inhibits apoptosis and Epstein-Barr virus reactivation. Cell 79:1137–1146

    Article  CAS  PubMed  Google Scholar 

  • Mannick JB, Miao XQ, Stamler JS (1997) Nitric oxide inhibits Fas-induced apoptosis. J Biol Chem 272:24125–24128

    Article  CAS  PubMed  Google Scholar 

  • Marletta MA, Yoon PS, Iyengar R, Leaf CD, Wishnok JS (1988) Macrophage oxidation of L-arginine to nitrite and nitrate: nitric oxide is an intermediate. Biochemistry 27:8706–8711

    Article  CAS  PubMed  Google Scholar 

  • Moncada S, Higgs A (1993) The L-arginine-nitric oxide pathway. N Engl J Med 329:2002–2012

    Article  CAS  PubMed  Google Scholar 

  • Moncada S, Palmer RM, Higgs EA (1991) Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol Rev 43:109–142

    CAS  PubMed  Google Scholar 

  • Nathan C, Xie QW (1994) Regulation of biosynthesis of nitric oxide. J Biol Chem 269:13725–13728

    CAS  PubMed  Google Scholar 

  • North AJ, Star RA, Brannon TS, Ujiie K, Wells LB, Lowenstein CJ, Snyder SH, Shaul PW (1994) Nitric oxide synthase type I and type III gene expression are developmentally regulated in rat lung. Am J Physiol 266:L635–L641

    CAS  PubMed  Google Scholar 

  • Nowicki MJ, Shi D, Cai Z, Bishop PR, May WL (2003) Developmental expression of endothelial nitric oxide synthase (eNOS) in the rat liver. Pediatr Res 54:732–738

    Article  CAS  PubMed  Google Scholar 

  • Sarih M, Souvannavong V, Adam A (1993) Nitric oxide synthase induces macrophage death by apoptosis. Biochem Biophys Res Commun 191:503–508

    Article  CAS  PubMed  Google Scholar 

  • Shaul PW (1997) Ontogeny of nitric oxide in the pulmonary vasculature. Semin Perinatol 21:381–392

    Article  CAS  PubMed  Google Scholar 

  • Townsend SF, Rudolph CD, Rudolph AM (1989) Changes in ovine hepatic circulation and oxygen consumption at birth. Pediatr Res 25:300–304

    Article  CAS  PubMed  Google Scholar 

  • Wink DA, Vodovotz Y, Laval J, Laval F, Dewhirst MW, Mitchell JB (1998) The multifaceted roles of nitric oxide in cancer. Carcinogenesis 19:711–721

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (WFZ, FAPESP grant nr 08/53003-9). Also, the authors are grateful to technicians from Lab of Pathology, Dept. of Biological Sciences, FOB-USP for excellent technical support.

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Authors and Affiliations

  1. Department of Biochemistry, Biology Institute, University of Campinas, Campinas, SP, Brazil

    W. F. Zambuzzi

  2. Department of Molecular Pathology, University of Sao Paulo, Bauru, SP, Brazil

    K. B. S. Paiva

  3. Department of Pathology, Federal University of Goias, Goiania, GO, Brazil

    A. C. Batista

  4. Department of Stomatology, Bauru Dental School, University of Sao Paulo, Bauru, SP, Brazil

    V. S. Lara

  5. Department of Cellular and Molecular Biology, Biology Institute, Federal Fluminense University, Niteroi, RJ, Brazil

    J. M. Granjeiro

  6. Division of Cell Signaling and Biossays, University of Campinas, Cidade Universitária “Zeferino Vaz”, Campinas, SP, 13083-970, Brazil

    W. F. Zambuzzi

Authors
  1. W. F. Zambuzzi
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  2. K. B. S. Paiva
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  3. A. C. Batista
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  4. V. S. Lara
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  5. J. M. Granjeiro
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Corresponding author

Correspondence to W. F. Zambuzzi.

Additional information

This work was conducted in the Bauru Dental School, University of Sao Paulo, Brazil.

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Zambuzzi, W.F., Paiva, K.B.S., Batista, A.C. et al. Immunohistochemical approach reveals involvement of inducible nitric oxide synthase in rat late development. J Mol Hist 40, 235–240 (2009). https://doi.org/10.1007/s10735-009-9225-2

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  • DOI: https://doi.org/10.1007/s10735-009-9225-2

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