- Research Article
- Published:
The molecular cloning of glial fibrillary acidic protein in Gekko japonicus and its expression changes after spinal cord transection
Cellular & Molecular Biology Letters volume 15, pages 582–599 (2010)
Abstract
The glial fibrillary acidic protein (GFAP) is an astrocyte-specific member of the class III intermediate filament proteins. It is generally used as a specific marker of astrocytes in the central nervous system (CNS). We isolated a GFAP cDNA from the brain and spinal cord cDNA library of Gekko japonicus, and prepared polyclonal antibodies against gecko GFAP to provide useful tools for further immunochemistry studies. Both the real-time quantitative PCR and western blot results revealed that the expression of GFAP in the spinal cord after transection increased, reaching its maximum level after 3 days, and then gradually decreased over the rest of the 2 weeks of the experiment. Immunohistochemical analyses demonstrated that the increase in GFAP-positive labeling was restricted to the white matter rather than the gray matter. In particular, a slight increase in the number of GFAP positive star-shaped astrocytes was detected in the ventral and lateral regions of the white matter. Our results indicate that reactive astrogliosis in the gecko spinal cord took place primarily in the white matter during a short time interval, suggesting that the specific astrogliosis evaluated by GFAP expression might be advantageous in spinal cord regeneration.
Abbreviations
- GFAP:
-
glial fibrillaryacidic protein
- IPTG:
-
isopropyl-β-D-thiogalactopyranoside
- RACE:
-
rapid amplification of cDNA ends
- SDS-PAGE:
-
sodium dodecyl sulfate polyacrylamide gel electrophoresis
References
Eng, L.F., Vanderheagen, J.J., Bignami, A. and Gerstl, B. An acidic protein isolated from fibrous astrocytes. Brain Res. 28 (1971) 351–354.
Bignami, A., Eng, L.F., Dahl, D. and Uyeda, C.T. Localization of the glial fibrillary acidic protein in astrocytes by immunofluorescence. Brain Res. 43 (1972) 429–435.
Reeves, S.A., Helman, L.J., Allison, A. and Israel, M.A. Molecular cloning and primary structure of human glial fibrillary acidic protein. Proc. Natl. Acad. Sci. 86 (1989) 5178–5182.
Bongcam-Rudloff, E., Nister, M., Betsholtz, C., Wang, J.L., Stenman, G., Huebner, K., Croce, C.M. and Westermark, B. Human glial fibrillary acidic protein: complementary DNA cloning, chromosome localization, and messenger RNA expression in human glioma cell lines of various phenotypes. Cancer Res. 51 (1991) 1553–1560.
Isaacs, A., Baker, M., Wavrant-De, Vrieze, F. and Hutton, M. Determination of the gene structure of human GFAP and absence of coding region mutations associated with frontotemporal dementia with parkinsonism linked to chromosome 17. Genomics 51 (1998) 152–154.
Eng, L.F., Ghirnikar, R.S. and Lee, Y.L. Glial fibrillary acidic protein: GFAP-31 years (1969–2000). Neurochem Res. 25 (2000) 1439–1451.
Nielsen, A.L., Holm, I.E., Johansen, M., Bonven, B., Jorgensen, P. and Jorgensen, A.L. A new splice variant of glial fibrillary acidic protein, GFAP epsilon, interacts with the presenilin proteins. J. Biol. Chem. 277 (2002) 29983–29991.
Geisler, N. and Weber, K. The amino acid sequence of chicken muscle desmin provides a common structural model for intermediate filament proteins. EMBO J. 1 (1982) 1649–1656.
Steinert, P.M. and Roop, D.R. Molecular and cellular biology of intermediate filaments. Annu. Rev. Biochem. 57 (1988) 593–625.
Parry, D.A.D. and Steinert, P.M. Intermediate filament structure. Curr. Opin. Cell Biol. 4 (1992) 94–98.
Liedtke, W., Edelmann, W., Bieri, P.L., Chiu, F.C., Cowan, N.J., Kucherlapati, R. and Raine, C.S. GFAP is necessary for the integrity of CNS white matter architecture and long-term maintenance of myelination. Neuron 17 (1996) 607–615.
Kimelberg, H.K. and Norenberg, M.D. Astrocytes. Sci. Am. 260 (1989) 66–76.
Bonni, A., Sun, Y., Nadal-Vicens, M., Bhatt, A., Frank, D.A., Rozovsky, I., Stahl, N., Yancopoulos, G.D. and Greenberg, M.E. Regulation of gliogenesis in the central nervous system by the JAK-STAT signaling pathway. Science 278 (1997) 477–483.
