Simultaneous monitoring of cerebral metal accumulation in an experimental model of Wilson’s disease by laser ablation inductively coupled plasma mass spectrometry

BMC Neuroscience

Ralf Weiskirchen¹  Wolfgang Stremmel⁴  Andreas Matusch³  Sabine Weiskirchen¹  Astrid Zimmermann²  Ricarda Uerlings¹  Uta Merle⁴  Sorina Georgiana Boaru¹ 

[1] Institute of Clinical Chemistry and Pathobiochemistry, RWTH Aachen University Hospital Aachen, Pauwelsstr. 30, D-52074 Aachen, Germany;Central Institute of Engineering, Electronic und Analytics (ZEA-3), Research Centre Jülich (FZJ), Jülich, Germany;Institute of Neuroscience and Medicine (INM-2), Research Centre Jülich (FZJ), Jülich, Germany;Department of Gastroenterology, Internal Medicine IV, University Clinics of Heidelberg, Heidelberg, Germany

关键词: ATPase7B;    Copper;    LA-ICP-MS;    Bio-imaging;    Wilson’s disease;   

Others  :  1091357
DOI  :  10.1186/1471-2202-15-98

 received in 2014-03-17, accepted in 2014-08-13,  发布年份 2014

【 摘 要 】

Background

Neuropsychiatric affection involving extrapyramidal symptoms is a frequent component of Wilson’s disease (WD). WD is caused by a genetic defect of the copper (Cu) efflux pump ATPase7B. Mouse strains with natural or engineered transgenic defects of the Atp7b gene have served as model of WD. These show a gradual accumulation and concentration of Cu in liver, kidneys, and brain. However, still little is known about the regional distribution of Cu inside the brain, its influence on other metals and subsequent pathophysiological mechanisms. We have applied laser ablation inductively coupled plasma mass spectrometry and performed comparative metal bio-imaging in brain sections of wild type and Atp7b null mice in the age range of 11–24 months. Messenger RNA and protein expression of a panel of inflammatory markers were assessed using RT-PCR and Western blots of brain homogenates.

Results

We could confirm Cu accumulation in brain parenchyma by a factor of two in WD (5.5 μg g⁻¹ in the cortex) vs. controls (2.7 μg g⁻¹) that was already fully established at 11 months. In the periventricular regions (PVR) known as structures of prominent Cu content, Cu was reduced in turn by a factor of 3. This corroborates the view of the PVR as efflux compartments with active transport of Cu into the cerebrospinal fluid. Furthermore, the gradient of Cu increasing downstream the PVR was relieved. Otherwise the architecture of Cu distribution was essentially maintained. Zinc (Zn) was increased by up to 40% especially in regions of high Cu but not in typical Zn accumulator regions, a side effect due to the fact that Zn is to some degree a substrate of Cu-ATPases. The concentrations of iron (Fe) and manganese (Mn) were constant throughout all regions assessed. Inflammatory markers TNF-α, TIMP-1 and the capillary proliferation marker α-SMA were increased by a factor of 2–3 in WD.

Conclusions

This study confirmed stable cerebral Cu accumulation in parenchyma and discovered reduced Cu in cerebrospinal fluid in Atp7b null mice underlining the diagnostic value of micro-local analytical techniques.

【 授权许可】

   
2014 Boaru et al.; licensee BioMed Central Ltd.

【 图 表 】

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