Cannabinoid and cholinergic systems interact during performance of a short-term memory task in the rat

It is now well established that cannabinoid agonists such as Δ⁹–tetrahydrocannabinol (THC), anandamide, and WIN 55,212-2 (WIN-2) produce potent and specific deficits in working memory (WM)/short-term memory (STM) tasks in rodents. Although mediated through activation of CB1 receptors located in memory-related brain regions such as the hippocampus and prefrontal cortex, these may, in part, be due to a reduction in acetylcholine release (i.e., cholinergic hypofunction). To determine the interaction between cannabinoid and cholinergic systems, we exposed rats treated with WIN-2 or cholinergic drugs to a hippocampal-dependent delayed nonmatch to sample (DNMS) task to study STM, and recorded hippocampal single-unit activity in vivo. WIN-2 induced significant deficits in DNMS performance and reduced the average firing and bursting rates of hippocampal principal cells through a CB1 receptor-mediated mechanism. Rivastigmine, an acetylcholinesterase inhibitor, reversed these STM deficits and normalized hippocampal discharge rates. Effects were specific to 1 mg/kg WIN-2 as rivastigmine failed to reverse the behavioral and physiological deficits that were observed in the presence of MK-801, an NMDA receptor antagonist. This supports the notion that cannabinoid-modulated cholinergic activity is a mechanism underlying the performance deficits in DNMS. Whether deficits are due to reduced nicotinic or muscarinic receptor activation, or both, awaits further analysis.Administration of both synthetic and phytocannabinoids, including Δ⁹–tetrahydrocannabinol (Δ⁹–THC), WIN 55,212-2 (WIN-2), and CP 55,940, impair working memory (WM) and short-term memory (STM) through a CB1 receptor-mediated mechanism in rats (Lichtman et al. 1995; Lichtman and Martin 1996; Hampson and Deadwyler 1998, 1999, 2000; Braida and Sala 2000; Egashira et al. 2002). This suggestive evidence for endocannabinoid involvement in memory formation was confirmed by Terranova and coworkers (1996), who demonstrated that the CB1 receptor antagonist rimonabant facilitated short-term olfactory memory, and this was partially reversed by the muscarinic receptor antagonist scopolamine. This suggests an interaction between cannabinoid and cholinergic systems such that endocannabinoid tone suppresses cholinergic transmission. Consequently, rats pretreated with eptastigmine, a second-generation cholinesterase inhibitor remained unaffected by the full CB1 receptor agonist CP 55,940 when tested in an eight arm radial maze (Braida and Sala 2000). And more recent evidence from Mishima and coworkers (2002) suggests that a block of cholinesterase with physostigmine and tetrahydroaminoacridine protects against WM impairments induced by Δ⁹–THC. These findings further support a potential role of the cholinergic system in cannabinoid-induced memory impairments.The exact mechanisms for this interaction still remain elusive, although cholinergic projection neurons from medial septum to hippocampus are likely to play an important role (Harkany et al. 2003, 2005; Fitz et al. 2008). However, the neuromodulatory action of pharmacologically active cannabinoids on septo-hippocampal cholinergic activity in vivo remains unexplored. Within the hippocampus, cannabinoids presynaptically inhibit the release of acetylcholine, possibly through the activation of CB1 receptors located on cholinergic nerve terminals given that these effects were blocked by rimonabant (Gifford and Ashby Jr. 1996; Gifford et al. 1997a, 2000; Kathmann et al. 2001a). Direct in vivo microdialysis studies in awake rats also showed cannabinoid-induced decreases in acetylcholine release in the hippocampus through a CB1 receptor-mediated mechanism (Gessa et al. 1997; Carta et al. 1998). High doses of rimonabant alone increase the amount of acetylcholine release in the hippocampus (Gessa et al. 1997, 1998) either by blocking the tonic inhibitory influence of endocannabinoids and/or through its inverse agonism at CB1 receptors. Such actions are in agreement with a 100% greater increase in electrically evoked hippocampal acetylcholine release in CB1^−/− mice (Kathmann et al. 2001b).In contrast, low doses of Δ⁹–THC (0.01–0.15 mg/kg), WIN-2 (0.01–0.5 mg/kg), and HU-210 (0.001–0.004 mg/kg) have been shown to enhance acetylcholine release (Acquas et al. 2000, 2001), indicating that cannabinoid modulation of acetylcholine release in the hippocampus is “biphasic.” This has been further supported by the work carried out by Tzavara and coworkers (2003), who demonstrated that low (0.5 mg/kg, intraperitoneally [i.p.]) and high (5 mg/kg, i.p.) doses of WIN-2 induce transient stimulation and prolonged inhibition of hippocampal acetylcholine efflux, respectively. This demonstrates that the dose of cannabinoids plays a key role in determining how much acetylcholine is released in the hippocampus.Such an interaction is likely to play an important role during the performance of a delayed nonmatch to sample (DNMS) task but has not been explored. Hence, a comprehensive pharmacological assessment was carried out here to (1) reveal the existence of such an interaction in terms of DNMS performance and (2) assess a possible cannabinoid-acetylcholine cross-talk on burst characteristics of hippocampal principal cells in CA3 and CA1.