Deletion of the prorenin receptor in the ureteric bud in mice inhibits Dot1/H3K79 pathway,Pediatric Research

当前位置： X-MOL 学术 › Pediatr. Res. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Deletion of the prorenin receptor in the ureteric bud in mice inhibits Dot1/H3K79 pathway
Pediatric Research ( IF 3.1 ) Pub Date : 2024-01-29 , DOI: 10.1038/s41390-024-03026-5
Renfang Song ₁ , Ihor V Yosypiv ₁

Affiliation

Background

The prorenin receptor (PRR) plays a critical role in ureteric bud (UB) branching morphogenesis. DOT1 Like (DOT1L), a histone methyltransferase specific for Histone 3 lysine 79 (H3K79), is important for differentiation of the UB-derived renal collecting duct cells. In this study, we tested whether DOT1L/H3 dimethyl K79 (H3m2K79) are regulated by PRR deletion in the UB and UB-derived collecting ducts in the embryonic mouse kidneys.

Methods

Mutant Hoxb7^Cre+/PRR^flox/flox (PRR^UB-/-) and control PRR^UB+/+, mice were studied on embryonic (E) day E17.5. DOT1L mRNA and protein expression in the kidney was examined by real-time qRT-PCR and immunohistochemistry, respectively. H3m2K79 protein expression was determined by immunohistochemistry and Western blot analysis.

Results

DOT1L mRNA levels were decreased in mutant compared to control mice (0.68 ± 0.06 vs. 1.0 ± 0.01, p < 0.01). DOT1L and H3m2K79 immunostaining was reduced in the mutant vs. control kidneys (Dot1: 0.62 ± 0.03 vs. 1.0 ± 0.01, p < 0.05; H3m2K79: 0.64 ± 0.04 vs.1.1 ± 0.01. p < 0.05.). Western blot analysis revealed decreased H3m2K79 protein levels in mutant compared to control kidneys (1.0 ± 0.06 vs. 1.5 ± 0.02, p < 0.05).

Conclusion

Targeted deletion of the PRR in the UB and UB-derived collecting ducts results in reduced DOT1L gene/protein and H3m2K79 protein expression in the embryonic mouse metanephroi in vivo.

Impact

The role of histone methylation in mediating the effect of the prorenin receptor on the ureteric bud branching (UB) morphogenesis and urine acidification during kidney development is unknown.
We demonstrate that histone H3 lysine (K) 79 dimethylation by methyltransferase Dot1 is reduced in the embryonic kidney of mice that lack the prorenin receptor in the UB lineage.

中文翻译：

小鼠输尿管芽中肾素原受体的缺失抑制Dot1/H3K79通路

背景

肾素原受体 (PRR) 在输尿管芽 (UB) 分支形态发生中起着至关重要的作用。 DOT1 Like (DOT1L) 是一种对组蛋白 3 赖氨酸 79 (H3K79) 具有特异性的组蛋白甲基转移酶，对于 UB 来源的肾集合管细胞的分化非常重要。在本研究中，我们测试了胚胎小鼠肾脏中 UB 和 UB 衍生集合管中PRR缺失是否调节 DOT1L/H3 二甲基 K79 (H3m2K79)。

方法

突变体Hoxb7 ^Cre+ / PRR ^flox/flox ( PRR ^{UB -/-} ) 和对照PRR ^UB+/+小鼠在胚胎 (E) 天 E17.5 进行研究。分别通过实时 qRT-PCR 和免疫组织化学检查肾脏中 DOT1L mRNA 和蛋白表达。通过免疫组织化学和蛋白质印迹分析测定H3m2K79蛋白表达。

结果

与对照小鼠相比，突变体小鼠的DOT1L mRNA 水平降低（0.68 ± 0.06 vs. 1.0 ± 0.01， p < 0.01）。与对照肾脏相比，突变肾脏中的 DOT1L 和 H3m2K79 免疫染色减少（Dot1：0.62 ± 0.03 与 1.0 ± 0.01， p < 0.05；H3m2K79：0.64 ± 0.04 与 1.1 ± 0.01。p < 0.05。）。蛋白质印迹分析显示，与对照肾脏相比，突变体肾脏中的 H3m2K79 蛋白水平降低（1.0 ± 0.06 与 1.5 ± 0.02， p < 0.05）。