Heteromers of amyloid precursor protein in cerebrospinal fluid

Molecular Neurodegeneration

Javier Sáez-Valero³  Jose-Luis Molinuevo¹  Kaj Blennow⁴  Gunnar Brinkmalm⁴  Alba Boix-Amorós²  Inmaculada Lopez-Font³  Inmaculada Cuchillo-Ibañez³ 

[1] Alzheimer’s Disease and Other Cognitive Disorders Unit, Neurology Service, Hospital Clinic, Barcelona, Spain;Instituto de Neurociencias de Alicante, Universidad Miguel Hernández-CSIC, Av. Ramón y Cajal s/n, Sant Joan d’Alacant, Spain;Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), Sant Joan d’Alacant, Spain;Clinical Neurochemistry Lab, Institute of Neuroscience and Physiology, University of Gothenburg, Mölndal Campus, Gothenburg, Sweden

关键词: ELISA;    Alzheimer’s disease;    Cerebrospinal fluid;    Heteromers;    sAPPβ;    sAPPα;   

Others  :  1138545
DOI  :  10.1186/1750-1326-10-2

 received in 2014-10-15, accepted in 2014-12-27,  发布年份 2015

【 摘 要 】

Background

Soluble fragments of the amyloid precursor protein (APP) generated by α- and β-secretases, sAPPα and sAPPβ, have been postulated as promising new cerebrospinal fluid (CSF) biomarkers for the clinical diagnosis of Alzheimer’s disease (AD). However, the capacity of these soluble proteins to assemble has not been explored and could be relevant. Our aim is to characterize possible sAPP oligomers that could contribute to the quantification of sAPPα and sAPPβ in CSF by ELISA, as well as to characterize the possible presence of soluble full-length APP (sAPPf).

Results

We employed co-immunoprecipitation, native polyacrylamide gel electrophoresis and ultracentrifugation in sucrose density gradients to characterize sAPP oligomers in CSF. We have characterized the presence of sAPPf in CSF from NDC and AD subjects and demonstrated that all forms, including sAPPα and sAPPβ, are capable of assembling into heteromers, which differ from brain APP membrane-dimers. We measured sAPPf, sAPPα and sAPPβ by ELISA in CSF samples from AD (n = 13) and non-disease subjects (NDC, n = 13) before and after immunoprecipitation with antibodies against the C-terminal APP or against sAPPβ. We demonstrated that these sAPP heteromers participate in the quantification of sAPPα and sAPPβ by ELISA. Immunoprecipitation with a C-terminal antibody to remove sAPPf reduced by ~30% the determinations of sAPPα and sAPPβ by ELISA, whereas immunoprecipitation with an APPβ antibody reduced by ~80% the determination of sAPPf and sAPPα.

Conclusions

The presence of sAPPf and sAPP heteromers should be taken into consideration when exploring the levels of sAPPα and sAPPβ as potential CSF biomarkers.

【 授权许可】

   
2015 Cuchillo-Ibañez et al.; licensee BioMed Central.

【 图 表 】

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