In 2004, the Pennsylvania Fish and Boat Commission implemented catch-and-release (CR) regulations on headwater stream systems to determine if eliminating angler harvest would result in an increase in the number of adult (≥100 mm) or large (≥175 mm) Brook Trout Salvelinus fontinalis. Under the CR regulations, angling was permitted on a year-round basis, no Brook Trout could be harvested at any time, and there were no tackle restrictions. A before-after–control-impact design was used to evaluate the experimental regulations. Brook Trout populations were monitored in 16 treatment (CR regulations) and 7 control streams (statewide regulations) using backpack electrofishing gear periodically for up to 15 years (from 1990 to 2003 or 2004) before the implementation of the CR regulations and over a 7–8-year period (from 2004 or 2005 to 2011) after implementation. We used Poisson mixed models to evaluate whether electrofishing catch per effort (CPE; catch/100 m²) of adult (≥100 mm) or large (≥175 mm) Brook Trout increased in treatment streams as a result of implementing CR regulations. Brook Trout CPE varied among sites and among years, and there was no significant effect (increase or decrease) of CR regulations on the CPE of adult or large Brook Trout. Results of our evaluation suggest that CR regulations were not effective at improving the CPE of adult or large Brook Trout in Pennsylvania streams. Low angler use, high voluntary catch and release, and slow growth rates in infertile headwater streams are likely the primary reasons for the lack of response.