It is generally accepted that the associative learning occurs when a behaviorally neutral conditioned stimulus (CS) is paired with an aversive unconditioned stimulus (US) in close temporal proximity. Eyeblink conditioning (EBC) is a simple form of associative learning for motor responses. Specific activation of a population of cells may be an effective and sufficient CS for establishing EBC. However, there has been no direct evidence to support this hypothesis. Here, we show in rats that optogenetic activation of the right caudal mPFC pyramidal neurons as a CS is sufficient to support the acquisition of delay eyeblink conditioning (DEC). Interestingly, the associative memory was not stably expressed during the initial period of daily conditioning session even after the CR acquisition reached the asymptotic level. Finally, the intensity and consistency of the CS were found to be crucial factors in regulating the retrieval of the associative memory. These results may be of importance in understanding the neural cellular mechanisms underlying associative learning and the mechanisms underlying retrieval process of memory.
