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Microglial expression of peptidylarginine deiminase 2 in the prenatal rat brain


Peptidylarginine deiminases (PADs) are Ca2t+-dependant post-translational modification enzymes that catalyze the citrullination of protein arginyl residues. PAD type 2 (PAD2) is thought to be involved in some processes of neurodegeneration and myelination in the central nervous system. In this study, we found PAD2-positive cells in rat cerebra in 19-to 21-day old embryos, i.e. at a developmental stage well before myelination begins. Most of the cells were microglial marker-positive cells found mainly in the prospective medulla, and others were microglial marker-negative cells found mainly in the prospective dentate gyrus of the hippocampus. The former seemed to be in an activated state as judged by morphological criteria. The specificity of the enzyme activity, immunoblotting and reverse transcriptase-polymerase chain reaction analyses revealed that these cells expressed PAD2 and not PAD1, PAD3 or PAD4. Our data is indicative of microglial expression of PAD2 in the prenatal developing cerebrum.



benzoyl-L-arginine ethyl ester






glial fibrillary acidic protein


kainic acid


microtubule associated protein 2


monoclonal antibody


myelin basic protein


peptydylarginine deiminase


peptidylarginine deiminase type 2


reverse transcriptase-polymerase chain reaction


sodium dodecyl sulfate


  1. Vossenaar, E.R., Zendman, A.J.W., van Venrooij, W.J. and Pruijn, G.J.M. PAD, a growing family of citrullinating enzymes: genes, features and involvement in disease. BioEssays 25 (2003) 1106–1118.

    PubMed  Article  CAS  Google Scholar 

  2. György, B., Tóth, E., Tarcsa, E., Falus, A. and Buzás, E.I. Citrullination: a posttranslational modification in health and disease. Int. J. Biochem. Cell. Biol. 38 (2006) 1662–1677.

    PubMed  Article  CAS  Google Scholar 

  3. Watanabe, K., Akiyama, K., Hikichi, K., Ohtsuka, R., Okuyama, A. and Senshu, T. Combined biochemical and immunochemical comparison of peptidylarginine deiminases present in various tissues. Biochim. Biophys. Acta 976 (1988) 375–383.

    Google Scholar 

  4. Asaga, H. and Senshu, T. Combined biochemical and immunocytochemical analyses of postmortem protein deimination in the rat spinal cord. Cell Biol. Int. 17 (1993) 525–532.

    PubMed  Article  CAS  Google Scholar 

  5. Asaga, H. and Ishigami, A. Protein deimination in the rat brain after kainate administration: citrulline-containing proteins as a novel marker of neurodegeneration. Neurosci. Lett. 299 (2001) 5–8.

    PubMed  Article  CAS  Google Scholar 

  6. Ishigami, A., Asaga, H., Ohsawa, T., Akiyama, K. and Maruyama, N. Peptidylarginine deiminase type I, type II, type III and type IV are expressed in rat epidermis. Biomed. Res. 22 (2001) 63–65.

    CAS  Google Scholar 

  7. Asaga, H. and Ishigami, A. Protein deimination in the rat brain: generation of citrulline-containing proteins in cerebrum perfused with oxygen-deprived media. Biomed. Res. 21 (2000) 197–205.

    CAS  Google Scholar 

  8. Vincent, S.R., Leung, E., and Watanabe, K. Immunohistochemical localization of peptidylarginine deiminase in the rat brain. J. Chem. Neuroanat. 5 (1992) 159–168.

    PubMed  Article  CAS  Google Scholar 

  9. Ishigami, A., Ohsawa, T., Hiratsuka, M., Taguchi H., Kobayashi, S., Saito, Y., Maruyama, S., Asaga, H., Toda, T., Kimura, N. and Mariyama, N. Abnormal accumulation of citrullinated proteins catalyzed by peptidylarginine deiminase in hippocampal extracts from patients with Alzheimer’s disease. J. Neurosci. Res. 80 (2005) 120–128.

    PubMed  Article  CAS  Google Scholar 

  10. Moscarello, M.A., Wood, D.D., Ackerley, C. and Boulias, C. Myelin in multiple sclerosis is developmentally immature. J. Clin. Invest. 94 (1994) 146–154.

    PubMed  CAS  Article  Google Scholar 

  11. Wood, D.D. and Moscarello, M.A. The isolation, characterization and lipid-aggregating properties of a citrulline containing myelin basic protein. J. Biol. Chem. 264 (1989) 5121–5127.

    PubMed  CAS  Google Scholar 

  12. Akiyama, K., Sakurai, Y., Asou, H. and Senshu, T. Localization of peptidylarginine deiminase type II in a stage-specific immature oligodendrocyte from rat cerebral hemisphere. Neurosci. Lett. 274 (1999) 53–55.

