Complementary stable and dynamic prelimbic ensembles encode learned threat value underlying generalization and discrimination.

Adaptive behavior requires assigning emotional value to sensory cues and generalizing to similar stimuli. The prelimbic cortex (PL) is critical for threat expression and discrimination, yet its neuronal ensembles undergo substantial turnover over time. How stable population codes emerge despite this reorganization remains unknown. Using longitudinal calcium imaging in freely moving mice, we tracked PL activity for 30 days during auditory discriminative fear learning and retrieval. Despite substantial ensemble turnover, stable graded representations persisted in PL populations. A generalized linear model showed that the tone threat value explained graded activity better than freezing, indicating that PL primarily encodes stimulus significance. Population similarity across stimulus presentations was highest for shock-associated and highly generalized tones, suggesting a stable population code underlying threat generalization. Stable neurons preserved graded gradients across sessions, whereas learning modulated the rate of dynamic frequency-selective neurons. Together, these populations enabled stable threat-value coding despite ongoing ensemble reorganization.