Abstract: SA-PO447
Embryonic Stage Adam10/Notch Pathway Excessive Activation Promotes Ectopic Proximal Tubules Formation and Kidney Fibrosis
Session Information
- Development and Regenerative Medicine
November 09, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Development, Stem Cells, and Regenerative Medicine
- 500 Development, Stem Cells, and Regenerative Medicine
Authors
- Li, Bingjue, Kidney Disease Center, the First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China
- Chen, Jianghua, Kidney Disease Center, the First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China
- Jiang, Hong, Kidney Disease Center, the First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China
Background
Chronic kidney disease (CKD) is an outstanding public health problem. It is important to elucidate the pathogenesis of CKD and researches on kidney development become a breakthrough. Studies have verified that Notch pathway plays a significant role in kidney development, it is widely believed that Notch promote the formation of proximal tubules. In addition to its role in kidney development, Notch was also found to be involved in kidney fibrosis. Here using prenatal chlorpyrifos (CPF) exposure mouse model we make a further study on the role of Notch in kidney development and fibrosis.
Methods
CPF 5mg/kg/d was administrated by subcutaneous injection in CPF-treated pregnant mice from gestation day 7.5-11.5 while the controls were injected with DMSO. RNA-seq was performed of E12.5, E14.5, E16.5 and E18.5 kidneys, RT-qPCR and WB were performed to verify gene expression. IF and IHC were performed to studied the protein levels and kidney structure changes of offspring mice (4 weeks, 8 weeks and 6 months). Masson staining and fibrosis factor detection were used to evaluate fibrosis. Kidney size and weight, Scr and Bun levels were measured to evaluate renal function.
Results
RNA-seq analysis revealed that Adam10/Notch and Aqp1 were increased in CPF group and nephron progenitor cell (NPC) marker Six2 was decreased. mRNA and protein levels were verified by RT-qPCR, WB, IF and IHC. IF and IHC showed the increasement of proximal tubules. Experiments of offspring mice kidneys showed abnormal embryonic kidney phenotypes persisted in adult kidneys. High activation of Notch also led to impaired kidney function and more severe renal fibrosis.
Conclusion
Excessive activation of Adam10/Notch pathway caused the depletion of SIX2+ NPCs and ectopic proximal tubules formation and aggravate kidney fibrosis.
Funding
- Government Support - Non-U.S.