Sex-dependent effects of in utero cannabinoid exposure on cortical function
- Aix Marseille University, INSERM, INMED, France
- Indiana University, United States
- University Roma Tre, Italy
- CHU Conception, Service de Psychiatrie, France
- CHU Timone Adultes, Service de Médecine Légale, France
Figures
Figure 1 with 1 supplement
Prenatal cannabinoid exposure induces sex-specific social deficits at adulthood.
(A–B) Adult male progeny from dams exposed during gestation to WIN had less contact (A): F(sex x treat)1,28=15.54, p<0.05, two-way ANOVA) and spent less time interacting with their congeners than …
Figure 1—figure supplement 1
Social isolation before testing did not induce sex-specific social changes in adult animals.
(A-B) Adult male and female naïve animals not isolated 24 hr before testing showed no difference in contact (A): F(sex x isolation)1,29=0.203, p>0.05, two-way ANOVA) and time to interact with their …
Figure 2
Prenatal cannabinoid exposure does not modify adult cognitive and anxious behaviors in both sexes.
(A–B) Prenatal cannabinoid exposure did not alter anxiety behavior in adult rats of either sex. No differences between WIN-exposed and SHAM animals were found in the elevated plus maze test as …
Figure 3
Prenatal cannabinoid exposure does not modify LTD in NAc of adult rats.
(A) Average time-courses of mean fEPSPs showing that low-frequency stimulation (indicated by arrow) induces LTD at accumbens synapses in both SHAM- (white circles, n = 5) and WIN- (green circles, n =…
Figure 4 with 3 supplements
Sexual divergent ablation of LTD in the PFC of adult rats prenatally exposed to WIN.
(A) Average time-courses of mea field EPSPs showing that low-frequency stimulation (10 min at 10 Hz, arrow) induced LTD of evoked fEPSPs recorded in the mPFC in SHAM (n = 7; white circles), but not …
Figure 4—figure supplement 1
mGlu2/3-LTD and long-term potentiation are spared by prenatal WIN exposure.
(A, B) Prenatal exposure to cannabinoids does not alter mGlu2/3-LTD. Average time of mean fEPSPs showing that mGlu2/3-agonist induced LTD (bath application of LY379268, 30 nM, 10 min) is identical …
Figure 4—figure supplement 2
Prenatal THC does not alter long-term potentiation.
(A) Average time-courses of mean fEPSPs showing that theta burst stimulation (indicated by arrow) induces LTP at layer V/VI mPFC synapses in both SHAM- (white circles, n = 8) and THC- (green …
Figure 4—figure supplement 3
Prenatal cannabinoid exposure does not modify synaptic CB1R potency or efficiency.
Dose-response curve for the cannabimimetic CP55,940 in SHAM (white symbols, n = 6–9 animals, EC50 = 0.240 μM, top value 42.01%, 95% CI for EC50 = 0.088–0.658) and WIN-exposed (green symbols, n = 5–6,…
Figure 5
Sexual divergent ablation of LTD in the PFC of adult rats prenatally exposed to THC.
(A) Average time-courses of men field EPSPs showing that low-frequency stimulation (10 min at 10 Hz, arrow) induced LTD of evoked fEPSPs recorded in the mPFC in SHAM (n = 4; white circles), but not …
Figure 6
Prenatal WIN exposure induces sex-specific alteration of intrinsic properties of layer five prefrontal pyramidal neurons in adult rats.
(A) Left, Current-voltage (I–V) curves recorded in SHAM (n = 19 cells/10 rats, white circles) and WIN-exposed rats (n = 23 cells/12 rats, green circles) showing altered membrane potentials in male …
Figure 7
Prenatal THC exposure induces sex-specific alteration of intrinsic properties of layer five prefrontal pyramidal neurons in adult rats.
(A) Current-voltage (I–V) curves recorded in SHAM (n = 8 cells/5 rats, white circles) and THC-exposed rats (n = 10 cells/5 rats, green circles) showing altered membrane potentials in male after an …
Figure 8
Prenatal cannabinoid treatment affects expression of endocannabinoid system components in a sex-dependent fashion.
