Abstract
Prediction and management of drug-induced renal injury (DIRI) rely on the knowledge of the mechanisms of drug insult and on the availability of appropriate animal models to explore it. Zebrafish (Danio rerio) offers unique advantages for assessing DIRI because the larval pronephric kidney has a high homology with its human counterpart and it is fully mature at 3.5 days post-fertilization. Herein, we aimed to evaluate the usefulness of zebrafish larvae as a model of renal tubular toxicity through a comprehensive analysis of the renal alterations induced by the lethal concentrations for 10% of the larvae for gentamicin, paracetamol and tenofovir. We evaluated drug metabolic profile by mass spectrometry, renal function with the inulin clearance assay, the 3D morphology of the proximal convoluted tubule by two-photon microscopy and the ultrastructure of proximal convoluted tubule mitochondria by transmission electron microscopy. Paracetamol was metabolized by conjugation and oxidation with further detoxification with glutathione. Renal clearance was reduced with gentamicin and paracetamol. Proximal tubules were enlarged with paracetamol and tenofovir. All drugs induced mitochondrial alterations including dysmorphic shapes (“donuts”, “pancakes” and “rods”), mitochondrial swelling, cristae disruption and/or loss of matrix granules. These results are in agreement with the tubular effects of gentamicin, paracetamol and tenofovir in man and demonstrate that zebrafish larvae might be a good model to assess functional and structural damage associated with DIRI.
Similar content being viewed by others
References
Arpagaus S, Rawyler A, Braendle R (2002) Occurrence and characteristics of the mitochondrial permeability transition in plants. J Biol Chem 277:1780–1787
Basile D, Anderson M, Sutton T (2012) Pathophysiology of acute kidney injury. Compr Physiol 2:1303–1353
Bonventre J, Yang L (2011) Cellular pathophysiology of ischemic acute kidney injury. J Clin Invest 121:4210–4221
Chevalier RL (2016) The proximal tubule is the primary target of injury and progression of kidney disease: role of the glomerulotubular junction. Am J Physiol Renal Physiol 311:F145–F161
Cianciolo Cosentino C, Skrypnyk NI, Brilli LL et al (2013) Histone deacetylase inhibitor enhances recovery after AKI. J Am Soc Nephrol 24:943–953
Clarot I, Chaimbault P, Hasdenteufel F et al (2004) Determination of gentamicin sulfate and related compounds by high-performance liquid chromatography with evaporative light scattering detection. J Chromatogr A 1031:281–287
Cook SF, King AD, van den Anker JN, Wilkins DG (2015) Simultaneous quantification of acetaminophen and five acetaminophen metabolites in human plasma and urine by high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry: method validation and application to a neonatal pharmacokinetic study. J Chromatogr B Analyt Technol Biomed Life Sci 1007:30–42
Drummond IA, Davidson AJ (2010) Zebrafish kidney development. In: Detrich HW, Westerfiled M, Zon LI (eds) Methods in cell biology. Elsevier Inc., Third Edit, pp 233–260
Galloway CA, Yoon Y (2012) Perspectives on: SGP symposium on mitochondrial physiology and medicine: what comes first, misshape or dysfunction? The view from metabolic excess. J Gen Physiol 139:455–463
Gemer O, Zaltztein E, Gorodischer R (1983) Absorption of orally administered gentamicin in infants with diarrhea. Pediatr Pharmacol (New York) 3:119–123
Goldstone JV, McArthur AG, Kubota A et al (2010) Identification and developmental expression of the full complement of Cytochrome P450 genes in Zebrafish. BMC Genomics 11:643
Graham GG, Scott KF (2005) Mechanism of action of paracetamol. Am J Ther 12:46–55
