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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Bellantuono, I. Find drugs that delay many diseases of old age. Nature 554, 293–295 (2018).
Tchkonia, T. & Kirkland, J. L. Aging, cell senescence, and chronic disease: emerging therapeutic strategies. JAMA 320, 1319–1320 (2018).
Martin-Montalvo, A. et al. Metformin improves healthspan and lifespan in mice. Nat. Commun. 4, 2192 (2013).
Neff, F. et al. Rapamycin extends murine lifespan but has limited effects on aging. J. Clin. Invest. 123, 3272–3291 (2013).
Zhu, Y. et al. The Achilles’ heel of senescent cells: from transcriptome to senolytic drugs. Aging Cell 14, 644–658 (2015).
Arriola Apelo, S. I., Pumper, C. P., Baar, E. L., Cummings, N. E. & Lamming, D. W. Intermittent administration of rapamycin extends the life span of female C57BL/6J mice. J. Gerontol. A Biol. Sci. Med. Sci. 71, 876–881 (2016).
Richardson, A. et al. Measures of healthspan as indices of aging in mice-a recommendation. J. Gerontol. A Biol. Sci. Med. Sci. 71, 427–430 (2016).
Kane, A. E., Keller, K. M., Heinze-Milne, S., Grandy, S. A. & Howlett, S. E. A murine frailty index based on clinical and laboratory measurements: links between frailty and pro-inflammatory cytokines differ in a sex-specific manner. J. Gerontol. A Biol. Sci. Med. Sci. 74, 275–282 (2019).
Whitehead, J. C. et al. A clinical frailty index in aging mice: comparisons with frailty index data in humans. J. Gerontol. A Biol. Sci. Med. Sci. 69, 621–632 (2014).
Hamm, R. J., Pike, B. R., O’Dell, D. M., Lyeth, B. G. & Jenkins, L. W. The rotarod test: an evaluation of its effectiveness in assessing motor deficits following traumatic brain injury. J. Neurotrauma 11, 187–196 (1994).
Wolff, B. S., Raheem, S. A. & Saligan, L. N. Comparing passive measures of fatigue-like behavior in mice. Sci. Rep. 8, 14238 (2018).
Schafer, M. J. et al. Exercise prevents diet-induced cellular senescence in adipose tissue. Diabetes 65, 1606–1615 (2016).
Halldorsdottir, S., Carmody, J., Boozer, C. N., Leduc, C. A. & Leibel, R. L. Reproducibility and accuracy of body composition assessments in mice by dual energy x-ray absorptiometry and time domain nuclear magnetic resonance. Int. J. Body Compos. Res. 7, 147–154 (2009).
Potter, P. K. et al. Novel gene function revealed by mouse mutagenesis screens for models of age-related disease. Nat. Commun. 7, 12444 (2016).
Xie, K. et al. Epigenetic alterations in longevity regulators, reduced life span, and exacerbated aging-related pathology in old father offspring mice. Proc. Natl Acad. Sci. USA 115, E2348–E2357 (2018).
Lamming, D. W. et al. Young and old genetically heterogeneous HET3 mice on a rapamycin diet are glucose intolerant but insulin sensitive. Aging Cell 12, 712–718 (2013).
Lamming, D. W. et al. Rapamycin-induced insulin resistance is mediated by mTORC2 loss and uncoupled from longevity. Science 335, 1638–1643 (2012).
Arriola Apelo, S. I. et al. Alternative rapamycin treatment regimens mitigate the impact of rapamycin on glucose homeostasis and the immune system. Aging Cell 15, 28–38 (2016).
Stout, M. B. et al. 17alpha-Estradiol alleviates age-related metabolic and inflammatory dysfunction in male mice without inducing feminization. J. Gerontol. A Biol. Sci. Med. Sci. 72, 3–15 (2017).
Alfaras, I. et al. Health benefits of late-onset metformin treatment every other week in mice. Npj. Aging Mech. Dis. 3, 16 (2017).
Diaz-Ruiz, A. et al.Overexpression of CYB5R3 and NQO1, two NAD(+) -producing enzymes, mimics aspects of caloric restriction. Aging Cell, e12767 (2018).
Mitchell, S. J. et al. Nicotinamide improves aspects of healthspan, but not lifespan, in mice. Cell Metab. 27, 667–676 e664 (2018).
