Aaron R. Best

Activity-Dependent Regulation of Synapses by Retrograde Messengers

Throughout the brain, postsynaptic neurons release substances from their cell bodies and dendrites that regulate the strength of the synapses they receive. Diverse chemical messengers have been implicated in retrograde signaling from postsynaptic neurons to presynaptic boutons. Here, we provide an overview of the signaling systems that lead to rapid changes in synaptic strength. We consider the capabilities, specializations, and physiological roles of each type of signaling system.

Coordinate synaptic mechanisms contributing to olfactory cortical adaptation.

Anterior piriform cortex (aPCX) neurons rapidly filter repetitive odor stimuli despite relatively maintained input from mitral cells. This cortical adaptation is correlated with short-term depression of afferent synapses, in vivo. The purpose of this study was to elucidate mechanisms underlying this nonassociative neural plasticity using in vivo and in vitro preparations and to determine its role in cortical odor adaptation. Lateral olfactory tract (LOT)-evoked responses were recorded in rat aPCX coronal slices. Extracellular and intracellular potentials were recorded before and after simulated odor stimulation of the LOT. Results were compared with in vivo intracellular recordings from aPCX layer II/III neurons and field recordings in urethane-anesthetized rats stimulated with odorants. The onset, time course, and extent of LOT synaptic depression during both in vitro electrical and in vivo odorant stimulation methods were similar. Similar to the odor specificity of cortical odor adaptation in vivo, there was no evidence of heterosynaptic depression between independent inputs in vitro. In vitro evidence suggests at least two mechanisms contribute to this activity-dependent synaptic depression: a rapidly recovering presynaptic depression during the initial 10-20 sec of the post-train recovery period and a longer lasting (∼120 sec) depression that can be blocked by the metabotropic glutamate receptor (mGluR) II/III antagonist (RS)-α-cyclopropyl-4-phosphonophenylglycine (CPPG) and by the β-adrenergic receptor agonist isoproterenol. Importantly, in line with the in vitro findings, both adaptation of odor responses in the β (15-35 Hz) spectral range and the associated synaptic depression can also be blocked by intracortical infusion of CPPG in vivo.

Inhibitory Regulation of Electrically Coupled Neurons in the Inferior Olive Is Mediated by Asynchronous Release of GABA

Inhibitory projection neurons in the deep cerebellar nuclei (DCN) provide GABAergic input to neurons of the inferior olive (IO) that in turn produce climbing fiber synapses onto Purkinje cells. Anatomical evidence suggests that DCN to IO synapses control electrical coupling between IO neurons. In vivo studies suggest that they also control the synchrony of IO neurons and play an important role in cerebellar learning. Here we describe the DCN to IO synapse. Remarkably, GABA release was almost exclusively asynchronous, with little conventional synchronous release. Synaptic transmission was extremely frequency dependent, with low-frequency stimulation being largely ineffective. However, due to the prominence of asynchronous release, stimulation at frequencies above 10 Hz evoked steady-state inhibitory currents. These properties seem ideally suited to transform the firing rate of DCN neurons into sustained inhibition that both suppresses the firing of IO cells and regulates the effective coupling between IO neurons.

Plasticity in the Olfactory System: Lessons for the Neurobiology of Memory

We are rapidly advancing toward an understanding of the molecular events underlying odor transduction, mechanisms of spatiotemporal central odor processing, and neural correlates of olfactory perception and cognition. A thread running through each of these broad components that define olfaction appears to be their dynamic nature. How odors are processed, at both the behavioral and neural level, is heavily dependent on past experience, current environmental context, and internal state. The neural plasticity that allows this dynamic processing is expressed nearly ubiquitously in the olfactory pathway, from olfactory receptor neurons to the higher-order cortex, and includes mechanisms ranging from changes in membrane excitability to changes in synaptic efficacy to neurogenesis and apoptosis. This review will describe recent findings regarding plasticity in the mammalian olfactory system that are believed to have general relevance for understanding the neurobiology of memory.

