Thanks Bill all it's it's really a
pleasure to come here by balls invitation

because so I've known Bill all since
we were training together at Yale and

I have to say that I've admired him since
basically the moment we met and because

your intelligence and your passion about
neuroscience is so clear so thank you for

inviting me thank you all for having me
here I'm going to talk today about some of

the work that my lab has done over the
last few years trying to understand how

the brain balances actions and habits and
then how these processes can be affected

by cocaine and if you can hear me please
just raise your hand there in the back.

OK So generally speaking my lab at Emory
is really interested in understanding

the long term effects of cocaine and
or stress hormone exposure and

we focus quite a bit on
adolescents because a lot of

epidemiological evidence will suggest
that if you are an adolescent and

you're exposed to cocaine you have worse
long term outcomes than individuals

who are exposed as adults so for example
you're less likely to seek treatment for

drug use disorders at any point in your
life span relative to individuals first

exposed to cocaine as adults and so there
seems to be some effects of cocaine and

other pathological insults like stress
on decision making process ease so

we're going to talk about that today and
within the brain we're going to focus

on the prefrontal cortex and in particular
on the orbital prefrontal cortex or

O.P.'s see so I'm highlighting it here
in purple on the kernel section from

the mouse brain atlas and we're interested
in the orbital cortex in my lab for

a lot of reasons for example it's hypo
active following cocaine exposure this is

an individuals who have had cocaine and
they're now off drugs even though they're

now drug abstinent they're still
showing hypo activity of the orbital

prefrontal cortex this region would
otherwise be really important for allowing

us to engage in inhibitory control and
to choose the best behaviors amongst many.

So if you're a rodent and I give you
cocaine we see the loss of dendritic

spines or the primary sites of synaptic
contacts within the orbital cortex and

this is very much unlike many other brain
regions where cocaine actually causes

dendritic spine proliferation if you're
an adolescent mouse and I give you cocaine

we actually see the last of your dendritic
trees within the orbital cortex and so

we see pretty gross changes in the near a
near anatomical composition of the orbital

cortex in individuals and
animals exposed to cocaine and

these changes are long lasting and so
we think about them quite a bit in

the context of how cocaine may have
long lasting behavioral effects so

we'll focus today on actions and habits
and I'll define these terms in a moment

and then we're going to hone in on
a potential molecular signaling mechanism

brain derived factor acting through
its high affinity receptor track B.

and then finally we're going to
try to take what we've learned

about how the orbital cortex balances
actions and habits how B.D.N.F.

may be involved in this process and we're
going to try to block habits that result

from cocaine exposure and
then in my last three slides I'd

like to show you some new data that we've
been collecting that has Take that the had

the data have taken us out of our favorite
brain region the orbital cortex and

unexpectedly into the prelim
that prefrontal cortex

which is also it turns out involved
in action outcome decision making.

All right so we're really excited in my
lab about that orbital cortex because it's

anatomically poised to really contribute
to complex decision making so

if we look at anterograde tracing studies
from my lab and from many other labs

we simply infuse into our great tracer
within the orbital cortex in the mouse so

we can see really nice terminal projection
patterns in the dorsal straight and

in the in that glow and this is really
exciting because these two regions

are important for goal directed action
as opposed to habits carry case so

I've mentioned this action versus
habit thing once or twice before so

let's pause and define these terms so
a goal directed action is exactly what it

sounds like it's engaging in some sort of
behavior with with the expectation that

that behavior will be reinforced so
you came here today with the expectation

that your action would be reinforced
with maybe a decent talk.

Often I get left there there you go
by contrast habits are different

they're stimulus elicited So these
are typically conceptualized as familiar

behaviors that we've engaged in so many
times that they're no longer linked to

the action outcome contingency but
instead instead are stimulus elicited So

one example from my life is has
to do with the fact that for

many years I wore glasses so I don't wear
glasses so much anymore but as those of

you who do are glasses you know when the
glasses fall down on the bridge of your

nose you have a hard time seeing because
you're not looking through the lens and so

you bring your hand up to adjust your
glasses so again I don't work last so

much anymore however when I have
a hard time seeing such as the lights

in the room dim for a lecture I
bring my hand up like this OK so

this is a classical habit there's
some sort of stimulus that is that I

have a hard time seeing and there's
some sort of motor behavior this so

you can imagine that habits in
many cases may be innocuous so

this makes me look kind of weird but it's
not hurting anybody however habits that go

arise so in the case of for example
drug seeking habits that we have a hard.

Haim inhibiting these kinds of habits
may contribute to true addiction and

indeed one of the prominent models of
addiction argues that addiction can begin

with occasional drug use it can then
proceed to have bitch will drug use and

then in some individuals
according to some environmental or

genetic circumstances that have been drug
use can then proceed to compulsive use and

relapse and so my lab is really interested
in understanding the mechanisms by

which habits form and and
the mechanisms by which we modify our

behaviors when our habits are not
advantageous and so to do this we use our

operant conditioning chambers here we have
a bunch of these chambers in the lab and

basically what you're seeing is
that our mouse can rear up on

our can explore the operant conditioning
chamber this animal is rearing up against

a nose put panel this nose put Powell
has three nose book recesses and

when he completes the the response
requirement he gets reinforced with

the food pellet that's delivered through
the dispenser appeared on the tube and

into the magazine so there's a mouse
waiting for his food pellet.

