Forebrain glycine transporter 1 deletion enhances sensitivity to CS–US discontiguity in classical conditioning

•A novel associative learning phenotype of forebrain neuronal GlyT1 is demonstrated.•The sensitivity to the CS–US contiguity in Pavlovian learning is enhanced.•This complements the previous report of increased sensitivity to CS pre-exposure.•Neuronal GlyT1 may weaken as well as strengthen the conditioned response.•The temporal expression of the conditioned response may also be modified.

The deletion of glycine transporter 1 (GlyT1) in forebrain neurons can apparently strengthen Pavlovian aversive conditioning, but this phenotype is not expressed if conditioning followed non-reinforced pre-exposures of the to-be-conditioned stimulus (CS). To examine whether GlyT1 disruption may only enhance aversive associative learning under conditions that most favour the formation of CS–US excitatory link, we evaluated the impact of GlyT1 disruption on the trace conditioning procedure whereby a trace interval between a tone CS and a shock US was introduced during conditioning. CS and US occurrences were thus rendered discontiguous, which was expected to impede conditioning compared with contiguous CS–US pairing. Conditioned freezing to the CS was measured in a retention test conducted 48 h after conditioning. The genetic disruption significantly modified the temporal dynamics of the freezing response over the course of the 8-min presentation of the CS, although the immediate conditioned response to the CS was unaffected. The separation between “trace” and “no-trace” conditions was augmented in the mutant mice, but this only became apparent in mid-session; and the augmentation can be attributed to the combined effects of (i) weaker conditioned freezing in the mutant relative to control subjects in the “trace” condition, and (ii) stronger conditioned freezing in mutants relative controls in the “no-trace” condition. The demonstrated increased sensitivity to the effect of CS–US temporal discontiguity further highlights the importance of GlyT1-dependent mechanisms in the regulation of associative learning.