    PubMed  Article  CAS  Google Scholar 

  13. Pritzker, L.B., Nguyen, T.A. and Moscarello, M.A. The developmental expression and activity of peptidylarginine deiminase in the mouse. Neurosci. Lett. 266 (1999) 161–164.

    PubMed  Article  CAS  Google Scholar 

  14. Pixley, S.K., Kobayashi, Y. and de Vellis, J. Monoclonal antibody to intermediate filament proteins in astrocytes. J. Neurosci. Res. 12 (1984) 525–541.

    PubMed  Article  CAS  Google Scholar 

  15. Izant, J.G. and McIntosh, J.R. Microtubule-associated proteins: a monoclonal antibody to MAP2 binds to differentiated neurons. Proc. Natl. Acad. Sci. USA 77 (1980) 4741–4745.

    PubMed  Article  CAS  Google Scholar 

  16. Streit, W.J. An improved staining method for rat microglial cells using the lectin from Griffonia simplicifolia (GSA I-B4). J. Histochem. Cytochem. 38 (1990) 1983–1686.

    Google Scholar 

  17. Lowry, O.H., Rosebrough, N.J., Farr, A.L. and Randall, R.J. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193 (1951) 265–275.

    PubMed  CAS  Google Scholar 

  18. Laemmli, U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227 (1970) 680–685.

    PubMed  Article  CAS  Google Scholar 

  19. Ishigami, A., Kuramoto, M., Yamada, M., Watanabe, K. and Senshu, T. Molecular cloning of two novel types of peptidylarginine deiminase cDNAs from retinoic acid-treated culture of a newborn rat keratinocyte cell line. FEBS Lett. 433 (1998) 113–118.

    PubMed  Article  CAS  Google Scholar 

  20. Watanabe, K. and Senshu, T. Isolation and characterization of cDNA clones encoding rat skeletal muscle peptidylarginine deiminase. J. Biol. Chem. 264 (1989) 15255–15260.

    PubMed  CAS  Google Scholar 

  21. Nishijyo, T., Kawada, A., Kanno, T., Shiraiwa, M. and Takahara, H. Isolation and molecular cloning of epidermal-and hair follicle-specific peptidylarginine deiminase (Type III) from rat. J. Biochem. 121 (1997) 868–875.

    PubMed  CAS  Google Scholar 

  22. Tso, J.Y., Sun, X.H., Kao, T.H., Reece, K.S. and Wu, R. Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: genomic complexity and molecular evolution of the gene. Nucleic Acids Res. 13 (1985) 2485–2502.

    PubMed  Article  CAS  Google Scholar 

  23. Asaga, H., Akiyama, K., Ohsawa, T. and Ishigami, A. Increased and type II-specific expression of peptidylarginine deiminase in activated microglia but not hyperplastic astrocytes following kainic acid-evoked neurodegeneration in the rat brain. Neurosci. Lett. 326 (2002) 129–132.

    PubMed  Article  CAS  Google Scholar 

  24. Joosten, E.A. and Gribnau, A.A. Astrocytes and guidance of outgrowing corticospinal tract axons in the rat. An immunocytochemical study using anti-vimentin and anti-glial fibrillary acidic protein. Neuroscience 31 (1989) 439–452.

    PubMed  Article  CAS  Google Scholar 

  25. Wright, P.W., Bolling, L.C., Calvert, M.E., Sarmento, O.F., Berkeley, E.V., Shea, M.C., Hao, Z., Jayes, F.C., Bush, L.A., Shetty, J., Shore, A.N., Reddi, P.P., Tung, K.S., Samy, E., Allietta, M.M., Sherman, N.E., Herr, J.C. and Coonrod, S.A. ePAD, an oocyte and early embryo-abundant peptidylarginine deiminase-like protein that localizes to egg cytoplasmic sheets. Dev. Biol. 56 (2003) 73–88.

    Google Scholar 

  26. Zhang, J., Dai, J., Zhao, E., Lin, Y., Zeng, L., Chen, J., Zheng, H., Wang, Y., Li, X., Ying, K., Xie, Y. and Mao, Y. cDNA cloning, gene organization and expression analysis of human peptidylarginine deiminase type VI. Acta Biochim. Pol. 51 (2004) 1051–1058.

    PubMed  CAS  Google Scholar 

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Correspondence to Hiroaki Asaga.

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Asaga, H., Ishigami, A. Microglial expression of peptidylarginine deiminase 2 in the prenatal rat brain. Cell Mol Biol Lett 12, 536–544 (2007).

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Key words

  • Protein deimination
  • Post-translational modification enzyme
  • Microglial cells
  • Central nervous system
  • Gene expression