Leptin Receptor-Expressing Lateral Hypothalamic Neurons

Gate Motivation
1
Anjali Gajendiran , Justin Siemian, Felipe Schiffino, Cara Borja, Sarah Sarsfield, Yeka Aponte
1
Centennial High School, Ellicott City, MD
Neuronal Circuits and Behavior Unit, Cellular Neurobiology Research Branch, NIDA IRP
Introduction Figure 1: Chemogenetic activation or inhibition of LHLEPR neurons alters Figure 4: Effects of LHLEPR neuron activity
• The lateral hypothalamus (LH) contains a diverse range of motivation for food. in the marble-burying test.
neuronal populations and is known to play a key role in the
control and regulation of motivated behaviors important to
survival.
• Leptin receptor-expressing neurons within the LH (LHLEPR)
are one subpopulation of γ-aminobutyric acid (GABA)ergic
neurons and may mediate feeding and reward.
• LHLEPR neurons project to several brain regions including the
ventral tegmental area (VTA), a region known to be important
for reward processing and motivated behaviors1.

Objective
• Determine whether LHLEPR neurons regulate motivation to
500 μm 500 μm
obtain food and water.
• The specific focuses of this study were:
o To investigate the effects of chemogenetic activation or
inhibition of LHLEPR neurons on progressive ratio (PR)
performance for food reward.
o To investigate the effects of optogenetic activation or
inhibition of LHLEPR neurons on PR performance for water
reward.
o To determine whether these behavioral effects were due to
generalized changes in locomotor activity or compulsive
behavior.

Methods n = 6-8/group; Paired t test, **p < 0.01

• Animals
o Two- to five-month-old male and female LEPRcre mice were
Conclusions
n = 8/group; Paired t-test, *p < 0.05, **p < 0.01
used. • Chemogenetic activation of LHLEPR neurons increased
Figure 2: Optogenetic activation or inhibition of LHLEPR neurons alters motivation for water. motivation for food in the PR task, whereas
• Surgeries chemogenetic inhibition of LHLEPR neurons decreased
o Adeno-associated viruses containing motivation to obtain food.
DIO-hM3D(Gq):mCherry, DIO-hM4D(Gi):mCherry,
• Optogenetic activation of LHLEPR neurons increased
FLEX-eGFP, FLEX-ChR2-tdTomato, or FLEX-ArchT-GFP
motivation to obtain water reinforcers, whereas
were injected bilaterally into the LH. For optogenetic
optogenetic inhibition of LHLEPR neurons decreased
studies, optic fibers were implanted bilaterally above the
motivation to obtain water.
LH.
• These effects did not appear to result from generalized
• Behavioral Experiments alterations in locomotor activity. The time spent in the
Progressive Ratio center of the open field was also unaffected.
o Food- or water-restricted mice were trained to press an • Marble-burying behavior was unaffected by
active lever for food pellet or 0.1 mL water reinforcers, optogenetic manipulation of LHLEPR neurons, but
respectively. Mice were initially trained on fixed ratio 1 increases in marble interaction time during LHLEPR
(FR1) and FR3 schedules prior to training on a PR schedule, neuron stimulation may suggest compulsive-related
in which the response requirement increased following behavioral effects.
each reinforcer based on the equation: Response Future Studies
Requirement = (5e 0.2 × Pellet Number) − 5. • Perform behavioral analyses during LHLEPR axon
o For inhibition studies, mice were tested in states of food- or terminal stimulation to assess the roles of their
water-restricted conditions. For excitation studies, mice projections to specific brain regions and better
were tested in food- or water-sated conditions. characterize LHLEPR neuron circuit function.
o For chemogenetic studies, clozapine N-oxide (CNO, 1 • Characterize inputs to LHLEPR neurons to understand
mg/kg) or vehicle was injected 20 min prior to the session. n = 6-8/group; Paired t-test, *p < 0.05, **p < 0.01
how these circuits modulate survival-related
For optogenetic studies, blue light (450 nm, 20 Hz, 10 ms behaviors.
pulses for 1 min every 10 min) or green light (520 nm, • Characterize the activity of LHLEPR neurons during
Figure 3: Effects of LHLEPR neuron activation or inhibition on open field locomotion.
constant, delivered every other second) were delivered various metabolic states such as hunger or thirst in
throughout the session until no reinforcer was earned for order to understand the role they play in eliciting
15 min or until 2 h had elapsed, whichever occurred first. survival-related behaviors.

Open Field References

o The effects of optogenetic activation or inhibition on LH LEPR 1
Nieh, E., Vander Weele, C., Matthews, G., Presbrey, K.,
mice was investigated. Photostimulation occurred during
Wichmann, R., Leppla, C., . . . Tye, K. (2016). Inhibitory
alternating 3 min epochs. The amount of time (in sec) spent
Input from the Lateral Hypothalamus to the Ventral
by the mice in the periphery or center of the box was
Tegmental Area Disinhibits Dopamine Neurons and
measured.
Promotes Behavioral Activation. Neuron, 90(6),
1286-1298. doi:10.1016/j.neuron.2016.04.035
Marble Burying 2
Yim, Y. S., Park, A., Berrios, J., Lafourcade, M.,
o 16 marbles were spaced in a 4 × 4 grid in a standard
Pascual, L. M., Soares, N., . . . Choi, G. B. (2017).
housing cage with ~5 cm deep bedding. Mice were allowed
Reversing behavioural abnormalities in mice exposed to
to freely explore the cage for 18 min, with laser stimulation
maternal inflammation. Nature, 549(7673), 482-487.
occurring during alternating 3 min epochs. An arbitrarily
doi:10.1038/nature23909
defined scoring standard was used2, where each fully
buried marble (>50%) resulted in a score of 1; each
half-buried marble (50%) resulted in a score of 0.5.
Acknowledgments
o Sessions were video recorded and the amount of time mice n = 6-8/group; Paired t test, *p < 0.05 • This work was supported by NIH/NIDA IRP.
spent interacting with the marbles was scored manually.