title: Demonstration of epithelial-mesenchymal transition in kidney. The contribution of a coupled histochemical and immunohistochemical staining with Periodic Acid-thionin Schiff.
abstract: 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.
authors.affiliation: Federal University of Espirito Santo School of Medicine, Santo, Brazil. tete@uvv.br
authors.forename: Maria Carmen Lopes Ferreira Silva
authors.initials: MC
authors.lastname: Santos
authors.forename: Carlos
authors.initials: C
authors.lastname: Musso
authors.forename: Venâncio Avancini Ferreira
authors.initials: VA
authors.lastname: Alves
authors.forename: Roberto
authors.initials: R
authors.lastname: Zatz
authors.forename: Clarice Kazue
authors.initials: CK
authors.lastname: Fujihara
authors.forename: Denise Maria Avancini Costa
authors.initials: DM
authors.lastname: Malheiros
mesh_terms: 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
chemical_list: D010640:Phenothiazines; C009469:thionine