So in the task that we're going
to talk about today if we zoom in

on the nose poke panel we're first going
to train the animals to respond to

nose book recesses of the outermost and
the innermost recess and we're going to

ask the animals to respond equally on both
recess and that very technical graphic I

just showed you there serves to remind
me to tell you thank you for laughing

to tell you that when we train the animals
we're training them to have equivalent

experience with both nose poked apertures
they're being reinforced on a fixed ratio

one schedule of reinforcement which simply
means that every nose poke is reinforced

and experience with each nose
poke is controlled such that

thirty pellets are associated with
each of those poker aperture so

over the course of training which takes
about a week we can we can generate

animals that are a very proficiency in
responding on both the left and their.

But B.
have equivalent experience with this

reinforcement.

And this is really important because it
allows us to then ask them is how they're

making decisions in their environment so
we're going to use what's called

an instrumental contingency degradation
procedure which is a fancy term for

something pretty simple actually so
as a reminder our mice have

have learned now that they can go thirty
reinforcers by nose poking on the left and

the right nose book aperture for a total
of sixty pellets across the session.

And now we're going to play a little
bit of a trick on the mouse so

we bring him into the conditioning chamber
on one day and we've included one of

the notebook recesses he has access to
this one let's say and now his responding

is reinforced but only on one nose poke so
he gets pellets just like he always did

on this nose poke recess he goes back to
the colony we bring him into the next

to the chambers the next day and now we're
going to degrade this contingency this

action outcome contingency and to do this
we're going to deliver food pellets for

free so the animal can no spoke as
little or as much as he wants and

rather than being reinforced food
pellets are going to be delivered

into that magazine at the same rate that
he earned them yesterday so if you let me

anthropomorphize just for a minute if you
are a mouse and you're in this task what

you should be thinking is wait a minute it
doesn't make sense for me to know spoke

over here because my nose poking no
longer results in reinforcement so

we can ask the mouse if he has learned of
this and if use utilizing the this new

information by simply bringing him back to
the conditioning chambers the next day for

a protest conducted in extinction this
is a very brief test we stick men for

five minutes and we're simply asking the
mouse where he wants to spend his time so

a typical mouse with moderate training and
you'll see a few examples of this in

a moment will preferentially and gauge
the response associated with the intact or

non degraded contingency and
heal by relatively speaking Hill

ignore the degraded response out
response outcome contingency so he's my.

To find his behavior previously
he went into the chambers and

responded equally on both those polka
pictures now he's modified his behavior so

he's preferentially engaging in behavior
that's likely to be reinforced so

by contrast if we generate mice that
have had a history of lots and lots and

lots of training we can engender habits
just like me in my classes adjusting

behavior and we have a mouse that will
run into the conditioning chambers and

he'll bang away on both those book
recesses just like he always did because

he's developed this left or
right nose poke habit OK So

we also appreciate in the literature
that if you expose mice or

rats to a lot and lots of cocaine or
him that Amine or alcohol we can engender

habit based behavior that forms before it
should so these are premature habits and

then additionally if we ablate or
otherwise in activate the orbital frontal

cortex we can also cause habit behavior
that fails to be modified and we think

it's because animals are not able to learn
new response outcome contingencies so

they defer back to what is familiar So
this paper by Grandma and

caustic came out in two thousand and
thirteen and it was really impactful and

continues to be impactful for
the field because they show using several

in activation techniques that in mice the
orbital cortex is necessary for learning

about response outcome contingencies
just as in monkeys Jane Taylor and

I had a paper out the same year
showing using one activation technique

the same thing but that was nice because
there was Can cordons but additionally we

were really interested in understand the
potential middle molecular mechanisms and

so one sort of obvious mechanism
to us anyway was beating enough.

So the brain derived nurture of a factor

is a nerve trophic factor that's
expressed throughout prenatal and

post into life you guys probably have
heard about it in the context of say X.

on guidance during early development but
good enough is also really important for

the stability of dendrites and
injured expires in adulthood and so

to ask whether B.D.N.F. was important for
the functions of the orbital cortex we

simply took mice that were homozygous for
a flock gene we infused bilaterally

into the orbital cortex and A.V. created
to reduce speed enough expression and

then we train the mice to perform the NO
spoke responses and then we ask them

if they can modify their behavior based on
changing response outcome contingencies

and what I think is pretty obvious is that
the mice with the knockdown could acquire

the nose poke responses but they generate
lower response rates that's pretty typical

of orbital lesions across lots of
species but what was more exciting for

us who is that when we modified the
response outcome contingencies are control

my smog of five their behavior so
instead of responding equivalently on both

nose pick up hers they prefer the newest
book that was likely to be reinforced

by contrast mice that big enough knock
down are unable to modify their behaviors.

So in two thousand and eleven as blow
mentioned I had moved to Emory and

I had this great student who was really
interested in understanding this effect

further So this is Kelsey zoom in here and
so the first thing that I asked Kelsey to

do was to replicate my original
experiment so at Emory we're obviously

geographically located in a different part
of the country we use different mice So

these mice are brought in a bulb see
as opposed to be six met background and

they're also using L.M.T. CRE from
the Emory core as opposed to an A.V.

