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Treatment with the snail peptide CGX-1007 reduces DNA damage and alters gene expression of c-fos and bcl-2 following focal ischemic brain injury in rats

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Abstract

Delayed cell death following ischemic brain injury has been linked to alterations in gene expression. In this study we have evaluated the upregulation of several genes associated with delayed cell death (c-fos, bax, and bcl-2) during the initial 24 h of transient middle cerebral artery occlusion (MCAo) in the rat and the effects of postinjury treatment with the NR2B subunit specific NMDA receptor antagonist CGX-1007 (Conantokin-G, Con-G). C-fos mRNA levels peaked at 1 h postinjury in both cortical and subcortical ischemic brain regions (30-fold increase), remained elevated at 4 h and returned to within normal, preinjury levels 24 h postinjury. The increase in mRNA levels correlated to increased protein expression in the entire ipsilateral hemisphere at 1 h. Regions of necrosis at 4 h were void of C-Fos immunoreactivity with continued upregulation in surrounding regions. At 24 h, loss of C-Fos staining was observed in the injured hemisphere except for sustained increases along the border of the infarct and in the cingulate cortex of vehicle treated rats. CGX-1007 treatment reduced c-fos expression throughout the infarct region by up to 50%. No significant differences were measured in either bcl-2 or bax mRNA expression between treatment groups. However, at 24 h postinjury CGX-1007 treatment was associated with an increase in Bcl-2 immunoreactivity that correlated to a reduction in DNA fragmentation. In conclusion, CGX-1007 effectively attenuated gene expression associated with delayed cell death as related to a neuroprotective relief of cerebral ischemia.

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Acknowledgements

The authors would like to thank Luisa Velarde, Brad Cunningham, and James Hart for their technical help in this study. We would also like to thank LTC Duane Belote for his assistance in the immunohistochemical analyses.

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Authors and Affiliations

  1. Walter Reed Army Institute of Research, Silver Spring, MD, USA

    A. J. Williams, R. Berti, J. R. Moffett, C. Yao, X. M. Lu, J. R. Dave & F. C. Tortella

  2. Uniformed Services University of Health Sciences, Bethesda, MD, USA

    A. J. Williams & G. Ling

  3. Department of Neuropharmacology and Molecular Biology, Division of Neurosciences, Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA

    A. J. Williams

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  1. A. J. Williams
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  2. G. Ling
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  3. R. Berti
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  4. J. R. Moffett
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  5. C. Yao
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  6. X. M. Lu
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  7. J. R. Dave
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  8. F. C. Tortella
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Correspondence to A. J. Williams.

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Research was conducted in compliance with the Animal Welfare Act and other federal statutes and regulations relating to animals and experiments involving animals and adheres to principles stated in the Guide for the Care and Use of Laboratory Animals, NRC Publication, 1996 edition. Opinions, interpretations, conclusions, and recommendations are those of the authors and are not necessarily endorsed by the US Army

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Williams, A.J., Ling, G., Berti, R. et al. Treatment with the snail peptide CGX-1007 reduces DNA damage and alters gene expression of c-fos and bcl-2 following focal ischemic brain injury in rats. Exp Brain Res 153, 16–26 (2003). https://doi.org/10.1007/s00221-003-1566-6

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  • DOI: https://doi.org/10.1007/s00221-003-1566-6

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