Wang, W. W.-Y.; Pethe, B.; Ward, A. P.; Wood, W.; Smith, C. T.; Frick, A. J.; Yates, S. S.; Gunderson, A. R.

1. Adaptive phenotypic plasticity can help organisms cope with global warming by increasing their heat tolerance, yet most research has focused on whole-organism traits. Much less is known about thermal plasticity at the gamete level despite increasing evidence that reproduction, and particularly sperm function, is especially sensitive to heat. 2. We tested for plasticity in sperm heat tolerance in two ways: via male thermal acclimation (pre-ejaculate stage) and via sperm cell heat hardening (post-ejaculate stage) in brown anole lizards (Anolis sagrei). We also measured plasticity in several other sperm and ejaculate traits in response to male thermal acclimation. 3. To test for effects of male thermal acclimation, adult males were exposed to one of two ecologically realistic, fluctuating temperature regimes that either mimic cool spring conditions or projected future summer conditions for eight weeks. Throughout this period, we repeatedly measured sperm heat tolerance (LT50), baseline motility, and sperm count. We also measured sperm morphology at the end of thermal acclimation. We predicted that males in the warmer treatment would produce more heat tolerant sperm. 4. To test for post-ejaculate heat hardening, we compared the heat tolerance of ejaculated sperm that either did or did not experience a high but non-lethal temperature prior to heat tolerance measurement. 5. We found no plasticity in sperm heat tolerance due to either male thermal acclimation or sperm cell heat shock. Long-term thermal acclimation of males did not increase sperm heat tolerance, nor change motility, sperm count, or sperm morphology. We also found no evidence for post-ejaculate heat hardening, as exposing sperm cells to mild sublethal heat shock did not enhance sperm heat tolerance. 6. Our results indicate that gametic traits have limited capacity for plastic adjustment to thermal stress, which is broadly consistent with the low levels of thermal plasticity found for whole-organism thermal tolerance across ectotherms. This highlights the vulnerability of reproductive traits to rising temperatures and the importance of evolutionary and behavioral responses to buffer organisms from climate change.