Skip to main content
SpringerLink
Log in

DNA Fragmentation and HSP72 Gene Expression by Adenovirus-Mediated Gene Transfer in Postischemic Gerbil Hippocampus and Ventricle

  • Published:
Metabolic Brain Disease Aims and scope Submit manuscript

Abstract

A replication defective adenoviral vector containing the E. coli lacZ gene (AdCMVnLacZ) was directly injected into right hippocampus and lateral ventricle immediately after 5 min of transient global ischemia in gerbils. The relations between the lacZ gene expression and DNA fragmentation or heat shock protein 72 (HSP72) immunoreactivity were examined up to 21 days post ischemia. The lacZ gene was transiently expressed at 1 day in the hippocampus except around the CA1 region, while a large number of the periventricular cells strongly expressed the lacZ gene from 8 h to 7 days. In CA1 layer terminal deoxynucleotidyl dUTP nick end labeling (TUNEL) positive cells, which were present only adjacent to the needle track at 8 h to 1 day, became more extensive in the whole CA1 layer at 3 to 7 days. TUNEL-positive cells were also detected around the DG at 1 day, around the needle track at 8 h to 3 days, and in the choroid plexus cells at 7 days HSP72 staining was detected in the subiculum at 1 to 3 days, the dentate granule cells at 8 h to 1 day, and in the CA3 or CA4 pyramidal cells at 1 to 3 days. Some lacZ expressing cells were double-positive with HSP72 in DG, while the majority of those were distinguished from the TUNEL-positive cells. Pyramidal neurons were almost completely lost in the CA1 sector at 7days after the ischemia. The present study demonstrates the successful LacZ gene transfer into the hippocampus and ventricle of postischemic gerbil brain except in the vulnerable CA1 layer by adenoviral vector injection. However adenovirus-mediated gene transfer may induce indirect apoptotic cell death in the DG and ventricle, in addition to direct traumatic injury around the needle track.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

REFERENCES

  • Abe, K., Aoki, M., Kawagoe, J., Yoshida, T., Hattori, A., Kogure, K. and Itoyama, Y. (1995). Ischemic delayed neuronal death: A mitochondrial hypothesis. Stroke 26:1478–1489.

    PubMed  Google Scholar 

  • Abe, K., Setoguchi, Y., Hayashi, T. and Itoyama, Y. (1997a). In vivo adenovirus-mediated gene transfer and the expression in ischemic and reperfused rat brain. Brain Res. 763:191–201.

    Article  PubMed  Google Scholar 

  • Abe, K., Setoguchi, Y., Hayashi, T. and Itoyama, Y. (1997b). Dissociative expression of adenoviral-mediated E.coli LacZ gene between ischemic and reperfused rat brains. Neurosci. Lett. 226:53–56.

    Article  PubMed  Google Scholar 

  • Abe, K., Tanzi, R.E. and Kogure, K. (1991). Induction of HSP70 mRNA after transient ischemia in gerbil brain. Neurosci. Lett. 125:166–168.

    Article  PubMed  Google Scholar 

  • Akli, S., Caillaud, C., Vigne, E., Stratford-Perricaudet, L.D., Poenaru, L., Perricaudet, M., Kahn, A. and Peschanski, M.R. (1993). Transfer of a foreign gene into the brain using adenovirus vectors. Nature Genet. 3:224–228.

    Article  PubMed  Google Scholar 

  • Bajocchi, G., Feldman, S.H., Crystal, R.G. and Mastrangeli, A. (1993). Direct in vivo gene transfer to ependymal cells in the central nervous system using recombinant adenovirus vectors. Nature Genet. 3:229–234.

    Article  PubMed  Google Scholar 

  • Betz, A.L., Yang, G.-Y. and Davidson, B.L. (1995). Attenuation of stroke size in rats using an adenoviral vector to induce overexpression of interleukin-1 receptor antagonist in brain. J. Cereb. Blood Flow Metab. 15:547–551.

    PubMed  Google Scholar 

  • Bilang-Bleuel, A., Revah, F., Colin, P., Locquet, I., Robert, J.-J., Mallet, J. and Horellou, P. (1997). Intrastriatal injection of an adenoviral vector expressing glial-cell-line-derived neurotrophic factor prevents dopaminergic neuron degeneration and behavioral impairment in a rat model of Parkinson disease. Proc. Natl. Acad. Sci. U.S.A. 94:8818–8823.

    Article  PubMed  Google Scholar 

  • Breakefield, X.O. (1993). Gene delivery into the brain using virus vectors. Nature Genet. 3:187–189.

    Article  PubMed  Google Scholar 

  • Byrnes, A.P., Rusby, J.E., Wood, M.J.A. and Charlton, H.M. (1995). Adenovirus gene transfer causes inflammation in the brain. Neuroscience 66:1015–1024.

