Unravelling the

Circuitry of Addiction
MOTIVATION
MOTIVATION

Addiction is a Problem
●

●
BACKGROUND
BACKGROUND

Stress Potentiates Cocaine Reward

What we know:
● Cocaine is rewarding
● Stress is aversive

Major finding:
● Stress makes cocaine
more rewarding
BACKGROUND

Stress Potentiates Cocaine Reward

STRESS
?? Cocaine
Reward
BACKGROUND

What Causes Stress-Induced Potentiation?

What we know:
● Stress releases dynorphin
● Dynorphin activates Kappa
Opioid Receptor (KOR)
A bit about KOR
Hypothesis: G-Protein Coupled Receptor
● KOR is responsible for 1. KOR activation is aversive
stress-induced potentiation 2. KOR activation is analgesic

Think “Fight or Flight”

BACKGROUND

Could KOR Cause Reward Potentiation?

Other Experiments:
● Knock out KOR -> no potentiation
● U50,488 (KOR agonist) + norBNI ->
no potentiation

What this means:

● KOR is required for
stress-induced reward
potentiation
*norBNI is a KOR selective opioid antagonist
BACKGROUND

Stress Potentiates Cocaine Reward

STRESS
Dynorphin
KOR
Activation ?? Cocaine
Release Reward
GOAL & HYPOTHESIS
GOAL & HYPOTHESIS

Goal
Determine the mechanism of KOR
mediated reward
KOR
Activation ?? Cocaine
Reward

Change in Baseline Hypothesis

KOR keeps dopamine neurons in
prolonged inhibitory state, lowering
baseline hedonic state.
EXPERIMENTAL DESIGN
EXPERIMENTAL DESIGN

Overview
➔ We know KOR activation causes stress-induced reward potentiation

➔ We want to test if this potentiation is a Baseline Effect

◆ Does KOR activation cause sustained DA inhibition
◆ Does sustained DA inhibition cause reward potentiation

Test:
1. Does non-KOR inhibition (via SwiChR) cause stress-induced reward potentiation?
2. How does SwiChR inhibition affect neuronal firing?
3. How does KOR activation affect neuronal firing?
EXPERIMENTAL DESIGN
Paradigm: Conditioned Place Preference (CPP)
Day 1: pretest + FSS Day 2: repeated FSS + conditioning

Cocaine Saline

30 min explore 15 min FSS 6 min X 4 rFSS

AM PM

Day 3: conditioning Day 4: Test

Cocaine Saline

AM PM Stressed Control

?
EXPERIMENTAL DESIGN
Brain Region Targeted
● Reward is associated with input from
dopamine neurons originating in the VTA

● Output is at NAc (reward processing)

● Stress causes KOR activation

KOR
● Blocking KOR reduces reward potentiation

?
● Activation of KOR → sustained inhibition
?
of dopamine neurons → reward
potentiation
EXPERIMENTAL DESIGN
Optogenetic Inhibition of DA Neurons
Does non-KOR inhibition (via SwiChR) cause stress-induced reward potentiation?

Implant cannula with light fiber

470nm Injection of AAV-DIO expressing SwiChR++ and eYFP
630nm Open
Close
connection to commutator

CPP with SwiChR++ on for 15 min in day 1 and 30 min in day 2

AAV - DIO
-SwiChR++ - eYFP Controls:
● No photostimulation
● AAV-eYFP injections + light
DAT-Cre mouse ● Wild Type
● Immunohistochemistry verification of injection’s targets
● Fiber placement verification
EXPERIMENTAL DESIGN
Optogenetic Inhibition of DA Neurons
How does SwiChR inhibition affect neuronal firing?

Implant cannula with optrode

470nm Injection of AAV-DIO expressing SwiChR++ and eYFP
630nm Open
Close
connection to commutator

CPP with SwiChR++ on for 15 min in day 1 and 30 min in day 2

AAV - DIO
-SwiChR++ - eYFP Controls:
● No photostimulation
● AAV-eYFP injections + light
DAT-Cre mouse ● Wild Type
● Immunohistochemistry verification of injection’s targets
● Fiber placement verification
EXPERIMENTAL DESIGN
SwiChR + in vivo Electrophysiology
How does SwiChR inhibition affect neuronal firing?

470nm We can investigate the mechanism through which DA neurons

630nm Open inhibition potentiate reward.
Close
● Ephys during inhibition, during conditioning, and during
posttest (memory recall)
AAV - DIO
-SwiChR++ - eYFP

DAT-Cre mouse
EXPERIMENTAL DESIGN
U50,488 + in vivo Fiber Photometry
How does KOR activation affect neuronal firing?
Light
Photometry
Implant cannula with photometry fiber

Injection of AAV-DIO expressing GCaMP

connection to commutator

AAV-DIO-
CPP with Intraperitoneal injection of U50,488 Kappa agonist for
GCaMP inhibition instead of optogenetics

Controls:
● No agonist injection
DAT-Cre mouse ● Wild Type
● Immunohistochemistry verification of injection’s targets
● Fiber placement verification
U50,488
FUTURE EXPERIMENTS
FUTURE EXPERIMENTS
Network Mapping
➔
◆
◆
◆

➔
◆
◆
◆