Analysis of microdissected human neurons by a sensitive ELISA reveals a correlation between elevated intracellular concentrations of Abeta42 and Alzheimer's disease neuropathology. - AlzForum Alzheimer Research Paper of the Week


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Home: Papers of the Week

Annotation

	Hashimoto M, Bogdanovic N, Volkmann I, Aoki M, Winblad B, Tjernberg LO. Analysis of microdissected human neurons by a sensitive ELISA reveals a correlation between elevated intracellular concentrations of Abeta42 and Alzheimer's disease neuropathology. Acta Neuropathol. 2010 May;119(5):543-54. PubMed Abstract

	Comments on Paper and Primary News

	Comment by: Paul Coleman, ARF Advisor
	Submitted 6 March 2010 \| Permalink	Posted 9 March 2010
	I recommend this paper The abstract leads one to believe the authors used an ELISA on either single or relatively few neurons captured by LCM? This would be a great technical advance. View all comments by Paul Coleman

	Comment by: Jurgen Goetz, ARF Advisor
	Submitted 9 March 2010 \| Permalink	Posted 10 March 2010
	I recommend this paper This is an interesting paper shedding light on the enigma of 'selective vulnerability' in AD. View all comments by Jurgen Goetz

	Comment by: Gunnar K. Gouras
	Submitted 10 March 2010 \| Permalink	Posted 10 March 2010
	I recommend this paper Biochemical analysis of intraneuronal Aβ in the brain is challenging and in contrast to the many immunohistochemical studies, there are very few biochemical studies on this topic. This very interesting study utilized laser capture microdissection microscopy and a new sensitive Aβ ELISA to demonstrate an increased Aβ42 to 40 ratio within neurons of sporadic AD compared to control brains. In addition, the authors show a marked increase in levels of Aβ in AD vulnerable (hippocampal CA1 pyramidal) compared to resistant (cerebellar Purkinje) neurons in AD brains. View all comments by Gunnar K. Gouras

	Comment by: Masakazu Hashimoto, Lars Tjernberg
	Submitted 11 March 2010 \| Permalink	Posted 11 March 2010
	I recommend this paper A brief reply to Dr. Coleman: We captured 500 neurons from each case. The final number of neurons analyzed by the ELISA was around 5 neurons per well. View all comments by Masakazu Hashimoto View all comments by Lars Tjernberg

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REAGENTS/MATERIAL:
Immunohistochemistry The paraffin-embedded sections were used to detect plaques in cerebellar sections using IHC. The section of frontal cortex from one Swe mutant case (APPswe-1) was used for positive staining. The primary antibodies (kindly provided by Dr. Jan Näslund) were adopted for detection of Ab40 and Ab42.
ELISA: The sandwich ELISA system from Wako Chemicals GmbH (Neuss, Germany) was adopted as a conventional ELISA (Ab40: Catalog No. 294-64701, Ab42: Catalog No. 292-64501). These ELISA systems include BNT77 as a capture antibody (anti human Ab4011–28 monoclonal antibody), and BA27 (Abx–40) or BC05 (Abx–42) for detection of Ab40 or Ab42, respectively.




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