Upregulation of tropoelastin (TE) gene expression in rat lung interstitial fibroblasts normally occurs during alveolar septation. TE message increases at the end of the first week of life, peaks on days 9-11, and returns to barely detectable levels over the next 7-10 days. Our previous in situ hybridization studies indicated that exposure of pups to > 95% oxygen from 3 to 13 days of age interfered with the increased in TE gene expression in interstitial fibroblasts normally seen during septation. However, when the pups were returned to room air, lung fibroblast TE message levels increased, exceeding levels seen in control lungs during the exposure. In addition, TE message levels remained elevated for a week after levels in control lungs had returned to background. A possible interpretation of these results was that the developmentally regulated increase in TE messenger RNA (mRNA) was downregulated by the hyperoxic exposure but resumed when the pups were returned to a normoxic environment. We report herein the results of a subsequent study conducted to determine whether continued hyperoxic exposure beyond day 13 would further delay the peak in TE mRNA. Rat pups were exposed to 95% O2 from 5 to 17 days of age. TE and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) message levels in lung interstitial fibroblasts were assessed by in situ hybridization. As observed in pups exposed from 3 to 13 days, hyperoxic exposure from days 5 to 17 also extended the period during which TE mRNA levels were elevated. After exposure, TE message levels were 99%, 262%, and 223% of controls on days 19, 21, and 23 respectively. In addition, delaying the exposure 2 days until the pups were 5 days old resulted in an upregulation of TE message, relative to control values, during the hyperoxic exposure. In hyperoxic pups, values for TE message expression were 105%, 152%, 168%, and 144% of control pups on days 9, 11, 13, and 16 respectively. The influence on peak TE message expression of postnatal age at the time of exposure was further explored to verify the results of the 3-13 and 5-17 day exposures. When pups were exposed continuously from 2, 3, 4, 5, or 6 days until 11 days of age, the results of both in situ hybridization and Northern blot analysis confirmed our previous observations, demonstrating that the postnatal age at which hyperoxic exposure is initiated influences TE message expression in the developing lung.