Table 1. P values of the graphs presented.
Figure 1. Anhedonia- and anxiety-like behaviour induced by
inflammatory pain in male and female rats. A) Timeline of the
experimental design B) Sucrose consumption during 48 hours in male,
expressed as mean ± SEM of mg/kg/24 hours C) Sucrose consumption during
48 hours in female, expressed as mean ± SEM of mg/kg/24hours D) Sucrose
consumption during 2 hours in male, expressed as mean ± SEM of mg/kg/2
hours E) Sucrose consumption during 2 hours in female, expressed as mean
± SEM of mg/kg/2 hours F) Time (seconds) in light chamber in the light
dark box test (LDB) in male, expressed as mean ± SEM G) Time (seconds)
in light chamber in the LDB in male, expressed as mean ± SEM H) Number
of transitions between chambers in the LDB in male, expressed as mean ±
SEM I) Number of transitions between chambers in the LDB in female,
expressed as mean ± SEM J) Mechanical sensitivity threshold in male,
expressed as mean ± SEM of paw withdrawal in grams K) Mechanical
sensitivity threshold in female, expressed as mean ± SEM of paw
withdrawal in grams. Black and red bars and symbols represent saline-
and CFA-treated male respectively, and grey and orange bars and symbols
represent saline- and CFA-treated female respectively. *, ** and ***
denotes significant differences between groups (two-way ANOVA for
repeated measures followed by Bonferroni multiple comparisons test,
p< 0.05, p< 0.01 and p< 0.001,
respectively), and #, ## and ### denotes significant differences
within groups compared with the basal measures (two-way ANOVA for
repeated measures followed by Bonferroni multiple comparisons test,
p< 0.05, p< 0.01 and p< 0.001,
respectively). Abbreviations: SPT, sucrose preference test; VFT, Von
Frey test; CFA, Complete Freund’s Adjuvant; SAL, saline; LDB, light-dark
box test.
Figure 2. Corticotropin-releasing factor system alterations
induced by inflammatory pain. A) Mean ± SEM of CRF mRNA relative gene
expression in NAc of male rats. B) Mean ± SEM of CRF mRNA relative gene
expression in Amy of male rats. C) Mean ± SEM of CRF1R mRNA relative
gene expression in NAc of male rats. D) Mean ± SEM of CRF1R mRNA
relative gene expression in Amy of male rats. E) Mean ± SEM of CRF1R
protein expression in NAc of male rats (CRF1R/GAPDH arbitrary units). F)
Mean ± SEM of CRF1R protein expression in Amy of male rats (CRF1R/GAPDH
arbitrary units). G) Plasma cortisol levels in male rats, expressed as
mean ± SEM of ng/mL. H) Mean ± SEM of CRF mRNA relative gene expression
in NAc of female rats I) Mean ± SEM of CRF mRNA relative gene expression
in Amy of male rats. C) Mean ± SEM of CRF1R mRNA relative gene
expression in NAc of female rats. J) CRF1R expression in NAc in
female, expressing as mean ± SEM of relative gene expression. K) Mean ±
SEM of CRF1R mRNA relative gene expression in Amy of female rats. L)
Mean ± SEM of CRF1R protein expression in NAc of female rats
(CRF1R/GAPDH arbitrary units). M) Mean ± SEM of CRF1R protein expression
in Amy of female rats (CRF1R/GAPDH arbitrary units). O) Plasma cortisol
levels in female rats, expressed as mean ± SEM of ng/mL. Black and red
bars represent saline- and CFA-treated male respectively, and grey and
orange bars represent saline- and CFA-treated female respectively. *
Denotes significant differences between groups (t-student for
independent samples or Welch’s t test p < 0.05).
