Nephrin and podocin mRNA detection in urine sediment of dogs with chronic kidney disease: preliminary observations Cover

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Nephrin and podocin mRNA detection in urine sediment of dogs with chronic kidney disease: preliminary observations

Journal of Veterinary Research

Volume 66 (2022): Issue 2 (June 2022)

By: Camilla de Souza, Mariana Coelho, Daniel Silva Antonelo, Danielle Passarelli, Arina Lázaro Rochetti, Heidge Fukumasu and Deise Carla Almeida Leite-Dellova

Open Access

|Apr 2022

Figures & Tables

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

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

Results for blood haematological parameters in healthy dogs and dogs with chronic kidney disease (CKD) at different stages

		CKD dogs

Parameter (reference range)	Healthy dogs (n = 10)	Stage 1 or 2 (n = 5)	Stage 3 or 4 (n = 9)
RBC (106/μL) (5.5–8.5)	6.7 ± 0.35	6.7 ± 0.49	4.7 ± 0.36**
HGB (g/dL) (12–18)	17.3 ± 0.83	16.7 ± 1.17	11.6 ± 0.88**
HCT (%) (37–55)	50.1 ± 2.49	49.7 ± 3.52	34.9 ± 2.63**
MCV (fL) (60–77)	75.2 ± 1.49	75.0 ± 2.11	75.2 ± 1.57
MCH (pg) (21–26)	26.0 ± 0.58	25.2 ± 0.82	25.0 ± 0.61
MCHC (%) (32–36)	34.6 ± 0.44	33.7 ± 0.62	33.2 ± 0.47
PLT (103/μL) (200–500)	307 ± 57.60	386 ± 39.7	378 ± 64.0
RBC morphology	normal	normal	normal
WBC (103/μL) (6–17)	7.1 ± 0.63	8.8 ± 1.35	7.5 ± 1.02
WBC morphology	normal	normal	normal

Results for blood biochemical parameters in healthy dogs and dogs with chronic kidney disease (CKD) at different stages

		CKD dogs

Parameter (reference range)	Healthy dogs (n = 10)	Stage 1 or 2 (n = 5)	Stage 3 or 4 (n = 9)
Albumin (g/dL) (2.3–3.8)	3.2 ± 0.16	2.8 ± 0.23	2.7 ± 0.20
Urea (mg/dL) (10–60)	39.4 ± 5.36	47.3 ± 13.48	259.4 ± 46.70*
Creatinine (mg/dL) (0.5–1.4)	1.1 ± 0.14	1.0 ± 0.17	7.4 ± 2.34*

Semi-quantitative analysis of detection of NPHS1 (nephrin mRNA) and NPHS2 (podocin mRNA) in urine sediment samples from healthy dogs and dogs with chronic kidney disease (CKD) at different stages

		CKD dogs

Parameter	Healthy dogs	Stage 1 or 2	Stage 3 or 4	P value
NPHS1 detection	4/10 (40%) ^a	3/5 (60%) ^b	1/9 (11%) ^c	0.0009
NPHS2 detection	2/10 (20%) ^a	4/5 (80%) ^b	1/9 (11%) ^a	0.0188

Results for routine urinary parameters in healthy dogs and dogs with chronic kidney disease (CKD) at different stages

		CKD dogs

Parameter (reference range)	Healthy dogs (n = 10)	Stage 1 or 2 (n = 5)	Stage 3 or 4 (n = 9)
USG (1.015–1.045)	1.037 ± 0.0040	1.022 ± 0.0074*	1.014 ± 0.0009**
Protein (mg/dL)	29.4 ± 2.44	210.6 ± 11.67***	124.0 ± 35.76*
Creatinine (mg/dL)	206.1 ± 24.00	94.4 ± 21.47**	70.0 ± 21.47**
UPC (< 0.5)	0.15 ± 0.01	2.25 ± 0.68*	1.76 ± 0.50*
Chemical analysis
(semiquantitative testing):
pH (6.0–7.5)	6.5 ± 0.37	6.4 ± 0.52	5.9 ± 0.39
Protein	between 0 and 1+	between 1 and 3+	between 1 and 2+
Glucose	negative	negative	negative
Ketones	negative	negative	negative
Bilirubin	negative	negative	negative
Urobilinogen	normal	normal	normal
Occult blood	negative	1+ in the urinary sediment of 1 dog	1+ in the urinary sediment of 2 dogs
Microscopic sediment
examination:
Epithelial cells/HPF (<3)	1.70 ± 0.26	1.00 ± 0.22	1.44 ± 0.43
Granular cast/HPF (<1)	rare in the urinary sediment of 1 dog	none	rare in the urinary sediment of 1 dog
Triple phosphate crystal	1+ in the urinary sediment of 2 dogs	2+ in the urinary sediment of 1 dog	none
Amorphous phosphate crystal	none	none	2+ in the urinary sediment of 1 dog
RBC/HPF (<5)	0.30 ± 0.13	0.30 ± 0.20	0.56 ± 0.49
WBC/HPF (<5)	0.50 ± 0.18	0.60 ± 0.25	0.83 ± 0.25
Bacteria	none	none	none

Semi-quantitative analysis of renal ultrasonographic abnormalities in dogs with chronic kidney disease (CKD) at different stages

	CKD dogs

Parameter	Stage 1 or 2	Stage 3 or 4	P value
Irregular contour	5/5 (100%)	6/9 (67%)	<0.0001
Loss of corticomedullary differentiation	4/5 (80%)	8/9 (89%)	0.1170
Increased cortical echogenicity	4/5 (80%)	8/9 (89%)	0.1170
Increased medullary echogenicity	0/5	5/9 (56%)	0.0197

DOI: https://doi.org/10.2478/jvetres-2022-0019 | Journal eISSN: 2450-8608

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Language: English

Page range: 281 - 288

Submitted on: Oct 27, 2021

Accepted on: Mar 29, 2022

Published on: Apr 22, 2022

Published by: National Veterinary Research Institute in Pulawy

In partnership with: Paradigm Publishing Services

Publication frequency: 4 issues per year

Keywords:

quantitative real-time PCR

Related subjects:

Molecular biology,

Microbiology and virology,

Life sciences, other,

Veterinary medicine

© 2022 Camilla de Souza, Mariana Coelho, Daniel Silva Antonelo, Danielle Passarelli, Arina Lázaro Rochetti, Heidge Fukumasu, Deise Carla Almeida Leite-Dellova, published by National Veterinary Research Institute in Pulawy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

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