- Email: [email protected]
Behavioral disorders in long-term alcohol exposed male rats’ offspring
S670
P.6.a. Addiction − Alcohol (basic)
P.6.a.004 Behavioral disorders in long-term alcohol exposed male rats’ offspring
their father consumed alcohol prior to conception. A direct relationship between paternal alcohol exposure and offspring health and behavior exists.
A. Voronina1 ° , G.M. Shayakhmetova2 , V.M. Kovalenko2 of Pharmacology and Toxicology NAMS of Ukraine, Kyiv, Ukraine; 2 Institute of Pharmacology and Toxicology NAMS of Ukraine, general toxicology, Kyiv, Ukraine
P.6.a.005 Administration of cannabidiol reduced reinforcement, motivation and relapse to alcohol
1 Institute
Introduction: It is evident that the human data on preconceptional alcohol consumption paternal effects are limited. And, what about the rodents’ data, they are insufficient and contradictory due to great diversity of animal species/strains and models of alcoholism. Due to these facts findings from studies in which males consume alcohol chronically are difficult to interpret, and they are potentially confounded. Moreover, multiple causal mechanisms for a paternal effect (including mutagenicity and epigenetic changes) have been suggested, but none seems satisfactory to explain all findings. Based on this information and considering that alcoholism is chronic disease highly prevalent in the world population, the present work reports the effects of long-term preconceptional 15% alcohol consumption by male Wistar rats on behavior of these males’ offspring. Materials and Methods: For the experimental (chronic alcoholism) model, reproducing male rats were selected according to the method for measuring voluntary alcohol self-administration in rats, which provides a continuous choice between an alcohol solution and water (two-bottle preference test). Following 150 days of 15% ethanol solution repeated self-administrations, the males from both groups were mated with virgin females in proestrus at the ratio 1 male : 2 females. Females were kept until natural delivery. The behavioral of offspring was studied on 21st day of postnatal development in the open-field and in hole board tests. In the open-field during a 2-min trial the number of sections crossed, vertical standing positions, defecations (fecal boli) and urinations were recorded. Results: In the table we represent the results of alcohol-addicted males’ offspring behavior in the open field in comparison with control group. Table 1. Effects of paternal alcohol exposure on offspring behavior in the open field (mean±S.E.M.; n = 24) Parameters
Group Control
Chronic alcoholism
Latent period, s Horizontal activity, number of sections crossed within 3 min peripheral sections central sections Vertical activity, number of rearing within 3 min Exploration activity, number of looking into holes within 3 min Grooming behavior, number of reactions within 3 min Urinations, number within 3 min Defecations, number within 3 min
1.29±0.20
1.03±0.30
92.83±4.60 20.50±2.60 11.41±0.94 9.95±0.78 1.25±0.17 1.0±0.18 1.25±0.25
62.09±6.90* 12.00±1.40* 8.40±1.01* 6.37±0.82* 1.68±0.37 0.90±0.13 1.12±0.19
Our findings suggest significant alteration of locomotor and emotional reactivity in offspring of chronically alcohol-treated males as compared with control. Particularly, in experimental group we recorded decrease of horizontal (number of crossed peripheral and central sections were less than in control 32% and 44% respectively), vertical (26%) and exploration activities (46%). Thus, our data provide support for relationship between offspring’s open field activity and paternal alcohol intake. Conclusions: Offspring not directly exposed to alcohol in utero may nevertheless be born with developmental abnormalities if
A. Viudez-Mart´ınez1 ° , M. Garc´ıa-Guti´errez1 , C. Navarr´on1 , M. Morales-Calero2 , F. Navarrete1 , A. Torres-Su´arez2 , J. Manzanares1 1 Instituto de Neurociencias, Laboratory of translational neuropsychopharmacology, Alicante, Spain; 2 Facuty of Pharmacy- Complutense University, Department of Pharmacy and Pharmaceutical Technology, Madrid, Spain Purpose of the study: Cannabidiol (CBD) is a phytocannabinoid constituent of Cannabis sativa that lacks the psychoactive effects of D9-tetrahydrocannabinol (THC). This compound has drawn increasing interest due to its broad spectrum pharmacological properties in neuropsychopharmacology as anxiolytic, antidepressant, antipsychotic, antiepileptic and neuroprotector [1,2]. Alongside, it has been shown that CBD also attenuates alcohol-induced