Analysis of β-amyloid-induced abnormalities on fibrin clot structure by spectroscopy and scanning electron microscopy

Pradeep K. Singh, Hanna E. Berk-Rauch, Nadine Soplop, Kunihiro Uryu, Sidney Strickland, Hyung Jin Ahn

Research output: Contribution to journalArticle

Abstract

This article presents methods for generating in vitro fibrin clots and analyzing the effect of beta-amyloid (Aβ) protein on clot formation and structure by spectrometry and scanning electron microscopy (SEM). Aβ, which forms neurotoxic amyloid aggregates in Alzheimer's disease (AD), has been shown to interact with fibrinogen. This Aβ-fibrinogen interaction makes the fibrin clot structurally abnormal and resistant to fibrinolysis. Aβ-induced abnormalities in fibrin clotting may also contribute to cerebrovascular aspects of the AD pathology such as microinfarcts, inflammation, as well as, cerebral amyloid angiopathy (CAA). Given the potentially critical role of neurovascular deficits in AD pathology, developing compounds which can inhibit or lessen the Aβ-fibrinogen interaction has promising therapeutic value. In vitro methods by which fibrin clot formation can be easily and systematically assessed are potentially useful tools for developing therapeutic compounds. Presented here is an optimized protocol for in vitro generation of the fibrin clot, as well as analysis of the effect of Aβ and Aβ-fibrinogen interaction inhibitors. The clot turbidity assay is rapid, highly reproducible and can be used to test multiple conditions simultaneously, allowing for the screening of large numbers of Aβ-fibrinogen inhibitors. Hit compounds from this screening can be further evaluated for their ability to ameliorate Aβ-induced structural abnormalities of the fibrin clot architecture using SEM. The effectiveness of these optimized protocols is demonstrated here using TDI-2760, a recently identified Aβ-fibrinogen interaction inhibitor.

Original languageEnglish (US)
Article numbere58475
JournalJournal of Visualized Experiments
Volume2018
Issue number141
DOIs
StatePublished - Nov 1 2018

Fingerprint

Fibrin
Amyloid
Electron Scanning Microscopy
Fibrinogen
Spectrum Analysis
Spectroscopy
Pathology
Scanning electron microscopy
Screening
Alzheimer Disease
Turbidity
Spectrometry
Assays
Cerebral Amyloid Angiopathy
Proteins
Serum Amyloid A Protein
Aptitude
Amyloid beta-Peptides
Fibrinolysis
Inflammation

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Keywords

  • Alzheimer's disease
  • Beta-amyloid
  • Biochemistry
  • Blood
  • Fibrinogen
  • Issue 141
  • Scanning electron microscopy
  • Spectroscopy

Cite this

Singh, Pradeep K. ; Berk-Rauch, Hanna E. ; Soplop, Nadine ; Uryu, Kunihiro ; Strickland, Sidney ; Ahn, Hyung Jin. / Analysis of β-amyloid-induced abnormalities on fibrin clot structure by spectroscopy and scanning electron microscopy. In: Journal of Visualized Experiments. 2018 ; Vol. 2018, No. 141.
@article{61f6186762a741da910afabe39b64679,
title = "Analysis of β-amyloid-induced abnormalities on fibrin clot structure by spectroscopy and scanning electron microscopy",
abstract = "This article presents methods for generating in vitro fibrin clots and analyzing the effect of beta-amyloid (Aβ) protein on clot formation and structure by spectrometry and scanning electron microscopy (SEM). Aβ, which forms neurotoxic amyloid aggregates in Alzheimer's disease (AD), has been shown to interact with fibrinogen. This Aβ-fibrinogen interaction makes the fibrin clot structurally abnormal and resistant to fibrinolysis. Aβ-induced abnormalities in fibrin clotting may also contribute to cerebrovascular aspects of the AD pathology such as microinfarcts, inflammation, as well as, cerebral amyloid angiopathy (CAA). Given the potentially critical role of neurovascular deficits in AD pathology, developing compounds which can inhibit or lessen the Aβ-fibrinogen interaction has promising therapeutic value. In vitro methods by which fibrin clot formation can be easily and systematically assessed are potentially useful tools for developing therapeutic compounds. Presented here is an optimized protocol for in vitro generation of the fibrin clot, as well as analysis of the effect of Aβ and Aβ-fibrinogen interaction inhibitors. The clot turbidity assay is rapid, highly reproducible and can be used to test multiple conditions simultaneously, allowing for the screening of large numbers of Aβ-fibrinogen inhibitors. Hit compounds from this screening can be further evaluated for their ability to ameliorate Aβ-induced structural abnormalities of the fibrin clot architecture using SEM. The effectiveness of these optimized protocols is demonstrated here using TDI-2760, a recently identified Aβ-fibrinogen interaction inhibitor.",
keywords = "Alzheimer's disease, Beta-amyloid, Biochemistry, Blood, Fibrinogen, Issue 141, Scanning electron microscopy, Spectroscopy",
author = "Singh, {Pradeep K.} and Berk-Rauch, {Hanna E.} and Nadine Soplop and Kunihiro Uryu and Sidney Strickland and Ahn, {Hyung Jin}",
year = "2018",
month = "11",
day = "1",
doi = "10.3791/58475",
language = "English (US)",
volume = "2018",
journal = "Journal of Visualized Experiments",
issn = "1940-087X",
publisher = "MYJoVE Corporation",
number = "141",

