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.

Keywords:
Nephrin, Glomerulosclerosis, Focal segmental, Podocytes , Nephrotic model, Antibody
1.
Kriz W, Shirato I, Nagata M, LeHir M, Lemley KV: The podocyte's response to stress: the enigma of foot process effacement. Am J Physiol Renal Physiol 2013;304:F333-F347.
2.
Liapis H, Romagnani P, Anders HJ: New insights into the pathology of podocyte loss: mitotic catastrophe. Am J Pathol 2013;183:1364-1374.
3.
Sethi S, Glassock RJ, Fervenza FC: Focal segmental glomerulosclerosis: towards a better understanding for the practicing nephrologist. Nephrol Dial Transplant 2015;30:375-384.
4.
Pippin JW, Brinkkoetter PT, Cormack-Aboud FC, Durvasula RV, Hauser PV, Kowalewska J, Krofft RD, Logar CM, Marshall CB, Ohse T, Shankland SJ: Inducible rodent models of acquired podocyte diseases. Am J Physiol Renal Physiol 2009;296:F213-F229.
5.
Kemper MJ, Wolf G, Müller-Wiefel DE: Transmission of glomerular permeability factor from a mother to her child. N Engl J Med 2001;344:386-387.
6.
Matalon A, Markowitz GS, Joseph RE, Cohen DJ, Saal SD, Kaplan B, D'Agati VD, Appel GB: Plasmapheresis treatment of recurrent FSGS in adult renal transplant recipients. Clin Nephrol 2001;56:271-278.
7.
Zimmerman SW: Increased urinary protein excretion in the rat produced by serum from a patient with recurrent focal glomerular sclerosis after renal transplantation. Clin Nephrol 1984;22:32-38.
8.
Lee VW, Harris DC: Adriamycin nephropathy: a model of focal segmental glomerulosclerosis. Nephrology (Carlton) 2011;16:30-38.
9.
Kaverina NV, Eng DG, Schneider RR, Pippin JW, Shankland SJ: Partial podocyte replenishment in experimental FSGS derives from nonpodocyte sources. Am J Physiol Renal Physiol 2016;310:F1397-F1413.
10.
Lichtnekert J, Kaverina NV, Eng DG, Gross KW, Kutz JN, Pippin JW, Shankland SJ: Renin-angiotensin-aldosterone system inhibition increases podocyte derivation from cells of renin lineage. J Am Soc Nephrol 2016;27:3611-3627.
11.
Pippin JW, Kaverina NV, Eng DG, Krofft RD, Glenn ST, Duffield JS, Gross KW, Shankland SJ: Cells of renin lineage are adult pluripotent progenitors in experimental glomerular disease. Am J Physiol Renal Physiol 2015;309:F341-F358.
12.
Roeder SS, Barnes TJ, Lee JS, Kato I, Eng DG, Kaverina NV, Sunseri MW, Daniel C, Amann K, Pippin JW, Shankland SJ: Activated ERK1/2 increases CD44 in glomerular parietal epithelial cells leading to matrix expansion. Kidney Int 2016;91:896-913.
13.
Zhang J, Pippin JW, Krofft RD, Naito S, Liu ZH, Shankland SJ: Podocyte repopulation by renal progenitor cells following glucocorticoids treatment in experimental FSGS. Am J Physiol Renal Physiol 2013;304:F1375-F1389.
14.
Zhang J, Pippin JW, Vaughan MR, Krofft RD, Taniguchi Y, Romagnani P, Nelson PJ, Liu ZH, Shankland SJ: Retinoids augment the expression of podocyte proteins by glomerular parietal epithelial cells in experimental glomerular disease. Nephron Exp Nephrol 2012;121:e23-e37.
15.
Zhang J, Yanez D, Floege A, Lichtnekert J, Krofft RD, Liu ZH, Pippin JW, Shankland SJ: ACE-inhibition increases podocyte number in experimental glomerular disease independent of proliferation. J Renin Angiotensin Aldosterone Syst 2014;16:234-248.
