Development of an Endothelial Cell-restricted Transgenic Reporter Rat: a Resource for Physiological Studies of Vascular Biology

Authors

Mikhail Alexeyev, University of South AlabamaFollow
Aron M. Geurts, Medical College of Wisconsin
Naga S. Annamdevula, University of South AlabamaFollow
C. Michael Francis, University of South Alabama
Silas Josiah Leavesley, University of South AlabamaFollow
Thomas C. Rich, University of South AlabamaFollow
Mark S. Taylor, University of South AlabamaFollow
Mike T. Lin, University of South AlabamaFollow
Ron Balczon, University of South AlabamaFollow
Jennifer Michelle Knighten, University of South Alabama
Diego F. Alvarez, Sam Houston State University
Troy Stevens, University of South AlabamaFollow

Document Type

Article

Publication Title

American Journal of Physiology-Heart and Circulatory Physiology

Abstract

Here, we report the generation of a Cre-recombinase (iCre) transgenic rat, where iCre is driven using a vascular endothelial-cadherin (CDH5) promoter. The CDH5 promoter was cloned from rat pulmonary microvascular endothelial cells and demonstrated ~60% similarity to the murine counterpart. The cloned rat promoter was 2,508 bp, it extended 79 bp beyond the transcription start site, and it was 22,923 bp upstream of the translation start site. The novel promoter was cloned upstream of codon-optimized iCre and subcloned into a Sleeping Beauty transposon vector for transpositional transgenesis in Sprague-Dawley rats. Transgenic founders were generated and selected for iCre expression. Crossing the CDH5-iCre rat with a tdTomato reporter rat resulted in progeny displaying endothelium-restricted fluorescence. tdTomato fluorescence was prominent in major arteries and veins, and it was similar in males and females. Quantitative analysis of the carotid artery and the jugular vein revealed that, on average, more than 50% of the vascular surface area exhibited strong fluorescence. tdTomato fluorescence was observed in the circulations of every tissue tested. The microcirculation in all tissues tested displayed homogenous fluorescence. Fluorescence was examined across young (6–7.5 mo), middle (14–16.5 mo), and old age (17–19.5 mo) groups. Although tdTomato fluorescence was seen in middle- and old-age animals, the intensity of the fluorescence was significantly reduced compared with that seen in the young rats. Thus, this endothelium-restricted transgenic rat offers a novel platform to test endothelial microheterogeneity within all vascular segments, and it provides exceptional resolution of endothelium within-organ microcirculation for application to translational disease models.

NEW & NOTEWORTHY The use of transgenic mice has been instrumental in advancing molecular insight of physiological processes, yet these models oftentimes do not faithfully recapitulate human physiology and pathophysiology. Rat models better replicate some human conditions, like Group 1 pulmonary arterial hypertension. Here, we report the development of an endothelial cell-restricted transgenic reporter rat that has broad application to vascular biology. This first-in-kind model offers exceptional endothelium-restricted tdTomato expression, in both conduit vessels and the microcirculations of organs.

First Page

H349

Last Page

H358

DOI

10.1152/ajpheart.00276.2020

Publication Date

8-1-2020

Department

College of Medicine

Comments

Copyright © 2020 the American Physiological Society

https://doi.org/10.1152/ajpheart.00276.2020

Recommended Citation

Alexeyev, M., Geurts, A. M., Annamdevula, N. S., Francis, C. M., Leavesley, S. J., Rich, T. C., ... & Stevens, T. (2020). Development of an endothelial cell-restricted transgenic reporter rat: A resource for physiological studies of vascular biology. American Journal of Physiology-Heart and Circulatory Physiology, 319(2), H349-H358.