Cannabis sativa is currently the most commonly-used, illicit drug in the United States. Cannabis use, particularly by adolescents, increases the risk of developing schizophrenia-like psychoses in later life. In addition, people with bipolar disorder have a 20-40% lifetime prevalence of cannabis use, compared to 6% in the general population. These and other epidemiological data demonstrate that cannabis use predisposes susceptible individuals to the development of psychiatric disorders. Glycogen synthase kinase 3 (GSK-3) is emerging as an important regulatory kinase in the limbic brain;over-activity of GSK-3 has been linked to both bipolar disorder and schizophrenia. GSK-3 is phosphorylated and inactivated by protein kinases, including Akt and is a down-stream component of several neurotransmitters involved in mood regulation and psychosis. Data presented in this proposal demonstrate that prolonged cannabinoid-1 receptor (CB1R) activation by the cannabis constituent, ?9-tetrahydrocannabinol (THC), significantly decreases GSK-3 phosphorylation in neurons. Since GSK-3 activity is reduced by phosphorylation, these data are consistent with enhanced GSK-3 activity following THC exposure. A second CB1R agonist, CP55940 shared these effects of THC. These preliminary data, together with data in the literature regarding D2 dopamine receptor signaling, were used to formulate the hypothesis that prolonged CB1R activation results in recruitment of ss-arrestin;ss-arrestin functions as a scaffold protein, bringing Akt in proximity with the protein phosphatase, PP2A. Akt is dephosphorylated and inactivated, resulting in dysinhibition of GSK-3 activity. Since GSK-3 over-activity is associated with mood dysregulation and psychosis, these data lead to the hypothesis that THC-mediated increase in GSK-3 activity contributes to the relationship between cannabis use and psychiatric disorders. The objective of the current project is to test the specific hypothesis that CB1R-mediated activation of GSK-3 occurs through ss-arrestin-mediated inhibition of Akt;is brain region specific and contributes to the anxiogenic and stress-enhancing effects of THC and other cannabinoid agonists.
The specific aims of this project are: (1) to determine the mechanism by which CB1R agonists alter the phosphorylation state of GSK-3 in primary neurons in culture;(2) the determine the effects of acute and chronic exposure of mice to CB1R agonists and antagonists on the phosphorylation and activities of Akt and GSK-3 in brain regions, particularly those of the limbic system: (3) to determine the role of ss-arrestin in the effects of CB1R agonists and antagonists in behavioral assays of anxiety and stress by comparing their effects in wild type and ss-arrestin-2 null mice. Successful completion of the studies outlined in this proposal will advance our long-term objective to determine the mechanisms by which cannabis exposure predisposes individuals to the development of psychiatric illness.

Public Health Relevance

Cannabis sativa is an annual plant that has been used by humans for thousands of years as a medicinal and recreational drug. While most individuals who use cannabis are not adversely affected, some users develop serious psychiatric disorders, including schizophrenia and bipolar disorder. The studies in this proposal will test the hypothesis that the primary psychoactive chemical of cannabis, ?9-tetrahydrocannabinol (THC) produces an over-activation of the enzyme glycogen synthase kinase in the brain and that this mechanism contributes to negative effects of THC on mood and cognition and is responsible for the increased incidence of psychiatric disorders in cannabis users.

Agency
National Institute of Health (NIH)
Institute
National Institute on Drug Abuse (NIDA)
Type
Research Project (R01)
Project #
5R01DA026996-03
Application #
8233541
Study Section
Molecular Neuropharmacology and Signaling Study Section (MNPS)
Program Officer
Wu, Da-Yu
Project Start
2010-03-01
Project End
2015-02-28
Budget Start
2012-03-01
Budget End
2013-02-28
Support Year
3
Fiscal Year
2012
Total Cost
$350,170
Indirect Cost
$119,795
Name
Medical College of Wisconsin
Department
Pharmacology
Type
Schools of Medicine
DUNS #
937639060
City
Milwaukee
State
WI
Country
United States
Zip Code
53226
Hillard, Cecilia J (2014) Stress regulates endocannabinoid-CB1 receptor signaling. Semin Immunol 26:380-8
Roberts, Christopher J; Stuhr, Kara L; Hutz, Michael J et al. (2014) Endocannabinoid signaling in hypothalamic-pituitary-adrenocortical axis recovery following stress: effects of indirect agonists and comparison of male and female mice. Pharmacol Biochem Behav 117:17-24
Liedhegner, Elizabeth Sabens; Vogt, Caleb D; Sem, Daniel S et al. (2014) Sterol carrier protein-2: binding protein for endocannabinoids. Mol Neurobiol 50:149-58
Liedhegner, Elizabeth Sabens; Sasman, Amy; Hillard, Cecilia J (2014) Brain region-specific changes in N-acylethanolamine contents with time of day. J Neurochem 128:491-506
McLaughlin, R J; Hill, M N; Dang, S S et al. (2013) Upregulation of CBýýý receptor binding in the ventromedial prefrontal cortex promotes proactive stress-coping strategies following chronic stress exposure. Behav Brain Res 237:333-7
Lee, Tiffany T-Y; Hill, Matthew N; Hillard, Cecilia J et al. (2013) Temporal changes in N-acylethanolamine content and metabolism throughout the peri-adolescent period. Synapse 67:4-10
Roberts, C J; Stuhr, K L; Hillard, C J (2012) Swim stress differentially affects limbic contents of 2-arachidonoylglycerol and 2-oleoylglycerol. Neuroscience 204:74-82
Hillard, C J; Weinlander, K M; Stuhr, K L (2012) Contributions of endocannabinoid signaling to psychiatric disorders in humans: genetic and biochemical evidence. Neuroscience 204:207-29
Hill, Matthew N; Hillard, Cecilia J; McEwen, Bruce S (2011) Alterations in corticolimbic dendritic morphology and emotional behavior in cannabinoid CB1 receptor-deficient mice parallel the effects of chronic stress. Cereb Cortex 21:2056-64