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. 2020 Sep;57(5):675-680.
doi: 10.1177/0300985820934111. Epub 2020 Jun 17.

Diagnosis of Collagen Type III Glomerulopathy Using Picrosirius Red and PASH/Masson's Trichrome Stain

Katelin L Davis  1   2 , Anne L Burnum  3 , Jessica A Beck  1   2 , Shannon G M Kirejczyk  3 , Margaret A Miller  1 , Mario F Sola  1 , Cathy A Brown  3
Affiliations

Diagnosis of Collagen Type III Glomerulopathy Using Picrosirius Red and PASH/Masson's Trichrome Stain

Katelin L Davis et al. Vet Pathol. 2020 Sep.

Abstract

Canine collagen type III glomerulopathy (Col3GP) is a rare juvenile nephropathy in which irregular type III collagen fibrils and fibronectin accumulate in glomerular capillary walls and the mesangium. Necropsy findings were reviewed from 5 puppies diagnosed with Col3GP at 6 to 18 weeks of age. Histologically, with hematoxylin and eosin stain, the glomerular capillary walls and mesangium were diffusely and globally expanded by homogeneous pale eosinophilic material. Ultrastructurally, the subendothelial zone and mesangium were expanded by fibronectin and cross-banded collagen type III fibrils, diagnostic of Col3GP. Two additional stains were employed to identify the material within glomeruli as fibrillar collagen using light microscopy. In all 5 cases, the material was red with picrosirius red and birefringent under polarized light, and was blue with periodic acid-Schiff/hematoxylin/trichrome (PASH/TRI), thereby identifying it as fibrillar collagen. Based on these unique staining characteristics with picrosirius red and PASH/TRI, Col3GP may be reliably diagnosed with light microscopy alone.

Keywords: collagen type III; collagenofibrotic glomerulopathy; dogs; kidney; picrosirius red; renal; trichrome.

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Conflict of interest statement

Declaration of Conflicting Interests

The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Figures

Figure 1.
Figure 1.. Collagen type III glomerulopathy, kidney, dogs.
Case 3. Glomerular capillary loops and mesangium are expanded by pale eosinophilic, homogeneous material. Bowman’s capsule is thickened. Hematoxylin and eosin.
Figure 2.
Figure 2.. Collagen type III glomerulopathy, kidney, dogs.
Case 2. Glomerular capillary loop. The subendothelial zone is markedly expanded by granular material and loosely arranged collagen fibrils (arrows). Podocyte foot processes are effaced (arrowhead). CL, capillary lumen; En, endothelial cell; asterisk, glomerular basement membrane. Bar = 500 nm. Inset: collagen fibrils have cross-striations, are 28 to 60 μm in diameter, and are often curved. Bar = 100 nm. Transmission electron microscopy. Uranyl acetate and lead citrate.
Figure 3a.
Figure 3a.. Collagen type III glomerulopathy, kidney, dogs.
Case 4, picrosirius red stain. The material expanding the glomerular capillary loops and mesangium stains red. Bowman’s capsule and the interstitium are also red.
Figure 3b.
Figure 3b.. Collagen type III glomerulopathy, kidney, dogs.
Case 4, picrosirius red stain under polarized light. The material expanding glomerular capillary loops and mesangium is yellow-red and green, consistent with fibrillar collagen. Bowman’s capsule and the interstitium are also yellow-red and green, consistent with fibrillar collagen.
Figure 4a.
Figure 4a.. Collagen type III glomerulopathy, kidney, puppy, case 1.
Periodic acid-Schiff and hematoxylin (PASH). The material expanding glomerular capillary loops and mesangium is weakly PAS-positive. Glomerular basement membranes are strongly PAS-positive.
Figure 4b.
Figure 4b.. Collagen type III glomerulopathy, kidney, puppy, case 1.
Masson’s trichrome (TRI). The material expanding glomerular capillary loops and mesangium as well as the glomerular basement membranes is dark blue, consistent with collagen.
Figure 4c.
Figure 4c.. Collagen type III glomerulopathy, kidney, puppy, case 1.
PASH/TRI combination. The material expanding capillary loops and mesangium is dark blue and differentiated from the glomerular basement membranes, which are purple.
Figure 5.
Figure 5.. Collagen type III glomerulopathy, kidney, puppy, case 1.
Normal glomerulus, control puppy (exact age unknown). PASH/TRI combination. Glomerular basement membranes are purple. Dark blue material is not observed within the capillary walls and mesangium.
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