Prenatal and early postnatal cannabis exposure interactions with adolescent chronic stress on anxiety-like, depression-like, and risk-taking behaviour methods
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Methods
Mouse dams orally consumed 5 mg/kg THC in whole cannabis oil daily from GD1-PD10. Offspring were exposed to chronic mild unpredictable stress throughout adolescence (PD28-56). From PD58, mice were challenged with a battery of tests to measure anxiety-like (elevated plus maze, open field test), stress coping (forced swim test, tail suspension test), anhedonia-like (sucrose preference), risk-taking behaviour (wire beam bridge), and social motivation (3 chamber sociability and social novelty task). Brain slices were taken 90 min after forced swim test to analyze c-Fos expression.
Methods
All protocols were in accordance with the ethical guidelines established by the Canadian Council for Animal Care and were approved by the University of Calgary Animal Care Committee. Adult (postnatal day (PD) 70 +) female and male C57Bl/6 mice used for breeding were from Charles River (Quebec, Canada). Animal studies are reported in compliance with the ARRIVE guidelines (Percie du Sert et al. ). They were housed in a 12 h/12 h light/dark cycle room (dark phase from 10:00 am to 10:00 pm) under controlled temperature (22 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\pm$$\end{document} 1 °C) and humidity conditions (40%) with free access to food and water. Dam vaginal openings were visually inspected...
Statistics
All results are represented as mean ± standard error of the mean (SEM) and were statistically analyzed and graphically presented using GraphPad PRISM® version 10 (GraphPad Inc., LaJolla, CA USA). The statistical tests used to analyse different data sets and the number of animals and litters used are indicated in each figure caption. Groups size was selected based on our previous studies (Sallam et al.; Peterson et al. ). Offspring from at least 4 different litters were included in each group. Normally distributed two group comparisons were made with a t-test. The proportion of mice crossing the wire beam bridge was tested with a Fisher's Exact Test first to examine sex differences, and then between the four treatment groups. Differences between specific treatment group pairs were made with a post-hoc Fisher's Exact Test with a Bonferroni correction. All ot...
Anxiety-like and risk-taking behaviour
The wire beam bridge task was previously used to evaluate risk-taking and risk assessment behaviour in rats exposed to THC during gestation via s.c. injections (Frau et al.; Traccis et al. ). Notably, mice remained on the apparatus for 10 min in our study as opposed to 3 min in theirs. Of the mice that attempted to cross the bridge at least once, 82% of them did so after 3 min, and therefore would have been counted as "non-crossers" in a shorter test. This likely explains the higher rate of bridge crossing in our animals. While the data from male and female rats were published separately and thus not directly compared (Frau et al.; Traccis et al. ), the same exposure procedure produced different results between sexes: in males, prenatal THC exposed animals, when acutely injected with THC, crossed the wire beam bridge more quickly and performed fewer stretched-a...
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Evidence quotes (4)
Mouse dams orally consumed 5 mg/kg THC in whole cannabis oil daily from GD1-PD10. Offspring were exposed to chronic mild unpredictable stress throughout adolescence (PD28-56). From PD58, mice were challenged with a battery of tests to measure anxiety-like (elevated plus maze, open field test), stress coping (forced swim test, tail suspension test), anhedonia-like (sucrose preference), risk-taking behaviour (wire beam bridge), and social motivation (3 chamber sociability and social novelty task). Brain slices were taken 90 min after forced swim test to analyze c-Fos expression.
All protocols were in accordance with the ethical guidelines established by the Canadian Council for Animal Care and were approved by the University of Calgary Animal Care Committee. Adult (postnatal day (PD) 70 +) female and male C57Bl/6 mice used for breeding were from Charles River (Quebec, Canada). Animal studies are reported in compliance with the ARRIVE guidelines (Percie du Sert et al. ). They were housed in a 12 h/12 h light/dark cycle room (dark phase from 10:00 am to 10:00 pm) under controlled temperature (22 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\pm$$\end{document} 1 °C) and humidity conditions (40%) with free access to food and water. Dam vaginal openings were visually inspected daily to determine the phases of the estrous cycle. Studies were designed to generate experimental groups of equal size using randomization. Pregnant female mice were randomly assigned to receive cannabis oil extract or vehicle. Dams received 5 mg/kg THC in whole cannabis extract (25 mg/mL THC,...
All results are represented as mean ± standard error of the mean (SEM) and were statistically analyzed and graphically presented using GraphPad PRISM® version 10 (GraphPad Inc., LaJolla, CA USA). The statistical tests used to analyse different data sets and the number of animals and litters used are indicated in each figure caption. Groups size was selected based on our previous studies (Sallam et al.; Peterson et al. ). Offspring from at least 4 different litters were included in each group. Normally distributed two group comparisons were made with a t-test. The proportion of mice crossing the wire beam bridge was tested with a Fisher's Exact Test first to examine sex differences, and then between the four treatment groups. Differences between specific treatment group pairs were made with a post-hoc Fisher's Exact Test with a Bonferroni correction. All other data were first analysed using a 3-Way ANOVA (repeated measures for weight data) with post-hoc Sidak's test when indicated by an interaction effect. In that case, post-hoc tests were performed between specific groups (for each sex where sex differences are noted): vehicle/control mice vs v...
The wire beam bridge task was previously used to evaluate risk-taking and risk assessment behaviour in rats exposed to THC during gestation via s.c. injections (Frau et al.; Traccis et al. ). Notably, mice remained on the apparatus for 10 min in our study as opposed to 3 min in theirs. Of the mice that attempted to cross the bridge at least once, 82% of them did so after 3 min, and therefore would have been counted as "non-crossers" in a shorter test. This likely explains the higher rate of bridge crossing in our animals. While the data from male and female rats were published separately and thus not directly compared (Frau et al.; Traccis et al. ), the same exposure procedure produced different results between sexes: in males, prenatal THC exposed animals, when acutely injected with THC, crossed the wire beam bridge more quickly and performed fewer stretched-attend postures. In contrast, female rats prenatally exposed to THC crossed the bridge significantly later than vehicle controls. Additionally, control animals also showed sex differences in bridge crossing: male rats rarely cross the bridge at baseline, whereas most female control rats crossed the b...
Machine-readable layer
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