Bettina Platt

Glutamate receptor function in learning and memory.

The contribution of glutamate to synaptic transmission, plasticity and development is well established; current evidence is based on diverse approaches to decipher function and malfunction of this principal transmitter. With respect to learning and memory, we are now able to identify more specifically the role played by the three main glutamate receptor classes in learning and memory: centre stage is clearly the NMDA receptor, with overwhelming evidence proving its involvement in the actual learning process (encoding), throughout the animal kingdom. This is discussed with respect to many different types of learning. Evidence for the contribution of the AMPA receptors (AMPARs) is less clear-cut due to the general problem of specificity: block of AMPARs will shutdown neuronal communication, and this will affect various components essential for learning. Therefore, the role of AMPARs cannot be established in isolation. Problems of interpretation are outlined and a specific involvement of AMPARs in the regulation of neuronal excitation related to learning is proposed. Metabotropic glutamate receptors (mGluRs) may contribute very little to the actual acquisition of new information. However, memory formation appears to require mGluRs, through the modulation of consolidation and/or recall. Overall, mGluR functions seem variable and dependent on brain structure and learning task.

The Cholinergic system and spatial learning

Acetlylcholine (ACh) in the central nervous system is critical for a multitude of functions. Here, we concentrate on declarative memory in humans, and its equivalent episodic-like memory in rodents and highlight current understanding of cholinergic system in these processes. Spatial memory formation represents a simple form of episodic-like memory in rodents that engages the basal forebrain cholinergic system and its target structures. In these, ACh exerts numerous functions. (1) During spatial acquisition learning, ACh efflux into the extracellular space is immediate in hippocampus and cortex; during consolidation of spatial reference memory, ACh levels are low. These requirements explain why ACh receptor blockade during acquisition blocks memory formation, and it is also consonant with the notion that an unspecific enhancement of cholinergic activity during consolidation is detrimental to memory formation. (2) Working and short-term memory for spatial locations engages the nucleus basalis – prefrontal cortex ACh system. ACh activity is trial related and maintained for some time post-training. (3) Striatal cholinergic activity is increased during stimulus–response learning and behavioural flexibility (reversal learning, extinction) providing a possible switch between different behavioural strategies. (4) At present, there is no clear difference between muscarinic and nicotinergic systems with respect to spatial learning. Antagonists of the respective receptors impair memory formation, agonists can reverse these deficits or may, under specific conditions act more like a general cognitive enhancers by way of improving attention. (5) Data reviewed here do not provide conclusive evidence for muscarinic or nicotinic receptors presenting as novel therapeutic targets, and there is no clear indication for ACh derived novel biomarkers for translational medicine. Unresolved and contradictory results are highlighted and discussed.

The cholinergic system and hippocampal plasticity.

Acetylcholine is an essential excitatory neurotransmitter in the central nervous system and undertakes a vital role in cognitive function. Consequently, there is ample evidence to suggest the involvement of both nicotinic and muscarinic acetylcholine receptors in the modulation of synaptic plasticity, which is believed to be the molecular correlate of learning and memory. In the hippocampus in particular, multiple subtypes of both nicotinic and muscarinic receptors are present at presynaptic and postsynaptic loci of both principal neurons and inhibitory interneurons, where they exert profound bi-directional influences on synaptic transmission. Further evidence points to a role for cholinergic activation in the induction and maintenance of synaptic plasticity, and key influences on hippocampal network oscillations. The present review examines these multiple roles of acetylcholine in hippocampal plasticity.

