TY - JOUR
T1 - Comparative Studies of Renin-Null Zebrafish and Mice Provide New Functional Insights
AU - Hoffmann, Scott
AU - Mullins, Linda
AU - Rider, Sebastien
AU - Brown, Cara
AU - Buckley, Charlotte B
AU - Assmus, Adrienne
AU - Li, Ziwen
AU - Sierra Beltran, Mariana
AU - Henderson, Neil
AU - Del Pozo, Jorge
AU - De Goes Martini, Alexandre
AU - Sequeira-Lopez, Maria Luisa S
AU - Gomez, R Ariel
AU - Mullins, John
N1 - Funding Information:
C. Brown, C.B. Buckley, S. Rider, and A. Assmus were supported by the British Heart Foundation Centre of Research Excellence Award (RE/08/001/23904); S. Hoffmann by MRC/EPSRC DTA OPTIMA EP/L016559/1; L. Mullins by the BHF CoRE and Kidney Research UK (RP_026_20180305). R.A. Gomez and M.L.S. Sequeira-Lopez are supported by National Institutes of Health Grants (DK 096373, DK 116718, DK 116196, and HL 148044).
Publisher Copyright:
© 2022 Lippincott Williams and Wilkins. All rights reserved.
PY - 2022/3
Y1 - 2022/3
N2 - BACKGROUND: The renin-angiotensin system is highly conserved across vertebrates, including zebrafish, which possess orthologous genes coding for renin-angiotensin system proteins, and specialized mural cells of the kidney arterioles, capable of synthesising and secreting renin.METHODS: We generated zebrafish with CRISPR-Cas9-targeted knockout of renin (ren-/-) to investigate renin function in a low blood pressure environment. We used single-cell (10×) RNA sequencing analysis to compare the transcriptome profiles of renin lineage cells from mesonephric kidneys of ren-/- with ren+/+ zebrafish and with the metanephric kidneys of Ren1c-/- and Ren1c+/+ mice.RESULTS: The ren-/- larvae exhibited delays in larval growth, glomerular fusion and appearance of a swim bladder, but were viable and withstood low salinity during early larval stages. Optogenetic ablation of renin-expressing cells, located at the anterior mesenteric artery of 3-day-old larvae, caused a loss of tone, due to diminished contractility. The ren-/- mesonephric kidney exhibited vacuolated cells in the proximal tubule, which were also observed in Ren1c-/- mouse kidney. Fluorescent reporters for renin and smooth muscle actin (tg(ren:LifeAct-RFP; acta2:EGFP)), revealed a dramatic recruitment of renin lineage cells along the renal vasculature of adult ren-/- fish, suggesting a continued requirement for renin, in the absence of detectable angiotensin metabolites, as seen in the Ren1YFP Ren1c-/- mouse. Both phenotypes were rescued by alleles lacking the potential for glycosylation at exon 2, suggesting that glycosylation is not essential for normal physiological function.CONCLUSIONS: Phenotypic similarities and transcriptional variations between mouse and zebrafish renin knockouts suggests evolution of renin cell function with terrestrial survival.
AB - BACKGROUND: The renin-angiotensin system is highly conserved across vertebrates, including zebrafish, which possess orthologous genes coding for renin-angiotensin system proteins, and specialized mural cells of the kidney arterioles, capable of synthesising and secreting renin.METHODS: We generated zebrafish with CRISPR-Cas9-targeted knockout of renin (ren-/-) to investigate renin function in a low blood pressure environment. We used single-cell (10×) RNA sequencing analysis to compare the transcriptome profiles of renin lineage cells from mesonephric kidneys of ren-/- with ren+/+ zebrafish and with the metanephric kidneys of Ren1c-/- and Ren1c+/+ mice.RESULTS: The ren-/- larvae exhibited delays in larval growth, glomerular fusion and appearance of a swim bladder, but were viable and withstood low salinity during early larval stages. Optogenetic ablation of renin-expressing cells, located at the anterior mesenteric artery of 3-day-old larvae, caused a loss of tone, due to diminished contractility. The ren-/- mesonephric kidney exhibited vacuolated cells in the proximal tubule, which were also observed in Ren1c-/- mouse kidney. Fluorescent reporters for renin and smooth muscle actin (tg(ren:LifeAct-RFP; acta2:EGFP)), revealed a dramatic recruitment of renin lineage cells along the renal vasculature of adult ren-/- fish, suggesting a continued requirement for renin, in the absence of detectable angiotensin metabolites, as seen in the Ren1YFP Ren1c-/- mouse. Both phenotypes were rescued by alleles lacking the potential for glycosylation at exon 2, suggesting that glycosylation is not essential for normal physiological function.CONCLUSIONS: Phenotypic similarities and transcriptional variations between mouse and zebrafish renin knockouts suggests evolution of renin cell function with terrestrial survival.
KW - actins
KW - glycosylation
KW - pericytes
KW - renin
KW - zebrafish
U2 - 10.1161/HYPERTENSIONAHA.121.18600
DO - 10.1161/HYPERTENSIONAHA.121.18600
M3 - Article
C2 - 35000430
SN - 0194-911X
VL - 79
SP - E56-E66
JO - Hypertension
JF - Hypertension
IS - 3
M1 - HYPERTENSIONAHA12118600
ER -