Intraperitoneal protein injection in the axolotl: the amphibian kidney as a novel model to study tubulointerstitial activation.
BACKGROUND
A substantial body of experimental evidence suggests that protein loading causes activation of proximal tubular epithelial cells with consecutive interstitial fibrosis. These studies have mostly been performed using mammalian in vivo models of glomerular damage or tissue cultures of mammalian tubulointerstitial cells. The kidney of the axolotl contains not only closed nephrons, but also nephrons with ciliated peritoneal funnels called nephrostomes that have access to the peritoneal fluid. Injection of protein into the peritoneal cavity fails to expose closed nephrons to a protein load, but causes selective uptake and transient storage of proteins in tubular epithelial cells of nephrons with nephrostomes. The purpose of the present study was to determine whether (a) the axolotl kidney can be used as a model to assess protein uptake by tubular cells in vivo in the absence of glomerular damage, and (b) this is accompanied by any evidence of tubular epithelial cell activation and interstitial fibrosis.


METHODS
Male and female axolotl (80 to 120 g of weight) were given a daily intraperitoneal injection of 1.5 mL endotoxin-free calf serum or saline as control. Kidneys were harvested after 4 or 10 days using perfusion fixation for light microscopy (fibrous tissue stain) and saline perfusion for immunohistochemistry (fibronectin, TGF-beta and collagen I).


RESULTS
The findings document selective storage of protein and lipids, progressive with time, in proximal tubular epithelial cells of nephrons draining the coelomic cavity. In addition, progressive focal accumulation of fibrous tissue was noted around protein-storing tubules. Immunohistochemical staining demonstrated the presence of fibronectin and TGF-beta in the tubular epithelial cells and interstitial cells.


CONCLUSIONS
The axolotl kidney provides a novel in vivo model to study tubulointerstitial activation and induction of interstitial fibrosis by protein loading. The findings are independent of alterations of glomerular function that may have potential confounding effects on peritubular hemodynamics, pO2, cell traffic, etc.
Gross
Marie Louise
ML
Department of Pathology, University of Heidelberg, Bergheimerstrasse 58, 69115 Heidelberg, Germany.
Hanke
Wilford
W
Koch
Andreas
A
Ziebart
Heike
H
Amann
Kerstin
K
Ritz
Eberhard
E
D000557:Ambystoma; D000818:Animals; D024042:Collagen Type I / Q000032:analysis; D004847:Epithelial Cells / Q000473:pathology; D005260:Female; D005353:Fibronectins / Q000032:analysis; D005355:Fibrosis; D007150:Immunohistochemistry; D007274:Injections, Intraperitoneal; D007687:Kidney Tubules, Proximal / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D008297:Male; D023421:Models, Animal; D011507:Proteinuria / Q000473:pathology*; D016212:Transforming Growth Factor beta / Q000032:analysis
D024042:Collagen Type I; D005353:Fibronectins; D016212:Transforming Growth Factor beta