Sex- and age-differences in cellular hallmarks of aging in a species with female-biased longevity and environmental sex determination

Marks, J. R.; Janzen, F. J.; Reinke, B. A.; Addis, E. A.; Adesioye, O.; Bock, S.; Clark, M.; Crowther, C.; Hoekstra, L. A.; Judson, J.; Krueger, C.; Sills, A. P.; Bronikowski, A. M.

doi:10.64898/2025.12.10.693506

bioRxiv · 10.64898/2025.12.10.693506

Sex- and age-differences in cellular hallmarks of aging in a species with female-biased longevity and environmental sex determination

Marks, J. R.·Janzen, F. J.·Reinke, B. A.·Addis, E. A.·Adesioye, O.·Bock, S.·Clark, M.·Crowther, C.·Hoekstra, L. A.·Judson, J.·Krueger, C.·Sills, A. P.·Bronikowski, A. M.

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Abstract

Cellular hallmarks of aging have been discovered and characterized in a number of model species for studying aging biology - such as humans, mice, fruit flies, and nematodes. Whether these canonical age-related changes to cellular physiology are present across diverse species that have variable rates of demographic aging remains less studied. Here, we tested whether several ubiquitous cellular hallmarks of aging - mitochondrial function, reactive oxygen species generation, and inducible DNA damage - change with age and in a sex-dependent manner in a species with indeterminate growth and reproduction (painted turtles, Chrysemys picta). A further feature of their biology that recommends them for an ecological model of vertebrate aging is their female-biased longevity, despite an absence of genotypic sex determination. Thus lifespan and aging may be reliable features of sex-specific life-histories. We measured aspects of mitochondrial health (cellular basal, maximal, and spare oxygen consumption rates), cellular levels of reactive oxygen species, and aspects of DNA damage and repair from exposure to UVB. We used these measures across several physiological axes as proxies for age-related physiological dysfunction. We further assessed our measures across several populations of painted turtles. We found that sex explained the largest proportion of variation, with males differing from females in mitochondrial function, reactive oxygen species production, and inducible DNA damage. In several cases, age significantly interacted with sex, but the effect size was small relative to sex alone. Thus, we found that sex, rather than age or size, was a consistent predictor of cellular aging physiological in this species with where females live longer and age slower.

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Marks, J. R., Janzen, F. J., Reinke, B. A., Addis, E. A., Adesioye, O., Bock, S., Clark, M., Crowther, C., Hoekstra, L. A., Judson, J., Krueger, C., Sills, A. P., Bronikowski, A. M.. 2025-12-13. Sex- and age-differences in cellular hallmarks of aging in a species with female-biased longevity and environmental sex determination. https://doi.org/10.64898/2025.12.10.693506

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