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58. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie e. V. (DGNC)

Deutsche Gesellschaft für Neurochirurgie (DGNC) e. V.

26. bis 29.04.2007, Leipzig

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Effects of focal ischemia on gene expression in cerebroarterial endothelial cells

Einfluss einer fokalen Ischämie auf die Genexpression in den Endothelzellen zerebraler Arterien

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  • corresponding author B. Brosch - Abteilung für Neurochirurgische Forschung, Universitätsklinikum Mannheim, Fakultät für Klinische Medizin Mannheim, Universität Heidelberg
  • N. Weinzierl - Abteilung für Neurochirurgische Forschung, Universitätsklinikum Mannheim, Fakultät für Klinische Medizin Mannheim, Universität Heidelberg
  • R. Erber - Abteilung für Neurochirurgische Forschung, Universitätsklinikum Mannheim, Fakultät für Klinische Medizin Mannheim, Universität Heidelberg
  • C. Heiser - Abteilung für Neurochirurgische Forschung, Universitätsklinikum Mannheim, Fakultät für Klinische Medizin Mannheim, Universität Heidelberg
  • L. Schilling - Abteilung für Neurochirurgische Forschung, Universitätsklinikum Mannheim, Fakultät für Klinische Medizin Mannheim, Universität Heidelberg

Deutsche Gesellschaft für Neurochirurgie. 58. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie e.V. (DGNC). Leipzig, 26.-29.04.2007. Düsseldorf: German Medical Science GMS Publishing House; 2007. DocP 117

The electronic version of this article is the complete one and can be found online at: http://www.egms.de/en/meetings/dgnc2007/07dgnc372.shtml

Published: April 11, 2007

© 2007 Brosch et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.en). You are free: to Share – to copy, distribute and transmit the work, provided the original author and source are credited.

Outline

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Objective: Transient focal brain ischemia in rats has been described to result in a de novo gene expression of the endothelin (ET)B receptor in the smooth muscle cells mediating contraction. In addition, there is an enhanced relaxation of the isolated middle cerebral artery (MCA) due to stimulation of ET(B) receptors located on the endothelium. The aim of the present study was to determine whether this enhanced relaxation is due to an upregulation of the ET(B) receptor-mRNA in the endothelial cells.

Methods: Adult male Sprague-Dawley rats underwent a 2 h MCA occlusion using the intraluminal filament technique followed by reperfusion. After sacrifice 2 days later, the brain was removed and the previously occluded MCA selectively perfused with an ice-cold buffer to disrupt the cell membranes. The total RNA was isolated from the perfusate and underwent reverse transcription followed by quantitative polymerase chain reaction (qPCR) analysis of the mRNA for ET(B), ET(A), ET-1, and endothelin converting enzyme (ECE) with elongation factor (EF)-1 serving as the house-keeping gene.

Results: After ischemia, the total mRNA content was threefold higher than in control vessels (p<0.05). The slopes of the dilution curves did not differ between the individual primers indicating similar efficiency of amplification. The expression of the ET(A) mRNA was not detectable under control conditions or after ischemia. The expression of ET(B) mRNA was not significantly altered after ischemia. However, the mRNA for ET-1 and ECE were significantly upregulated (p<0.05 vs. control vessels).

Conclusions: The results indicate a marked upregulation of the ET-1 and ECE gene expression in the cerebroarterial endothelial cells after occlusion-reperfusion injury. Furthermore, there appears to be a differential alteration of gene expression in the endothelial and smooth muscle cells (at least with respect to the ET(B) receptor). In conclusion, the present results further emphasize the importance of the vascular ET-system in the pathophysiology of postischemic events.