Serotonin Evokes Endocannabinoid Release and Retrogradely Suppresses Excitatory Synapses

5-HT2-type serotonin receptors (5-HT2Rs) are widely expressed throughout the brain and mediate many of the modulatory effects of serotonin. It has been thought that postsynaptic 5-HT2Rs act primarily by depolarizing neurons and thereby increasing their excitability. However, it is also known that 5-HT2Rs are coupled to Gq/11-type G-proteins and that some other types of Gq/11-coupled receptors can regulate synapses by evoking endocannabinoid release and activating presynaptic cannabinoid-type 1 receptors (CB1Rs). Here, we examine whether activation of 5-HT2Rs can regulate synapses through such a mechanism by studying excitatory synapses onto cells in the inferior olive (IO). These cells express 5-HT2Rs on their soma and dendrites, and the IO receives extensive serotonergic input. We find that the excitatory synaptic inputs onto IO cells are strongly suppressed by serotonin receptor agonists as well as release of endogenous serotonin. Both 5-HT2Rs and 5-HT1BRs contribute to this modulation by decreasing the probability of glutamate release from presynaptic boutons. The suppression by 5-HT2Rs is of particular interest because it is prevented by CB1R antagonists, and 5-HT2Rs are thought to be located only postsynaptically on IO cells. Our results indicate that serotonin activates 5-HT2Rs on IO neurons, thereby releasing endocannabinoids that act retrogradely to suppress glutamate release by activating presynaptic CB1Rs. These findings establish a link between serotonin signaling and endocannabinoid signaling. Based on the extensive distribution of 5-HT2Rs and CB1Rs, it seems likely that this mechanism could mediate many of the actions of 5-HT2Rs throughout the brain.

Sustained Elevation of Dendritic Calcium Evokes Widespread Endocannabinoid Release and Suppression of Synapses onto Cerebellar Purkinje Cells

Endocannabinoids can act as retrograde messengers that allow postsynaptic cells to regulate the strength of their synaptic inputs. In the cerebellum, Purkinje cells (PCs) release endocannabinoids through two mechanisms. Synaptic activation evokes local endocannabinoid release that relies on a pathway that involves the metabotropic glutamate receptor mGluR1 and phospholipase-C (PLC). In contrast, depolarization evokes endocannabinoid release from the entire dendritic arbor. This leads to depolarization-induced suppression of inhibitory (DSI) and excitatory (DSE) synapses by a mechanism that does not involve mGluR1 or PLC. This latter mechanism of endocannabinoid release has only been observed under artificial conditions that transiently elevate postsynaptic calcium to >5 μm. Here, we tested the possibility that this mechanism could lead to retrograde inhibition in response to more realistic calcium signals. At both climbing fiber and inhibitory synapses onto PCs, we found that prolonging the elevation of calcium significantly lowered the peak calcium required to evoke PLC-independent endocannabinoid release. This suggests that the mechanism of endocannabinoid release involved in DSI and DSE is likely to evoke endocannabinoid release in response to physiologically relevant levels of calcium. When dendritic calcium was elevated to 0.4–1 μm for 15 s or more, endocannabinoid release from PCs selectively suppressed inhibitory synapses. This suggests that inhibitory synapses are more sensitive to prolonged calcium increases. Thus, in contrast to localized retrograde inhibition evoked by synaptic activation, modest but sustained calcium elevation could globally suppress inhibitory synapses onto PCs.

Cortical Metabotropic Glutamate Receptors Contribute to Habituation of a Simple Odor-Evoked Behavior

Defining the circuits that are involved in production and cessation of specific behaviors is an ultimate goal of neuroscience. Short-term behavioral habituation is the response decrement observed in many behaviors that occurs during repeated presentation of non-reinforced stimuli. Within a number of invertebrate models of short-term behavioral habituation, depression of a defined synapse has been implicated as the mechanism. However, the synaptic mechanisms of short-term behavioral habituation have not been identified within mammals. We have shown previously that a presynaptic metabotropic glutamate receptor (mGluR)-dependent depression of synapses formed by olfactory bulb afferents to the piriform (olfactory) cortex significantly contributes to adaptation of cortical odor responses. Here we show that blockade of mGluRs within the olfactory cortex of awake, behaving rats diminishes habituation of a simple odor-induced behavior, strongly implicating a central mechanism for sensory gating in olfaction.

