Ferric Chloride-Induced Arterial Thrombosis and Sample Collection for 3D Electron Microscopy Analysis

Authors

Smita Joshi, University of KentuckyFollow
Alexis N. Smith, University of Kentucky
Kanakanagavalli Shravani Prakhya, University of KentuckyFollow
Hammodah Rawhi Hammodah Alfar, University of KentuckyFollow
Joshua Lykins, University of KentuckyFollow
Ming Zhang, University of Kentucky
Irina D. Pokrovskaya, University of Arkansas for Medical Sciences
Maria A. Aronova, National Institutes of Health
Richard D. Leapman, National Institutes of Health
Brian Storrie, University of Arkansas for Medical SciencesFollow
Sidney W. Whiteheart, University of KentuckyFollow

Abstract

Cardiovascular diseases are a leading cause of mortality and morbidity worldwide. Aberrant thrombosis is a common feature of systemic conditions like diabetes and obesity, and chronic inflammatory diseases like atherosclerosis, cancer, and autoimmune diseases. Upon vascular injury, usually the coagulation system, platelets, and endothelium act in an orchestrated manner to prevent bleeding by forming a clot at the site of the injury. Abnormalities in this process lead to either excessive bleeding or uncontrolled thrombosis/insufficient antithrombotic activity, which translates into vessel occlusion and its sequelae. The FeCl3-induced carotid injury model is a valuable tool in probing how thrombosis initiates and progresses in vivo. This model involves endothelial damage/denudation and subsequent clot formation at the injured site. It provides a highly sensitive, quantitative assay to monitor vascular damage and clot formation in response to different degrees of vascular damage. Once optimized, this standard technique can be used to study the molecular mechanisms underlying thrombosis, as well as the ultrastructural changes in platelets in a growing thrombus. This assay is also useful to study the efficacy of antithrombotic and antiplatelet agents. This article explains how to initiate and monitor FeCl3-induced arterial thrombosis and how to collect samples for analysis by electron microscopy.

Document Type

Article

Publication Date

3-2023

Notes/Citation Information

Copyright © 2023 JoVE Journal of Visualized Experiments

Digital Object Identifier (DOI)

https://dx.doi.org/10.3791/64985

Funding Information

The work was supported by grants from the NIH, NHLBI (HL56652, HL138179, and HL150818), and a Department of Veterans Affairs Merit Award to S.W.W., R01 HL 155519 to B.S., and NIBIB intramural program grant to R.D.L.

Repository Citation

Joshi, Smita; Smith, Alexis N.; Prakhya, Kanakanagavalli Shravani; Alfar, Hammodah Rawhi Hammodah; Lykins, Joshua; Zhang, Ming; Pokrovskaya, Irina D.; Aronova, Maria A.; Leapman, Richard D.; Storrie, Brian; and Whiteheart, Sidney W., "Ferric Chloride-Induced Arterial Thrombosis and Sample Collection for 3D Electron Microscopy Analysis" (2023). Saha Cardiovascular Research Center Faculty Publications. 81.
https://uknowledge.uky.edu/cvrc_facpub/81