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. 2016 Jun 1;78(5):895-9.
doi: 10.1292/jvms.15-0722. Epub 2016 Feb 6.

Expression of phospholipase A2 receptor in primary cultured podocytes derived from dog kidneys

Go Sugahara  1 , Junichi Kamiie, Ryosuke Kobayashi, Takayuki Mineshige, Kinji Shirota
Affiliations

Expression of phospholipase A2 receptor in primary cultured podocytes derived from dog kidneys

Go Sugahara et al. J Vet Med Sci. .

Abstract

Phospholipase A2 receptor (PLA2R) expressed in human podocytes has been highlighted as a causative autoantigen of human idiopathic membranous nephropathy. However, its expression was found to be minimal or absent in murine and rat podocytes. In this study, immunofluorescence revealed the expression of PLA2R in the glomerular podocytes in the kidney tissue sections of dogs. We then attempted to culture canine podocytes and investigate the expression of PLA2R in these cells. Glomeruli were isolated from dog kidneys and cultured to obtain podocytes using nylon mesh-based isolation method as followed for isolating rat podocytes. The cultured cells expressed PLA2R mRNA and protein in addition to other podocyte markers (synaptopodin, podocin and nephrin). These results indicate that the canine podocytes express PLA2R.

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Figures

Fig. 1.
Fig. 1.
Expression of phospholipase A2 receptor (PLA2R) in the canine glomerulus. The expression of PLA2R (red) was mainly observed in podocytes cell body and cell processes and also in parietal epithelial cells. Intrinsic fluorescence of red blood cells was demonstrated as yellow. Bar=30 µm.
Fig. 2.
Fig. 2.
Phase-contrast microscopy of isolated canine glomeruli and cellular growth of podocytes. Phase-contrast microscopy of isolated glomeruli from canine renal tissue (A) and their cellular outgrowths after 2 days (B) and 3 days (C and D) in culture. *Isolated glomerulus, Bar=100 µm.
Fig. 3.
Fig. 3.
Expression of phospholipase A2 receptor (PLA2R) and podocyte markers in primary cultured canine podocytes. Immunofluorescence for podocyte-specific proteins (synaptopodin, podocin and nephrin) (A) and PLA2R (B), reverse transcriptase polymerase chain reaction (RT-PCR) for podocyte-specific markers (NPHS2, SYNPO and NPHS1), PLA2R1 and GAPDH(C) and Western blotting for PLA2R protein (D) in canine primary cultured podocytes. Mouse IgG: negative control. Arrowheads: expression on marginal region. Asterisk: Isolated glomerulus. Bar=25 µm.
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