16.
Greka A, Mundel P: Cell biology and pathology of podocytes. Annu Rev Physiol 2012;74:299-323.
17.
Kestila M, Lenkkeri U, Mannikko M, Lamerdin J, McCready P, Putaala H, Ruotsalainen V, Morita T, Nissinen M, Herva R, Kashtan CE, Peltonen L, Holmberg C, Olsen A, Tryggvason K: Positionally cloned gene for a novel glomerular protein-nephrin-is mutated in congenital nephrotic syndrome. Mol Cell 1998;1:575-582.
18.
Lee YK, Kwon T, Kim DJ, Huh W, Kim YG, Oh HY, Kawachi H: Ultrastructural study on nephrin expression in experimental puromycin aminonucleoside nephrosis. Nephrol Dial Transplant 2004;19:2981-2986.
19.
Kuusniemi AM, Qvist E, Sun Y, Patrakka J, Ronnholm K, Karikoski R, Jalanko H: Plasma exchange and retransplantation in recurrent nephrosis of patients with congenital nephrotic syndrome of the Finnish type (NPHS1). Transplantation 2007;83:1316-1323.
20.
Li X, He JC: An update: the role of nephrin inside and outside the kidney. Sci China Life Sci 2015;58:649-657.
21.
Jones N, Blasutig IM, Eremina V, Ruston JM, Bladt F, Li H, Huang H, Larose L, Li SS, Takano T, Quaggin SE, Pawson T: Nck adaptor proteins link nephrin to the actin cytoskeleton of kidney podocytes. Nature 2006;440:818-23.
22.
Orikasa M, Matsui K, Oite T, Shimizu F: Massive proteinuria induced in rats by a single intravenous injection of a monoclonal antibody. J Immunol 1988;141:807-814.
23.
Putaala H, Soininen R, Kilpeläinen P, Wartiovaara J, Tryggvason K: The murine nephrin gene is specifically expressed in kidney, brain and pancreas: inactivation of the gene leads to massive proteinuria and neonatal death. Hum Mol Genet 2001;10:1-8.
24.
Hansen DT, Robida MD, Craciunescu FM, Loskutov AV, Dörner K, Rodenberry JC, Wang X, Olson TL, Patel H, Fromme P, Sykes KF: Polyclonal antibody production for membrane proteins via genetic immunization. Sci Rep 2016;6:21925.
25.
Aoyama T, Kamata K, Yamanaka N, Takeuchi Y, Higashihara M, Kato S: Characteristics of polyclonal anti-human nephrin antibodies induced by genetic immunization using nephrin cDNA. Nephrol Dial Transplant 2006;21:1073-1081.
26.
Naito S, Kamata K, Aoyama T, Yamanaka N, Okamoto T, Tazaki H: Antigen-binding abilities of anti-nephrin antibody are prescribed by signal sequence of expression vector in genetic immunization. Clin Exp Nephrol 2011;15:339-345.
27.
Logar CM, Brinkkoetter PT, Krofft RD, Pippin JW, Shankland SJ: Darbepoetin alfa protects podocytes from apoptosis in vitro and in vivo. Kidney Int 2007;72:489-498.
28.
Naito S, Pippin JW, Shankland SJ: The glomerular parietal epithelial cellʼs responses are influenced by SM22 alpha levels. BMC Nephrol 2014;15:174.
29.
Holzman LB, St John PL, Kovari IA, Verma R, Holthofer H, Abrahamson DR: Nephrin localizes to the slit pore of the glomerular epithelial cell. Kidney Int 1999;56:1481-1491.
30.
Gollner D, Kawachi H, Oite T, Oka M, Nagase M, Shimizu F: Strain variation in susceptibility to the development of monoclonal antibody 5-1-6-induced proteinuria in rats. Clin Exp Immunol 1995;101:341-345.
31.