Crieff from roughly and so there
are a lot of differences in experimental

details here nevertheless I hope you
can appreciate that Kelsey pretty

readily replicated my finding So
first if she knocked down between.

Effort in the orbital cortex are mice
could learn to perform the nose poke

responses but the knocked on my head lower
overall response rates more importantly

when we modify the response outcome
contingencies are mice with big enough

knock down are unable to keep up and so
they defer back to a familiar approach and

so this tells us that beating off within
the orbital cortex is really important for

the functions of the orbital cortex
in terms of modifying behavior or

engaging in behavioral flexibility one
response outcome contingencies change and

in the absence of that we might instead
defer back to a familiar habit.

So Kelcey really wanted to ask what which
circuits might be important for these

kinds of behaviors and then can we learn
can we use what we've learned to block

habits that me emerged due to for example
cocaine or stress hormone exposure.

And so using if to take on the first
question we used what's called

a disconnection strategy and so Kelsey and
another student of mindless pits

were really interested in
understanding whether the eating F.

expressing projections from the orbital
cortex to the Amidala were important for

response outcome learning and memory and
so again to test the strategy or

to test the potential connection
the use of disconnection design and

what we're doing is we're taking advantage
of the strong evidence that in the mouse

the venture lateral over to
cortical projections to the and

they are it's a lateral And so
what that means is that the left or

Bell cortex is talking to the left and
the lower and the right orbital cortex is

talking to the right to make dua And so
what we can do then is reduce B.D.N.F.

within one hemisphere of the orbital
cortex and in this case came a genetically

silence that the L A in
the contra lateral hemisphere and

when we do this we're causing
a disconnection because R.B.T. enough to

fish an orbital neurons are trying to talk
to a healthy and they and are healthy or.

World neurons are trying to
talk to a silenced I think.

So in these designs you always have
course have viral vectors that are less

expressing Flora for as your control but
additionally you can infuse your

your creeks pressing viral doctors in
your dreads within one hemisphere and

in this case you're knocking
out one connection so

knocking out let's say the left
orbital cortex talking to the left and

then but
you have your right now at work intact so

we also went into these experiments
with the hypothesis that B.D.N.F.

dependent interactions between the orbital
cortex and the would be particularly

impactful during the consolidation or the
retention of new learning and memory so we

took our mice through all of the training
and testing that I've described to you so

far but we didn't in activate until
immediately following the contingency

degradation day just as a reminder this is
the day that the my scott free pellets so

this is the day they should
be learning something new and

that is well wait a minute this nose poke
response over here doesn't matter anymore

I don't I don't need to generate this it
doesn't yield food reinforcement and so

immediately after that session we pulled

myself out of the conditioning chambers
and we inactivity that the L.A.

basin lateral and below we bring them back
to the testing chambers the next day for

a protest and

again we're simply asking the mice
where they want to spend their time.

And so what you can see is that in both
male and female mice our typical animal or

control expressing control of viral vector
expressing animals can preferentially

engage the response it's likely to be
reinforced and they collect the response

that's unlikely to be reinforced the same
is true of the mice that have one

connection spared by contrast if we have
our contra lateral groups we see that

these mice are unable to update their
response strategies so they differ back

to what is familiar and that is to
engage both responses equally So

what this tells us is that beating enough
dependent interactions between the orbital

cortex and the base lateral MacDill are
really important for laying down this new

response outcome memory which then
allows you to engage the behavioral flex

again engage in behavioral
flexibility later on Any Questions.

OK so we were really excited
a that this timing worked right so

this this tells us something really
critical is tells us what phase of

the learning is being affected by at least
the deficient or the deficient orbital

to make the interactions that we're
impacting when we knock down B.D.N.F. and

so we capitalized on this finding by
trying to block have it based behavior

that resulted from bilateral P.D.F.
knockdown and so now we're going

back to scenario where we're reducing
B.D.N.F. bilaterally within the orbital

cortex of our mice we're training them
to perform to know spoke responses and

then we modify the likelihood that one
response will be reinforced immediately

following that session we pull the animals
out of the conditioning chambers and

half of the mice receive an experimental
novel compound called seventy

if it's a seventy putatively has
agonistic activities or track B.

which is the hyphen in the receptors for
Vienna and

after they've had this injection they
go back to the colony and we bring them

back to the chambers the next day when
they're drug free and we ask the mice

where they want to spend their time so
as I showed you a few slides ago B.D.N.F.

knock down causes this deferral to have
it based behavior however if we intervene

with this track be agonist we can
rescue the decision making strategies.

Now Kelsey wanted to ask whether this
track the agonist could block habit

based behavior that occurs under more
naturalistic circumstances and so she took

intact typical mice and she trained them
every day for a month using escalating

random interval schedules of reinforcement
and the reason this is important is

because as you can see it generates really
high rates of responding that cause havoc

based behavior in our mice again just
like me with this glasses behavior

that I engage in because I've done it like
a million times before our mice generate

habit based behavior because they've
no SPOKE TO million times before so

again immediately after this contingency
degradation procedure Kelsey pulled

the mice out of the conditioning chambers
she injected half with vehicle half.

