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Ticks are hematophagous ectoparasites that transmit a wide range of pathogens. The lone star tick, Amblyomma americanum, is one of the most widely distributed ticks in the Midwest and Eastern United States. Lone star ticks, as other three-host ixodid ticks, can survive in harsh environments for extended periods without a blood meal. Physiological mechanisms that allow them to survive during hot and dry seasons include thermal tolerance and water homeostasis. Dermal fluid secretions have been described in metastriate ticks including A. americanum. We hypothesized that tick dermal secretion in the unfed tick plays a role in thermoregulation, as described in other hematophagous arthropods during blood feeding. In this study, we found that physical contact with a heat probe at 45 °C or high environmental temperature at ∼50 °C can trigger dermal secretion in A. americanum and other metastriate ticks in the off-host period. We demonstrated that dermal secretion plays a role in evaporative cooling when ticks are exposed to high temperatures. We find that type II dermal glands, having paired two cells and forming large glandular structures, are the source of dermal secretion. The secretion was triggered by an injection of serotonin, and the serotonin-mediated secretion was suppressed by a pretreatment with ouabain, a Na/K-ATPase blocker, implying that the secretion is controlled by serotonin and the downstream Na/K-ATPase.