A new approach to reduce the number of animals used in experimental focal cerebral ischemia models
| Autor: | Martin Dichgans, Martin Liebetrau, Gerhard F. Hamann, Dorothe Burggraf, Helge K. Martens, Milan R. Vosko |
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| Rok vydání: | 2005 |
| Předmět: |
Male
Pathology medicine.medical_specialty Central nervous system Blotting Western Ischemia Infarction Biology Brain Ischemia Basal ganglia medicine Animals Rats Wistar General Neuroscience Histological Techniques Brain medicine.disease Immunohistochemistry Rats Disease Models Animal medicine.anatomical_structure Coronal plane Sample Size Basal lamina Perfusion |
| Zdroj: | Neuroscience letters. 386(2) |
| ISSN: | 0304-3940 |
| Popis: | We describe a novel experimental set-up that allows biochemical, immunohistochemical and morphometric recording of multiple parameters from a single rat brain. The whole brain was cut (coronal sectioning) in a volumetric manner, and 100 cryo-sections (10 microm) were collected from the region of infarction. By use of a scalpel to dissect the cryosection, crude brain material was obtained from the cortical and basal ganglia areas of ischemic and non-ischemic hemispheres. Material from four 10 microm thick sections of the same animal was pooled. About 30 microg protein lysate was extracted per four sections with various lysis buffers; this sufficed for one biochemical or enzymatic test called "micro-Western-blots" or "micro-zymographies". Scraping brain material from cryosections allows the detection of up to 25 parameters from adjacent brain sections of one single rat brain. Different analysis are possible, we have chosen, e.g. to compare factors affecting the basal lamina of cerebral microvessels like the content of the metalloproteinases-2/-9, their tissue inhibitors, the plasminogen activators, collagen type IV, parameters to test the blood-brain barrier: hemoglobin and the protein of the perfusion solution BSA and the infarction volume. On the basis of these parameters it was possible to compare the interactions of the complex processes in the ischemic brain in the same animal in adjacent sections. Thus, this method increases the validity of data comparisons and reduces significantly the number of animals needed in various experimental settings. |
| Databáze: | OpenAIRE |
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