Eng, L.F. and Ghirnikar, R.S. GFAP and astrogliosis. Brain Pathol. 4 (1994) 229–237.
Ransom, B., Behar, T. and Nedergaard, M. New roles for astrocytes (stars at last). Trends Neurosci. 26 (2003) 520–522.
May, P.C., Boggs, L.N., Fuson, K.S., Bender, M., Li, W., Miller, F.D., Hyslop, P., Calligaro, D., Seubert, P., Johnson-Wood, K., Chen, K., Games, D. and Schenk, D. GFAP as a marker of plaque pathology in PDAPP transgenic mouse. Soc. Neurosci. Abstr. 23 (1997) 1638.
Canady, K.S. and Rubel, E.W. Rapid and reversible astrocytic reaction to afferent activity blockade in chick cochlear nucleus. J. Neurosci. 12 (1992) 1001–1009.
Bignami, A. and Dahl, D. The astrocytic response to stabbing. Immunofluorescence studies with antibodies to astrocytic-specific protein (GFAP) in mammalian and submammalian vertebrates. Neuropathol. Appl. Neurobiol. 2 (1976) 99–110.
Silver, J. and Miller, J.H. Regeneration beyond the glial scar. Nat. Rev. Neurosci. 5 (2004) 146–156.
Wang, X., Messing, A. and David, S. Axonal and nonneuronal cell responses to spinal cord injury in mice lacking glial fibrillary acidic protein. Exp. Neurol. 148 (1997) 568–576.
Menet, V., Prieto, M., Privat, A. and Gimenezy, Ribotta, M. Axonal plasticity and functional recovery after spinal cord injury in mice deficient in both glial fibrillary acidic protein and vimentin genes. Proc. Natl. Acad. Sci. 100 (2003) 8999–9004.
Faulkner, J.R., Herrmann, J.E., Woo, M.J., Tansey, K.E., Doan, N.B. and Sofroniew, M.V. Reactive astrocytes protect tissue and preserve function after spinal cord injury. J. Neurosci. 24 (2004) 2143–2155.
Okada, S., Nakamura, M., Katoh, H., Miyao, T., Shimazaki, T., Ishii, K., Yamane, J., Yoshimura, A., Iwamoto, Y., Toyama, Y. and Okano, H. Conditional ablation of Stat3 or Socs3 discloses a dual role for reactive astrocytes after spinal cord injury. Nat. Med. 12 (2006) 829–834.
Cristino, L., Pica, A., Della, Corte, F. and Bentivoglio, M. Plastic changes and nitric oxide synthase induction in neurons which innervated the regenerated tail of the lizard Gekko gecko II. The response of dorsal root ganglion cells to tail amputation and regeneration. Brain Res. 871 (2000) 83–93.
Pixley, S.K. and De, Vellis, J. Transition between immature radial glia and mature astrocytes studied with a monoclonal antibody to vimentin. Brain Res. 317 (1984) 201–209.
Voigt, T. Development of glial cells in the cerebral walls of ferrets: direct tracing of their transformation from radial glia into astrocytes. J. Comp. Neurol. 289 (1989) 74–88.
Elmquist, J.K., Swanson, J.J., Sakaguchi, D.S., Ross, L.R. and Jacobson, C.D. Developmental distribution of GFAP and vimentin in the Brazilian opossum brain. J. Comp. Neurol. 344 (1994) 283–296.
Lazzari, M. and Franceschini, V. Intermediate filament immunohistochemistry of astroglial cells in the leopard gecko, Eublepharis macularius. Anat. Embryol. 210 (2005) 275–286.
Kalman, M. and Pritz, M.B. Glial fibrillary acidic protein-immunopositive structures in the brain of acrocodilian, Caiman crocodilus, and its bearing on the evolution of astroglia. J. Comp. Neurol. 431 (2001) 460–480.
Liu, Y., Ding, F., Liu, M., Jiang, M., Yang, H., Feng, X. and Gu, X. EST-based identification of genes expressed in brain and spinal cord of Gekko japonicus, a species demonstrating intrinsic capacity of spinal cord regeneration. J. Mol. Neurosci. 29 (2006) 21–28.
Rehm, B.H. Bioinformatic tools for DNA/protein sequence analysis, functional assignment of genes and protein classification. Appl. Micro-Biol. Biotechnol. 57 (2001) 579–592.
Altschul, S.F., Madden, T.L., Schaffer, A.A., Zhang, J., Zhang, Z., Miller, W. and Lipman, D.J. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 25 (1997) 3389–3402.
Felsenstein, J. PHYLIP (phylogeny inference package). version 3.6. Department of Genome Sciences, University of Washington, Seattle (2004).
