Understanding how populations adapt to different thermal environments is an important issue for biodiversity conservation in the context of recent global warming. To test the hypothesis that populations from southern region are more sensitive to climate change than northern region in cold-water species, we determined the thermal tolerance of two geographical populations of a cold-water fish, Rhynchocypris oxycephalus: the Hangzhou population from southern region and the Gaizhou population from northern region, then compared their transcriptomic responses between a control and a high temperature treatment. The results showed that the thermal tolerance range and thermal tolerance polygon area of Hangzhou population were narrower than the Gaizhou population, indicating populations from southern region were possibly more vulnerable. Further transcriptomic analysis revealed that the Gaizhou population expressed more temperature responding genes than the Hangzhou population (583 VS. 484), corresponding with their higher thermal tolerance, while some of these genes (e.g. heat shock protein) showed higher expression in the Hangzhou population under control condition, suggesting individuals from southern region possibly have already responded to the present higher environmental temperature pressure. Therefore, these results confirm the prediction that populations from southern region are more sensitive to global warming, and will be important for their future conservation.