Repetitive negative thinking (RNT) is a transdiagnostic risk factor for internalizing psychopathology, and theoretical models suggest that RNT may maintain symptoms by interfering with psychophysiological habituation. The present study therefore examined associations between RNT and habituation within and between study sessions. Community members (N=86) completed a habituation task involving exposure to acoustic probes at up to five sessions spaced 7 days apart on average. Eyeblink startle response was measured using the electromyography startle magnitude. Self-reported anxiety was assessed before and after the habituation task at each session. Multilevel growth curve modeling indicated that RNT was associated with a higher “floor” (i.e., asymptote) of startle responding as evidenced by reduced within-session startle habituation at later sessions. Results suggest that RNT may disrupt startle habituation and are consistent with theoretical models proposing that RNT sustains physiological activation to support avoidance of negative emotional contrasts or perceived future threats.