Presentation 2 (1)Presentation 2 (1)

1. FLUOXITINE
TREATMENT IS
EFFECTIVE IN A RAT
MODEL OF
CHILDHOOD-INDUCED
POST-TRAUMATIC
STRESS DISORDER
Presented by: Yashu Sharma
Study Type: Randomized Controlled
Trial
Published in: Journal of Child
Psychology and Psychiatry, 2019
(Assumed source; adjust if needed)

2. Introduction
Q. Can a common antidepressant
treat the PTSD?”
Q. What is Fluoxetine?
Q. Why it is used in PTSD?
Q. Does fluoxetine actually work for
core symptoms of PTSD, or are we
chasing a placebo?
Q. What is PTSD (Post Traumatic
Stress Disorder)?

3. What is PTSD
(Post Traumatic
Stress Disorder) ?
• Post-Traumatic Stress Disorder
(PTSD): Mental health condition
following exposure to trauma
• Core symptoms: Re-
experiencing, avoidance, negative
mood and cognition,
hyperarousal
• Affects combat veterans, abuse
survivors, accident victims, and
more
• High rates of chronic impairment

4. Treatment
for PTSD ?
• First-line: Cognitive
Behavioral Therapy (CBT),
particularly trauma-focused
• Pharmacotherapy: SSRIs
(fluoxetine, sertraline,
paroxetine) approved by
FDA
• Limited efficacy in some
subpopulations; response is
variable

5. Fluoxitine :
• A Selective Serotonin Reuptake Inhibitor (SSRI)
• Widely used to treat depression, OCD, and
anxiety
• Although SSRIs are first-line treatment for
post-traumatic stress disorder (PTSD)
Patients
• Fluoxitine dose varies for defferent age groups
and stages
• It is generelly used for treat MDD (Major
Dipressive Disorder) - 20mg/day , OCD
(Obssesive Compulsive Disorder) 20mg/day ,
Bullimia Nervosa 60mg/day, Panic Disorder
and PDD (Premenstrual Disphoric Disorder) -
10mg/day

6. Goals and Study objectives
 Mixed results in previous PTSD trials
 Need for well-controlled data
specific to fluoxetine
 Goal: evaluate efficacy and safety in
individuals with diagnosed PTSD
Primary Aim: Evaluate fluoxetine's
efficacy in reducing PTSD symptom
severity
Secondary Aim: Assess its impact on
functional outcomes and adverse
events

7. Process overview
1. Creating Trauma (Juvenile Stress - JVS)
•
Rats were exposed to stressful experiences during their
early life:
Day 1: Forced to swim for 10 minutes.
Day 2: Placed on a high platform for 30 minutes, three
times.
Day 3: Restrained in a small box for 2 hours.
2. Treatment with Fluoxetine
FLXjuv (early): Fluoxetine in the drinking water for one
month immediately after juvenile stress (postnatal days 30–
63).
FLXadlt (late): Same dosage of fluoxetine, but delivered for
one month starting in adulthood (days 64–93).
Control groups: No stress + water, stress + water, and stress
+ early or late SSRI.
Fluoxetine was given in drinking water for about a month..

8. 3. Testing Anxiety (Elevated Plus Maze - EPM)
Later, all rats were tested using a maze:
• The maze has open arms and closed arms.
• Rats that are more anxious spend more time in
the closed arms.
• Their movement and behavior were tracked.
Q. Why This Matters ?
Childhood is a sensitive time: Trauma during this period can
cause long-term mental health issues.
But it’s also a good time for treatment: Giving fluoxetine
early can prevent PTSD symptoms.
Waiting until adulthood might be too late or less effective.

9. Summary of Materials and Methods:
The study followed NIH guidelines and was
approved by the University of Haifa Ethics
Committee. A total of 116 male Sprague Dawley
rats (PND 22, 35–49 g) were used. After arriving
from Harlan, Jerusalem, the rats were
acclimated for 5 days. They were housed in
groups of 3–4 per cage in a temperature-
controlled room (22 ± 2 °C) with a 12-hour
light/dark cycle. They had ad libitum access to
standard rodent chow pellets and water. The
sample size was chosen based on prior
experience and standard practices in the field.

