Characterization of the Two Inducible Cre Recombinase-Based Mouse Models NG2-CreER™ and PDGFRb-P2A-CreERT2 for Pericyte Labeling in the Retina
D Mayr, J Preishuber-Pflügl, A Koller… - Current Eye …, 2022 - Taylor & Francis
Current Eye Research, 2022•Taylor & Francis
Abstract Purpose Pericytes (PCs), located abluminal of endothelial cells on capillaries, are
essential for vascular development and stability. They display a heterogeneous morphology
depending on organ localization, differentiation state, and function. Consequently, PCs
show a diverse gene expression profile, impeding the usage of a unique PC marker and
therefore the distinct identification of PCs. Inducible reporter mouse models represent an
important tool for investigating the fate of PCs under physiological and pathophysiological …
essential for vascular development and stability. They display a heterogeneous morphology
depending on organ localization, differentiation state, and function. Consequently, PCs
show a diverse gene expression profile, impeding the usage of a unique PC marker and
therefore the distinct identification of PCs. Inducible reporter mouse models represent an
important tool for investigating the fate of PCs under physiological and pathophysiological …
Purpose
Pericytes (PCs), located abluminal of endothelial cells on capillaries, are essential for vascular development and stability. They display a heterogeneous morphology depending on organ localization, differentiation state, and function. Consequently, PCs show a diverse gene expression profile, impeding the usage of a unique PC marker and therefore the distinct identification of PCs. Inducible reporter mouse models represent an important tool for investigating the fate of PCs under physiological and pathophysiological conditions. PC-specific expression efficiency of the fluorescence reporter tdTomato following tamoxifen induction was analyzed and compared in two inducible Cre recombinase-expressing mouse models under control of the NG2 and PDGFRb promotor.
Methods
The NG2-CreER™-tdTomato and the PDGFRb-P2A-CreERT2-tdTomato mice were treated with tamoxifen at three defining time points of retinal vascular development: post-natal days (P)5, P10/11/12, and P48/49/50/51. TdTomato reporter induction efficiency was determined by analyzing retinal whole mounts utilizing confocal microscopy, using the antibodies Anti-neural/glial antigen 2 (PCs), Anti-Collagen IV (basement membrane), and Anti-Glutamine Synthetase (Müller glial cells).
Results
Tamoxifen induction at the three different time points resulted in PC-specific expression of tdTomato in both reporter models. In the NG2-CreER™-tdTomato mouse, the induction efficiency ranged from 21.9 to 35.5%. In the PDGFRb-P2A-CreERT2-tdTomato mouse, an induction efficiency between 78.9 and 94.1% was achieved. TdTomato expression in the retina was restricted to PCs and vascular smooth muscle cells in the NG2-CreER™-tdTomato mouse, however, in the PDGFRb-P2A-CreERT2-tdTomato mouse, tdTomato was also expressed in Müller glial cells.
Conclusion
Both reporter mouse models represent promising tools for fate-mapping studies of PCs. While the NG2-CreER™-tdTomato mouse reveals very specific labeling of PCs in the retina, its induction efficiency is lower compared to the PDGFRb-P2A-CreERT2-tdTomato mouse. Although the latter revealed a high percentage of tdTomato-positive PCs in the retina, additional labeling of Müller cells potentially hampers analysis of reporter-positive PCs.
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