A toolbox for the longitudinal assessment of healthspan in aging mice


The number of people aged over 65 is expected to double in the next 30 years. For many, living longer will mean spending more years with the burdens of chronic diseases such as Alzheimer’s disease, cardiovascular disease, and diabetes. Although researchers have made rapid progress in developing geroprotective interventions that target mechanisms of aging and delay or prevent the onset of multiple concurrent age-related diseases, a lack of standardized techniques to assess healthspan in preclinical murine studies has resulted in reduced reproducibility and slow progress. To overcome this, major centers in Europe and the United States skilled in healthspan analysis came together to agree on a toolbox of techniques that can be used to consistently assess the healthspan of mice. Here, we describe the agreed toolbox, which contains protocols for echocardiography, novel object recognition, grip strength, rotarod, glucose tolerance test (GTT) and insulin tolerance test (ITT), body composition, and energy expenditure. The protocols can be performed longitudinally in the same mouse over a period of 4–6 weeks to test how candidate geroprotectors affect cardiac, cognitive, neuromuscular, and metabolic health.

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Fig. 1: A graphic representation of the experimental design for healthspan assessment in response to a geroprotector of choice in C57BL6/J using the recommended toolbox.
Fig. 2: Representative images from high-resolution echocardiography systems.
Fig. 3: Novel object recognition (NOR) memory enhancement in exercised male mice.
Fig. 4: Assessment of muscle strength and neuromuscular function.
Fig. 5: Effects of dietary protein level on energy balance.
Fig. 6: Glucose and insulin tolerance tests.

Data availability

All data shown in this paper are available from the authors upon reasonable request.


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We thank K. R. McGreevy from the Cajal Institute for her assistance with Fig. 3, and D. E. Cohen for enabling D.W.L. to attend the 2017 MouseAGE annual meeting. This article is based on work from COST Action (BM1402: MouseAGE), supported by COST (European Cooperation in Science and Technology; I.B., P.K.P., D.E., L.J.T.). Part of this work has been funded by the European Union Research and Innovation Program Horizon 2020 (grant agreement number 730879; I.B.). The Lamming laboratory is supported in part by the NIH (AG051974, AG056771 and AG062328) and by the US Department of Veterans Affairs (I01-BX004031), and this work was supported using facilities and resources from the William S. Middleton Memorial Veterans Hospital. D.E. was supported by a grant from the Helmholtz-Future Topic ‘Aging and Metabolic Programming’ (AMPro), the Mayo clinic is funded by Robert and Arlene Kogod, the Connor Group, and partially by NIA grant AG13925. R.dC., C.D.G., and I.N. are supported by the Intramural Research Program of the NIA/NIH. This work does not necessarily represent the official views of the NIH, the Department of Veterans Affairs or the United States Government.

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I.B., R.d.C., D.E., C.D.G., I.N.-E., S.M., P.K.P., T.T., J.L.T., A.L. and D.W.L. contributed equally to the development of the protocols. I.B. and D.L. wrote the manuscript. All other authors edited and approved the final manuscript.

Correspondence to I. Bellantuono or D. W. Lamming.

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Peer review information Nature Protocols thanks Erin R. Hascup, Susan Howlett and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Bellantuono, I., de Cabo, R., Ehninger, D. et al. A toolbox for the longitudinal assessment of healthspan in aging mice. Nat Protoc (2020). https://doi.org/10.1038/s41596-019-0256-1

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