Abstract
The mouse Agouti gene encodes a paracrine signaling factor which promotes melanocytes to produce yellow instead of black pigment. It has been reported that Agouti mRNA is confined to the dermal papilla after birth in various mammalian species. In this study, we created and characterized a knockin mouse strain in which Cre recombinase was expressed in-frame with endogenous Agouti coding sequence. The Agouti–Cre mice were bred with reporter mice (Rosa26-tdTomato or Rosa26-ZsGreen) to trace the lineage of Agouti-expressing cells during development. In skin, the reporter was detected in some dermal fibroblasts at the embryonic stage and in all dermal fibroblasts postnatally. It was also expressed in all mesenchymal lineage cells in other organs/tissues, including eyes, tongue, muscle, intestine, adipose, prostate and testis. Interestingly, the reporter expression was excluded from epithelial cells in the above organs/tissues. In brain, the reporter was observed in the outermost meningeal fibroblasts. Our work helps to illustrate the Agouti expression pattern during development and provides a valuable mouse strain for conditional gene targeting in mesenchymal lineage cells in multiple organs.
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Acknowledgements
We thank Rongyin Gui at Shanghai University for helping to breed the mice.
Funding
National Key Research and Development Program of China, Grant/Award Number: 2020YFA01130002; National Natural Science Foundation of China, Grant/Award Number: 81972563 and 81903054.
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XHL: Conceptualization, Methodology, Supervision, Data curation, Writing. RS, LPG: Conceptualization, Methodology, Supervision. XRS, HLZ, XBZ, YGW, XYT, Investigation, Data curation.
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Dr. Ruilin Sun is the vice general manager of Shanghai Model Organisms Center, Inc. which provides animal model services. The other authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
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Supplementary Fig. S1
Agouti expression in different populations of skin cells. A Schematic of back skin at embryonic day 14.5 (E14.5). B Agouti mRNA expression in all cell populations of the skin at E14.5 based on RNA-seq data reported previously (Sennett et al. 2015). C Schematic of back skin at postnatal day 5 (P5) (Rezza et al. 2016). D Agouti mRNA expression in all cell populations of the skin at P5 based on RNA-seq data. E Agouti mRNA expression in all cell populations of the skin at P4 based on DNA microarray data (Rendl et al. 2005) (TIF 7422 KB)
Supplementary Fig. S2
Agouti–Cre induces recombination in mesenchymal lineage cells in multiple organs at embryonic stage and in newborns. The whole embryo at E12.5 or organs at E12.5 (N = 3), E17.5 (N = 3) and P0 (N = 3) from Agouti–Cre, Rosa26-ZsGreen mice were harvested, fixed, sectioned, stained for nuclei by Hoechst and observed under the microscope. A Whole embryo. B Eye. The left panel is the enlarged view from the area indicated within the red dashed line in the right panel. C Tongue. D Smooth muscle. E Small intestine. F. Testis. The dash line represents the interface between epithelial and mesenchymal compartments. Scale bar, 100 μm (TIF 40907 KB)
Supplementary File 1
Full Report for the establishment of Agouti–Cre knockin mouse model (PDF 1137 KB)
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Shen, XR., Zhang, HL., Zhao, XB. et al. A Cre knockin mouse reveals specific expression of Agouti gene in mesenchymal lineage cells in multiple organs and provides a unique tool for conditional gene targeting. Transgenic Res 32, 143–152 (2023). https://doi.org/10.1007/s11248-023-00334-0
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DOI: https://doi.org/10.1007/s11248-023-00334-0