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. 2023 Nov 22;34(2):273–290. doi: 10.1681/ASN.2022010050

Figure 4.

Figure 4

Increased rate of duplex kidneys and renal hypodysplasia in Arhgef6-deficient mice. Heterozygous Arhgef6+/− were mated with hemizygous Arhgef6−/Y and litters sacrificed at day 4 postnatally. Kidneys and urinary tract have been evaluated macroscopically and dissected on suspected CAKUT phenotype. Specimens have then been processed for Masson's trichrome (MT) staining and subsequent blinded evaluation of multiple sections of each specimen. (A) Two exemplary kidney pairs from Arhgef6−/− and Arhgef6−/Y mice, respectively, are shown demonstrating duplex kidneys. Red arrowhead points to fissure: whole organ overview (upper) and MT-stained section (lower). (B) One exemplary kidney pair of an Arhgef6−/Y mouse is shown demonstrating a left hypoplastic kidney—whole organ overview. (C) One exemplary kidney of an Arhgef6−/Y mouse is shown demonstrating a dysplastic kidney—MT-stained section. (D) Graph shows relative frequency of three different observed CAKUT phenotypes (duplex kidney, hypodysplasia, and hydronephrosis) for wild-type Arhgef6+/Y and hemizygous Arhgef6−/Y animals from macroscopic evaluation. Unpaired t test; *P<0.05. (E) Graph shows relative frequency of total observed CAKUT for wild-type Arhgef6+/Y and hemizygous Arhgef6−/Y animals from macroscopic evaluation. Unpaired t test; *, P<0.05. (F) In a second evaluation, whole litters from Arhgef6+/− × Arhgef6−/Y matings were evaluated at 4 weeks of age independent of macroscopic CAKUT. Kidneys were processed for histology, and parenchyma was scored for dysplastic features (cystic structures, very heterogeneous parenchyma, loss of zonal compartmentalization). Bar graph shows relative frequency of dysplasia in wild-type Arhgef6+/Y and hemizygous Arhgef6−/Y animals. Unpaired t test; *P<0.05. Nephron quantity was estimated by counting the number of glomeruli in longitudinal midline sections of kidneys. Scatter dot plot shows numbers of glomeruli for wild-type Arhgef6+/Y and hemizygous Arhgef6−/Y animals. Each animal is represented by two data points, one for each kidney. Mann–Whitney test; ***P<0.001.