And she brought them back to
the chambers the next day drug free and

asked them where they wanted
to spend their time and so

our control mice develop habits as
expected by contrast are mice that were

given seventy if they are able to
really nicely flexibly modify their

behavior suggesting excuse me that
suggesting that this track be agonist

is still attaining the functions
of the orbital cortex in his task.

So I'm not going to show you the data for
a time but just as a side note since

this initial paper came out showing
that seventy has four causes

phosphorylation attractive this finding
has been questioned quite a bit and so

in this experiment we actually we
replicated this whole experiment but

using a two by two design where mice
some mice received both seventy D.H.

off the track the agonist but they were
pretreated with twelve which is a trick

the antagonist and that was able to
block the effects of the seventy D.H.

off the track the putative
track the agonist so

this is all just to say that although
seven if you are familiar with it it

may indeed have an intended off
target effects we think that at least

this effect I'm showing you here can
be attributed to its activity or trick.

OK so so summarize here is you know
a lot of work couple of years of a Ph D.

thesis but ultimately what we want to do
is get to the point where we can develop

new strategies to block habit based
behavior that results from say cocaine or

stress or exposure and
we were particularly interested in

adolescent cocaine exposure and I had sort
of alluded to this earlier but the reason

is that when individuals are exposed to
cocaine during adolescence there are long

term impacts on decision making process
sees And additionally we know from rodent

studies you have a long term impact on
the anatomy of the orbital cortex So

for example if we expose cocaine mice to
cocaine from posting all day thirty one to

thirty five this is a pretty brief period
that would correspond to early adolescence

or ages twelve thirteen fourteen in humans
and then allow the animals to be drug free

for three weeks and then look at their
orbital neurons we see that the basal

rates are actually feed in layer five and
then additionally these

neurons are less complex which is what
you're seeing initial analysis here

on top of all of all of this we
see a loss of spines in the same.

Can And so we're seeing again these
long term consequences this long term

simple fixation of orbital neurons and
we think that it may be associated

with some of the decisionmaking deficits
I'm going to talk about on the next slide.

And this I'm jumping my own gun
here just to confirm that this same

cocaine exposure procedure can cause havoc
based behavior in our mice so again if we

expose cocaine to mice during adolescence
and we use the exact same task that

I've been discussing so far in adulthood
we find that our cocaine exposed behavior

really flexible they're less they're less
able to modify their behaviors when action

outcome contingencies change and I'll show
you an example of this in a moment but

really you know the end goal here is to
correct these habits right to improve

the animal's ability to to engage
in behavioral flexibility and

so my student Liz sort of combined
these two projects and she asked

whether we could block cocaine induced
habits by this track the agonist seventy.

And so she used exactly the same
timing that I mentioned previously so

we're giving my scope cane for five
days from P thirty one thirty five we're

letting them grow up without any more drug
exposure we're taking it through exactly

the same conditioning procedure that I've
discussed so far and just as I promised

you are my set have had a history of
cocaine exposure develop habit by a C.S.

as adults so they're less able to engage
in behavioral flexibility based on action

consequence associations However if
Liz pairs seventy distract the agonist

with the opportunity for new learning she
can rescue the animals decision making so

the animals can come into the tap into
the operant conditioning chambers and

preferentially engage a response
that's likely to be reinforced

as opposed to one that's
unlikely to be reinforced.

So as I mentioned earlier to
earlier whether seventy has

tracked the agonist properties has been
questioned and so did her homework.

Here and she systemically administered
this struck the agonist and

took brains from the mice at the same time
point that we think this new memory is

being laid down and
when she extracts that we're told her so

medial prefrontal cortical tissue
she sees a main effect of drugs on.

The phosphorylation but I think any
of us could look at this graph and

recognize that it's likely being driven
by the signal in the orbital cortex and

so why this track the agonist would
preferentially penetrate the orbital

cortex we don't know yet but it may be
something that is advantageous to us and

in the future.

And so so that was really exciting but
what is sort of less exciting about

seventy and beating extract
the approaches in general is that B.

is really important for pain systems and
so giving us systemic administration of

a track The Agonist would be very much
expected to heighten pain sensitivity so

obviously is a major drawback if we're
thinking about clinical applications

right it's very unlikely that this
would be really translatable and

so Liz To her credit started to think
a little bit outside of the box here and

tried to find drugs that would really
stimulate the systems preferentially and

in the cortex as opposed to other
parts of the nervous system and so

the list upon M.D.M.A. and yes it's
that M.D.M.A. that you're thinking of so

it is we're talking about M.D.M.A. the
club drogue M.D.M.A. turns out at very low

doses so doses that are not
reinforcing to you to us or Hue mice.

Cause B.D.N.F. stimulation in the cortex
media release excuse me and so so

Liz first wanted to ask whether M.D.M.A.
could block cocaine induced habits and

indeed she she was successful so here we
see that in a separate cohort of mice

if the animals were exposed to cocaine
as adolescents they have a hard time

adjusting their behaviors in adulthood
However if she pairs low dose M.D.M.A.

with this opportunity for a new learning.

