Purified high molecular weight synthetic Aβ(1-42) and biological Aβ oligomers are equipotent in rapidly inducing MTT formazan exocytosis

Adam M. Weidner; Molly Housley; M. Paul Murphy; Harry LeVine

doi:10.1016/j.neulet.2011.03.082

Purified high molecular weight synthetic Aβ(1-42) and biological Aβ oligomers are equipotent in rapidly inducing MTT formazan exocytosis

Adam M. Weidner, Molly Housley, M. Paul Murphy, Harry LeVine

Research output: Contribution to journal › Article › peer-review

5 Scopus citations

Abstract

Synthetic soluble Aβ oligomers are often used as a surrogate for biologic material in a number of model systems. We compared the activity of Aβ oligomers (synthetic and cell culture media derived) on the human SH-SY5Y neuroblastoma and C2C12 mouse myoblast cell lines in a novel, modified MTT assay. Separating oligomers from monomeric peptide by size exclusion chromatography produced effects at peptide concentrations approaching physiologic levels (10-100nM). Purified oligomers, but not monomers or fibrils, elicited an increase of a detergent-insoluble form of MTT formazan within 2h as opposed to a control toxin (H₂O₂). This effect was comparable for biological and synthetic peptide in both cell types. Monomeric Aβ attenuated the effect of soluble oligomers. This study suggests that the activities of biological and synthetic oligomers are indistinguishable during early stages of Aβ oligomer-cell interaction.

Original language	English
Pages (from-to)	1-5
Number of pages	5
Journal	Neuroscience Letters
Volume	497
Issue number	1
DOIs	https://doi.org/10.1016/j.neulet.2011.03.082
State	Published - Jun 15 2011

Bibliographical note

Funding Information:
The authors would like to acknowledge the contribution of Dr. Susan Catalano of Cognition Therapeutics (Pittsburgh, PA) for studies with primary rat neuronal cultures. We also acknowledge the support of NIH/NIA grants NS058382 (M.P.M., A.M.W.), AG005119 (M.P.M., H.L.) and the Alzheimer's Association IIRG-06-27275 (A.M.W., H.L.). M.H. was supported by an NSF REU Site grant ( DBI-0648233 ).

Keywords

C2C12 myoblast
ELISA
H4 neuroglioma
SH-SY5Y neuroblastoma
Size exclusion chromatography

ASJC Scopus subject areas

Neuroscience (all)

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10.1016/j.neulet.2011.03.082

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title = "Purified high molecular weight synthetic Aβ(1-42) and biological Aβ oligomers are equipotent in rapidly inducing MTT formazan exocytosis",

abstract = "Synthetic soluble Aβ oligomers are often used as a surrogate for biologic material in a number of model systems. We compared the activity of Aβ oligomers (synthetic and cell culture media derived) on the human SH-SY5Y neuroblastoma and C2C12 mouse myoblast cell lines in a novel, modified MTT assay. Separating oligomers from monomeric peptide by size exclusion chromatography produced effects at peptide concentrations approaching physiologic levels (10-100nM). Purified oligomers, but not monomers or fibrils, elicited an increase of a detergent-insoluble form of MTT formazan within 2h as opposed to a control toxin (H2O2). This effect was comparable for biological and synthetic peptide in both cell types. Monomeric Aβ attenuated the effect of soluble oligomers. This study suggests that the activities of biological and synthetic oligomers are indistinguishable during early stages of Aβ oligomer-cell interaction.",

keywords = "C2C12 myoblast, ELISA, H4 neuroglioma, SH-SY5Y neuroblastoma, Size exclusion chromatography",

author = "Weidner, {Adam M.} and Molly Housley and Murphy, {M. Paul} and Harry LeVine",

note = "Funding Information: The authors would like to acknowledge the contribution of Dr. Susan Catalano of Cognition Therapeutics (Pittsburgh, PA) for studies with primary rat neuronal cultures. We also acknowledge the support of NIH/NIA grants NS058382 (M.P.M., A.M.W.), AG005119 (M.P.M., H.L.) and the Alzheimer's Association IIRG-06-27275 (A.M.W., H.L.). M.H. was supported by an NSF REU Site grant ( DBI-0648233 ). ",

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AU - LeVine, Harry

N1 - Funding Information: The authors would like to acknowledge the contribution of Dr. Susan Catalano of Cognition Therapeutics (Pittsburgh, PA) for studies with primary rat neuronal cultures. We also acknowledge the support of NIH/NIA grants NS058382 (M.P.M., A.M.W.), AG005119 (M.P.M., H.L.) and the Alzheimer's Association IIRG-06-27275 (A.M.W., H.L.). M.H. was supported by an NSF REU Site grant ( DBI-0648233 ).

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N2 - Synthetic soluble Aβ oligomers are often used as a surrogate for biologic material in a number of model systems. We compared the activity of Aβ oligomers (synthetic and cell culture media derived) on the human SH-SY5Y neuroblastoma and C2C12 mouse myoblast cell lines in a novel, modified MTT assay. Separating oligomers from monomeric peptide by size exclusion chromatography produced effects at peptide concentrations approaching physiologic levels (10-100nM). Purified oligomers, but not monomers or fibrils, elicited an increase of a detergent-insoluble form of MTT formazan within 2h as opposed to a control toxin (H2O2). This effect was comparable for biological and synthetic peptide in both cell types. Monomeric Aβ attenuated the effect of soluble oligomers. This study suggests that the activities of biological and synthetic oligomers are indistinguishable during early stages of Aβ oligomer-cell interaction.

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Purified high molecular weight synthetic Aβ(1-42) and biological Aβ oligomers are equipotent in rapidly inducing MTT formazan exocytosis

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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