10. Fluoxetine treatment through drinking water (FLX)
Baseline fluid intake
– Measured 24-h water consumption per cage (1 g = 1 ml) for 3 days before treatment
Dose and delivery
– Target: 10 mg/kg/day FLX, a level shown to yield clinically relevant plasma levels, lower cortical
5- HT turnover, and reduce depression-like behavior in rats
– FLX stock: 3 mg/ml (Vetmarket, Israel) diluted into drinking water
Preparation & adjustment
– Fresh FLX solutions made twice weekly
– Concentration recalculated from each cage’s average daily intake and each rat’s body weight
(fluid intake measured twice/week; weight once/week)
Light protection & controls
– Amber, opaque bottles to prevent FLX photodegradation
– Control animals drank plain water
•Next steps often include verifying plasma FLX levels, tracking behavior in tests like forced swim
or sucrose preference, and exploring neurochemical endpoints (e.g., hippocampal BDNF).

11. Experimental Design
Rats were randomly assigned to groups; no blinding was performed.
• After habituation in the vivarium, each rat either underwent
juvenile variable stress (JVS) on postnatal days (PND) 27–29 or
remained unexposed.
• From PND 30 onward, rats received one of three drinking-water
regimens for 30–33 days:
• Regular water (no fluoxetine)
• Fluoxetine throughout the juvenile window (FLXjuv: PND 30–63)
• Fluoxetine during adulthood (FLXadlt: PND 64–93)
• On PND 93, all animals were tested in the elevated plus maze (EPM)
to assess anxiety-like behavior.
• The full experimental timeline is depicted in Figure 1.

13. Behaviour profiling
•Define “normal” behavior using 30 control
animals on seven EPM measures (distance and
duration metrics).
• For each measure, calculate the 20th percentile
(lower or upper, depending on what indicates
abnormality).
• Test each animal’s seven scores against these
cutoffs.
• Label an animal “affected” if 4 of its measures
≥
fall beyond the 20th-percentile thresholds.
• Animals with fewer than four out-of-range
measures are deemed “unaffected.”
• Figure 2 shows the representative performances
of an “affected” animal.

14. Statistical Analysis
Animal weight over time: repeated‐measures two‐way ANOVA
• Factors: stress (control vs. JVS) and drug treatment (no FLX,
FLXjuv, FLXadlt)
Fluid intake during each treatment window (PND 30–63 and
PND 64–93): repeated‐measures two‐way ANOVA
• Factors: stress (control vs. JVS) and fluid type (water vs. FLX)
elevated plus‐maze behavior: two‐way ANOVA
• Factors: stress (control vs. JVS) and drug (no FLX, FLXjuv,
FLXadlt)
• Bonferroni post hoc comparisons for drug effects when the
ANOVA was significant
Individual behavioral profiling (affected vs. unaffected): Chi-
squared goodness‐of‐fit test
All analyses checked for statistical assumptions and run in IBM
SPSS Statistics v19

16. Results overview
JVS led to increased anxiety-like behavior and
a higher proportion of affected rats.
Fluoxetine treatment during juvenility
(FLXjuv) significantly reduced the proportion
of affected rats to levels similar to controls.
Fluoxetine treatment during adulthood
(FLXadlt) was not effective and even showed
anxiogenic effects in some measures.

17. Results
Weight
 No significant differences across stress, drug, or their interaction (all F’s 1.07, NS).
≤
 Interpretation: Neither early‐life stress nor fluoxetine (FLX) treatment altered d rats’
body weight.
Fluid intake in juveniles (FLX vs. water)
 Stress: F(1,40)=3.12, NS
 Drug (FLX): F(1,40)=0.07, NS
 Stress×Drug: F(1,40)=0.01, NS
 Interpretation: FLX and stress had no effect on drinking when given during the
juvenile period.
Fluid intake in adults (FLX vs. water)
Stress: F(1,43)=3.64, NS
 Drug (FLX): F(1,43)=10.28, p<0.01
 Stress×Drug: F(1,43)=2.74, NS
 nterpretation: Adult rats on FLX drank significantly less fluid than water-only controls;
stress had no impact.
(See Fig. 3 for the adult FLX–water intake comparison.)

