Yosefa Avraham

Low dose anandamide affects food intake, cognitive function, neurotransmitter and corticosterone levels in diet-restricted mice

This investigation reports the possible role of the endocannabinoid anandamide on modulating the behavioral and neurochemical consequences of semi-starvation. We studied the effect of very low dose anandamide (0.001 mg/kg) administration on food intake, cognitive function and catecholaminergic and serotonergic pathways in two murine brain areas concerned with appetite (hypothalamus) and learning (hippocampus), and the peripheral corticosterone response to the stress of 40% diet restriction. Anandamide-treated mice consumed 44% more food (P<0.05) during 1 week of 2.5-h feeding each day. In the hypothalamus, there were significantly increased concentrations of norepinephrine (P<0.01), dopamine (P<0.05) and 5-hydroxytryptamine (5-HT) (P<0.001). In the hippocampus, anandamide increased significantly norepinephrine and dopamine, but decreased 5-HT (all at P<0.001). Diet restriction was accompanied in both areas by a significant decrease in all neurotransmitter concentrations that were partially restored by anandamide for dopamine and 5-HT, but not for norepinephrine. In animals on diet restriction, anandamide significantly improved impaired maze performance. Norepinephrine turnover and plasma corticosterone levels were also raised significantly by anandamide. The fact that low dose anandamide improved food intake, cognitive function and reversed some of the neurotransmitter changes caused by diet restriction, might have implications for the treatment of cachexia associated with acquired immunodeficiency syndrome (AIDS) and cancer, for mood changes sometimes associated with dieting, and in the extreme case, of patients with anorexia.

Short-term fasting and prolonged semistarvation have opposite effects on 2-AG levels in mouse brain

2-Arachidonoyl glycerol (2-AG) levels in whole mouse brain and two of its regions-hippocampus and hypothalamus-were determined after diet restriction (between 60 and 40%) lasting 12 days. The diet restriction lowered the level of 2-AG, which in the hypothalamus depended on the severity of the diet restriction, while the level in the hippocampus was not dependent on the diet regimen. As these observations differ from previously published data showing elevation of 2-AG levels in rat brain after 24 h of severe food restriction, we measured 2-AG levels in whole mouse brain after a comparable period of full starvation (fasting). We confirmed the elevation of 2-AG levels. It seems possible that these time-dependent variations of 2-AG levels may be of importance as a general coping strategy by animals during periods of starvation.

Nutritional Status, Cognition, and Survival A NEW ROLE FOR LEPTIN AND AMP KINASE

Although adequate nutrition is essential for optimal neural activity and survival, mild energy restriction may improve cognition and prolong longevity. Energy status is monitored by the cellular AMP-activated protein kinase (AMPK) system, whereas leptin regulates total energy balance. We investigated the roles of AMPK and leptin in cognition and survival under diet restriction (DR). Hippocampal AMPK activity increases with energy restriction. Modest activation (DR to 60%) induces neurogenesis and improves cognition. However, DR to 40% augmented AMPK activity, reduced cognition and catecholamines, and increased neural apoptosis and mortality. Leptin signaling is preserved only in DR to 60%, countering the effects of AMPK “overactivation” by preventing neuroapoptosis, restoring noradrenergic activity and behavioral performance, and increasing longevity. The balance between leptin and AMPK is crucial in determining neuronal fate, cognitive ability, and survival. Should these findings extend to Man, then controlled activation of AMPK may improve neurodegenerative diseases, and leptin may have a new role in treating stress-associated malnutrition.

Cannabidiol ameliorates cognitive and motor impairments in bile-duct ligated mice via 5-HT1A receptor activation

Background and purpose:  We aimed to demonstrate the involvement of 5‐HT1A receptors in the therapeutic effect of cannabidiol, a non‐psychoactive constituent of Cannabis sativa, in a model of hepatic encephalopathy induced by bile‐duct ligation (BDL) in mice.

Animal models in the investigation of anorexia.

Anorexia nervosa (AN) is an eating disorder of unknown origin that most commonly occurs in women and usually has its onset in adolescence. Patients with AN invariably have a disturbed body image and an intense fear of weight gain. There is currently no definitive treatment for this disease, which carries a 20% mortality over 20 years. Development of an appropriate animal model of AN has been difficult, as the etiology of this eating disorder likely involves a complex interaction between genetic, environmental, social, and cultural factors. In this review, we focus on several possible rodent models of AN. In our laboratory, we have developed and studied three different mouse models of AN based on clinical profiles of the disease; separation stress, activity, and diet restriction (DR). In addition, we discuss the spontaneous mouse mutation anx/anx and several mouse gene knockout models, which have resulted in an anorexic phenotype. We highlight what has been learned from each of these models and possibilities for future models. It is hoped that a combination of the study of such models, together with genetic and clinical studies in patients, will lead to more rational and successful prevention/treatment of this tragic, and often fatal, disease.

Endocannabinoids affect neurological and cognitive function in thioacetamide-induced hepatic encephalopathy in mice

Endocannabinoids function as neurotransmitters and neuromodulators in the central nervous system via specific receptors and apparently have a neuroprotective role. We assumed that the endocannabinoid system could be involved in the pathogenesis of hepatic encephalopathy (HE), a neuropsychiatric syndrome due to liver disease. We used a mouse model of a thioacetamide induced fulminant hepatic failure. We found that the levels of the endocannabinoid 2-arachidonoyl-glycerol (2-AG) were elevated in the brain. Treatment with either 2-AG or with the CB1 receptor antagonist, SR141716A, improved a neurological score, activity and cognitive function. Activation of the CB2 receptor by a selective agonist, HU308, also improved the neurological score. 2-AG activity could be blocked with the specific CB2 receptor antagonist SR144528A. The CB1 receptor agonist noladin ether was inactive. We conclude that the endocannabinoid system may play an important role in the pathogenesis of HE. Modulation of this system either by exogenous agonists specific for the CB2 receptors or possibly also by antagonists to the CB1 receptors may have therapeutic potential.

