Effects of hM4Di activation in CamKII basolateral amygdala neurons and CNO treatment on sensory-specific vs. general PIT: refining PIT circuits and considerations for using CNO

Psychopharmacology (Berl). 2020 May;237(5):1249-1266. doi: 10.1007/s00213-020-05453-8. Epub 2020 Jan 24.


Background: Pavlovian stimuli can influence instrumental behaviors via phenomena such as Pavlovian-to-instrumental transfer (PIT). PIT arises via dissociable processes as sensory-specific PIT (SS-PIT) and general PIT. The basolateral amygdala (BLA) mediates SS-PIT, but not general PIT. However, the specific BLA neuronal populations involved are unknown.

Aims: To determine the contribution of glutamatergic BLA neurons to the expression of SS-PIT and to the recall of sensory-specific properties of stimulus-outcome associations.

Methods: BLA neurons were transduced with virus containing either GFP or hM4Di, driven by the CamKII promoter. Rats were then tested for SS and general PIT and subsequently for expression of Pavlovian outcome devaluation effects and conditioned taste aversion following injections of vehicle or clozapine-N-oxide (CNO, the hM4Di agonist).

Results: CNO selectively blocked SS-PIT in the hM4Di-expressing group, but not controls, without altering expression of Pavlovian outcome devaluation or sensory-specific taste aversion in either group. Unexpectedly, CNO disrupted general PIT in both groups.

Conclusions: CamKII BLA neurons mediate the expression of SS-PIT by enabling Pavlovian stimuli to trigger recall of the correct action-outcome associations rather than by mediating recall of the sensory-specific properties of the stimulus-outcome association. Separately, our data demonstrate that CNO alone is sufficient to disrupt affective, but not sensory-specific processes, an effect that was not due to generalized motor disruption. This non-specific effect on general PIT may be related to CNO-induced shifts in internal state. Together, these data identify BLA CamKII neurons as critical for the expression of SS-PIT and reveal important considerations for using CNO to study general affective motivation.

Keywords: Appetitive learning; Cue-triggered food seeking; DREADDS; Incentive motivation; Pavlovian motivation; Pavlovian-to-instrumental transfer; Reward; hM4Di.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Basolateral Nuclear Complex / drug effects
  • Basolateral Nuclear Complex / metabolism*
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2 / metabolism*
  • Clozapine / administration & dosage
  • Clozapine / analogs & derivatives*
  • Conditioning, Classical / drug effects
  • Conditioning, Classical / physiology*
  • Feeding Behavior / drug effects
  • Feeding Behavior / physiology
  • Feeding Behavior / psychology
  • Female
  • Male
  • Motivation / drug effects
  • Motivation / physiology
  • Neurons / drug effects
  • Neurons / metabolism*
  • Piperazines / metabolism*
  • Rats
  • Rats, Sprague-Dawley


  • DREADD agonist compound 21
  • Piperazines
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2
  • Clozapine
  • clozapine N-oxide