High-Frequency Oscillations Are Not Necessary for Simple Olfactory Discriminations in Young Rats

Individual olfactory bulb mitral/tufted cells respond preferentially to groups of molecularly similar odorants. Bulbar interneurons such as periglomerular and granule cells are thought to influence mitral/tufted odorant receptive fields through mechanisms such as lateral inhibition. The mitralgranule cell circuit is also important in the generation of the odor-evoked fast oscillations seen in the olfactory bulb local field potentials and hypothesized to be an important indicator of odor quality coding. Infant rats, however, lack a majority of these inhibitory interneurons until the second week of life. It is unclear if these developmental differences affect olfactory bulb odor coding or behavioral odor discrimination. The following experiments are aimed at better understanding odor coding and behavioral odor discrimination in the developing olfactory system. Single-unit recordings from mitral/tufted cells and local field-potential recordings from both the olfactory bulb and anterior piriform cortex were performed in freely breathing urethane-anesthetized rats (postnatal day 7 to adult). Age-dependent behavioral odor discrimination to a homologous series of ethyl esters was also examined using a cross-habituation paradigm. Odorants were equated in all experiments for concentration (150 ppm) using a flow dilution olfactometer. In concordance with the reduced interneuron population, local field potentials in neonates lacked detectable odor-evoked γ-frequency oscillations that were observed in mature animals. However, mitral/tufted cell odorant receptive fields and behavioral odor discrimination did not significantly change, despite known substantial changes in local circuitry and neuronal populations, over the age range examined. The results suggest that high-frequency local field-potential oscillations do not reflect processes critical for simple odor discrimination.

Trans-neuronal modification of anterior piriform cortical circuitry in the rat

Long-Evans rats received unilateral naris closure on postnatal day 1 (PN1) or sham surgery. On PN30, brains were processed for anterograde horseradish peroxidase (HRP) labeling of lateral olfactory tract (LOT) fibers in anterior piriform cortex (aPCX) Layer Ia, Timm staining of association/commissural fibers in aPCX Layer Ib, or Golgi staining for reconstruction of aPCX semilunar cell dendrites. The results demonstrate that the width of aPCX Layer Ia was reduced ipsilateral to the sealed naris compared to undeprived controls. No significant change in Layer Ib was detected. Furthermore, semilunar cell dendrites were reduced by unilateral deprivation compared to undeprived controls. The reduction in dendritic tree size was localized to distal dendritic segments, roughly corresponding to Layer Ia. These results suggest an activity-dependent change in both the distribution of cortical afferents and in the dendritic field of their target cells. While these results are similar to those reported for other sensory systems, the relatively simple architecture and laminated organization of bilateral inputs to the aPCX make the olfactory system an ideal model system to examine experience-dependent synaptic reorganization and its functional consequences.

A postnatal sensitive period for plasticity of cortical afferents but not cortical association fibers in rat piriform cortex

Male and female rats underwent unilateral naris occlusion or sham surgery on either post-natal day (PN) 1 or after PN30. Following at least 30 days of unilateral olfactory deprivation, rats were urethane anesthetized and recordings were made from anterior piriform cortex (aPCX). Shock stimulation of afferent fibers (lateral olfactory tract) and association/commissural fibers evoked field potentials in aPCX that were analyzed across groups and between ages. The results demonstrate that early-onset unilateral olfactory deprivation depresses field potentials evoked by stimulation of the deprived cortical afferent, while late-onset deprivation did not. In contrast, intracortical association fiber mediated field potentials in the deprived cortex were enhanced after both early-onset and late-onset deprivation. These results suggest differential developmental plasticity of afferent and association fiber pathways in paleocortex that mirrors that previously described in neocortical sensory systems.