Allard, J. S. et al. Prolonged metformin treatment leads to reduced transcription of Nrf2 and neurotrophic factors without cognitive impairment in older C57BL/6J mice. Behav. Brain. Res. 301, 1–9 (2016).
Lee, S. et al. Assessment of cognitive impairment in a mouse model of high-fat diet-induced metabolic stress with touchscreen-based automated battery system. Exp. Neurobiol. 27, 277–286 (2018).
Benice, T. S. & Raber, J. Object recognition analysis in mice using nose-point digital video tracking. J. Neurosci. Methods 168, 422–430 (2008).
Bettis, T. & Jacobs, L. F. Sex differences in object recognition are modulated by object similarity. Behav. Brain Res. 233, 288–292 (2012).
Prendergast, B. J., Onishi, K. G. & Zucker, I. Female mice liberated for inclusion in neuroscience and biomedical research. Neurosci. Biobehav. Rev. 40, 1–5 (2014).
Clayton, J. A. & Collins, F. S. Policy: NIH to balance sex in cell and animal studies. Nature 509, 282–283 (2014).
McCullough, L. D., McCarthy, M. M. & de Vries, G. J. NIH policy: status quo is also costly. Nature 510, 340 (2014).
Brooks, S. P., Pask, T., Jones, L. & Dunnett, S. B. Behavioural profiles of inbred mouse strains used as transgenic backgrounds. II: Cogn. tests. Genes Brain Behav. 4, 307–317 (2005).
Feridooni, H. A., Sun, M. H., Rockwood, K. & Howlett, S. E. Reliability of a frailty index based on the clinical assessment of health deficits in male C57BL/6J mice. J. Gerontol. A Biol. Sci. Med. Sci. 70, 686–693 (2015).
Sukoff Rizzo, S. J. et al. Assessing healthspan and lifespan measures in aging mice: optimization of testing protocols, replicability, and rater reliability. Curr. Protoc. Mouse Biol. 8, e45 (2018).
Palmer, A. K. et al. Targeting senescent cells alleviates obesity-induced metabolic dysfunction. Aging Cell, e12950 (2019).
Xu, M. et al. Senolytics improve physical function and increase lifespan in old age. Nat. Med. 24, 1246–1256 (2018).
Mitchell, S. J. et al. Effects of sex, strain, and energy intake on hallmarks of aging in mice. Cell Metab. 23, 1093–1112 (2016).
Cummings, N. E. et al. Restoration of metabolic health by decreased consumption of branched-chain amino acids. J. Physiol. 596, 623–645 (2018).
Xie, K. et al. Every-other-day feeding extends lifespan but fails to delay many symptoms of aging in mice. Nat. Commun. 8, 155 (2017).
McGreevy, K. R. et al. Intergenerational transmission of the positive effects of physical exercise on brain and cognition. Proc. Natl Acad. Sci. USA (2019).
Lamming, D. W. et al. Depletion of Rictor, an essential protein component of mTORC2, decreases male lifespan. Aging Cell 13, 911–917 (2014).
Zhu, Y. et al. New agents that target senescent cells: the flavone, fisetin, and the BCL-XL inhibitors, A1331852 and A1155463. Aging (Albany NY) 9, 955–963 (2017).
Yousefzadeh, M. J. et al. Fisetin is a senotherapeutic that extends health and lifespan. EbioMedicine 36, 18–28 (2018).
Xu, M. et al. Targeting senescent cells enhances adipogenesis and metabolic function in old age. eLife 4, e12997 (2015).
Mercken, E. M. et al. SRT2104 extends survival of male mice on a standard diet and preserves bone and muscle mass. Aging Cell 13, 787–796 (2014).
Mitchell, S. J. et al. The SIRT1 activator SRT1720 extends lifespan and improves health of mice fed a standard diet. Cell Rep. 6, 836–843 (2014).
Minor, R. K. et al. SRT1720 improves survival and healthspan of obese mice. Sci. Rep. 1, 70 (2011).
Kane, A. E. et al. Impact of longevity interventions on a validated mouse clinical frailty index. J. Gerontol. A Biol. Sci. Med. Sci. 71, 333–339 (2016).
Kane, A. E. et al. A comparison of two mouse frailty assessment tools. J. Gerontol. A Biol. Sci. Med. Sci. 72, 904–909 (2017).