Cannabidiol Targets Mitochondria to Regulate Intracellular Ca2+ Levels

Cannabinoids and the endocannabinoid system have attracted considerable interest for therapeutic applications. Nevertheless, the mechanism of action of one of the main nonpsychoactive phytocannabinoids, cannabidiol (CBD), remains elusive despite potentially beneficial properties as an anti-convulsant and neuroprotectant. Here, we characterize the mechanisms by which CBD regulates Ca2+ homeostasis and mediates neuroprotection in neuronal preparations. Imaging studies in hippocampal cultures using fura-2 AM suggested that CBD-mediated Ca2+ regulation is bidirectional, depending on the excitability of cells. Under physiological K+/Ca2+ levels, CBD caused a subtle rise in [Ca2+]i, whereas CBD reduced [Ca2+]i and prevented Ca2+ oscillations under high-excitability conditions (high K+ or exposure to the K+ channel antagonist 4AP). Regulation of [Ca2+]i was not primarily mediated by interactions with ryanodine or IP3 receptors of the endoplasmic reticulum. Instead, dual-calcium imaging experiments with a cytosolic (fura-2 AM) and a mitochondrial (Rhod-FF, AM) fluorophore implied that mitochondria act as sinks and sources for CBDs [Ca2+]i regulation. Application of carbonylcyanide-p-trifluoromethoxyphenylhydrazone (FCCP) and the mitochondrial Na+/Ca2+ exchange inhibitor, CGP 37157, but not the mitochondrial permeability transition pore inhibitor cyclosporin A, prevented subsequent CBD-induced Ca2+ responses. In established human neuroblastoma cell lines (SH-SY5Y) treated with mitochondrial toxins, CBD (0.1 and 1 μm) was neuroprotective against the uncoupler FCCP (53% protection), and modestly protective against hydrogen peroxide- (16%) and oligomycin- (15%) mediated cell death, a pattern also confirmed in cultured hippocampal neurons. Thus, under pathological conditions involving mitochondrial dysfunction and Ca2+ dysregulation, CBD may prove beneficial in preventing apoptotic signaling via a restoration of Ca2+ homeostasis.

Aluminium toxicity in the rat brain: histochemical and immunocytochemical evidence

Although the neurotoxic actions of aluminium (Al) have been well documented, its contribution to neurodegenerative diseases such as Alzheimers disease remains controversial. In the present study, we applied histochemical techniques to identify changes induced by intracerebroventricular Al injections (5.4 microg in 5.5 microl, daily over a period of 5 successive days) in the adult rat brain after survival periods of either 1 or 6 weeks. For both Al- and saline-infused controls, no major signs of gross histological changes were evident in cresyl violet-stained sections. Al (as indicated by the fluorescent Morin staining) was concentrated in white matter of the medial striatum, corpus callosum, and cingulate bundle. Immunoreactivity of astrocytes and phagocytic microglia based on glial fibrillary acidic protein and ED1 markers, respectively, revealed a greater inflammatory response in Al-injected animals compared to controls. Damage of the cingulate bundle in Al-treated animals led to a severe anterograde degeneration of cholinergic terminals in cortex and hippocampus, as indicated by acetylcholinesterase labelling. Our data suggest that the enhancement of inflammation and the interference with cholinergic projections may be the modes of action through which Al may cause learning and memory deficits, and contribute to pathological processes in Alzheimers disease.

Aluminum impairs hippocampal long-term potentiation in rats in vitro and in vivo.

Although aluminum (Al) contributes to a variety of cognitive dysfunctions and mental diseases, the underlying mechanisms of Al interactions with the nervous system are still unknown. We have studied the action of Al on synaptic transmission and long-term potentiation (LTP) by performing electrophysiological recordings both in vivo, using freely moving animals, and in vitro, using hippocampal slices. In vivo recordings of the population spikes (PSs) of dentate gyrus granule cells in response to medial perforant path stimulation were performed on both acutely and chronically (Al each day for 5 days) intraventricularly injected animals. Acute Al-infusion (calculated brain concentrations of 0.27, 0.68, and 2.7 micrograms/ml) had no influence on baseline values. Al at 0.27 microgram/ml did not alter the induction and maintenance of LTP, but 0.68 and especially 2.7 micrograms/ml Al lead to a reduction in LTP, and the potentiation declined to baseline within 2 h. In chronic animals their neuronal responsiveness was reduced and in 30% of the rats the PS was completely lost. High-frequency tetanization failed to induce LTP. In slices, field potentials were evoked stimulating Schaffer collaterals and recording pyramidal cells of the CA1 region. Bath application of 0.68 microgram/ml Al increased the baseline amplitude of the PS slightly, whereas 2.7 micrograms/ml decreased the amplitude and concentrations > 5.4 micrograms/ml blocked the PS completely. Induction of LTP in the presence of 0.68 microgram/ml Al led to a smaller increase of the PS amplitude compared to controls, but the duration of LTP was not affected. In the presence of 2.7 micrograms/ml Al LTP was further reduced and declined to baseline levels within 60 min. Given that LTP is a form of synaptic plasticity underlying some forms of learning, our data suggest that both preparations are suitable models for investigating actions of Al-induced neurotoxicity.