    Article  PubMed  Google Scholar 

  • Charriaut-Marlangue, C., Margaill, I., Represa, A., Popovici, T., Plotkine, M. and Ben-Ari, Y. (1996). Apoptosis and necrosis after reversible focal ischemia: an in situ DNA fragmentation analysis. J. Cereb. Blood Flow Metab. 16:186–194.

    Article  PubMed  Google Scholar 

  • Chen, J., Graham, S.H., Nakayama, M., Zhu, R.L., Jin, K., Stetler, R.A. and Simon, R.P. (1997). Apoptosis repressor genes bcl-2 and bcl-x-long are expressed in the rat brain following global ischemia. J. Cereb. Blood Flow Metab. 17:2–10.

    Article  PubMed  Google Scholar 

  • Davidson, B.L., Allen, E.D., Kozarsky, K.F., Wilson, J.M. and Roessler, B.J. (1993). A model system for in vivo gene transfer into the central nervous system using an adenoviral vector. Nature Genet. 3:219–223.

    Article  PubMed  Google Scholar 

  • Deshpande, J., Bergstedt, K., Linden, T., Kalimo, H. and Wieloch, T. (1992). Ultrastructural changes in the hippocampal CA1 region following transient cerebral ischemia: evidence against programmed cell death. Exp. Brain Res. 88:91–105.

    PubMed  Google Scholar 

  • Gething, M.-J. and Sambrook, J. (1992). Protein folding in the cell. Nature 355:33–45.

    Article  PubMed  Google Scholar 

  • Heistad, D.D. and Faraci, F.M. (1996). Gene therapy for cerebral vascular disease. Stroke 27:1688–1693.

    PubMed  Google Scholar 

  • Honkaniemi, J., Massa, S.M., Breckinridge, M. and Sharp, F.R. (1996). Global ischemia induces apoptosis-associated genes in hippocampus. Mol. Brain Res. 42:79–88.

    Article  PubMed  Google Scholar 

  • Iwai, T., Hara, A., Niwa, M., Nozaki, M., Uematu, T., Sakai, N. and Yamada, H. (1995). Temporal profile of nuclear DNA fragmentation in situ in gerbil hippocampus following transient forebrain ischemia. Brain Res. 671:305–308.

    Article  PubMed  Google Scholar 

  • Kawagoe, J., Abe, K., Sato, S., Nagano, I., Nakamura, S. and Kogure, K. (1992). Distributions of heat shock protein-70 mRNAs and heat shock cognate protein-70 mRNAs after transient global ischemia in gerbil brain. J. Cereb. Blood Flow Metab. 12:794–801.

    PubMed  Google Scholar 

  • Kihara, S., Shiraishi, T., Nakagawa, S., Toda, K. and Tabuchi, K. (1994). Visualization of DNA double strand breaks in the gerbil hippocampal CA1 following transient ischemia. Neurosci. Lett. 175:133–136.

    Article  PubMed  Google Scholar 

  • Kirino, T. (1982). Delayed neuronal death in the gerbil hippocampus following ischemia. Brain Res. 239:57–69.

    Article  PubMed  Google Scholar 

  • La Salle, G.L.G., Robert, J.J., Berrard, S., Ridoux, V., Stratford-Perricaudet, L.D., Perricaudet, M. and Mallet, J. (1993). An adenovirus vector for gene transfer into neurons and glia in the brain. Science 259:988–990.

    PubMed  Google Scholar 

  • Lee, T.-H., Abe, K., Aoki, M., Nakamura, M., Kogure, K. and Itoyama, Y. (1994). The protective effect of L-threo-3,4-dihydroxyphenylserine on ischemic hippocampal neuronal death in gerbils. Stroke 25:1425–1432.

    PubMed  Google Scholar 

  • Li, Y., Chopp, M., Jiang, N., Yao, F. and Zaloga, C. (1995). Temporal profile of in situ DNA fragmentation after transient middle cerebral artery occlusion in the rat. J. Cereb. Blood Flow Metab. 15:389–397.

    PubMed  Google Scholar 

  • MacManus, J.P., Buchan, A.M., Hill, I.E., Rasquinha, I. and Preston, E. (1993). Global ischemia can cause DNA fragmentation indicative of apoptosis in rat brain. Neurosci. Lett. 164:89–92.

    Article  PubMed  Google Scholar 

  • MacManus, J.P., Hill, I.E., Preston, E., Rasquinha, I., Walker, T. and Buchan, A.M. (1995). Differences in DNA fragmentation following transient cerebral or decapitation ischemia in rats. J. Cereb. Blood Flow Metab. 15:728–737.

    PubMed  Google Scholar 

  • Newman, K.D., Dunn, P.F., Owens, J.W., Schulick, A.H., Virmani, R., Sukhova, G., Libby, P. and Dichek, D.A. (1995). Adenovirus-mediated gene transfer into normal rabbit arteries results in prolonged vascular cell activation, inflammation, and neointimal hyperplasia. J. Clin. Invest. 96:2955–2965.

