期刊
JOURNAL OF NEUROSCIENCE METHODS
卷 180, 期 1, 页码 34-42出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jneumeth.2009.02.015
关键词
ELISA; Tau; Three repeat tau; Four repeat tau; Progressive supranuclear palsy and tauopathies; Alzheimer's disease
资金
- Progressive Supranuclear Palsy Association
- Cure PSP
- Reta Lila Weston Trust for Medical Research
- UK Medical Research Council [G0501560]
- MRC [G0501560] Funding Source: UKRI
- Alzheimers Research UK [ART-PhD2007-2] Funding Source: researchfish
- Medical Research Council [G0501560] Funding Source: researchfish
Tau protein plays an important role in stabilising and assembling neuronal microtubules. Pathological changes in expression and aggregation of tau isoforms containing three (3R-tau) and four (4R-tau) microtubule-binding repeat domains are associated with several tauopathies. This paper describes novel sandwich ELISAs for quantification of 3R- and 4R-tau in brain. The assays are constructed using well-characterised isoform-specific antibodies (RD3 and RD4) as capture antibodies and an affinity-purified HRP-anti-tau peptide antibody and biotin-tyramide amplification for detection. For 3R-tau, we achieved a minimal detection limit in buffer of 460 pg mL(-1) and a recovery of 81.0% using 500 pg mL(-1) recombinant 3R-tauspiked in diluted brain homogenate. Mean intra- and inter-assay variation of the 3R-tau ELISA was 8.8 and 10.5%, respectively. For 4R-tau, the detection limit was 780 pg; mL(-1) and the recovery of 5 ng mL(-1) spiked recombinant 4R-tau was 86.0% and the mean intra- and inter-assay variation was 10.4 and 15.6%, respectively. With these assays, we showed that in progressive suprainuclear palsy (PSP) brains, 4R-tau is significantly increased in frontal cortex and caudate, the two regions that are usually associated with 4R-tau-dominant pathology. This increase was not observed in occipital lobe, a region that is spared of tau inclusions. No differences in 3R-tau levels were found between PSP and control brains in all regions tested. With this, we have for the first time developed ELISAs for quantification of 3R- and 4R-tau isoforms in pathological samples. These could prove useful in the pathological investigation and differential diagnosis of tauopathies. (C) 2009 Elsevier B.V. All rights reserved.
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