Abstract
Seizures commonly occur in glioma patients, but their pathogenesis is poorly understood, in part due to a lack of valid and versatile experimental models. We have established a new model that enables comprehensive neuropathological and neurophysiological analysis on identical tissue preparations. Rat C6 glioma cells stably transfected with a green fluorescence protein (GFP) gene are transplanted into rat neocortex, giving rise to diffusely invading gliomas histologically resembling human glioblastomas. After 2 weeks, 500-µm-thick cerebral slices are prepared, stained with the voltage-sensitive dye RH795, and fluorescence changes associated with origin and spread of abnormal bioelectric activity upon washout of Mg2+ are detected by a 464-element photodiode array at a rate of 785 frames/s. GFP fluorescence promotes identification of tumor cells during electrophysiological experiments and in neuropathological analyses using frozen and paraffin-embedded tissue sections. By performing subsequent histological analysis of the slices examined neurophysiologically, origin and spread of abnormal activity can be correlated with structural and molecular (immunohistochemical) features. Specifically, we found that ictaform activity was initiated in cortical areas diffusely invaded by single tumor cells. This model is useful for further elucidating the electrophysiological, molecular and structural basis of glioma-associated epileptogenesis.
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Acknowledgements
We thank Dr. Anke Hahnenkamp, Institute of Physiological Chemistry, University Hospital Muenster, for providing the plasmid pMPSGGFP, and Professor Ulrich Mußhoff, Institute of Physiology, University Hospital Muenster, for advice in confocal microscopy. We are grateful to Mrs. Birgit Herrenpoth and Mrs. Andrea Wagner for excellent technical assistance. This study was supported by grant KO 1779/4-1 from Deutsche Forschungsgemeinschaft (DFG).
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V.S. and R.K. contributed equally to this study
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Senner, V., Köhling, R., Püttmann-Cyrus, S. et al. A new neurophysiological/neuropathological ex vivo model localizes the origin of glioma-associated epileptogenesis in the invasion area. Acta Neuropathol 107, 1–7 (2004). https://doi.org/10.1007/s00401-003-0771-5
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DOI: https://doi.org/10.1007/s00401-003-0771-5