Chuang PY, He JC: Signaling in regulation of podocyte phenotypes. Nephron Physiol 2009;111:p9-p15.
32.
Nagata M: Podocyte injury and its consequences. Kidney Int 2016;89:1221-1230.
33.
Meyer TN, Schwesinger C, Wahlefeld J, Dehde S, Kerjaschki D, Becker JU, Stahl RA, Thaiss F: A new mouse model of immune-mediated podocyte injury. Kidney Int 2007;72:841-852.
34.
Matsusaka T, Sandgren E, Shintani A, Kon V, Pastan I, Fogo AB, Ichikawa I: Podocyte injury damages other podocytes. J Am Soc Nephrol 2011;22:1275-1285.
35.
Wharram BL, Goyal M, Wiggins JE, Sanden SK, Hussain S, Filipiak WE, Saunders TL, Dysko RC, Kohno K, Holzman LB, Wiggins RC: Podocyte depletion causes glomerulosclerosis: diphtheria toxin-induced podocyte depletion in rats expressing human diphtheria toxin receptor transgene. J Am Soc Nephrol 2005;16:2941-2952.
36.
Chaudhuri A, Kambham N, Sutherland S, Grimm P, Alexander S, Concepcion W, Sarwal M, Wong C: Rituximab treatment for recurrence of nephrotic syndrome in a pediatric patient after renal transplantation for congenital nephrotic syndrome of Finnish type. Pediatr Transplant 2012;16:E183-E187.
37.
Li X, Chuang PY, D'Agati VD, Dai Y, Yacoub R, Fu J, Xu J, Taku O, Premsrirut PK, Holzman LB, He JC: Nephrin preserves podocyte viability and glomerular structure and function in adult kidneys. J Am Soc Nephrol 2015;26:2361-2377.
38.
Kitching AR, Hutton HL: The Players: cells involved in glomerular disease. Clin J Am Soc Nephrol. 2016;11:1664-1674.
39.
Han Y, Ma FY, Tesch GH, Manthey CL, Nikolic-Paterson DJ: Role of macrophages in the fibrotic phase of rat crescentic glomerulonephritis. Am J Physiol Renal Physiol. 2013;304:F1043-F1053.
40.
Schießl IM, Bardehle S, Castrop H: Superficial nephrons in BALB/c and C57BL/6 mice facilitate in vivo multiphoton microscopy of the kidney. PLoS One. 2013;8:e52499.
41.
Li M, Armelloni S, Ikehata M, Corbelli A, Pesaresi M, Calvaresi N, Giardino L, Mattinzoli D, Nisticò F, Andreoni S, Puliti A, Ravazzolo R, Forloni G, Messa P, Rastaldi MP: Nephrin expression in adult rodent central nervous system and its interaction with glutamate receptors. J Pathol 2011;225:118-128.
42.
Palmén T, Ahola H, Palgi J, Aaltonen P, Luimula P, Wang S, Jaakkola I, Knip M, Otonkoski T, Holthöfer H: Nephrin is expressed in the pancreatic beta cells. Diabetologia 2001;44:1274-1280.
43.
Tsuchiya Y, Minami Y, Umemura Y, Watanabe H, Ono D, Nakamura W, Takahashi T, Honma S, Kondoh G, Matsuishi T, Yagita K: Disruption of MeCP2 attenuates circadian rhythm in CRISPR/Cas9-based Rett syndrome model mouse. Genes Cells 2015;20:992-1005.
44.
Yoo HY, Song YH, Foo M, Seo E, Hwang DS, Seo JH: Recombinant mussel proximal thread matrix protein promotes osteoblast cell adhesion and proliferation. BMC Biotechnol 2016;16:16.
45.
Zhang T, Yin Y, Liu H, Du W, Ren C, Wang L, Lu H, Zhang Z: Generation of VDR knock-out mice via zygote injection of CRISPR/Cas9 system. PLoS One 2016;11:e0163551.
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