PROJECT SUMMARY
Research demonstrates that substance-use disorder, including opiate use disorder, affects one's decision
making and reward processing resulting in impaired goal-directed behaviors. However, there remains a lack of
understanding of the neurobiology that underlies persistent reward-seeking despite value changes that would
normally alter goal-directed behavior. The neuropeptide S (NPS) system, made up of the peptide and its receptor
(NPSR1), drives reward-seeking behaviors, however the underlying mechanism is not understood. We
generated both NPS-Cre and NPSR1-Cre driver mouse lines for accessing and examining the key circuit
components of NPS/NPSR1-mediated behaviors. I isolated a population of NPS-containing cells adjacent to the
locus coeruleus (LC) and found that these neurons project to the orbitofrontal cortex (OFC), a region with dense
expression of NPSR1. I found that these neurons respond to delivery of reward-predictive cues as well as
consumption, but to a lesser extent. I then preliminarily tested the effects of oral fentanyl self-administration on
this population and uncovered a bidirectional response to delivery of the conditioned stimulus (enhanced
activity), and fentanyl reward (quiescence). In this proposal, I first aim to determine the dynamics of OFCNPSR1
neuronal ensembles during natural- and drug-seeking behaviors (Aim 1) using two-photon microscopy.
This aim seeks to further investigate OFCNPSR1 activity during self-administration of fentanyl reward as well as
how devaluation of either a natural or drug reward is modulated by OFCNPSR1 activity. My second aim is to
examine how hindbrain NPS neuron activity and neuropeptide release impact OFC encoding during
reward-seeking behaviors (Aim 2). This aim serves to close this gap in knowledge by using multi-site fiber
photometry to simultaneously image hindbrain NPS, and OFC neuron activity during reward-seeking behaviors.
Finally, in the R00 portion of this grant, I will expand the scope of this study by determining the sufficiency of
the OFCNPSR1-Basolateral Amygdala (BLA) projection in driving drug-seeking behaviors (Aim 3). I found
that OFCNPSR1 neurons are connected with the BLA. In addition, the BLA expresses NPSR1, however, the source
of endogenous NPS to the BLA is unknown as well as any modulation the OFCNPSR1 neurons may provide to this
region. This aim serves to uncover the source of BLA NPS as well as investigate the role of BLANPSR1 neurons in
reward-seeking behavior and how this activity may be affected by OFCNPSR1 projections. I will utilize a series of
complimentary cutting-edge neuroscience techniques to dissect the role of the OFCNPSR1 neurons in drug-
seeking behavior. The central goal is to aid in future investigation of whether NPS signaling represents an
important avenue for altering drug-seeking behavior. This proposal provides substantial new training in SLM, in
vivo calcium imaging of network and single-cell activity, neuropharmacological and molecular viral manipulations,
and career development skills for transition as an independent academic investigator.