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. 2021 Mar 31;11:7251. doi: 10.1038/s41598-021-86629-w

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Renin-expressing cells arise during mid-gestation as B-1 progenitor cells and persist throughout adult life in the peritoneal cavity as B-1 B cells. (a) Renin progenitors appear in the yolk sac at E11.5 as individual green fluorescent protein positive (GFP⁺) cells within the yolk sac tissue of Ren1^dcre/+;mTmG mice. (b) Semi-quantitative RT-PCR confirmed renin expression in the yolk sac at E11.5. Newborn kidneys and E11.5 livers were used as positive and negative controls respectively. (c) Flow cytometry representative plots show that renin-lineage cells (GFP⁺, x-axis) appear in the fetal liver and spleen at E15.5. With advancing gestational age (E15.5–P0, left to right), there is an increase in the percentage of GFP⁺ cells in both the fetal liver (top panel) and fetal spleen (bottom panel) (Here, GFP represents cells that have expressed renin and red fluorescent protein (RFP) represents cells that have not expressed renin). (d) Percentage of renin progenitors from E15.5 (− 4) to P0 using flow cytometry. (e) Renin lineage (GFP⁺) cells are present in the spleen within the white pulp in newborn mice. (f) Renin-expressing cells in the fetal tissues show a B-1 progenitor phenotype (CD19⁺, B220^dim, CD43⁺, and Lin⁻). (g) The percentage of renin-lineage cells decreases in the bone marrow and spleen over time but persists in the peritoneal cavity throughout adult life. (h) Renin lineage (GFP⁺) cells in the peritoneal cavity are B-1b B cells (B220⁺, CD23⁻, CD11b⁺, CD5⁻) and B-1a B cells (B220⁺, CD23⁻, CD11b⁺, CD5⁺). (i) IMAGESTREAM flow cytometry shows that renin-lineage (GFP⁺) peritoneal cells express B220, CD11b, and + /− CD5. Cells that express CD5 are B-1a B cells, and cells that do not express CD5 are B-1b B cells.