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. 2022 Apr 22;66(2):281-288.
doi: 10.2478/jvetres-2022-0019. eCollection 2022 Jun.

Nephrin and Podocin mRNA Detection in Urine Sediment of Dogs with Chronic Kidney Disease: Preliminary Observations

Camilla de Souza  1 , Mariana Coelho  1 , Daniel Silva Antonelo  2 , Danielle Passarelli  1 , Arina Lázaro Rochetti  1 , Heidge Fukumasu  1 , Deise Carla Almeida Leite-Dellova  1
Affiliations

Nephrin and Podocin mRNA Detection in Urine Sediment of Dogs with Chronic Kidney Disease: Preliminary Observations

Camilla de Souza et al. J Vet Res. .

Abstract

Introduction: Dogs with chronic kidney disease (CKD) may have alterations in the glomerular filtration barrier, including podocyte loss. Detection of podocyte mRNA in urine could be useful for assessing podocyturia in dogs with kidney disease. The objective of this study was to evaluate the presence of nephrin mRNA (NPHS1) and podocin mRNA (NPHS2) in urine sediments of dogs with naturally occurring CKD and healthy dogs.

Material and methods: Twenty-four dogs, 14 with CKD and 10 as healthy controls, underwent clinical evaluation. The dogs with CKD were divided into two groups, according to the International Renal Interest Society criteria: stage 1 or 2 CKD (n = 5) and stage 3 or 4 CKD (n = 9). Urine was collected by catheterisation or free catch and RNA isolation from the urine sediments was optimised using glycogen as a co-precipitant. Detection of NPHS1 and NPHS2 in the sediment samples was performed using quantitative real-time PCR.

Results: Both types of mRNA were detected in samples from all groups, but the percentages of detection were higher in the group of dogs with stage 1 or 2 CKD and lower in the group of dogs with stage 3 or 4 disease.

Conclusion: Physiological podocyturia was observed in healthy dogs, and the results suggest differential podocyturia in dogs with CKD, according to the stage of the disease, i.e. an increase in podocyturia in dogs at stage 1 or 2 and a reduction in podocyturia in dogs at stage 3 or 4.

Keywords: NPHS1; NPHS2; dog; podocyturia; quantitative real-time PCR.

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Conflict of interest statement

Conflict of Interest Conflict of Interests Statement: The authors declare that there is no conflict of interests regarding the publication of this article.

Figures

Fig. 1
Fig. 1
Electrophoresis of total RNA in agarose gel (2%). Column 1 – total RNA isolated from mouse lung cells; Columns 2 and 3 – total RNA isolated from urine sediment of subject dogs. Ribosomal RNA bands (28S and 18S) were observed in all samples, indicating adequate RNA isolation. The genomic DNA band appeared in samples 1 and 2 because DNAse had not yet been used in the reaction
Fig. 2
Fig. 2
cDNA product electrophoresis in 1.5% agarose gel. Neg. – negative control; Column 1 – sample from a dog with stage 3 chronic kidney disease; Column 2 – sample from a dog of the control group; Column 3 – sample from a dog with stage 1 chronic kidney disease. The length of 18S was 80 bp, that of NPHS1 was 156 bp, that of NPHS2 was 140 bp, and the DNA ladder was 100 bp long
See this image and copyright information in PMC

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