neurodegeneration in the cortico-limbic pathway by reducing the number of Fluoro-Jade B positive cells in the entorhinal cortex in a modified Majchrowicz alcohol binge model in mice. This result suggests the neuroprotective activity of CBD [3]. The aim of this study was to evaluate the effect of CBD in alcohol reinforcement, motivation and relapse. Methods used: To evaluate the effects of CBD on ethanolmediated physiological, reinforcing and motivational actions, different experimental paradigms were carried out in male C57BL/6J mice. CBD-mediated modulation of time-related and dose-response effects of ethanol on rectal temperature (3 g/kg, p.o.), handling-induced convulsions (HIC; 4 g/kg, i.p.) and blood ethanol concentrations (BEC; 3 g/kg p.o.) were analyzed. Voluntary ethanol consumption in a two-bottle choice paradigm was performed to study the effect of CBD (30, 60 and 120 mg/kg[NRF1] /day, i.p.) on ethanol intake and preference. In addition, oral ethanol self-administration experiment was performed to evaluate the effect of CBD on the reinforcement and motivation, for this experiment poly-e-caprolactone spherical microparticles with small pores which provided a CBD continuous controlled release were used (30 mg/kg, s.c.). A further oral self-administration experiment was carried out to study CBD effects on relapse (60 and 120 mg/kg/day, i.p.) induced by ethanol. Real time-PCR experiments [NRF2] were performed to analyze the gene expression of tyrosine hydroxylase (TH) in the ventral tegmental area (VTA), and m-opioid receptor gene expression in the nucleus accumbens (NAcc). Results: The administration of CBD blocked ethanol-induced hypothermia and convulsions compared to control group (Twoway RM ANOVA; p < 0.001). There was no effect on ethanol absorption produced by CBD administration compared to vehicle group. CBD significantly reduced ethanol intake and preference in the two-bottle choice model and ethanol consumption and motivation in the oral ethanol self-administration paradigm compared to vehicle-treated mice (Two-way RM ANOVA; p < 0.05). Interestingly, CBD significantly reduced ethanol-induced relapse (Two-way RM ANOVA; p = 0.006). In addition, CBD significantly decreased TH gene expression in the VTA (40%) compared to the vehicle group (Student’s t-test, p < 0.05) and reduced m-opioid
P.6.a. Addiction − Alcohol (basic)
P.6.a.004 Behavioral disorders in long-term alcohol exposed male rats’ offspring
their father consumed alcohol prior to conception. A direct relationship between paternal alcohol exposure and offspring health and behavior exists.
A. Voronina1 ° , G.M. Shayakhmetova2 , V.M. Kovalenko2 of Pharmacology and Toxicology NAMS of Ukraine, Kyiv, Ukraine; 2 Institute of Pharmacology and Toxicology NAMS of Ukraine, general toxicology, Kyiv, Ukraine
P.6.a.005 Administration of cannabidiol reduced reinforcement, motivation and relapse to alcohol
1 Institute
Introduction: It is evident that the human data on preconceptional alcohol consumption paternal effects are limited. And, what about the rodents’ data, they are insufficient and contradictory due to great diversity of animal species/strains and models of alcoholism. Due to these facts findings from studies in which males consume alcohol chronically are difficult to interpret, and they are potentially confounded. Moreover, multiple causal mechanisms for a paternal effect (including mutagenicity and epigenetic changes) have been suggested, but none seems satisfactory to explain all findings. Based on this information and considering that alcoholism is chronic disease highly prevalent in the world population, the present work reports the effects of long-term preconceptional 15% alcohol consumption by male Wistar rats on behavior of these males’ offspring. Materials and Methods: For the experimental (chronic alcoholism) model, reproducing male rats were selected according to the method for measuring voluntary alcohol self-administration in rats, which provides a continuous choice between an alcohol solution and water (two-bottle preference test). Following 150 days of 15% ethanol solution repeated self-administrations, the males from both groups were mated with virgin females in proestrus at the ratio 1 male : 2 females. Females were kept until natural delivery. The behavioral of offspring was studied on 21st day of postnatal development in the open-field and in hole board tests. In the open-field during a 2-min trial the number of sections crossed, vertical standing positions, defecations (fecal boli) and urinations were recorded. Results: In the table we represent the results of alcohol-addicted males’ offspring behavior in the open field in comparison with control group. Table 1. Effects of paternal alcohol exposure on offspring behavior in the open field (mean±S.E.M.; n = 24) Parameters
