Journal of Molecular Neuroscience

, Volume 43, Issue 3, pp 268–274 | Cite as

Glial Amyloid Precursor Protein Expression is Restricted to Astrocytes in an Experimental Toxic Model of Multiple Sclerosis

  • Tim Clarner
  • Jan Philipp Buschmann
  • Cordian Beyer
  • Markus Kipp
Article

Abstract

The amyloid precursor protein is rapidly induced in reactive glia in response to pathological stimuli and inflammation. In this study, we investigated its expression in an experimental multiple sclerosis animal model, the cuprizone mouse model which reveals massive myelin loss. Cuprizone intoxication for 5 weeks induced immense demyelination of the corpus callosum and resulted in hypertrophic and hyperplastic astrocytosis accompanied by microglia/macrophage invasion. Using double-immunofluorescence, real-time quantitative PCR and Western Blot, we observed that activated astrocytes are the main source of amyloid precursor protein during demyelination. In order to rule out astrocytes, in general, responding to inflammatory and toxic compounds by amyloid precursor protein expression, neonatal astroglia cultures were exposed to various stimuli. Under control conditions, astroglial amyloid precursor protein was only moderately expressed. None of the treatments had a significant effect on its expression in vitro. Our results suggest that amyloid precursor protein is specifically up-regulated under cuprizone-induced demyelination. It remains to be further elucidated whether amyloid precursor protein-positive astrocytes are directly implicated in the pathological mechanism of demyelination.

Keywords

Amyloid precursor protein Astroglia Microglia Cuprizone Demyelination 

Abbreviations

AD

Alzheimers disease

APP

Amyloid precursor protein

CC

Corpus callosum

cDNA

Complementary DNA

Cu

Cuprizone

EAE

Experimental Autoimmune Encephalitis

GFAP

Glial fibrillary acidic protein

H2O2

Hydrogen peroxide

HPRT

Hypoxanthine guanine phosphoribosyltransferase

Iba1

Ionized calcium-binding adaptor molecule 1

M-MLV

Moloney Murine Leukemia Virus

MS

Multiple sclerosis

OD

Optical density

PLB-TCEP

Protein loading buffer Tris (2-carboxyethyl)phosphine Hydrochloride

PLP

Proteolipid protein

Real-time RT-PCR

Real-time reverse transcriptase quantitative polymerase chain reaction

SDS-PAGE

Sodium dodecylsulfate polyacrylamide gel electrophoresis

SEM

Standard error of the mean

TAQ Polymerase

Thermus aquaticus DNA polymerase

TBS

Tris buffered saline

Th1 cells

T helper 1 cells

TNF-α

Tumor necrosis factor alpha

Notes

Acknowledgements

This project was supported by the START-Program (M.K.) of the Faculty of Medicine, RWTH Aachen University and by the Hertie-Foundation (M.K.). We would like to thank H. Helten for excellent technical support.

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Tim Clarner
    • 1
  • Jan Philipp Buschmann
    • 1
  • Cordian Beyer
    • 1
  • Markus Kipp
    • 1
  1. 1.Institute of Neuroanatomy, Faculty of MedicineRWTH Aachen UniversityAachenGermany

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