Hentschel DM, Park KM, Cilenti L et al (2005) Acute renal failure in zebrafish: a novel system to study a complex disease. Am J Physiol Ren Physiol 288:F923–F929
Herlitz LC, Mohan S, Stokes MB et al (2010) Tenofovir nephrotoxicity: acute tubular necrosis with distinctive clinical, pathological, and mitochondrial abnormalities. Kidney Int 78:1171–1177
Hill A, Mesens N, Steemans M et al (2012) Comparisons between in vitro whole cell imaging and in vivo zebrafish-based approaches for identifying potential human hepatotoxicants earlier in pharmaceutical development. Drug Metab Rev 44:127–140
Huang SM, Xu F, Lam SH et al (2013) Metabolomics of developing zebrafish embryos using gas chromatography- and liquid chromatography-mass spectrometry. Mol BioSyst 9:1372–1380
Kersten S, Arjona FJ (2017) Ion transport in the zebrafish kidney from a human disease angle: possibilities, considerations, and future perspectives. Am J Physiol Renal Physiol 312:F172–F189
Kramer-Zucker AG, Wiessner S, Jensen AM, Drummond IA (2005) Organization of the pronephric filtration apparatus in zebrafish requires nephrin, podocin and the FERM domain protein Mosaic eyes. Dev Biol 285:316–329
Kremer JR, Mastronarde DN, McIntosh JR (1996) Computer visualization of three-dimensional image data using IMOD. J Struc Biol. 116(1):71–76
Kumai Y, Bernier NJ, Perry SF (2014) Angiotensin-II promotes Na+ uptake in larval zebrafish, Danio rerio, in acidic and ion-poor water. J Endocrinol 220:195–205
Kurmi M, Golla VM, Kumar S et al (2016) Stability behaviour of antiretroviral drugs and their combinations. 4: characterization of degradation products of tenofovir alafenamide fumarate and comparison of its degradation and stability behaviour with tenofovir disoproxil fumarate. J Pharm Biomed Anal 131:146–155
Levi L, Ziv T, Admon A et al (2012) Insight into molecular pathways of retinal metabolism, associated with vitellogenesis in zebrafish. AJP Endocrinol Metab 302:E626–E644
Lopez-Novoa JM, Quiros Y, Vicente L et al (2011) New insights into the mechanism of aminoglycoside nephrotoxicity: an integrative point of view. Kidney Int 79:33–45
Luckenbach T, Fischer S, Sturm A (2014) Current advances on ABC drug transporters in fish. Comp Biochem Physiol Part C Toxicol Pharmacol 165:28–52
Mastronarde DN (1997) Dual-axis tomography: an approach with alignment methods that preserve resolution. J Struc Biol 120:343–352
Mazer M, Perrone J (2008) Acetaminophen-induced nephrotoxicity: pathophysiology, clinical manifestations, and management. J Med Toxicol 4:2–6
McGrath P, Li C-Q (2008) Zebrafish: a predictive model for assessing drug-induced toxicity. Drug Discov Today 13:394–401
Mihaljevic I, Popovic M, Zaja R, Smital T (2016) Phylogenetic, syntenic, and tissue expression analysis of slc22 genes in zebrafish (Danio rerio). BMC Genomics 17:626
Nevedomskaya E, Mayboroda OA, Deelder AM (2011) Cross-platform analysis of longitudinal data in metabolomics. Mol BioSyst 7:3214–3222
Olson H, Betton G, Robinson D et al (2000) Concordance of the toxicity of pharmaceuticals in humans and in animals. Regul Toxicol Pharmacol 32:56–67
Pacchiarotta T, Hensbergen PJ, Wuhrer M et al (2012) Fibrinogen alpha chain O-glycopeptides as possible markers of urinary tract infection. J Proteomics 75:1067–1073
Pannicke U, Hönig M, Hess I et al (2009) Reticular dysgenesis (aleukocytosis) is caused by mutations in the gene encoding mitochondrial adenylate kinase 2. Nat Genet 41:101–105
Peng H-C, Wang Y-H, Wen C-C et al (2010) Nephrotoxicity assessments of acetaminophen during zebrafish embryogenesis. Comp Biochem Physiol C Toxicol Pharmacol 151:480–486