19. What they did ?
• Used a Chi-square “goodness of fit” test to see if each group’s split of “affected” vs. “non-affected” rats
deviated from an expected ratio.
• Baseline (control) expectation:
• 20% of rats “affected,” 80% “non-affected.”
• Control groups:
• Untreated controls
• Controls + fluoxetine in juvenility (FLXjuv)
• Controls + fluoxetine in adulthood (FLXadlt)
• → None of these deviated from the 20:80 split (i.e. fluoxetine alone doesn’t change anything).
• Impact of juvenile variable stress (JVS):
• JVS alone: significantly more rats became “affected” than the 20:80 expectation.
• JVS + FLXadlt: still a significant jump in “affected” rats.
• JVS + FLXjuv: NO significant jump—rats look just like controls.
• Zooming in on JVS groups only:
• Here the “normal” JVS split was actually closer to 35% affected, 65% non-affected.
• JVS + FLXjuv: dropped “affected” rats back toward control levels (significantly different from 35:65).
• JVS + FLXadlt: stayed at roughly 35:65 (no rescue).

20. Conclusion:
Early (juvenile) fluoxetine protects against the behavior-altering
effects of chronic stress, whereas starting fluoxetine in adulthood
does not.
Beyond these stats, you might wonder:
• What brain circuits are being “re-set” by juvenile fluoxetine?
• Could a shorter or lower-dose juvenile regimen work just as well?
• How long does this protective effect last into adulthood?
• Might there be a window before juvenility where it’s even more (or
less) effective?
Poking into those questions could reveal not only how fluoxetine
shields young brains from stress, but also guide smarter, age-tailored
treatments down the line.

21. Discussion
• Early Juvenile Variable Stress (JVS) triggers long‐lasting anxiety‐like
behavior in adulthood, confirming juvenility as a window of
vulnerability.
• Averaged EPM scores suggest fluoxetine (FLX) treatment appears
anxiogenic, but averaging masks important individual differences.
• Behavioral profiling reveals juvenile FLX significantly reduces the
proportion of JVS-affected rats to control levels, whereas adult FLX does
not.
• The contrasting FLX effects may stem from timing: adult rats tested
immediately post‐treatment versus juveniles tested after a month.
• Early interventions (pharmacological or enriched environment) during
juvenility reverse behavioral, endocrine, and neurobiological alterations
more effectively than adulthood treatments.

22.  In humans, trauma-focused psychotherapies show strong
childhood PTSD efficacy, and early family-based programs can
normalize HPA‐axis function.
 Pharmacological treatments in childhood PTSD remain under-
studied; SSRIs yield mixed results across anxiety and mood
disorders.
 Future research should identify early predictors of PTSD risk to
target interventions to vulnerable children and compare juvenile vs.
adult treatment outcomes.
 Our animal model supports juvenility as a sensitive period for both
vulnerability and effective intervention; human studies must verify
optimal timing and strategies.
• In humans, trauma-focused psychotherapies show strong
childhood PTSD efficacy, and early family-based programs can
normalize HPA‐axis function.
• Pharmacological treatments in childhood PTSD remain under-
studied; SSRIs yield mixed results across anxiety and mood
disorders.
• Future research should identify early predictors of PTSD risk to
target interventions to vulnerable children and compare juvenile
vs. adult treatment outcomes.
• Our animal model supports juvenility as a sensitive period for
both vulnerability and effective intervention; human studies
must verify optimal timing and strategies.

24. THANK YOU
Refrences:
Ariel L, Inbar S, Edut S, Richter-Levin G. Fluoxetine treatment is effective in a rat model of childhood-
induced post-traumatic stress disorder. Transl Psychiatry. 2017 Nov 30;7(11):1260. doi:
10.1038/s41398-017-0014-5. PMID: 29187754; PMCID: PMC5802710.​