Cannabidiol ameliorates cognitive and motor impairments in mice with bile duct ligation

BACKGROUND/AIMS The endocannabinoid system in mice plays a role in models of human cirrhosis and hepatic encephalopathy (HE), induced by a hepatotoxin. We report now the therapeutic effects of cannabidiol (CBD), a non-psychoactive constituent of Cannabis sativa, on HE caused by bile duct ligation (BDL), a model of chronic liver disease. METHODS CBD (5mg/kg; i.p.) was administered over 4weeks to mice that had undergone BDL. RESULTS Cognitive function in the eight arm maze and the T-maze tests, as well as locomotor function in the open field test were impaired by the ligation and were improved by CBD. BDL raised hippocampal expression of the TNF-alpha-receptor 1 gene, which was reduced by CBD. However, BDL reduced expression of the brain-derived neurotrophic factor (BDNF) gene, which was increased by CBD. The effects of CBD on cognition, locomotion and on TNF-alpha receptor 1 expression were blocked by ZM241385, an A(2)A adenosine receptor antagonist. BDL lowers the expression of this receptor. CONCLUSIONS The effects of BDL apparently result in part from down-regulation of A(2)A adenosine receptor. CBD reverses these effects through activation of this receptor, leading to compensation of the ligation effect.

Alterations in septohippocampal cholinergic innervations and related behaviors after early exposure to heroin and phencyclidine

Mice were exposed to diacetylmorphine (heroin) or phencyclidine (PCP) prenatally or neonatally. At a later age, they were tested for hippocampus-related behavioral deficits and concomitant alterations in the septohippocampal cholinergic innervations. Actually, this is an application of the previously established phenobarbital neuroteratogenicity model to heroin and PCP. Prenatal exposure was accomplished transplacentally by injecting the mother 10 mg/kg heroin or PCP on gestation days 9-18. Neonatal administrations were applied directly by injections of 10 mg/kg of either drug to the pups between neonatal days 2-21. At the age of 50 days, mice exposed to heroin and PCP prenatally exhibited a 107% and 159% increase in their muscarinic cholinergic receptors Bmax, respectively. Neonatal exposure to heroin or PCP caused an 83% and 76% increase in the receptors respectively. On the behavioral level, both prenatal and neonatal exposure to heroin or PCP reduced performance in the hippocampus related eight-arm maze and Morris mazes. Depending on the drug, the test and the period of drug administration, the reduction ranged between 10% and 75%. The results suggest that heroin and PCP induce alterations in the septohippocampal cholinergic innervations and in related behavioral performance. Further studies are necessary in order to connect the biochemical and behavioral events in causal relationships.

Behavioral and neurochemical alterations caused by diet restriction - the effect of tyrosine administration in mice

We have investigated the effect of tyrosine administration on the cognitive and neurochemical alterations caused by diet restriction (DR) in mice, as a possible model for some of the behavioral symptoms of patients with anorexia nervosa. Young female mice were fed to 100, 60, and 40% of the calculated daily nutritional requirements for a period of up to 18 days. Cognitive function was evaluated using a modified eight-arm maze with water as a reward. Animals fed to 60% of controls showed significantly improved maze performance while this was impaired in animals on DR to 40%. However, in these animals, injections of tyrosine (100 mg/kg/day) restored performance. Improved maze performance in the 60% DR and 40% DR + tyrosine animals was related to increased beta:alpha tone in the hippocampus- an area, together with the septum, responsible for spatial learning. This was associated with changes in alpha- and beta-receptor density (Bmax), without affecting affinity (Kd); and increased norepinephrine (NE) in the 40% DR + tyrosine group, and methoxyhydroxyphenylglycol (MHPG) in both groups. In the hypothalamus, the brain area responsible for energy metabolism, there was a progressive increase in alpha:beta tone with increasing DR associated with changes in Bmax. Tyrosine treatment reversed these alterations. Tyrosine improves some of the neurobiological disturbances of DR without causing an increase in body weight. Such a strategy might have important implications for the possible treatment of patients with anorexia nervosa.

Cannabinoids ameliorate cerebral dysfunction following liver failure via AMP-activated protein kinase

Hepatic encephalopathy (HE) is a neuropsychiatry disorder of complex pathogenesis caused by acute or chronic liver failure. We studied the etiology of cerebral dysfunction in a murine model of HE induced by either bile duct ligation or thioacetamide administration. We report that stimulation of cerebral AMP‐activated protein kinase (AMPK), a major intracel‐lular energy sensor, is a compensatory response to liver failure. This function of AMPK is regulated by endo‐cannabinoids. The cannabinoid system controls systemic energy balance via the cannabinoid receptors CB‐1 and CB‐2. Under normal circumstances, AMPK activity is mediated by CB‐1 while CB‐2 is barely detected. However, CB‐2 is strongly stimulated in response to liver failure. Administration of ∆9‐tetrahy‐drocannabinol (THC) augmented AMPK activity and restored brain function in WT mice but not in their CB‐2 KO littermates. These results suggest that HE is a disease of energy flux. CB‐2 signaling is a cerebral stress response mechanism and makes AMPK a promising target for its treatment by modulating the canna‐binoid system.—Dagon, Y., Avraham, Y., Ilan, Y., Mechoulam, R., Berry, E. M. Cannabinoids ameliorate cerebral dysfunction following liver failure via AMP‐activated protein kinase. FASEB J. 21, 2431–2441 (2007)