Yu, D. et al. Short-term methionine deprivation improves metabolic health via sexually dimorphic, mTORC1-independent mechanisms. FASEB J. 32, 3471–3482 (2018).
Ramos, F. J. et al. Rapamycin reverses elevated mTORC1 signaling in lamin A/C-deficient mice, rescues cardiac and skeletal muscle function, and extends survival. Sci. Transl. Med. 4, 144ra103 (2012).
Darcy, J. et al. Increased environmental temperature normalizes energy metabolism outputs between normal and Ames dwarf mice. Aging 10, 2709–2722 (2018).
Cohen, D. E., Supinski, A. M., Bonkowski, M. S., Donmez, G. & Guarente, L. P. Neuronal SIRT1 regulates endocrine and behavioral responses to calorie restriction. Genes Dev. 23, 2812–2817 (2009).
Bitto, A. et al. Transient rapamycin treatment can increase lifespan and healthspan in middle-aged mice. eLife 5, https://doi.org/10.7554/eLife.16351 (2016).
Chen, C., Liu, Y., Liu, Y. & Zheng, P. mTOR regulation and therapeutic rejuvenation of aging hematopoietic stem cells. Sci. Signal 2, ra75 (2009).
Harrison, D. E. et al. Rapamycin fed late in life extends lifespan in genetically heterogeneous mice. Nature 460, 392–395 (2009).
Dai, D. F. et al. Altered proteome turnover and remodeling by short-term caloric restriction or rapamycin rejuvenate the aging heart. Aging Cell 13, 529–539 (2014).
Flynn, J. M. et al. Late-life rapamycin treatment reverses age-related heart dysfunction. Aging Cell 12, 851–862 (2013).
Hou, Y. et al. NAD+ supplementation normalizes key Alzheimer’s features and DNA damage responses in a new AD mouse model with introduced DNA repair deficiency. Proc. Natl Acad. Sci. USA 115, E1876–E1885 (2018).
Spilman, P. et al. Inhibition of mTOR by rapamycin abolishes cognitive deficits and reduces amyloid-beta levels in a mouse model of Alzheimer’s disease. PLoS ONE 5, e9979 (2010).
Baur, J. A. et al. Resveratrol improves health and survival of mice on a high-calorie diet. Nature 444, 337–342 (2006).
Fontana, L., Kennedy, B. K., Longo, V. D., Seals, D. & Melov, S. Medical research: treat ageing. Nature 511, 405–407 (2014).
Mitchell, S. J. et al. Daily fasting improves health and survival in male mice independent of diet composition and calories. Cell Metab. 29, 221–228 e223 (2019).
Wilkinson, J. E. et al. Rapamycin slows aging in mice. Aging Cell 11, 675–682 (2012).
Turturro, A. et al. Growth curves and survival characteristics of the animals used in the Biomarkers of Aging Program. J. Gerontol. A Biol. Sci. Med. Sci. 54, B492–501 (1999).
Dumas, S. N. & Lamming, D. W. Next generation strategies for geroprotection via mTORC1 inhibition. J. Gerontol. A Biol. Sci. Med. Sci., https://doi.org/10.1093/gerona/glz056 (2019).
Harrison, D. E. et al. Acarbose, 17-alpha-estradiol, and nordihydroguaiaretic acid extend mouse lifespan preferentially in males. Aging Cell 13, 273–282 (2014).
Lamming, D. W. Diminished mTOR signaling: a common mode of action for endocrine longevity factors. SpringerPlus 3, 735 (2014).
Strong, R. et al. Longer lifespan in male mice treated with a weakly estrogenic agonist, an antioxidant, an alpha-glucosidase inhibitor or a Nrf2-inducer. Aging Cell 15, 872–884 (2016).
Miller, R. A. et al. Rapamycin-mediated lifespan increase in mice is dose and sex dependent and metabolically distinct from dietary restriction. Aging Cell 13, 468–477 (2014).
Fontana, L. et al. Decreased consumption of branched-chain amino acids improves metabolic health. Cell Rep. 16, 520–530 (2016).
De Leon, E. R. et al. Age-dependent protection of insulin secretion in diet induced obese mice. Sci. Rep. 8, 17814 (2018).
Gill, J. F., Santos, G., Schnyder, S. & Handschin, C. PGC-1alpha affects aging-related changes in muscle and motor function by modulating specific exercise-mediated changes in old mice. Aging Cell 17, e12697 (2018).
