This study analyzed the association between heatstroke incidence and daily maximum wet bulb globe temperature (WBGT) for all 47 prefectures in Japan by age group and severity using time-series analysis, controlling for confounders, such as seasonality and long-term trends. With the obtained association, the relative risk between the reference WBGT (defined as the value at which heatstroke starts to increase) and the daily maximum WBGT at 30 °C (RR(wbgt30)) of each prefecture were calculated. For the heatstroke data, the daily number of heatstroke patients transported by ambulance at the prefecture level, provided by the Fire and Disaster Management Agency, was utilized. The analysis was conducted for age groups of 7-17 y, 18-64 y, and ≥65 y, and for severity of Deceased, Severe, Moderate (combined as DSM), and Mild. The analysis period was set from May 1 to September 30, 2015-2019. Finally, the correlation between RR(wbgt30) and the average daily maximum WBGT during the analysis period (aveWBGTms) of each prefecture was analyzed to examine the regionality of heatstroke incidence. The result showed that RR(wbgt30) is negatively correlated with aveWBGTms for the age group 18-64 y and ≥65 y (except for the age group 7-17 y) and for severity. The natural logarithm of the RR(wbgt30) of all 47 prefectures ranged from 2.0 to 8.2 for the age group 7-17 y, 1.1 to 4.0 for the age group 18-64 y, 1.8 to 6.0 for the age group ≥65 y, and 1.0 to 3.6 for DSM, and 0.9 to 4.0 for Mild. This regionality can be attributed to the effects of heat adaptation, where people in hotter regions are accustomed to implementing measures against hot environments and are more heat acclimatized than people in cooler regions.