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DNA Methylation Shifted as Namibian Male Cheetahs Became Territorial

Adult male cheetah standing in Kruger National Park
Research: Molecular biology & biochemistry, Genetics & genomics

DNA methylation changed extensively when free-ranging male cheetahs (Acinonyx jubatus jubatus) in Namibia shifted from wide-ranging non-territorial “floaters” to territory holders. Researchers sampled the same ten adult males twice — first as floaters and later after they had become territorial. DNA methylation is an epigenetic modification that can influence gene regulation without changing the underlying DNA sequence.

Widespread methylation changes within the same males

The paired design allowed the researchers to compare each male with himself, separating the tactic shift from fixed genetic differences among animals. Using MBD-seq, which enriches methylated DNA fragments before sequencing to compare methylation patterns across the genome, they found widespread changes with a predominance of hypermethylation, or increased methylation, in territorial males.

Differentially methylated regions (DMRs) overlapped genes associated with anatomical and head development, brain and neurodevelopment, cAMP biosynthesis — part of intracellular signalling — and ion-channel transport. Several individual genes illustrate the biological links the authors discuss. MYO18B is involved in muscle-specific gene regulation and actin organisation, PLXNA1 in neuronal guidance and skeletal development, and PTPRS in brain development and regulation of neurotrophic signalling. These overlaps make the genes plausible candidates for processes accompanying territoriality, but do not show that methylation of these genes caused the phenotypic changes.

The molecular pattern matched a major phenotypic transition

The molecular patterns corresponded to the striking phenotypic transition already known in male cheetahs. After acquiring territories, males gain muscle mass and larger heads, become more aggressive and bold, and reduce their movements from large roaming ranges to much smaller territories centred on marking trees.

The methylation was measured in peripheral blood mononuclear cells (PBMCs), however. DMRs linked to brain-, muscle- or head-development genes therefore do not mean that the same methylation changes were demonstrated in brain, muscle or cranial tissue. MBD-seq also measures methylation patterns rather than gene expression itself, so the study identifies an epigenetic signature associated with the transition rather than demonstrating which genes were turned up or down in the relevant tissues.

Association does not establish cause and effect

The study cannot show that DNA methylation caused territoriality. The researchers accounted for age, and age-associated methylation did not reproduce the same coherent functional pattern, but they lacked repeated samples from males that remained floaters and samples taken during the transition. The epigenetic changes could therefore precede the shift, accompany it or arise as a consequence of hormonal, metabolic and behavioural changes after territory acquisition.

Cheetahs have unusually low genetic diversity, making this within-individual regulatory flexibility especially interesting. The results show a coordinated epigenetic signature associated with phenotypic plasticity in a wild mammal, but the authors emphasise that further transcriptomic, hormonal and functional work is needed before the association can be turned into a causal mechanism.

Sources and image credit

  1. Vullioud, C., Geweiler, D., Melzheimer, J., Heinrich, S., Fickel, J., Wachter, B., & Weyrich, A. (2026). Epigenetic Changes Associated With a Phenotypic Transition in Namibian Cheetahs. Molecular Ecology, 35(17), e70527.Primary open-access article first published 7 September 2026; full text checked. Ten free-ranging adult male cheetahs in Namibia were sampled first as non-territorial floaters and later as territory holders. The paired design found widespread DNA-methylation changes, predominantly hypermethylation in territorial males. DMRs overlapped genes associated with anatomical and head development, neurodevelopment, cAMP biosynthesis and ion-channel transport. Methylation was measured in PBMCs and does not demonstrate the same changes in brain, muscle or cranial tissue; MBD-seq did not directly measure gene expression. The study cannot determine whether methylation changes preceded, accompanied or resulted from the transition.

Image: Adult male cheetah on the S126 road south of Satara, Kruger National Park, South Africa, 3 June 2022. Photo: Bernard DUPONT / Wikimedia Commons. CC BY-SA 2.0. Representative image; not one of the Namibian study animals. · CC BY-SA 2.0 · Source

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Molecular biology & biochemistry

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