Group Control
Chronic alcoholism
Latent period, s Horizontal activity, number of sections crossed within 3 min peripheral sections central sections Vertical activity, number of rearing within 3 min Exploration activity, number of looking into holes within 3 min Grooming behavior, number of reactions within 3 min Urinations, number within 3 min Defecations, number within 3 min
1.29±0.20
1.03±0.30
92.83±4.60 20.50±2.60 11.41±0.94 9.95±0.78 1.25±0.17 1.0±0.18 1.25±0.25
62.09±6.90* 12.00±1.40* 8.40±1.01* 6.37±0.82* 1.68±0.37 0.90±0.13 1.12±0.19
Our findings suggest significant alteration of locomotor and emotional reactivity in offspring of chronically alcohol-treated males as compared with control. Particularly, in experimental group we recorded decrease of horizontal (number of crossed peripheral and central sections were less than in control 32% and 44% respectively), vertical (26%) and exploration activities (46%). Thus, our data provide support for relationship between offspring’s open field activity and paternal alcohol intake. Conclusions: Offspring not directly exposed to alcohol in utero may nevertheless be born with developmental abnormalities if
A. Viudez-Mart´ınez1 ° , M. Garc´ıa-Guti´errez1 , C. Navarr´on1 , M. Morales-Calero2 , F. Navarrete1 , A. Torres-Su´arez2 , J. Manzanares1 1 Instituto de Neurociencias, Laboratory of translational neuropsychopharmacology, Alicante, Spain; 2 Facuty of Pharmacy- Complutense University, Department of Pharmacy and Pharmaceutical Technology, Madrid, Spain Purpose of the study: Cannabidiol (CBD) is a phytocannabinoid constituent of Cannabis sativa that lacks the psychoactive effects of D9-tetrahydrocannabinol (THC). This compound has drawn increasing interest due to its broad spectrum pharmacological properties in neuropsychopharmacology as anxiolytic, antidepressant, antipsychotic, antiepileptic and neuroprotector [1,2]. Alongside, it has been shown that CBD also attenuates alcohol-induced neurodegeneration in the cortico-limbic pathway by reducing the number of Fluoro-Jade B positive cells in the entorhinal cortex in a modified Majchrowicz alcohol binge model in mice. This result suggests the neuroprotective activity of CBD [3]. The aim of this study was to evaluate the effect of CBD in alcohol reinforcement, motivation and relapse. Methods used: To evaluate the effects of CBD on ethanolmediated physiological, reinforcing and motivational actions, different experimental paradigms were carried out in male C57BL/6J mice. CBD-mediated modulation of time-related and dose-response effects of ethanol on rectal temperature (3 g/kg, p.o.), handling-induced convulsions (HIC; 4 g/kg, i.p.) and blood ethanol concentrations (BEC; 3 g/kg p.o.) were analyzed. Voluntary ethanol consumption in a two-bottle choice paradigm was performed to study the effect of CBD (30, 60 and 120 mg/kg[NRF1] /day, i.p.) on ethanol intake and preference. In addition, oral ethanol self-administration experiment was performed to evaluate the effect of CBD on the reinforcement and motivation, for this experiment poly-e-caprolactone spherical microparticles with small pores which provided a CBD continuous controlled release were used (30 mg/kg, s.c.). A further oral self-administration experiment was carried out to study CBD effects on relapse (60 and 120 mg/kg/day, i.p.) induced by ethanol. Real time-PCR experiments [NRF2] were performed to analyze the gene expression of tyrosine hydroxylase (TH) in the ventral tegmental area (VTA), and m-opioid receptor gene expression in the nucleus accumbens (NAcc). Results: The administration of CBD blocked ethanol-induced hypothermia and convulsions compared to control group (Twoway RM ANOVA; p < 0.001). There was no effect on ethanol absorption produced by CBD administration compared to vehicle group. CBD significantly reduced ethanol intake and preference in the two-bottle choice model and ethanol consumption and motivation in the oral ethanol self-administration paradigm compared to vehicle-treated mice (Two-way RM ANOVA; p < 0.05). Interestingly, CBD significantly reduced ethanol-induced relapse (Two-way RM ANOVA; p = 0.006). In addition, CBD significantly decreased TH gene expression in the VTA (40%) compared to the vehicle group (Student’s t-test, p < 0.05) and reduced m-opioid