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A new mouse model for renal lesions produced by intravenous injection of diphtheria toxin A-chain expression plasmid
Shingo Nakamura1, Masuo Terashima, Natsuko Kikuchi
1Division of Moleculer Nephrology and Bioartificial Organs, The Institute of Medical Sciences, Tokai University, Bohseidai, Isehara, Kanagawa 259-1193, Japan. snaka@me.ndmc.ac.jp
Background:
Various animal models of renal failure have been produced and used to investigate mechanisms underlying renal disease and develop therapeutic drugs. Most methods available to produce such models appear to involve subtotal nephrectomy or intravenous administration of antibodies raised against basement membrane of glomeruli. In this study, we developed a novel method to produce mouse models of renal failure by intravenous injection of a plasmid carrying a toxic gene such as diphtheria toxin A-chain (DT-A) gene. DT-A is known to kill cells by inhibiting protein synthesis.
Methods:
An expression plasmid carrying the cytomegalovirus enhancer/chicken beta-actin promoter linked to a DT-A gene was mixed with lipid (FuGENE trade mark 6) and the resulting complexes were intravenously injected into adult male B6C3F1 mice every day for up to 6 days. After final injection, the kidneys of these mice were sampled on day 4 and weeks 3 and 5.
Results:
H-E staining of the kidney specimens sampled on day 4 revealed remarkable alterations in glomerular compartments, as exemplified by mesangial cell proliferation and formation of extensive deposits in glomerular basement membrane. At weeks 3 and 5, gradual recovery of these tissues was observed. These mice exhibited proteinuria and disease resembling sub-acute glomerulonephritis.
Conclusions:
Repeated intravenous injections of DT-A expression plasmid DNA/lipid complex caused temporary abnormalities mainly in glomeruli of mouse kidney. The disease in these mice resembles sub-acute glomerulonephritis. These DT-A gene-incorporated mice will be useful as animal models in the fields of nephrology and regenerative medicine.
Insights
Researchers developed a new method to create mouse models of renal failure using diphtheria toxin A-chain (DT-A) gene injection. This novel approach offers a valuable tool for studying kidney disease and regenerative medicine.
Area of Science:
- Nephrology
- Molecular Biology
- Animal Models
Background:
- Existing animal models for renal failure often involve invasive procedures like subtotal nephrectomy or antibody administration.
- There is a need for novel, less invasive methods to generate reliable animal models of kidney disease.
Purpose of the Study:
- To develop a novel method for creating mouse models of renal failure.
- To utilize the diphtheria toxin A-chain (DT-A) gene for inducing kidney damage in mice.
Main Methods:
- An expression plasmid containing the DT-A gene was complexed with lipid (FuGENE 6).
- These complexes were administered intravenously to adult male B6C3F1 mice daily for up to 6 days.
- Kidney tissues were analyzed at different time points post-injection (day 4, weeks 3, and 5).
Main Results:
- Histological analysis revealed significant glomerular alterations, including mesangial cell proliferation and GBM deposits.
- Mice exhibited proteinuria and developed a condition resembling sub-acute glomerulonephritis.
- Gradual recovery of kidney tissue was observed by weeks 3 and 5.
Conclusions:
- Repeated intravenous injections of the DT-A expression plasmid/lipid complex induce transient glomerular abnormalities in mice.
- The induced kidney disease closely mimics sub-acute glomerulonephritis.
- These DT-A gene-modified mice serve as a valuable model for nephrology and regenerative medicine research.