Graber, T. G., Ferguson-Stegall, L., Liu, H. & Thompson, L. V. Voluntary aerobic exercise reverses frailty in old mice. J. Gerontol. A Biol. Sci. Med. Sci. 70, 1045–1058 (2015).
Chellappa, K. et al. Hypothalamic mTORC2 is essential for metabolic health and longevity. Aging Cell 18, e13014 (2019).
Yu, D. et al. Calorie-restriction-induced insulin sensitivity is mediated by adipose mTORC2 and not required for lifespan extension. Cell Rep. 29, 236–248.e3 (2019).
Nadon, N. L., Strong, R., Miller, R. A. & Harrison, D. E. NIA interventions testing program: investigating putative aging intervention agents in a genetically heterogeneous mouse model. EbioMedicine 21, 3–4 (2017).
Yuan, R. et al. Genetic coregulation of age of female sexual maturation and lifespan through circulating IGF1 among inbred mouse strains. Proc. Natl Acad. Sci. USA 109, 8224–8229 (2012).
Ryan, D. P. et al. A paternal methyl donor-rich diet altered cognitive and neural functions in offspring mice. Mol. Psychiatry 23, 1345–1355 (2018).
Fernandez-Twinn, D. S., Constancia, M. & Ozanne, S. E. Intergenerational epigenetic inheritance in models of developmental programming of adult disease. Semin. Cell Dev. Biol. 43, 85–95 (2015).
Martin-Gronert, M. S. & Ozanne, S. E. Early life programming of obesity. Med. Wieku Rozwoj. 17, 7–12 (2013).
Menting, M. D. et al. Maternal obesity in pregnancy impacts offspring cardiometabolic health: Systematic review and meta-analysis of animal studies. Obes. Rev. 20, 675–685 (2019).
Dearden, L., Bouret, S. G. & Ozanne, S. E. Sex and gender differences in developmental programming of metabolism. Mol. Metab. 15, 8–19 (2018).
Evered, L., Scott, D. A. & Silbert, B. Cognitive decline associated with anesthesia and surgery in the elderly: does this contribute to dementia prevalence? Curr. Opin. Psychiatry 30, 220–226 (2017).
Schulte, P. J. et al. Association between exposure to anaesthesia and surgery and long-term cognitive trajectories in older adults: report from the Mayo Clinic Study of Aging. Br. J. Anaesth. 121, 398–405 (2018).
Steinmetz, J. & Rasmussen, L. S. Anesthesia and the risk of dementia in the elderly. Presse Med. 47, e45–e51 (2018).
Butterfield, N. N., Graf, P., Ries, C. R. & MacLeod, B. A. The effect of repeated isoflurane anesthesia on spatial and psychomotor performance in young and aged mice. Anesth. Analg. 98, 1305–1311 (2004).
Xu, H. et al. Smaller sized inhaled anesthetics have more potency on senescence-accelerated prone-8 mice compared with senescence-resistant-1 mice. J. Alzheimers Dis. 39, 29–34 (2014).
Li, X. M. et al. Disruption of hippocampal neuregulin 1-ErbB4 signaling contributes to the hippocampus-dependent cognitive impairment induced by isoflurane in aged mice. Anesthesiology 121, 79–88 (2014).
Li, R. L. et al. Postoperative impairment of cognitive function in old mice: a possible role for neuroinflammation mediated by HMGB1, S100B, and RAGE. J. Surg. Res. 185, 815–824 (2013).
Campbell, J. E. & Drucker, D. J. Pharmacology, physiology, and mechanisms of incretin hormone action. Cell Metab. 17, 819–837 (2013).
Linnemann, A. K. et al. Glucagon-like peptide-1 regulates cholecystokinin production in beta-cells to protect from apoptosis. Mol. Endocrinol. 29, 978–987 (2015).
Renner, S., Blutke, A., Streckel, E., Wanke, R. & Wolf, E. Incretin actions and consequences of incretin-based therapies: lessons from complementary animal models. J. Pathol. 238, 345–358 (2016).
Leiter, E. H., Premdas, F., Harrison, D. E. & Lipson, L. G. Aging and glucose homeostasis in C57BL/6J male mice. FASEB J. 2, 2807–2811 (1988).
