The distribution and concentration of selected elements by PIXE method and DNA damage using comet assay in brains of 1st instars of grasshoppers Chorthippus brunneus from unpolluted (Pilica) and polluted (Olkusz) site, additionally exposed to various doses of zinc during diapause or after hatching, were measured. We tried to assess the degree of possible pre-adaptation of the insects to heavy metals and evaluate the utility of these parameters in estimation of insect exposure to industrial pollutants. Additionally, the mechanism of zinc toxicity for grasshopper brains was discussed. We observed the correlation between experimental zinc dose, zinc contents in the brain and DNA damage in neuroblasts, but only in groups exposed to lower zinc concentration. For higher zinc concentration the amount of the metal in brain and DNA damage remained at the control level. Some site-related differences in DNA damage between grasshoppers from Pilica and Olkusz were observed during short-term exposure (after hatching). Significant increase in the calcium contents in the brain, proportional to zinc concentration in sand, was also observed, especially in the offsprings from Olkusz. The results may be the basis for further searching for molecular mechanisms of defense against heavy metals in insects living in polluted habitats.