Fear conditioning-induced time- and subregion-specific increase in expression of mGlu5 receptor protein in rat hippocampus

Memory formation involves encoding, consolidation and retention. These processes have been the subjects of considerable research, but physiological mechanisms underlying consolidation have proved difficult to dissociate experimentally. Previous reports have indicated a role for metabotropic glutamate receptors (mGluRs) in memory formation, and we here examined the specific role of mGluRs in the consolidation phase of memory formation. Particular weight was given to the hippocampus due to a high expression level for group I mGluRs and its outstanding role in spatial learning. Rats were first trained in a combined context and cue conditioning paradigm. Then, ex vivo analysis of neuronal tissue taken from hippocampal CA1, CA3 or dentate gyrus of behaviourally trained animals showed a 3-fold hyper-expression of mGluR5 protein in CA3 one day after acquisition training. This increase was transient and greatly diminished within ten days. The decline was paralleled by an increase in mGluR5 protein expression in CA1 and, to a lesser extent, in dentate gyrus, ten days posttraining. Overexpression in CA1 was also obtained after 9 days of extinction training. These data provide new insight into the role of the hippocampus and its subregions in memory consolidation. They support the notion that mGluRs in CA3 may play a part in short-term, and those in CA1 may play a part in long-term consolidation of memory.

Synthetic and plant-derived cannabinoid receptor antagonists show hypophagic properties in fasted and non-fasted mice

Background and purpose:  Obesity is a severe health problem in the modernized world and understanding the central nervous mechanisms underlying food‐seeking behaviour and reward are at the forefront of medical research. Cannabinoid receptors have proven an efficient target to suppress hunger and weight gain by their pharmacological inactivation.

The cholinergic system, EEG and sleep

Acetylcholine is a potent excitatory neurotransmitter, crucial for cognition and the control of alertness and arousal. Vigilance-specific recordings of the electroencephalogram (EEG) potently reflect thalamo-cortical and brainstem-cortical cholinergic activity that drives theta rhythms and task-specific cortical (de-synchronisation. Additionally, cholinergic projections from the basal forebrain act as a relay centre for the brainstem-cortical arousal system, but also directly modulate cortical activity, and thus promote wakefulness or rapid-eye movement (REM) sleep. Disease states such as sleep disorders, dementia and certain types of epilepsy are a further reflection of the potent cholinergic impact on CNS physiology and function, and highlight the relevance and inter-dependence of sleep and EEG. With novel technologies and computational tools now becoming available, advanced mechanistic insights may be gained and new avenues explored for diagnostics and therapeutics.

Abnormal Cognition, Sleep, EEG and Brain Metabolism in a Novel Knock-In Alzheimer Mouse, PLB1

Late-stage neuropathological hallmarks of Alzheimers disease (AD) are β-amyloid (βA) and hyperphosphorylated tau peptides, aggregated into plaques and tangles, respectively. Corresponding phenotypes have been mimicked in existing transgenic mice, however, the translational value of aggressive over-expression has recently been questioned. As controlled gene expression may offer animal models with better predictive validity, we set out to design a transgenic mouse model that circumvents complications arising from pronuclear injection and massive over-expression, by targeted insertion of human mutated amyloid and tau transgenes, under the forebrain- and neurone-specific CaMKIIα promoter, termed PLB1Double. Crossing with an existing presenilin 1 line resulted in PLB1Triple mice. PLB1Triple mice presented with stable gene expression and age-related pathology of intra-neuronal amyloid and hyperphosphorylated tau in hippocampus and cortex from 6 months onwards. At this early stage, pre-clinical 18FDG PET/CT imaging revealed cortical hypometabolism with increased metabolic activity in basal forebrain and ventral midbrain. Quantitative EEG analyses yielded heightened delta power during wakefulness and REM sleep, and time in wakefulness was already reliably enhanced at 6 months of age. These anomalies were paralleled by impairments in long-term and short-term hippocampal plasticity and preceded cognitive deficits in recognition memory, spatial learning, and sleep fragmentation all emerging at ∼12 months. These data suggest that prodromal AD phenotypes can be successfully modelled in transgenic mice devoid of fibrillary plaque or tangle development. PLB1Triple mice progress from a mild (MCI-like) state to a more comprehensive AD-relevant phenotype, which are accessible using translational tools such as wireless EEG and microPET/CT.