Ennaceur, A. & Delacour, J. A new one-trial test for neurobiological studies of memory in rats. 1: behavioral data. Behav. Brain Res. 31, 47–59 (1988).
Leger, M. et al. Object recognition test in mice. Nat. Prot. 8, 2531–2537 (2013).
Servick, K. Mouse microbes may make scientific studies harder to replicate. Science, https://doi.org/10.1126/science.aah7199 (2016).
Cheng, C. J., Gelfond, J. A. L., Strong, R. & Nelson, J. F. Genetically heterogeneous mice exhibit a female survival advantage that is age- and site-specific: results from a large multi-site study. Aging Cell, e12905, https://doi.org/10.1111/acel.12905 (2019).
Kalueff, A. V., Keisala, T., Minasyan, A., Kuuslahti, M. & Tuohimaa, P. Temporal stability of novelty exploration in mice exposed to different open field tests. Behav. Process. 72, 104–112 (2006).
Heyser, C. J. & Chemero, A. Novel object exploration in mice: not all objects are created equal. Behav. Process. 89, 232–238 (2012).
Ennaceur, A. One-trial object recognition in rats and mice: methodological and theoretical issues. Behav. Brain Res. 215, 244–254 (2010).
Raber, J. Novel images and novel locations of familiar images as sensitive translational cognitive tests in humans. Behav. Brain Res. 285, 53–59 (2015).
Frick, K. M. & Gresack, J. E. Sex differences in the behavioral response to spatial and object novelty in adult C57BL/6 mice. Behav. Neurosci. 117, 1283–1291 (2003).
Prut, L. & Belzung, C. The open field as a paradigm to measure the effects of drugs on anxiety-like behaviors: a review. Eur. J. Pharmacol. 463, 3–33 (2003).
Klein, S. M. et al. Noninvasive in vivo assessment of muscle impairment in the mdx mouse model—a comparison of two common wire hanging methods with two different results. J. Neurosci. Methods 203, 292–297 (2012).
Karp, N. A., Segonds-Pichon, A., Gerdin, A. K., Ramirez-Solis, R. & White, J. K. The fallacy of ratio correction to address confounding factors. Lab. Anim. 46, 245–252 (2012).
Gregg, T. et al. Pancreatic beta-cells from mice offset ageaAssociated mitochondrial deficiency with reduced KATP channel activity. Diabetes 65, 2700–2710 (2016).
Schnelle, M. et al. Echocardiographic evaluation of diastolic function in mouse models of heart disease. J. Mol. Cell. Cardiol. 114, 20–28 (2018).
Majumder, S. et al. Lifelong rapamycin administration ameliorates age-dependent cognitive deficits by reducing IL-1beta and enhancing NMDA signaling. Aging Cell 11, 326–335 (2012).
Lin, A. L. et al. Chronic rapamycin restores brain vascular integrity and function through NO synthase activation and improves memory in symptomatic mice modeling Alzheimer’s disease. J. Cereb. Blood Flow. Metab. 33, 1412–1421 (2013).
Ozcelik, S. et al. Rapamycin attenuates the progression of tau pathology in P301S tau transgenic mice. PLoS ONE 8, e62459 (2013).
Caccamo, A., Majumder, S., Richardson, A., Strong, R. & Oddo, S. Molecular interplay between mammalian target of rapamycin (mTOR), amyloid-beta, and Tau: effects on cognitive impairments. J. Biol. Chem. 285, 13107–13120 (2010).
Lee, A. S. et al. Aged skeletal muscle retains the ability to fully regenerate functional architecture. Bioarchitecture 3, 25–37 (2013).
van Norren, K. et al. Behavioural changes are a major contributing factor in the reduction of sarcopenia in caloric-restricted ageing mice. J. Cachexia Sarcopenia Muscle 6, 253–268 (2015).
Ge, X. et al. Grip strength is potentially an early indicator of age-related decline in mice. Pathobiol. Aging Age Relat. Dis. 6, 32981 (2016).
Hirsch, C. H., Buzkova, P., Robbins, J. A., Patel, K. V. & Newman, A. B. Predicting late-life disability and death by the rate of decline in physical performance measures. Age Ageing 41, 155–161 (2012).
Bogue, M. A. et al. Accessing data resources in the mouse phenome database for genetic analysis of murine life span and health span. J. Gerontol. A Biol. Sci. Med. Sci. 71, 170–177 (2016).
