Immature mosquito development and survival of adults are highly sensitive to environmental temperature, which can alter gene expression during the mosquito life-cycle. To further understand how heat shock proteins are developmentally expressed in mosquitoes, we subjected first instar larvae, 16-h old pupae and female of Aedes aegypti (L.) (Diptera: Culicidae) to heat shock treatment for 0, 15, 30, 60, and 180 min at 23 and 42°C. The heat shock protein genes AeaHsp26, AeaHsp83, and AeaHsc70 were examined by comparing relative transcript expression levels at 42°C compared with 23°C. Upregulated transcripts from heat shock treatment at 42°C and control were further confirmed and quantified by quantitative real-time polymerase chain reaction. Data revealed that first instar larvae were more sensitive to heat shock treatment than pupae and adults (i.e., relative AeaHsp26 expression levels in larvae were 10-fold greater than in the females. AeaHsp83 expression levels in larvae, pupae and adults were upregulated 2- to 50-fold greater by heat shock treatment at 42°C compared with 23°C. AeaHsc70 expression levels in larvae, pupae and adults, however, were upregulated less than AeaHsp26 and AeaHsp83 at the higher temperature. Statistical analysis indicated that AeaHsp26 and AeaHsp83 genes were significantly upregulated in Ae. aegypti larvae and pupae after 15, 30, 60, and 180 min exposure to high temperature (42°C). The current study has shown that AeaHsp26 and AeaHsp83 are important markers of stress and may function as critical proteins to protect and enhance survival of Ae. aegypti larvae and pupae.