CB1, TRPV1, mGlu5, and DAGLα mRNA levels in medial prefrontal cortex at PND90 were determined by real-time PCR in male (A, C, E, and G) and female (B, D, F, and H) rats using Taqman probes and a …
Figure 9
Positive allosteric modulation of mGlu5 restores LTD and normalizes social interaction in male rats prenatally exposed to WIN.
(A) Average time-courses of mean field EPSPs showing that, in WIN-exposed males (WIN, n = 17, green circle), LTD can be restored with the mGlu5positive allosteric modulator CDPPB (10 μM, n = 4, blue …
Figure 10
In prenatally exposed female, TRPV1 receptors mediate LTD, not CB1R.
(A) Average time course of mean fEPSPs showing that LTD (arrow) is inhibited by the selective TRPV1 antagonist capsazepine (CPZ, black circles, n = 5) but not the CB1R antagonist SR141617A (SR, blue …
Figure 11 with 1 supplement
Sex-specific substrates of LTD in adult mPFC.
(A) Time course of normalized field EPSPs recordings from layer 5 mPFC after 10 Hz stimulation protocol (10 min, arrow) in presence of CB1R antagonist SR141716 (SR, n = 6, blue circles) or TRPV1 …
Figure 11—figure supplement 1
Sex-specific substrates of LTD in naïve adult mPFC.
(A) Time course of normalized field EPSPs recordings from layer 5 mPFC after 10 Hz stimulation protocol (10 min, arrow) in presence of the selective CB1R antagonist NESS0327 (n = 7, blue circles) or …
Figure 12
FAAH inhibition restores LTD and social interaction in male rats prenatally exposed to cannabinoids.
(A) Average tie-courses of mean fEPSPs after 10 Hz LTD protocol (1à min, arrow) in WIN-exposed mPFC slices preincubated for 45 min in 2 μM URB597, a FAAH inhibitor. (B) Individual experiments (grey) …
Tables
Table 1
Statistical significance of different behavioral tasks in SHAM and Naive animals.
Prenatal treatment from GD 5 to GD 20 with vehicle solution in control dams (SHAM group) did not alter sociability (social interaction), anxiety (elevated plus maze), and cognition (temporal order …
| Behavior | Parameter | Treatment | Sex | n | Mean | SEM | P value (Unpaired t test) |
|---|---|---|---|---|---|---|---|
| Social Interaction | # of contacts | NAIVE | Male | 5 | 216.200 | 28.420 | 0.439 |
| SHAM | 10 | 235.800 | 10.550 | ||||
| NAIVE | Female | 4 | 211.800 | 21.600 | 0.85 | ||
| SHAM | 7 | 215.100 | 5.713 | ||||
| Elevated Plus Maze | % Time Open | NAIVE | Male | 6 | 25.900 | 2.988 | 0.665 |
| SHAM | 7 | 24.270 | 2.245 | ||||
| NAIVE | Female | 9 | 25.530 | 2.464 | 0.359 | ||
| SHAM | 9 | 22.540 | 1.978 | ||||
| % Open Entries | NAIVE | Male | 6 | 27.740 | 3.248 | 0.164 | |
| SHAM | 7 | 33.950 | 2.678 | ||||
| NAIVE | Female | 9 | 33.160 | 2.520 | 0.475 | ||
| SHAM | 9 | 30.230 | 3.111 | ||||
| Temporal order | Discrimination index | NAIVE | Male | 8 | 0.320 | 0.085 | 0.702 |
| SHAM | 7 | 0.366 | 0.078 | ||||
| NAIVE | Female | 8 | 0.258 | 0.132 | 0.896 | ||
| SHAM | 7 | 0.277 | 0.038 | ||||
| Object Recognition | Discrimination Index | NAIVE | Male | 10 | 0.547 | 0.087 | 0.792 |
| SHAM | 8 | 0.580 | 0.086 | ||||
| NAIVE | Female | 7 | 0.323 | 0.126 | 0.481 | ||
| SHAM | 7 | 0.421 | 0.045 |
Table 2
Statistical significance of gene expression changes in adult rat medial prefrontal cortex after in utero WIN treatment.
