Document Detail

Triple fluorescence labelling of neuronal, glial and vascular markers revealing pathological alterations in various animal models.
MedLine Citation:
PMID:  19028564     Owner:  NLM     Status:  MEDLINE    
The simultaneous detection of glia, vessels and neurons facilitates insights into the complex chemoarchitecture of the central nervous system. Here, we present a simple, robust and versatile approach for the carbocyanine triple fluorescence labelling of neuronal, vascular and glial markers. The usefulness of this procedure is shown for rat brain tissue under physiological conditions, after traumatic brain injury caused by controlled cortical impact injury, and after stroke following middle cerebral artery occlusion. Moreover, the versatility of the method is verified by its application to sections from old triple transgenic mice with age-dependent beta-amyloidosis and tau hyperphosphorylation in the hippocampus, modelling neuropathological alterations in Alzheimer's disease. To exemplify the usefulness of the approach for analysis of the enteric nervous system, it was applied to whole mounts from the horse intestine. The biotinylated lectin from potato (Solanum tuberosum) is presented as an excellent tool to detect both vessels and microglia. Furthermore, this lectin revealed macrophages after experimental insults, and senile plaques in aged triple transgenic mice. A large portion of astroglia was demonstrated by immunolabelling of glial fibrillary acidic protein. Neurons were detected by monoclonal antibodies directed against neuronal nuclei and, in horse tissues, mouse-anti-HuC/D recognizing a conserved nuclear protein. Confocal laser-scanning microscopy elucidated spatial relationships of the relevant markers and their pathological alterations after experimental insults and in transgenic mice with Alzheimer-like lesions.
Wolfgang Härtig; Andreas Reichenbach; Cornelia Voigt; Johannes Boltze; Larysa Bulavina; Martin U Schuhmann; Johannes Seeger; Gerald F Schusser; Christiane Freytag; Jens Grosche
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Publication Detail:
Type:  Journal Article     Date:  2008-11-06
Journal Detail:
Title:  Journal of chemical neuroanatomy     Volume:  37     ISSN:  1873-6300     ISO Abbreviation:  J. Chem. Neuroanat.     Publication Date:  2009 Mar 
Date Detail:
Created Date:  2009-02-02     Completed Date:  2009-06-22     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8902615     Medline TA:  J Chem Neuroanat     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  128-38     Citation Subset:  IM    
Paul Flechsig Institute for Brain Research, University of Leipzig, Jahnallee 59, 04109 Leipzig, Germany.
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MeSH Terms
Biological Markers / analysis,  metabolism
Blood Vessels / metabolism,  pathology*
Brain / pathology*,  physiopathology
Brain Diseases / pathology*,  physiopathology
Disease Models, Animal
Enteric Nervous System / cytology,  metabolism
Mice, Transgenic
Microscopy, Confocal / methods
Microscopy, Fluorescence
Neuroglia / metabolism,  pathology*
Neurons / metabolism,  pathology*
Plant Lectins / diagnostic use*
Rats, Inbred SHR
Rats, Sprague-Dawley
Staining and Labeling / methods
Reg. No./Substance:
0/Biological Markers; 0/Plant Lectins; 0/potato lectin

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