title: Semiautomated quantitative image analysis of glomerular immunohistochemistry markers desmin, vimentin, podocin, synaptopodin and WT-1 in acute and chronic rat kidney disease models.
abstract: Five different glomerular immunohistochemistry markers were evaluated and compared in four different acute and chronic rat kidney disease models. Progression of glomerular or podocyte damage was shown in the puromycin aminonucleoside nephrosis (PAN) and Zucker fatty/spontaneously hypertensive heart failure F1 hybrid (ZSF1) rat model. Progression and prevention of glomerular damage was demonstrated in the Zucker diabetic fatty (ZDF) and Dahl salt-sensitive (Dahl SS) rat. Immunohistochemistry was performed for desmin, vimentin, podocin, synaptopodin and Wilms tumor protein-1 (WT-1), and evaluation of glomerular immunohistochemistry markers was done by semiautomated quantitative image analysis. We found desmin and WT-1 as the most sensitive markers for podocyte damage in both acute and chronic glomerular damage followed by vimentin, podocin and synaptopodin. We were able to demonstrate that early podocyte damage as shown by increased desmin and vimentin staining together with either a phenotypic podocyte change or podocyte loss (reduced numbers of WT-1-stained podocytes) drives the progression of glomerular damage. This is followed by a reduction in podocyte-specific proteins such as podocin and synaptopodin. Our report describes the different sensitivity of glomerular or podocyte markers and gives future guidance for the selection of the most sensitive markers for efficacy testing of new drugs as well as for the selection of tissue-based toxicity markers for glomerular or podocyte injury. In addition to functional clinical chemistry markers, desmin and WT-1 immunohistochemistry offers reliable and valuable data on the morphologic state of podocytes.
authors.affiliation: Roche Pharmaceutical Research & Early Development, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, 4070, Basel, Switzerland. juergen.funk@roche.com.
authors.forename: J
authors.initials: J
authors.lastname: Funk
authors.affiliation: Roche Pharmaceutical Research & Early Development, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: V
authors.initials: V
authors.lastname: Ott
authors.affiliation: Roche Pharmaceutical Research & Early Development, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: A
authors.initials: A
authors.lastname: Herrmann
authors.affiliation: Roche Pharmaceutical Research & Early Development, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: W
authors.initials: W
authors.lastname: Rapp
authors.affiliation: Roche Pharmaceutical Research & Early Development, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: S
authors.initials: S
authors.lastname: Raab
authors.affiliation: Roche Pharmaceutical Research & Early Development, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: W
authors.initials: W
authors.lastname: Riboulet
authors.affiliation: Roche Pharmaceutical Research & Early Development, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: A
authors.initials: A
authors.lastname: Vandjour
authors.affiliation: Roche Pharmaceutical Research & Early Development, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: E
authors.initials: E
authors.lastname: Hainaut
authors.affiliation: Roche Pharmaceutical Research & Early Development, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: A
authors.initials: A
authors.lastname: Benardeau
authors.affiliation: Roche Pharmaceutical Research & Early Development, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: T
authors.initials: T
authors.lastname: Singer
authors.affiliation: Roche Pharmaceutical Research & Early Development, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: B
authors.initials: B
authors.lastname: Jacobsen
mesh_terms: D000208:Acute Disease; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002908:Chronic Disease; D003893:Desmin / Q000032:analysis*; D004195:Disease Models, Animal; D007150:Immunohistochemistry; D047908:Intracellular Signaling Peptides and Proteins / Q000032:analysis*; D007674:Kidney Diseases / Q000378:metabolism*; D008297:Male; D008565:Membrane Proteins / Q000032:analysis*; D008840:Microfilament Proteins / Q000032:analysis*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014746:Vimentin / Q000032:analysis*; D025721:WT1 Proteins / Q000032:analysis*
chemical_list: D015415:Biomarkers; D003893:Desmin; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D008840:Microfilament Proteins; C407223:NPHS2 protein; C503969:Synpo protein, rat; D014746:Vimentin; D025721:WT1 Proteins
keywords: Automated image analysis; Chronic kidney disease; Desmin; Immunohistochemistry; Podocyte damage; WT-1