Peterson RT, MacRae CA (2012) Systematic approaches to toxicology in the zebrafish. Annu Rev Pharmacol Toxicol 52:433–453
Pluskal T, Castillo S, Villar-Briones A, Oresic M (2010) MZmine 2: modular framework for processing, visualizing, and analyzing mass spectrometry-based molecular profile data. BMC Bioinformatics 11:395
Price VR (2002) ATP depletion of tubular cells causes dissociation of the zonula adherens and nuclear translocation of—catenin and LEF-1. J Am Soc Nephrol 13:1152–1161
Rafelski SM (2013) Mitochondrial network morphology: building an integrative, geometrical view. BMC Biol 11:71
Randhawa MA (2009) Calculation of LD50 values from the method of Miller and Tainter, 1944. J Ayub Med Coll Abbottabad 21:184–185
Rider SA, Tucker CS, Del-Pozo J et al (2012) Techniques for the in vivo assessment of cardio-renal function in zebrafish (Danio rerio) larvae. J Physiol 590:1803–1809
Rider SA, Mullins LJ, Verdon RF et al (2015) Renin expression in developing zebrafish is associated with angiogenesis and requires the notch pathway and endothelium. Am J Physiol Ren Physiol 309:F531–F539
Saad M, Cavanaugh K, Verbueken E et al (2016) Xenobiotic metabolism in the zebrafish: a review of the spatiotemporal distribution, modulation and activity of Cytochrome P450 families 1 to 3. J Toxicol Sci 41:1–11
Schieber NL, Nixon SJ, Webb RI et al (2010) Modern approaches for ultrastructural analysis of the zebrafish embryo. Methods Cell Biol 96:425–442
Schindelin J, Arganda-Carreras I, Frise E et al (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9:676–682
Schindelin J, Rueden CT, Hiner MC, Eliceiri KW (2015) The ImageJ ecosystem: an open platform for biomedical image analysis. Mol Reprod Dev 82:518–529
Smith KY, Patel P, Fine D et al (2009) Randomized, double-blind, placebo-matched, multicenter trial of abacavir/lamivudine or tenofovir/emtricitabine with lopinavir/ritonavir for initial HIV treatment. AIDS 23:1547–1556
Tourret J, Deray G, Isnard-Bagnis C (2013) Tenofovir effect on the kidneys of HIV-infected patients: a double-edged sword? J Am Soc Nephrol 24:1519–1527
Westhoff JH, Giselbrecht S, Schmidts M et al (2013) Development of an automated imaging pipeline for the analysis of the zebrafish larval kidney. PLoS ONE 8:1–13
Wingert RA, Davidson AJ (2008) The zebrafish pronephros: a model to study nephron segmentation. Kidney Int 73(10):1120–1127
Wingert RA, Selleck R, Yu J et al (2007) The cdx genes and retinoic acid control the positioning and segmentation of the zebrafish pronephros. PLoS Genet 3:1922–1938
Youle RJ, van der Bliek AM (2012) Mitochondrial fission, fusion, and stress. Science 337:1062–1065
Acknowledgments
We would like to thank Maysa Franco and Ana Cristina Borges from the Fish Facility of the Gulbenkian Institute of Science.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Funding
This work was supported by the Calouste Gulbenkian Foundation, Gulbenkian Professorship 121986/2012; the Foundation for Science and Technology through the grant ANR/BEX-BID/0153/2012, contract IF/00951/2012 (to SSL), fellowship PD/BD/52420/2013 (to RJ) and travel ship SFRH/BSAB/114291/2016 (to JM); iNOVA4Health Research Unit, LISBOA-01-0145-FEDER-007344.
Ethical statement
All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.
Conflict of interest
The authors declare that they have no conlficts of interest.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Gorgulho, R., Jacinto, R., Lopes, S.S. et al. Usefulness of zebrafish larvae to evaluate drug-induced functional and morphological renal tubular alterations. Arch Toxicol 92, 411–423 (2018). https://doi.org/10.1007/s00204-017-2063-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00204-017-2063-1