Expression of hsp70 in the third-instar larval tissues of transgenic Drosophila melanogaster (hsp70-lacZ) following dietary exposure to organophosphate insecticide chlorpyrifos for various time intervals was investigated. Effect of the chemical on different developmental stages of the fly was also evaluated by looking at survivorship, hatchability, emergence, fecundity, fertility, and reproductive performance. Our results showed that the toxicant evokes profound cytotoxic effect as evidenced by dark blue staining in salivary gland, proventriculus, brain ganglia, and midgut in the lowest concentration of the chemical following 24 and 48 h of exposure. On the other hand, a significant increase (61%) in hsp70 expression in the above larval tissues was observed in the next higher concentration of the toxicant after 6 h exposure when compared with that of the lowest dose. A further increase in exposure time caused 100% larval mortality. Similarly, larvae exposed to higher concentrations of the toxicant, exhibited 100% mortality within 1 h of treatment. The insecticide caused a delay in emergence and a severe reduction in survivorship of the flies in a dose-dependent manner with 100% mortality within a day of exposure in the highest dosed group. A delay in emergence by 3 days was evident even in the lowest concentration of the chemical. A drastic effect of the chemical on hatchability was found in the highest dosed group with 100% embryonic mortality at post-16 stage of the embryo. Reproductive performance was significantly affected even in the lowest dosed group. The present study suggests that certain larval tissues of Drosophila, a nontarget organism, are vulnerable to chlorpyrifos as evidenced by hsp70 expression. Further, the adverse effect of the toxicant is reflected on various stages of development of the fly including reproductive performance.