Supplementary Material for: New Anti-Nephrin Antibody Mediated Podocyte Injury Model Using a C57BL/6 Mouse Strain

Background: Focal segmental glomerulosclerosis (FSGS)
is considered a subset of the podocytopathies. The molecular
pathogenesis of podocytopathy is still unknown. There has not been an
experimental animal model of isolated podocytopathy induced by antibody
in C57BL/6 strain, which is widely used as the genetic background.
Nephrin is closely associated with the slit diaphragm of the glomerular
podocyte, and has recently received attention as a potential therapeutic
target. The function of nephrin, especially its role in FSGS
development via podocytopathy, is being elucidated. We report our
experience with a C57BL/6 FSGS model induced by polyclonal rabbit
anti-mouse nephrin antibody (α-mNep Ab). Methods: α-mNep
Ab, which was generated by genetic immunization, was administered into
C57BL/6 mice at once, intravenously. Urinary protein excretion, the
development of glomerulosclerosis and the number of podocyte in mouse
kidney were evaluated. Results: The α-mNep Ab-induced FSGS
was associated with massive proteinuria and nephrotic syndrome. In
periodic acid-Schiff staining, FSGS was observed from day 7 after
antibody injection. Podocyte numbers and podocyte marker (anti-Wilms
tumor 1 and anti-synaptopodin)-positive areas were clearly decreased.
These results suggest that this FSGS mouse model reliably reproduces the
human nephrotic syndrome and FSGS. Conclusion: We
succeeded in making the nephrotic syndrome model mice induced by α-mNep
Ab using C57BL/6. This model may be useful for studying the mechanisms
of podocytopathy.