Figure 3. Kappa opioid receptor/Dynorphinergic system
alterations induced by inflammatory pain in male and female rats. A)
Mean ± SEM of pDYN mRNA relative gene expression in NAc of male rats. B)
Mean ± SEM of pDYN mRNA relative gene expression in Amy of male rats. C)
Mean ± SEM of KOR mRNA relative gene expression in NAc of male rats. D)
Mean ± SEM of KOR mRNA relative gene expression in Amy of male rats. E)
Mean ± SEM of DYN protein expression (ug/tissue) in NAc of male rats. F)
Mean ± SEM of DYN protein expression (ug/tissue) in Amy of male rats. G)
Mean ± SEM of KOR protein expression in NAc of male rats (KOR/GAPDH
arbitrary units). H) Mean ± SEM of KOR protein expression in Amy of male
rats (KOR/GAPDH arbitrary units). I) Mean ± SEM of pDYN mRNA relative
gene expression in NAc of female rats. J) Mean ± SEM of pDYN mRNA
relative gene expression in Amy of female rats. K) Mean ± SEM of KOR
mRNA relative gene expression in NAc of female rats. L) Mean ± SEM of
KOR mRNA relative gene expression in Amy of female rats. M) Mean ± SEM
of DYN protein expression (ug/tissue) in NAc of female rats. N) Mean ±
SEM of DYN protein expression (ug/tissue) in Amy of female rats. O) Mean
± SEM of KOR protein expression in NAc of female rats (KOR/GAPDH
arbitrary units). P) Mean ± SEM of KOR protein expression in Amy of
female rats (KOR/GAPDH arbitrary units). * Denotes significant
differences between groups (t-student for independent samples or Welch’s
t test p < 0.05).
Figure 4. Anhedonia- and anxiety-like behaviour induced by
inflammatory pain in female rats is blocked by NorBNI intra-NAc
administration. A) Timeline of the experimental design. B) Sucrose
consumption during 48 hours in female rats, expressed as mean ± SEM of
mg/kg/24 hours mean ± SEM. C) Sucrose consumption during 2 hours in
female rats, expressed as mean ± SEM of mg/kg/2 hours. D) Time in light
chamber (seconds) in the light-dark box test (LDB). E) Number of
transitions between chambers in the LDB, expressed as mean ± SEM. F)
Mechanical sensitivity threshold (g) in female rats, expressed as mean ±
SEM of paw withdrawal threshol. Grey and orange bars represent saline-
and CFA-treated female rats respectively, and empty and full bars
represents the intracranial treatments of aCSF and NorBNI respectively.
*, ** and *** denotes significant differences between groups (three-way
ANOVA for repeated measures followed by Bonferroni multiple comparisons
test, p < 0.05, p < 0.01 and P < 0.001,
respectively), and #, ## and ### denotes significant differences
within groups compared with the basal measures (two-way ANOVA for
repeated measures followed by Bonferroni multiple comparisons test, p
< 0.05, p < 0.01 and P < 0.001,
respectively). Abbreviation: SPT, sucrose preference test; VFT, Von Frey
test; CFA, Complete Freund’s Adjuvant; SAL, saline; LDB, light-dark box
test; NorBNI, Norbinaltorphimine.
.
Figure S1. Daily calendar of the behavioural protocol. CFA,
complete Freund adjuvant; SAL, saline; LDB, light–dark box test; SPT,
sucrose preference test; VFT, von Frey test.
Figure S2. Schematic of the brain areas dissection. A) Area of
the NAc dissection B) Area of the amygdala dissection.
Figure S3. Daily calendar of the behavioural protocol of the
Experiment 2 and intracranial administration verification. A) Daily
calendar of the behavioural protocol. B) Dots represent the site where
aCSF and NorBNI were injected: Black represent saline/aCSF treated
female rats, grey represent saline/NorBNI treated female rats, red
represents CFA/aCSF treated female rats, and orange represents
CFA/NorBNI treated female rats. Abbreviatures: CFA, complete Freund
adjuvant; SAL, saline; LDB, light–dark box test; SPT, sucrose
preference test; VFT, von Frey test.