The levels of mRNA’s of interest were determined from medical prefrontal cortex punches using qPCR (See Materials and methods for details). Statistical significance was determined using the unpaired …
| MALE | Female | |||||
|---|---|---|---|---|---|---|
| Targets | Treatment | N | p-Value (Unpaired t-test) | Treatment | N | p-Value (Unpaired t-test) |
| CB1 | SHAM | 8 | 0.3279 | SHAM | 7 | 0.0643 |
| WIN | 8 | WIN | 8 | |||
| CB2 | SHAM | 7 | 0.9379 | SHAM | 6 | 0.1427 |
| WIN | 7 | WIN | 7 | |||
| TRPV1 | SHAM | 7 | 0.2281 | SHAM | 8 | 0.0098 |
| WIN | 8 | WIN | 7 | |||
| mGlu5 | SHAM | 7 | 0.0273 | SHAM | 8 | 0.0092 |
| WIN | 7 | WIN | 7 | |||
| mGlu1 | SHAM | 8 | 0.0400 | SHAM | 7 | 0.3910 |
| WIN | 8 | WIN | 8 | |||
| DAGL alpha | SHAM | 8 | 0.4370 | SHAM | 8 | 0.0024 |
| WIN | 8 | WIN | 8 | |||
| DAGL beta | SHAM | 7 | 0.4218 | SHAM | 6 | 0.3783 |
| WIN | 7 | WIN | 7 | |||
| NAPE-PLD | SHAM | 7 | 0.4258 | SHAM | 7 | 0.7518 |
| WIN | 7 | WIN | 7 | |||
| MAGL | SHAM | 8 | 0.5097 | SHAM | 8 | 0.2147 |
| WIN | 8 | WIN | 8 | |||
| FAAH | SHAM | 8 | 0.5505 | SHAM | 8 | 0.4096 |
| WIN | 8 | WIN | 7 | |||
| ABHD6 | SHAM | 7 | 0.2437 | SHAM | 6 | 0.6014 |
| WIN | 7 | WIN | 7 | |||
| CRIP1a | SHAM | 7 | 0.6414 | SHAM | 7 | 0.1887 |
| WIN | 7 | WIN | 7 | |||
Table 3
Reproduction data and pup weight after in utero WIN exposure.
Dam weight gain was calculated from GD 1 to GD 21 for n = 10 dams per treatment group. Pup weight at different postnatal days (PND) was calculated for n = 9–10 male and female pups from different …
| p-Value (ANOVA) | |||||
|---|---|---|---|---|---|
| Naive | SHAM | WIN | |||
| mean ± SEM | mean ± SEM | mean ± SEM | |||
| Dam weight gain (%) | |||||
| 33.8 ± 0.93 | 34.3 ± 1.91 | 33.1 ± 1.42 | 0.745 | ||
| Pregnancy length (days) | |||||
| Reproduction data | 22.4 ± 0.14 | 22.6 ± 0.29 | 22.6 ± 0.17 | 0.923 | |
| Litter size | 13 ± 0.82 | 12.9 ± 0.78 | 12.7 ± 0.67 | 0.854 | |
| PND 1 | 6.8 ± 0.2 | 6.7 ± 0.1 | 6.8 ± 0.2 | 0.965 | |
| PND 10 | 23.5 ± 0.89 | 24.6 ± 0.65 | 22.6 ± 0.72 | 0.207 | |
| Pup weight (grams) | PND 13 | 31.8 ± 0.62 | 30.3 ± 0.82 | 30.5 ± 0.75 | 0.339 |
| PND 25 | 64.5 ± 2.24 | 65.6 ± 0.77 | 66.7 ± 1.55 | 0.647 | |
| PND 90 | 452.7 ± 12.28 | 484.2 ± 12.52 | 481.7 ± 17.33 | 0.261 |
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.36234.022