Animals can later on it shoes can
engage in behavioral flexibility and

preferentially engage the action that's
most likely to be reinforced so again

Liz did her homework here she systemically
injected M.D.M.A. took the brains and

confirmed that indeed we see an elevation
in mature enough within the cortex

as you can see right there
Additionally we see an elevation

phosphorylation of urc which is one of
the downstream substrates of correct B.

and then confirming other studies we
actually see a loss of beating up within

the dorsal straight i'm And interestingly
no effects within the medial prefrontal

cortex or in Manila and so I would say
while this profile is really advantageous

to us one really important future question
will be to understand why I'm D.N.A.

has such different effects throughout
different brain regions on earth trophic

systems so
in all of the work that I've shown you so

far we've been reducing beating enough
selectively within the orbital cortex

using viral vector strategies and
then I've been stimulating track be using

systemic approaches so
seven or M.D.M.A. and

so these these data have all really nicely
come together to argue that beating F.

within the orbital cortex is
really really important for

our military to flexibly adjust our
behavior based on action consequences.

What it doesn't tell us is where Track
be binding is important so again B.

is a high affinity receptor for P.D.F. so
this might seem like a silly question to

you is like if if if if in the record
tax is important obviously track B.

and over cortex is really important but

that's not necessarily the case because
B.D.N.F. is subject to anterograde and

retrograde transport so it may be
that when we knock down beating F.

within the orbital cortex we deprive
let's say the Amidala of B.D.N.F.

and it's really binding within the image
to live that's important and so

we wanted to ascertain where Track be
binding in the brain was important and

to do this we teamed with Kerry wrestler
who had created a lead to a viral

director that over expresses track
a truncated form of traffic so

the track becomes in two flavors
one is the full length ice

a form that has intracellular tail and one
is a truncated ice a form that doesn't and

so one track the binds it has to
dime arises out of phosphor late so

to be an active track bigger scepter you
need both of those full length tails so

here and then for
example you can phosphor really irk.

Can be as well however if you have
only truncated ice of forms or

one truncated ice a form
you are an active and so

by over expressing
a truncated form of track B.

where it were essentially competing
with and dodginess full length track B.

for B.D.N.F. and so that again
this is their viral director here

that's our friend Carrie there and so.

This experiment Liz actually
infused two volumes of

the the trying to track the viral vector
so some mice receive the point five

micro leader volume other mice receive
the one point five micro leader volume and

what you're seeing here is histological
representations of the infusion spread and

what I hope you're thinking is Shannon
that looks exactly the same it does

when we cut when we quantitatively
measure the viral vector spread we

actually don't see any differences based
on the volume and that's important for

reasons I'll show you know a minute but
it's also not not surprising so if any of

you have worked with plenty viral doctors
you know that they're really really sticky

so they tend to stay where you put them
which is really handy if you're working in

a very small mouse brain like us and so
what we're really intending to do by

infusing two volumes of this viral vector
was not to generate a larger spread but

to generate a greater degree of knock down
and so to as a surrogate for this approach

as a surrogate for this measure one of
the things that we did was we measured or

quantified the tag on the viral vector and
indeed what you're seeing is in what we're

calling the half or the low dose viral
vector we see a nice and I signal but

it's much much much stronger in the sort
of want to quote unquote high dose viral

vector and so all I'm trying
to get at is that we took some

principles from pharmacology where you
might expect dose dependent responses and

we apply them to viral vector strategies
where we would expect here to see dose

dependent effects and
that was indeed the case so

when mice are Win Lose tested her animals

you can see that all of the mice again
with biological infusions of the orbital

cortex could acquire
those book responses but

are mice with a full dose quote unquote of
the truncated track be you were unable to

generate as robust response rates as
again as is typical of orbital damage and

then more importantly when we modify these
response outcome contingencies are mice

with the full dose of the truncated tract
the viral Dr what we're unable to keep.

So this looks a lot like beating F.

knocked down and it suggests that beating
after knock down binding to local track be

in the orbital cortex is at least as
some water or rather quite important or

engaging in response
outcome of contingencies or

behavioral flexibility and I'm sort
of hedging there because I think that

it's very likely that the systems
in other brain regions

may also be implicated in this kind of
behavioral flexibility but certainly our

data would suggest that the orbital cortex
is is a primary a primary place where B.

B. B. is really important again for

behaving based on action
consequence contingencies.

And then just for the interest of time
I'm not going to show you the data but

we were able to block the behavioral
effects of M.D.M.A. And

so again this suggests that M.D.M.A.
requires track be

within the orbital cortex to have
these pro cognitive effects.

So if we can just pause for
one minute what I showed you today so

far was that B.D.N.F. track be within
the orbital cortex is really important for

selecting actions based on anticipated
consequences OK we think this is occurring

via local orbital track the binding and
interactions with the in the villa.

And that conversely cocaine weakens
memory for consequences and

this occurs I should mention in both
rodents as I showed you today but

also humans so Karen Esha had a paper in
Science a couple of years ago now showing

using a contingency degradation of Crete
procedure just like the one that I've been

talking about that humans with a history
of cocaine abuse are more biased towards

making stimulus dependent habit based
responses in an operant conditioning task

as opposed to quote unquote goal directed
action outcome decision decisions and

so these the findings I'm showing
you here have been directly

translated into human experiments.

