Abstract
The use of murine models to mimic human kidney disease is becoming increasingly common. Our research is focused on the assessment of glomerular function in diabetic nephropathy and podocyte-specific VEGF-A knock-out mice; therefore, this protocol describes the full kidney work-up used in our lab to assess these mouse models of glomerular disease, enabling a vast amount of information regarding kidney and glomerular function to be obtained from a single mouse. In comparison to alternative methods presented in the literature to assess glomerular function, the use of the method outlined in this paper enables the glomerular phenotype to be fully evaluated from multiple aspects. By using this method, the researcher can determine the kidney phenotype of the model and assess the mechanism as to why the phenotype develops. This vital information on the mechanism of disease is required when examining potential therapeutic avenues in these models. The methods allow for detailed functional assessment of the glomerular filtration barrier through measurement of the urinary albumin creatinine ratio and individual glomerular water permeability, as well as both structural and ultra-structural examination using the Periodic Acid Schiff stain and electron microscopy. Furthermore, analysis of the genes dysregulated at the mRNA and protein level enables mechanistic analysis of glomerular function. This protocol outlines the generic but adaptable methods that can be applied to all mouse models of glomerular disease.
Highlights
The use of murine models to mimic human kidney disease is becoming increasingly common
This protocol describes a method in which the glomeruli are isolated from the mouse kidney cortex, and the protein/RNA are isolated. This allows specific analysis of the protein/mRNA dysregulation in the glomeruli of the disease model. This protocol describes a full kidney work-up that should be carried out in mouse models of glomerular disease, enabling a vast amount of information regarding kidney and glomerular function to be obtained from a single mouse
Upon measurement of the urinary albumin creatinine ratio at weeks 0, 4, 10, and 14 after doxycycline induction of vascular endothelial growth factor A (VEGF-A) KO, VEGF-A KO mice developed progressive albuminuria by 10 weeks compared to wild type (WT) littermate controls
Summary
The use of murine models to mimic human kidney disease is becoming increasingly common. The rationale behind the use of this method is that it enables the glomerular phenotype to be fully evaluated from multiple aspects This includes assessing the glomerular permeability, both to protein and to water, the glomerular structural abnormalities, and changes in the expression/ splicing of mRNAs and proteins essential for normal glomerular function. By using this method, the researcher is able to determine the kidney phenotype of the model and assess the mechanism as to why the phenotype develops. Determine the creatinine concentration of each sample by reading the plate at an absorbance of 490 nm before and after the addition of the acid solution (CAUTION, corrosive; avoid contact with skin). Normalize the data to the baseline value of each mouse for graphical representation
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
