Demonstration of epithelial-mesenchymal transition in kidney. The contribution of a coupled histochemical and immunohistochemical staining with Periodic Acid-thionin Schiff.
Traditional Periodic Acid Schiff has been extensively used, coupled with immunohistochemistry for epithelia or mesenchymal cells, to highlight renal tubular basement membrane (TBM). We recently tried to perform such technique in a 5/6 nephrectomy model of progressive renal fibrosis to demonstrate TBM disruption as an evidence for epithelial-mesenchymal transdifferentiation. Despite excellent basement membrane staining with traditional fuchsin-Periodic Acid Schiff, the interface between epithelial and mesenchymal cells was frequently blurred when revealed with 3'3 diaminobenzidine tetrachloride-peroxidase. Also, it was inadequate when revealed with alkaline phosphatase-fast red. We devised a triple staining method with Periodic Acid-Thionin Schiff to highlight basement membrane in blue, after double immunostaining for epithelium and mesenchymal cells. Blue basement membrane rendered a brisk contrast and highlighted boundaries between epithelial-mesenchymal interfaces. This method was easy to perform and useful to demonstrate the TBM, yield a clear demonstration of the very focal TBM disruption found in this model of progressive renal fibrosis.
Santos
Maria Carmen Lopes Ferreira Silva
MC
Federal University of Espirito Santo School of Medicine, Santo, Brazil. tete@uvv.br
Musso
Carlos
C
Alves
Venâncio Avancini Ferreira
VA
Zatz
Roberto
R
Fujihara
Clarice Kazue
CK
Malheiros
Denise Maria Avancini Costa
DM
D000818:Animals; D001485:Basement Membrane / Q000473:pathology*; D004847:Epithelial Cells / Q000473:pathology; D005355:Fibrosis; D006651:Histocytochemistry / Q000379:methods*; D007150:Immunohistochemistry / Q000379:methods*; D007684:Kidney Tubules / Q000473:pathology*; D008297:Male; D008648:Mesoderm / Q000473:pathology; D010503:Periodic Acid-Schiff Reaction; D010640:Phenothiazines; D051381:Rats
D010640:Phenothiazines; C009469:thionine