And then I showed you today that
seventy can both and D.N.A.

can both restore action in
consequence decision making and

then just as a shameless plug we also
showed that seventy eight V.H.F.

can restore action consequence decision
making following stress from one exposure

OK so if we take a step back we think the
orbital cortex is doing in general is that

some sort of learning related plasticity
within this brain region is enabling us to

retain action in consequence memories and
thus engage in the most optimal

behaviors later on and we think I say
retention here because of this study for

example that has parsed out these
different phases of learning and

memory using in activation techniques so
if any of you in the room

are familiar with all of this action
consequence action outcome literature you

might have been wondering this whole time
why am I not talking about the prelim big

cortex which is situated here in yellow So
the reason you're thinking about that is

maybe you're thinking about the work
of Bernard Blaine that has shown since.

The late ninety's that the prelim because
Texas is also important for action

consequence decision making so the prelim
a court tax appears to be important for

the acquisition of those and also the
consolidation of those associations and

when I was just starting my law when you
know we tried really really really hard

to determine well with to find a role for
B.D.N.F.

in these processes within the prelim
it court techs and despite a lot

of our efforts we really couldn't so
it doesn't seem like beating F.

is important for the function of
the prelim the cortex in acquiring action

outcome associations However in a During
the same time I start to become interested

in three kinase and in my last three
slides I want to show you some of the new

data that we've been generating showing
that peer three kinase may actually be

important for
the functions of the prelim that cortex so

P I three kinase says it is a signaling
complex that helps us transducer signals

from the cell surface to air pathway and
one of the reasons that I

that it sort of came on to my radar
is that it's upward by cocaine and

the effects are really long lasting So
again if you expose an animal to cocaine

and euthanize it three weeks later you
can see an abbreviation of a Q T M two or

phosphorylation within
the medial prefrontal cortex.

So it's involved in snappy Genesis and
indeed expanded Genesis

somewhat unlike that B.D.N.F. tract
the pathway which might be more associated

with injured explained stabilisation it's
composed of P.D. five if you intend sudden

units of which there are eight so
this really is a signaling complex and

there are lots of different bits and
pieces that we can manipulate but

we care about in the context of again
these next few slides ten data so

people in ten Beta is a catalyst
subunit and it's downstream of G.

protein coupled receptors and
I can i was interested in ten data and

I became interested thanks to my colleague
Gary the cell so Gary studies Fragile X.

Syndrome and he contacted me and
like you know twenty twelve and

was like Hey Shannon we got these
def Marwan knock on my switcher or

as you guys probably know a model for
Fragile X.

syndrome and we find this crazy thing.

Is way way way up regulated I mean
there's like way too much of it and

so we think that potentially if we could
reduce P one ten beta and knock it down

we might be able to normalize some of the
average decision making that these mice

engage in putatively Gary doesn't do a lot
of doesn't do much behavioral work and

so he said we've got the mice we've
got the virus you've got the boxes

let's get together and I said OK And so we
start you know we stuck this viral vector

that reduces P one ten beta into
the prefrontal cortex of our fragile X.

model mice and we just wanted the
prefrontal cortex with this viral vector

and we were able to indeed rescue some of
their decision making impairments and.

If you're interested this is the paper so

all this time we're thinking about
Will Gary sing about Fragile X.

I think about cocaine and I think about
cocaine because of this paper that came

out way back in two thousand and two and
this paper show that if you infuse into

the ventricles Pancho I three kinase
inhibitor you can block the expression

of cocaine induced locomotor sensitization
and so the reason this was really really

exciting even though it lacked bring
region specificity for example was that

the investigators were able to block this
plasticity that occurred as a result

of cocaine so the animals are already
way experience with cocaine lots and

lots of drug and they block able to block
sensitization and so this is a lot like

people right so if you're an individual
who has developed cocaine addiction

it's then that you're seeking treatment
you're not coming in before you know

the cocaine exposure starts to Pete and
been Perrier your life activities and so

we were wondering whether we can inhibit
the P one ten beta in particular

to block some of the long term
consequences of cocaine so

we raised a little money from children's
and actually didn't know this but

we're we're in like a children's
building right now us as children so

that's very fitting so we raise a little
bit of money and off we went so first we

needed to confirm that P one ten data was
nicely poised within the prefrontal cortex

throughout adolescent development to
potentially regularly cortical dependent

behaviors so we can see really nice
expression patterns really robust

expression patterns throughout the medial
and lateral orbital peripheral cortex

throughout personal development and
then we infused the viral vectors

selectively in the mediocre frontal
cortex to knock down pure intent data and

I have to credit I mean a gross for
making his viral vectors and so

in one of the experiments we did we
really wanted to sort of draw inspiration

from that two thousand and two paper and
block some of the behavioral consequences

of cocaine exposure and so
in this experiment we implanted mice with.

Indwelling juggler catheters that allowed
the animals to instead of self administer

food in the conditioning chambers
actually sell them Mr cocaine so

instead of going into the conditioning
chambers and nose picking and

to recesses for food they instead
knows book an essential recess for

an infusion of a Coke of cocaine
reinforcer given directly.