Engvall, E. and Perlman, P. Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of immunoglobulin G. Immunochemistry 8 (1971) 871–874.
Rataboul, P., Faucon, Biguet, N., Vernier, P., De, Vitry, F., Boularand, S., Privat, A. and Mallet, J. Identification of a human glial fibrillary acidic protein cDNA: a tool for the molecular analysis of reactive gliosis in the mammalian central nervous system. J. Neurosci. Res. 20 (1988) 165–175.
Hozumi, I., Chiu, F.C. and Norton, W.T. Biochemical and immunocytochemical changes in glial fibrillary protein after stab wounds. Brain Res. 524 (1990) 64–71.
Goncalves, C.A., Leite, M.C. and Nardin, P. Biological and methodological features of the measurement of S100B, a putative marker of brain injury. Clin. Biochem. 41 (2008) 755–763.
Norenberg, M.D. Distribution of glutamine synthetase in the rat central nervous system. J. Histochem. Cytochem. 27 (1979) 756–762.
Inagaki, K., Gonda, T., Nishizawa, K., Kitamura, S., Sato, C., Ando, S., Tanabe, K., Kikuchi, K., Tsuiki, S. and Nishi, Y. Phosphorylation sites linked to glial filament disassembly in vitro locate in a non-alpha-helical head domain. J. Biol. Chem. 265 (1990) 4722–4729.
Takemura, M., Gomi, H., Colucci-Guyon, E. and Itohara, S. Protectiverole of phosphorylation in turnover of glial fibrillary acidic protein in mice. J. Neurosci. 22 (2002) 6972–6979.
Chen, W.J. and Liem, R.K. The endless story of the glial fibrillary acidic protein. J. Cell Sci. 107 (1994) 2299–2311.
Bock, E. Nervous system specific proteins. J. Neurochem. 30 (1978) 7–14.
Nesic, O., Lee, J., Johnson, K.M., Ye, Z., Xu, G.Y., Unabia, G.C., Wood, T.G., McAdoo, D.J., Westlund, K.N., Hulsebosch, C.E. and Regino, Perez-Polo, J. Transcriptional profiling of spinal cord injury-induced central neuropathic pain. J. Neurochem. 95 (2005) 998–1014.
Tian, D.S., Dong, Q., Pan, D.J., He, Y., Yu, Z.Y., Xie, M.J. and Wang, W. Attenuation of astrogliosis by suppressing of microglial proliferation with the cell cycle inhibitor olomoucine in rat spinal cord injury model. Brain Res. 1154 (2007) 206–214.
Ritz, M.F. and Hausmann, O.N. Effect of 17beta-estradiol on functional outcome, release of cytokines, astrocyte reactivity and inflammatory spreading after spinal cord injury in male rats. Brain Res. 1203 (2008) 177–188.
Huang, X., Kim, J.M., Kong, T.H., Park, S.R., Ha, Y., Kim, M.H., Park, H., Yoon, S.H., Park, H.C., Park, J.O., Min, B.H. and Choi, B.H. GM-CSF inhibits glial scar formation and shows long-term protective effect after spinal cord injury. J. Neurol. Sci. 277 (2009) 87–97.
Pekny, M., Wilhelmsson, U., Bogestål, Y.R. and Pekna, M. The role of astrocytes and complement system in neural plasticity. Int. Rev. Neurobiol. 82 (2007) 95–111.
Morin-Richaud, C., Feldblum, S. and Privat, A. Astrocytes and oligodendrocytes reactions after a total section of the rat spinal cord. Brain Res. 783 (1998) 85–101.
Collins, G.H. and West, N.R. Glial activity during axonal regrowth following cryogenic injury of rat spinal cord. Brain Res. Bull. 22 (1989) 71–79.
Soriede, A.J. Variations in the perineural glial changes after different types of nerve lesion: light and electron microscopic investigations on the facial nucleus of the rat. Neuropathol. Appl. Neurobiol. 7 (1981) 195–204.
Alonso, G. and Privat, A. Reactive astrocytes involved in the formation of lesional scars differ in the mediobasal hypothalamus and in other forebrain regions. J. Neurosci. Res. 34 (1993) 510–522.
Sofroniew, M.V. and Vinters, H.V. Astrocytes: biology and pathology. Acta Neuropathol. 119 (2010) 7–35.
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Gao, D., Wang, Y., Liu, Y. et al. The molecular cloning of glial fibrillary acidic protein in Gekko japonicus and its expression changes after spinal cord transection. Cell Mol Biol Lett 15, 582–599 (2010). https://doi.org/10.2478/s11658-010-0029-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.2478/s11658-010-0029-x