Xu, M. et al. JAK inhibition alleviates the cellular senescence-associated secretory phenotype and frailty in old age. Proc. Natl Acad. Sci. USA 112, E6301–6310 (2015).
Peterson, R. L., Parkinson, K. C. & Mason, J. B. Manipulation of ovarian function significantly influenced sarcopenia in postreproductive-age mice. J. Transplant. 2016, 4570842 (2016).
Graber, T. G., Ferguson-Stegall, L., Kim, J. H. & Thompson, L. V. C57BL/6 neuromuscular healthspan scoring system. J. Gerontol. A Biol. Sci. Med. Sci. 68, 1326–1336 (2013).
Simmons, D. A. et al. A small molecule TrkB ligand reduces motor impairment and neuropathology in R6/2 and BACHD mouse models of Huntington’s disease. J. Neurosci. 33, 18712–18727 (2013).
Jeong, H. et al. Sirt1 mediates neuroprotection from mutant huntingtin by activation of the TORC1 and CREB transcriptional pathway. Nat. Med. 18, 159–165 (2011).
Zhang, Y. et al. Rapamycin extends life and health in C57BL/6 mice. J. Gerontol. A Biol. Sci. Med. Sci. 69, 119–130 (2014).
Cummings, N. E. & Lamming, D. W. Regulation of metabolic health and aging by nutrient-sensitive signaling pathways. Mol. Cell. Endocrinol. 455, 13–22 (2017).
Lamming, D. W. & Anderson, R. M. Metabolic effects of caloric restriction in eLS. (John Wiley & Sons, Ltd, 2014).
Malloy, V. L. et al. Methionine restriction decreases visceral fat mass and preserves insulin action in aging male Fischer 344 rats independent of energy restriction. Aging Cell 5, 305–314 (2006).
Green, C. L. & Lamming, D. W. Regulation of metabolic health by essential dietary amino acids. Mech. Ageing Dev. 177, 186–200 (2018).
Fang, Y. et al. Duration of rapamycin treatment has differential effects on metabolism in mice. Cell Metab. 17, 456–462 (2013).
Brewer, R. A., Gibbs, V. K. & Smith, D. L. Jr. Targeting glucose metabolism for healthy aging. Nutr. Healthy Aging 4, 31–46 (2016).
Tschop, M. H. et al. A guide to analysis of mouse energy metabolism. Nat. Methods 9, 57–63 (2011).
Lusk, G. ANIMAL CALORIMETRY: Twenty-Fourth Paper. ANALYSIS OF THE OXIDATION OF MIXTURES OF CARBOHYDRATE AND FAT. J. Biol. Chem. 59, 41–42 (1924).
Goodpaster, B. H. & Sparks, L. M. Metabolic flexibility in health and disease. Cell Metab. 25, 1027–1036 (2017).
Solon-Biet, S. M. et al. Dietary protein to carbohydrate ratio and caloric restriction: comparing metabolic outcomes in mice. Cell Rep. 11, 1529–1534 (2015).
U.S. Department of Health and Human Services. Health, United States, 2016 With Chartbook on Long-term Trends in Health (National Center for Heath Statistics, 2016).
Taffet, G. E., Pham, T. T. & Hartley, C. J. The age-associated alterations in late diastolic function in mice are improved by caloric restriction. J. Gerontol. A Biol. Sci. Med. Sci. 52, B285–290 (1997).
Casaclang-Verzosa, G., Enriquez-Sarano, M., Villaraga, H. R. & Miller, J. D. Echocardiographic approaches and protocols for comprehensive phenotypic characterization of valvular heart disease in mice. J. Vis. Exp. https://doi.org/10.3791/54110 (2017).
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.
The authors declare no competing interests.
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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Key references using this protocol
Martin-Montalvo, A. et al. Nat. Commun. 4, 2192 (2013): https://doi.org/10.1038/ncomms3192.
Neff, F. et al. J. Clin. Invest. 123, 3272–3291 (2013): https://doi.org/10.1172/JCI67674.
Zhu, Y. et al. Aging Cell 14, 644–658 (2015): https://doi.org/10.1111/acel.12344.
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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 15, 540–574 (2020). https://doi.org/10.1038/s41596-019-0256-1
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