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Anea et al. Vessel Plus 2018;2:16 Vessel Plus
DOI: 10.20517/2574-1209.2018.46

Original Article Open Access

Immunohistochemistry of the circadian clock in
mouse and human vascular tissues

Ciprian B. Anea , Ana M. Merloiu , David J. R. Fulton , Vijay Patel , R. Dan Rudic 1
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1 Department of Pharmacology & Toxicology, Medical College of Georgia at Augusta University, Augusta, GA 30912, USA.
2 Department of Surgery, Medical College of Georgia at Augusta University, Augusta, GA 30912, USA.
Correspondence to: Dr. R. Dan Rudic, Department of Pharmacology & Toxicology, Medical College of Georgia at Augusta
University, Augusta, GA 30912, USA. E-mail: rrudic@augusta.edu

How to cite this article: Anea CB, Merloiu AM, Fulton DJR, Patel V, Rudic RD. Immunohistochemistry of the circadian clock in
mouse and human vascular tissues. Vessel Plus 2018;2:16. http://dx.doi.org/10.20517/2574-1209.2018.46

Received: 14 Jun 2018 First Decision: 3 Jul 2018 Revised: 10 Jul 2018 Accepted: 11 Jul 2018 Published: 20 Jul 2018
Science Editor: Alexander D. Verin Copy Editor: Jun-Yao Li Production Editor: Cai-Hong Wang

Abstract
Aim: The circadian clock is a molecular network that controls the body physiological rhythms. In blood vessels, the
circadian clock components modulate vascular remodeling, blood pressure, and signaling. The goal in this study was to
determine the pattern of expression of circadian clock proteins in the endothelium, smooth muscle, and adventitia of the
vasculature of human and mouse tissues.

Methods: Immunohistochemistry was performed in frozen sections of mouse aorta, common carotid artery, femoral
artery, lung, and heart at 12 AM and 12 PM for Bmal1, Clock, Npas2, Per and other clock components. Studies of
expression were also assessed in human saphenous vein both by immunoblotting and immunohistochemistry.

Results: In this study, we identified the expression of Bmal1, Clock, Npas, Per1, Cry1, and accessory clock
components by immunohistochemical staining in the endothelium, smooth muscle and adventitia of the mouse
vasculature with differing temporal and cellular profiles depending on vasculature and tissue analyzed. The human
saphenous vein also exhibited expression of clock genes that exhibited an oscillatory pattern in Bmal1 and Cry by
immunoblotting.

Conclusion: These studies show that circadian clock components display differences in expression and localization
throughout the cardiovascular system, which may confer nuances of circadian clock signaling in a cell-specific manner.

Keywords: Circadian blood vessel, vascular endothelium, smooth muscle, Clock, Bmal1, aorta, human, mouse, Per, Cry

© The Author(s) 2018. Open Access This article is licensed under a Creative Commons Attribution 4.0
International License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use,
sharing, adaptation, distribution and reproduction in any medium or format, for any purpose, even commercially, as long
as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license,
and indicate if changes were made.

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