So what you can see is that the mice
can acquire the correct poke response

they neglect the nose poke response
that is unreinforced over the course of

several sessions we then take them through
extinction which is a pretty common

technique used in my field so
you sort of mimic therapy for better or

worse and then after all of this only
after all of this exposure do we infuse

the viral doctors which reduced P one ten
beta in the medial prefrontal cortex and

we brought the mice back for
a read reinstatement test and so

here we're simply playing the mice
cocaine associated cues and

we're asking our mice Why does
this make you want to respond and

our scrambled control mice generate
lots of responses as you might expect

by contrast this is precisely mitigated by
the selective knockdown of P one ten beta

so it's pretty hard not to get excited
about these data so we ran a whole bunch

of other experiments and to summarize here
we can block the reinstatement of cocaine

seeking under multiple conditions
including context induced reinstatement

we can also block the expression
of locomotor sensitization So

this fully recapitulate the two
thousand and two paper that used a pan

inhibitor in the ventricles right so we're
honing in quite a bit relative to that

first finding And we also mitigate
cocaine induced elevations in fossil M.

TOR which is what you would
expect based on the signaling.

And of course you're wondering what about
our habits and so we took the mice through

exactly the same procedure that I've
been explaining for the last you know X.

number of minutes now and

we again expose the animals to
cocaine before we infuse the viral.

Dr So these animals have had a lot of
cocaine exposure in this case we used to

procedure from Schoenbaum And so low this
is the first evidence in rodents that

cocaine causes habits again as in people
and then we have a washout period and

then we infuse the viral vector and then
finally we train the mice to respond for

food and we take them through
the contingency degradation procedure so

you can see during acquisition mice
with a history of cocaine exposure

generally higher response rates
this is pretty characteristic and

had been seen in previous studies
what's more important is that our mice

that had a history of cocaine exposure
generate habit based behavior just like

you would expect by contrast we can
mitigate those habits and engender

behavioral flexibility by knocking down P
one ten data within the medial prefrontal

cortex these same data can be shown
in a slightly different way by simply

generating a response preference ratio so
in this case you have the non degraded.

Response rate over the degraded one is
a habit it's no preference and so our

cocaine expose my surgeon or eating habit
based behavior like really expects and

we can rescue that pretty nicely with
the Q In ten Beta inhibitory viral

vector and So to summarize this catalytic
subunit P one ten beta is expressed

really nicely throughout the postnatal
prefrontal cortex and in the absence

of cocaine it really doesn't do very much
that does it doesn't do much at all and so

for example this is a whole list of
behavioral assays that these mice

that mice that were cocaine naive that
had the viral Lector were tested and

we really didn't see any phenotypes
However when we layer on cocaine

we start to see that Q industry and
statement for

example can be blocked this is the fields
primary test for relapse like behavior.

We can block locomotor sensitization and
cocaine induced habits and so

we're trying to just in general in my lab
we're trying to hone in on these maybe

more subtle therapeutic approaches such
as single payer through kind of sub unit.

Or maybe a low dose M.D.M.A.

paired with the opportunity for
new learning to try to help facilitate.

Behavioral flexibility in a digital
ZX that are struggling with drug use

disorders.

And so in terms of our next steps some of
the things that we're thinking about a lot

are whether habits are associated with or

preceding addiction like behaviors This
may seem like an exciting question but

it's actually really important because it
could shed light on see whether habits for

example could be predictive of addiction
like behavior or whether looking I

have a bias sees it's too or whether you
know might be too late at that point.

And because have a bias these can
be tested in humans this has direct

translational appeal so one of the reasons
we care a lot about being F A Q three

kinases at the effect neuron structure
as I mentioned throughout my talk and

we think that this is really important for

the behavioral consequences
of these manipulations.

And the reason we think this is for

example if you simply look at
typical healthy mice and you count

dendritic spine densities in a particular
mushroom shape spines these are the spines

that are likely to contain synapses within
the orbital cortex you get a really nice

predictor of how the animals will respond
in our operant conditioning chamber so

willing and gender action outcome
based decision making strategies or

will they rely on habits and so
you can see here a correlation between

mushroom shaped spines and that response
preference where anything greater than one

is a goal directed strategy and
anything at one is a habit based strategy.

And we think that the effects
are the effects of these compounds

on dendritic spines maybe
brain regions specific and

also more importantly the effects
on dendritic spine plasticity

on behavior is going to be highly
brain region specific So for

example within the prelim that cortex we
just published that printing of preview of

the cortical dendritic spines was really
important for enabling the animals to git.

Right Action outcome decision making and
I want to thank Haley here who actually

helped quite a bit with this project so
thank you.

And then finally we're trying to
understand how all of these molecules in

the circuits are actually coordinating so
why would the prelim the cortex rely on

more of teeth carry three
kinase to be enough for

example in acquiring action
outcome can contingencies and so

these are questions that we're trying to
tackle using perching quantification and

direct manipulation of
specific circuits and

specific proteins so I'm going to stop
there I would like to thank my team

at Emory we have a lot of great graduate
students in the Emory neuroscience program

some of you may actually be
an Emory nurse science program and

so you know these are this is the team
that was largely in place when these

viral when these studies were conducted
we have other folks who preceded and

came and came later and then things
to cure a wrestler of a cell for

essentially agents and
collaboration thank you also to you and

I'd be happy answering questions thank.

