Abstract
The lymphatic system plays a key role in tissue fluid homeostasis, immune cell trafficking, and fat absorption. We previously reported a bacterial artificial chromosome (BAC)-based lymphatic reporter mouse, where EGFP is expressed under the regulation of the Prox1 promoter. This reporter line has been widely used to conveniently visualize lymphatic vessels and other Prox1-expressing tissues such as Schlemm’s canal. However, mice have a number of experimental limitations due to small body size. By comparison, laboratory rats are larger in size and more closely model the metabolic, physiological, and surgical aspects of humans. Here, we report development of a novel lymphatic reporter rat using the mouse Prox1-EGFP BAC. Despite the species mismatch, the mouse Prox1-EGFP BAC enabled a reliable expression of EGFP in Prox1-expressing cells of the transgenic rats and allowed a convenient visualization of all lymphatic vessels, including those in the central nervous system, and Schlemm’s canal. To demonstrate the utility of this new reporter rat, we studied the contractile properties and valvular functions of mesenteric lymphatics, developed a surgical model for vascularized lymph node transplantation, and confirmed Prox1 expression in venous valves. Together, Prox1-EGFP rat model will contribute to the advancement of lymphatic research as a valuable experimental resource.
Highlights
In mammals, the blood and the lymphatic systems are necessary for proper circulation
We report the successful generation of a transgenic rat model using the engineered mouse Prox1-EGFP bacterial artificial chromosome (BAC) that was previously used to create the Prox1-EGFP mouse model[9, 17]
To generate a Prox1-EGFP reporter rat model, we initially asked whether the mouse Prox1-EGFP BAC clone, which was previously used to create the Prox1-EGFP mouse model[9, 17], would be functional in rat tissues
Summary
To generate a Prox1-EGFP reporter rat model, we initially asked whether the mouse Prox1-EGFP BAC clone, which was previously used to create the Prox1-EGFP mouse model[9, 17], would be functional in rat tissues. The mouse BAC clone clearly expressed EGFP when transfected in non-mouse cells, such as SW620 (human colon carcinoma)[18], CHO (Chinese hamster ovary cells)[19], and L6 (rat myoblasts)[20] (Supplemental Fig. 1B) These primary data encouraged us to use the mouse BAC clone for generation of Prox1-EGFP reporter rat. With the larger size of the rat, it is technically easier to perform sophisticated surgeries and translational investigations Taken together, this new animal model would enable the convenient, direct, and possibly non-invasive visualization of lymphatic vessels and lymphoid organs, and provide unique opportunities for the anatomic, physiological and surgical studies of the lymphatics
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