    PubMed  Google Scholar 

  • Nitatori, T., Sato, N., Waguri, S., Karasawa, Y., Araki, H., Shibanai, K., Kominami, E. and Uchiyama, Y. (1995). Delayed neuronal death in the CA1 pyramidal cell layer of the gerbil hippocampus following transient ischemia is apoptosis. J. Neurosci. 15:1001–1011.

    PubMed  Google Scholar 

  • Nowak, T.S. (1991). Localization of 70 kDa stress protein mRNA induction in gerbil brain after ischemia. J. Cereb. Blood Flow Metab. 11:432–439.

    PubMed  Google Scholar 

  • Petito, C.K., Feldmann, E., Pulsinelli, W.A. and Plum, F. (1987). Delayed hippocampal damage in humans following cardiorespiratory arrest. Neurology 37:1281–1286.

    PubMed  Google Scholar 

  • Petito, C.K., Torres-Munoz, J., Roberts, B., Olarte, J.-P., Nowak, T.S. and Pulsinelli, W.A. (1997). DNA fragmentation follows delayed neuronal death in CA1 neurons exposed to transient global ischemia in the rat. J. Cereb. Blood Flow Metab. 17:967–976.

    Article  PubMed  Google Scholar 

  • Pulsinelli, W.A., Brierley, L.B. and Plum, F.C. (1982). Temporal profile of neuronal damage in a model of transient ischemia. Ann. Neurol. 11:491–498.

    PubMed  Google Scholar 

  • Rosenfeld, M.A., Siegfried, W., Yoshimura, K., Yoneyama, K., Fukayama, M., Stier, L.E. et al. (1991). Adenovirus-mediated transfer of a recombinant alphal-antitrypsin gene to the lung epithelium in vivo. Science 252:431–434.

    PubMed  Google Scholar 

  • Sei, Y., Von Lubitz, D.K.J.E., Basile, A.S., Borner, M.M., Lin, R.C.-S., Skolnick, P. and Fossom, L.H. (1994). Internucleosomal DNA fragmentation in gerbil hippocampus following forebrain ischemia. Neurosci. Lett. 171:179–182.

    Article  PubMed  Google Scholar 

  • Setoguchi, Y., Danel, C. and Crystal, R.G. (1994). Stimulation of erythropoiesis by in vivo gene therapy: Physiologic consequences of transfer of the human erythropoietin gene to experimental animals using an adenovirus vector. Blood 84:2946–2953.

    PubMed  Google Scholar 

  • States, B.A., Honkaniemi, J., Weinstein, P.R. and Sharp, F.R. (1996). DNA fragmentation and HSP70 protein induction in hippocampus and cortex occurs in separate neurons following permanent middle cerebral artery occlusions. J. Cereb. Blood Flow Metab. 16:1165–1175.

    Article  PubMed  Google Scholar 

  • Thilmann, R., Xie, Y., Kleihues, P. and Kiessling, M. (1986). Persistent inhibition of protein synthesis precedes delayed neuronal death in postischemic gerbil hippocampus. Acta Neuropathol. (Berl.) 71:88–93.

    Google Scholar 

  • Vass, K., Welch, W.J. and Nowak, T.S. (1988). Localization of 70-kDa stress protein induction in gerbil brain after ischemia. Acta Neuropathol. (Berl.) 77:128–135.

    Google Scholar 

  • Verma, I.M. and Somia, N. (1997). Gene therapy-promises, problems and prospects. Nature 389:239–242.

    Article  PubMed  Google Scholar 

  • Yang, G.-Y., Zhao, Y.-J., Davidson, B.L. and Betz, A.L. (1997). Overexpression of interleukin-1 receptor antagonist in the mouse brain reduces ischemic brain injury. Brain Res. 751:181–188.

    Article  PubMed  Google Scholar 

  • Yoshidomi, M., Hayashi, T., Abe, K. and Kogure, K. (1989). Effect of a new calcium channel blocker, KB-2796, on protein synthesis of the CA1 pyramidal cell and delayed neuronal death following transient forebrain ischemia. J. Neurochem. 53:137–142.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Neurology, Okayama University Medical School, Okayama, Japan

    H. Kitagawa & K. Abe

  2. Tokushima New Drug Research Institute, Otsuka Pharmaceutical Co., Ltd., Tokushima, Japan

    H. Kitagawa, Y. Mitsumoto, N. Koga & T. Mori

  3. Department of Respiratory Medicine, Juntendo University School of Medicine, Tokyo, Japan

    Y. Setoguchi & Y. Fukuchi

Authors
  1. H. Kitagawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Y. Setoguchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Y. Fukuchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Y. Mitsumoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. N. Koga
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. T. Mori
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. K. Abe
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kitagawa, H., Setoguchi, Y., Fukuchi, Y. et al. DNA Fragmentation and HSP72 Gene Expression by Adenovirus-Mediated Gene Transfer in Postischemic Gerbil Hippocampus and Ventricle. Metab Brain Dis 13, 211–223 (1998). https://doi.org/10.1023/A:1023224025407

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1023224025407

Search

Navigation