Podocyte developmental defects caused by adriamycin in zebrafish embryos and larvae: a novel model of glomerular damage.
The zebrafish pronephros is gaining popularity in the nephrology community, because embryos are easy to cultivate in multiwell plates, allowing large number of experiments to be conducted in an in vivo model. In a few days, glomeruli reach complete development, with a structure that is similar to that of the mammalian counterpart, showing a fenestrated endothelium and a basement membrane covered by the multiple ramifications of mature podocytes. As a further advantage, zebrafish embryos are permeable to low molecular compounds, and this explains their extensive use in drug efficacy and toxicity experiments. Here we show that low concentrations of adriamycin (i.e. 10 and 20 µM), when dissolved in the medium of zebrafish embryos at 9 hours post-fertilization and removed after 48 hours (57 hpf), alter the development of podocytes with subsequent functional impairment, demonstrated by onset of pericardial edema and reduction of expression of the podocyte proteins nephrin and wt1. Podocyte damage is morphologically confirmed by electron microscopy and functionally supported by increased clearance of microinjected 70 kDa fluorescent dextran. Importantly, besides pericardial edema and glomerular damage, which persist and worsen after adriamycin removal from the medium, larvae exposed to adriamycin 10 and 20 µM do not show any myocardiocyte alterations nor vascular changes. The only extra-renal effect is a transient delay of cartilage formation that rapidly recovers once adriamycin is removed. In summary, this low dose adriamycin model can be applied to analyze podocyte developmental defects, such as those observed in congenital nephrotic syndrome, and can be taken in consideration for pharmacological studies of severe early podocyte injury.
Department of Medical, Surgery and Health Sciences, Università degli Studi di Trieste, Trieste, Italy; Renal Research Laboratory, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico & Fondazione D'Amico per la Ricerca sulle Malattie Renali, Milano, Italy.
Cristina
C
Zennaro
Department of Biomedical, Surgical and Dental Sciences, University of Milano & IRCCS Orthopedic Institute, Milano, Italy.
Massimo
M
Mariotti
Department of Medical, Surgery and Health Sciences, Università degli Studi di Trieste, Trieste, Italy.
Michele
M
Carraro
Gruppo Ospedaliero San Donato Foundation, Milano, Italy.
Sara
S
Pasqualetti
Renal Research Laboratory, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico & Fondazione D'Amico per la Ricerca sulle Malattie Renali, Milano, Italy; Department of Cardiovascular Research- Unit of Bio-imaging, Mario Negri Institute for Pharmacological Research, Milano, Italy.
Alessandro
A
Corbelli
Renal Research Laboratory, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico & Fondazione D'Amico per la Ricerca sulle Malattie Renali, Milano, Italy.
Silvia
S
Armelloni
Renal Research Laboratory, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico & Fondazione D'Amico per la Ricerca sulle Malattie Renali, Milano, Italy.
Min
M
Li
Renal Research Laboratory, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico & Fondazione D'Amico per la Ricerca sulle Malattie Renali, Milano, Italy.
Masami
M
Ikehata
Azienda Ospedaliero-Universitaria Ospedali Riuniti di Trieste, Trieste, Italy.
Milan
M
Clai
Azienda Ospedaliero-Universitaria Ospedali Riuniti di Trieste, Trieste, Italy.
Mary
M
Artero
Division of Nephrology, Dialysis, and Renal Transplant, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milano, Italy.
Piergiorgio
P
Messa
Azienda Ospedaliero-Universitaria Ospedali Riuniti di Trieste, Trieste, Italy.
Giuliano
G
Boscutti
Renal Research Laboratory, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico & Fondazione D'Amico per la Ricerca sulle Malattie Renali, Milano, Italy.
Maria Pia
MP
Rastaldi
D000818:Animals; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000494:pharmacology*; D047108:Embryonic Development / Q000187:drug effects*; D006321:Heart / Q000187:drug effects / Q000196:embryology; D007678:Kidney Glomerulus / Q000187:drug effects* / Q000196:embryology* / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D007814:Larva; D010539:Permeability / Q000187:drug effects; D050199:Podocytes / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D015027:Zebrafish / Q000196:embryology*
D004317:Doxorubicin