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Iohexol plasma clearance, a simple and reliable method to measure renal function in conscious mice

Pflügers Archiv - European Journal of Physiology volume 468, pages 1587–1594 (2016)Cite this article

Abstract

In mice, renal function evaluated by serum creatinine has limitations. Gold standard methods using radioactive markers are cumbersome. We aimed to develop the iohexol plasma clearance as a simple assessment of renal function in conscious mice. We used two groups of mice: testing and validation, formed by 16 animals (8 male and 8 female) each. Iohexol was injected intravenously into the tail vein (6.47 mg), and tail tip blood samples were collected at 1, 3, 7, 10, 15, 35, 55, and 75 min. Iohexol plasma clearances were calculated in two ways: (1) two-compartment model (CL2) using all time points and (2) one-compartment model (CL1) using only the last four points. In the testing group, CL1 overestimated the true clearance (CL2). Therefore, CL1 was recalculated applying a correction factor calculated as the ratio between CL2/CL1. The latter was considered as the simplified method. CL2 averaged 223.3 ± 64.3 μl/min and CL1 252.4 ± 76.4 μl/min, which lead to a CF of 0.89. Comparable results for CL2, CL1, and simplified method were observed in the validation group. Additionally, we demonstrated the capacity of the simplified method to quantitatively assess different degrees of renal function in three mouse models: hyperoxaluric-CKD (87.4 ± 28.3 μl/min), heminephrectomized (135–0 ± 50.5 μl/min), and obese (399.6 ± 112.1 μl/min) mice. We have developed a simple and reliable method to evaluate renal function in conscious mice under diverse clinical conditions. Moreover, the test can be repeated in the same animal, which makes the method useful to examine renal function changes over time.

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Acknowledgments

SLL is a research fellow supported by the Instituto de Salud Carlos III (Grants for Río Hortega specialized healthcare post-training contracts), ISCIII CM15/00214, Spain.

The authors thank the support from (a) Instituto de Salud Carlos III for the following grant: PI13/00342 and Fondos FEDER for the following grants: PI 07/0732, the REDINREN RD/0021/0008 and PI10/02428 (b) Fundación Caja Canarias for the following grant: DIAB05 (c) We thank the IMBRAIN project (FP7-RE6-POT-2012-CT2012–31637-IMBRAIN) funded under the 7th Frameworks Programme capacities.

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    Affiliations

    1. Nephrology Department, University Hospital of the Canary Islands, La Laguna, Tenerife, Spain

      Sergio Luis-Lima

    2. University of La Laguna, Instituto Tecnologías Biomédicas (ITB), Tenerife, Spain

      Sergio Luis-Lima & Esteban Porrini

    3. Research Unit, University Hospital of the Canary Islands, La Laguna, S/C Tenerife, Spain

      Ana Elena Rodríguez-Rodríguez

    4. Department of Pathology, Centre for Biomedical Research on Rare Diseases (CIBERER), La Laguna, S/C Tenerife, Spain

      Cristina Martín-Higueras & Eduardo Salido

    5. Department of Physiology and Institute of Biomedical Technologies, University of La Laguna, Tenerife, Spain (D.A.d.l.R.), La Laguna, S/C Tenerife, Spain

      Catalina Sierra-Ramos & Diego Alvarez de la Rosa

    6. IRCCS- Istituto di Ricerche Farmacologiche Mario Negri, Clinical Research Center for Rare Diseases ‘Aldo & Cele Daccò’, Ranica, BG, Italy

      Fabiola Carrara & Flavio Gaspari

    7. Pharmacology Unit, Medical School, University of La Laguna, La Laguna, S/C Tenerife, Spain

      María Rosa Arnau

    Authors
    1. Sergio Luis-Lima
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    2. Ana Elena Rodríguez-Rodríguez
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    10. Esteban Porrini
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    Correspondence to Esteban Porrini.

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    All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. 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.

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    Sergio Luis-Lima and Ana Elena Rodríguez-Rodríguez contributed equally to this work.

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    Luis-Lima, S., Rodríguez-Rodríguez, A.E., Martín-Higueras, C. et al. Iohexol plasma clearance, a simple and reliable method to measure renal function in conscious mice. Pflugers Arch - Eur J Physiol 468, 1587–1594 (2016). https://doi.org/10.1007/s00424-016-1843-4

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    • DOI: https://doi.org/10.1007/s00424-016-1843-4

    Keywords

    • Glomerular filtration rate
    • Iohexol plasma clearance
    • Conscious mice
    • One-compartment model
    • Gold standard technique