}

Analysis of β-amyloid-induced abnormalities on fibrin clot structure by spectroscopy and scanning electron microscopy. / Singh, Pradeep K.; Berk-Rauch, Hanna E.; Soplop, Nadine; Uryu, Kunihiro; Strickland, Sidney; Ahn, Hyung Jin.

In: Journal of Visualized Experiments, Vol. 2018, No. 141, e58475, 01.11.2018.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Analysis of β-amyloid-induced abnormalities on fibrin clot structure by spectroscopy and scanning electron microscopy

AU - Singh, Pradeep K.

AU - Berk-Rauch, Hanna E.

AU - Soplop, Nadine

AU - Uryu, Kunihiro

AU - Strickland, Sidney

AU - Ahn, Hyung Jin

PY - 2018/11/1

Y1 - 2018/11/1

N2 - This article presents methods for generating in vitro fibrin clots and analyzing the effect of beta-amyloid (Aβ) protein on clot formation and structure by spectrometry and scanning electron microscopy (SEM). Aβ, which forms neurotoxic amyloid aggregates in Alzheimer's disease (AD), has been shown to interact with fibrinogen. This Aβ-fibrinogen interaction makes the fibrin clot structurally abnormal and resistant to fibrinolysis. Aβ-induced abnormalities in fibrin clotting may also contribute to cerebrovascular aspects of the AD pathology such as microinfarcts, inflammation, as well as, cerebral amyloid angiopathy (CAA). Given the potentially critical role of neurovascular deficits in AD pathology, developing compounds which can inhibit or lessen the Aβ-fibrinogen interaction has promising therapeutic value. In vitro methods by which fibrin clot formation can be easily and systematically assessed are potentially useful tools for developing therapeutic compounds. Presented here is an optimized protocol for in vitro generation of the fibrin clot, as well as analysis of the effect of Aβ and Aβ-fibrinogen interaction inhibitors. The clot turbidity assay is rapid, highly reproducible and can be used to test multiple conditions simultaneously, allowing for the screening of large numbers of Aβ-fibrinogen inhibitors. Hit compounds from this screening can be further evaluated for their ability to ameliorate Aβ-induced structural abnormalities of the fibrin clot architecture using SEM. The effectiveness of these optimized protocols is demonstrated here using TDI-2760, a recently identified Aβ-fibrinogen interaction inhibitor.

AB - This article presents methods for generating in vitro fibrin clots and analyzing the effect of beta-amyloid (Aβ) protein on clot formation and structure by spectrometry and scanning electron microscopy (SEM). Aβ, which forms neurotoxic amyloid aggregates in Alzheimer's disease (AD), has been shown to interact with fibrinogen. This Aβ-fibrinogen interaction makes the fibrin clot structurally abnormal and resistant to fibrinolysis. Aβ-induced abnormalities in fibrin clotting may also contribute to cerebrovascular aspects of the AD pathology such as microinfarcts, inflammation, as well as, cerebral amyloid angiopathy (CAA). Given the potentially critical role of neurovascular deficits in AD pathology, developing compounds which can inhibit or lessen the Aβ-fibrinogen interaction has promising therapeutic value. In vitro methods by which fibrin clot formation can be easily and systematically assessed are potentially useful tools for developing therapeutic compounds. Presented here is an optimized protocol for in vitro generation of the fibrin clot, as well as analysis of the effect of Aβ and Aβ-fibrinogen interaction inhibitors. The clot turbidity assay is rapid, highly reproducible and can be used to test multiple conditions simultaneously, allowing for the screening of large numbers of Aβ-fibrinogen inhibitors. Hit compounds from this screening can be further evaluated for their ability to ameliorate Aβ-induced structural abnormalities of the fibrin clot architecture using SEM. The effectiveness of these optimized protocols is demonstrated here using TDI-2760, a recently identified Aβ-fibrinogen interaction inhibitor.

KW - Alzheimer's disease

KW - Beta-amyloid

KW - Biochemistry

KW - Blood

KW - Fibrinogen

KW - Issue 141

KW - Scanning electron microscopy

KW - Spectroscopy

UR - http://www.scopus.com/inward/record.url?scp=85058027234&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85058027234&partnerID=8YFLogxK

U2 - 10.3791/58475

DO - 10.3791/58475

M3 - Article

C2 - 30582601

AN - SCOPUS:85058027234

VL - 2018

JO - Journal of Visualized Experiments

JF - Journal of Visualized Experiments

SN - 1940-087X

IS - 141

M1 - e58475

ER -