A.

Lot Thank you.

Yeah great.

Yes So we we have thought about sex
differences in a couple of contexts So

firstly we have looked at individual
differences in cocaine self ministration

during adolescence and then how animals
make decisions in adulthood and

we find that in females in females
that are resilient to cocaine So

in other words the males that can develop
really behaviorally flexible strategies in

adulthood even though they've had cocaine
females generate larger dendritic spines

heads within the orbital cortex so they're
able to recruit seems these neurons to

help be resilient to the drug whereas
we just don't see this in males and

we also see that females we see a larger
population of our females generating

what we would consider resilience so
that's that's one thing to consider but

then in terms of looking at or working
in the field of actions and habits one

always has to consider sex because there
are some really really nice studies using

the fork or geo type mice from our
Arnold showing that females develop.

Food reinforced habits more rapidly than
males and this is sex hormone dependent so

it's not going to addle sex dependent and
so that's something we have to consider

when we design our studies and
the converse Lee Jackie Barker and

others have shown that when the reinforcer
is alcohol males develop the habits more

rapidly than females and so
certainly you know although I think and

I would like us just to like dump
males and females together and

pretend like there are no sex differences
when our writing and how we go writing our

grant we just can't we just can't because
the propensity to develop habits is

fundamentally different so I have to
answer your question is a good enough.

Yes.

Yeah.

Wow.

You.

Know.

You're great.

Life picture yeah.

But you're.

Yes So I think the question that you're
asking is when we're talking about habits

here are we talking about habits that
result from say lots of experience and

struggle dependent action verses habits
that are maybe default behaviors

that are due to a failure in updating our
response strategies because we fail to

learn that new response tragedies
would be warranted Is that fair so

I think that what we're studying
throughout all of this is the latter So

we're we see the animals are deferring to
what I'm calling a habit based strategy

and many of us in the field do use
that term even though it's it doesn't

differentiate between the two types of
habits right so you have overtraining

habits this me with the glasses thing or
this morning I was in the ladies room and

a gentleman walked in I suspect that
was his habit going arrive right he and

his habit is to turn right here just on
the incorrect floor so so that would be

a pen and overtraining have it versus
have it by default where we engage in

familiar behavior because we fail to learn
that something different was better and

so I think because that new learning is
preferred a quarter call dependent and

we're mucking up the prefrontal cortex
that's really what we're studying here and

we can confirm that by do you
by using by taking the mice

after the protest and then continuing to
reinforce both nose poke responses and

then taking them through another
cycle of contingency degradation and

the reason that this helps us quite a bit
is that if you have developed a habit

through your straddle dependent processes
taking over your behavior then you're

going to continue to be
insensitive to that contingency

degradation because we reinforce that
habit by continuing to train you on

both notebook recesses However if you just
fail to learn that your actions produce

consequences that additional training
actually improves your ability

to then surmount the contingency that.

Ration procedure and so

in virtually all of the experiments that
I've shown you today we did take those

extra steps we retrain the mice using a
fixed ratio schedule of reinforcement for

four five six sessions take them through
another cycle of contingency degradation

procedures and in all the cases in cocaine
exposure in the case of big enough knock

down trying to track the over expression
our mice can quote unquote catch up so

I think what we're seeing is failures and
prefrontal dependent learning and

memory that caused this deferral to we
what we call a habit these behavior so

I think and this language is something
that we struggle with in the field so

my colleague Andrew Holmes distinguishes
in his studies between early habits and

late habits and
I think that's sort of maybe.

Imperfectly you get that
the idea that yes it is

you know we call these these
behaviors operationally habits but

you can certainly arrive at these habits
through at least two different avenues.

Thank you yeah

yeah.

OK.

Souls.

Cool.

Yeah right so
that's the question we've been able to so

block some of the plasticity that would be
associated with new learning using things

like Le truculent A which blocks
effect in Asian and so we mostly got

in using pharmacological approaches
that are not cell type specific and

so one of the next steps for
my lab and you know may they and

I each smile on me is to start to look
at cell type specificity Yeah I mean

that's essential right we it's
the clear next step Yeah.

It's.

All.

The rule.

All.

In one.

Yeah yeah so another thing that I
would really like to do again May and

I smile is that is look at habits that
persist despite negative consequences and

we can do this very easily
in rodents by simply.

Equipping our conditioning
chambers with foot shocks that

deliver shock if the animal
continues to seek drug or

continues to seek reinforcement and so
a handful of labs in my field to do this

in the context of a cocaine itself
ministration that is cued and so it makes

it really difficult to understand whether
the cocaine seeking is habit based because

there's a explicit probably in Q All we
would need to do is strip out that Q.

and then we can look at whether
these cocaine seeking habits indeed

are a bitch will and
be resistant to punishment.

Thank you.

One of my Tech students you guys know.

These guys took up a class of mine so
I was wondering you know.

I won a name.

OK thanks guys thank you.