Daniel Ulrich

Pattern-Specific Associative Long-Term Potentiation Induced by a Sleep Spindle-Related Spike Train

Spindles are non-rapid eye movement (non-REM) sleep EEG rhythms (7-14 Hz) that occur independently or in association with slow oscillations (0.6-0.8 Hz). Despite their proposed function in learning and memory, their role in synaptic plasticity is essentially unknown. We studied the ability of a neuronal firing pattern underlying spindles in vivo to induce synaptic plasticity in neocortical pyramidal cells in vitro. A spindle stimulation pattern (SSP) was extracted from a slow oscillation upstate that was recorded in a cat anesthetized with ketamine-xylazine, which is known to induce a sleep-like state. To mimic the recurrence of spindles grouped by the slow oscillation, the SSP was repeated every 1.5 s (0.6 Hz). Whole-cell patch-clamp recordings were obtained from layer V pyramidal cells of rat somatosensory cortex with infrared videomicroscopy, and composite EPSPs were evoked within layers II-III. Trains of EPSPs and action potentials simultaneously triggered by the SSP induced an NMDA receptor-dependent short-term potentiation (STP) and an L-type Ca2+ channel-dependent long-term potentiation (LTP). The number of spindle sequences affected the amount of STP-LTP. In contrast, spindle trains of EPSPs alone led to long-term depression. LTP was not consistently induced by a regular firing pattern, a mirrored SSP, or a randomized SSP; however, a synthetic spindle pattern consisting of repetitive spike bursts at 10 Hz reliably induced STP-LTP. Our results show that spindle-associated spike discharges are efficient in modifying excitatory neocortical synapses according to a Hebbian rule. This is in support of a role for sleep spindles in memory consolidation.

GABAB receptor-mediated responses in GABAergic projection neurones of rat nucleus reticularis thalami in vitro.

1. Whole‐cell voltage‐clamp recordings were obtained from GABAergic neurones of rat nucleus reticularis thalami (NRT) in vitro to assess pre‐ and postsynaptic GABAB receptor‐mediated responses. Presynaptic inhibition of GABA release was studied at terminals on local axon collaterals within NRT as well as on projection fibres in the somatosensory relay nuclei. 2. The GABAB receptor agonist (R)‐baclofen (10 microM) reduced monosynaptically evoked GABAA‐mediated inhibitory postsynaptic currents (IPSCs) in NRT and somatosensory relay cells to 11 and 12% of control, respectively. 3. Action potential‐independent miniature IPSCs (mIPSCs) were observed in both cell types. Mean mIPSC amplitude was 20 pA in both NRT and relay cells at a holding potential of 0 mV. The mean mIPSC frequencies were 0.83 and 2.2 Hz in NRT and relay cells, respectively. Baclofen decreased mIPSP frequency by about half in each cell type without affecting amplitude. 4. Paired‐burst inhibition of evoked IPSCs was studied in relay and NRT cells by applying pairs of 100 Hz stimulus bursts separated by 600 ms. The mean ratio of second to first peak IPSC amplitudes was 0.77. 5. In NRT cells baclofen induced a linear postsynaptic conductance increase of 0.82 nS with an associated reversal potential of ‐121 mV. A small (0.14 nS) GABAB component of the evoked IPSC was detected in only a minority of NRT cells (3 of 18). 6. All pre‐ and postsynaptic effects of baclofen, as well as PBI, were largely reversed by the specific GABAB receptor antagonist CGP 35348 (0.5 mM). 7. We conclude that activation of GABAB receptors in NRT leads to presynaptic autoinhibition of IPSCs in both NRT and relay cells, and to direct activation of a small linear K+ conductance. In addition our experiments suggest that reciprocal connectivity within NRT can be partially mediated by a small GABAB inhibitory event.

Purinergic inhibition of GABA and glutamate release in the thalamus: implications for thalamic network activity

Adenosine is a CNS depressant with both pre- and postsynaptic actions. Presynaptically, adenosine decreases neurotransmitter release in the hippocampus but only at excitatory terminals. In the thalamus, however, we show that, in addition to its actions at excitatory synapses, adenosine strongly suppresses monosynaptic inhibitory currents both in relay cells of the thalamic ventrobasal complex (VB) and in inhibitory neurons of the nucleus reticularis thalami (nRt). A concomitant increase in transmission failures and results coefficient of variation analysis are both consistent with a presynaptic mechanism. Pharmacological manipulations support an A1 receptor-mediated process. Slow thalamic oscillations induced in vitro by extracellular stimulation and recorded with extracellular multiunit electrodes in VB and nRt are dampened by adenosine without affecting their periodicity. We conclude that adenosine can presynaptically down-regulate inhibitory postsynaptic responses in thalamus and exert robust antioscillatory effects, likely by synergistic depression of both excitatory and inhibitory neurotransmitter release.

Firing Mode-Dependent Synaptic Plasticity in Rat Neocortical Pyramidal Neurons

Pyramidal cells in the mammalian neocortex can emit action potentials either as series of individual spikes or as distinct clusters of high-frequency bursts. However, why two different firing modes exist is largely unknown. In this study, we report that in layer V pyramidal cells of the rat somatosensory cortex, in vitro associations of EPSPs with spike bursts delayed by +10 msec led to long-term synaptic depression (LTD), whereas pairings with individual action potentials at the same delay induced long-term potentiation. EPSPs were evoked extracellularly in layer II-III and recorded intracellularly in layer V neurons with the whole-cell or nystatin-based perforated patch-clamp technique. Bursts were evoked with brief somatic current injections, resulting in three to four action potentials with interspike frequencies of ∼200 Hz, characteristic of intrinsic burst firing. Burst-firing-associated LTD (Burst-LTD) was robust over a wide range of intervals between -100 and +200 msec, and depression was maximal (∼50%) for closely spaced presynaptic and postsynaptic events. Burst-LTD was associative and required concomitant activation of low voltage-activated calcium currents and metabotropic glutamate receptors. Conversely, burst-LTD was resistant to blockade of NMDA receptors or inhibitory synaptic potentials. Burst-LTD was also inducible at already potentiated synapses. We conclude that intrinsic burst firing represents a signal for resetting excitatory synaptic weights.

Removal of Synaptic Ca2+-Permeable AMPA Receptors during Sleep

There is accumulating evidence that sleep contributes to memory formation and learning, but the underlying cellular mechanisms are incompletely understood. To investigate the impact of sleep on excitatory synaptic transmission, we obtained whole-cell patch-clamp recordings from layer V pyramidal neurons in acute slices of somatosensory cortex of juvenile rats (postnatal days 21–25). In animals after the dark period, philanthotoxin 74 (PhTx)-sensitive calcium-permeable AMPA receptors (CP-AMPARs) accounted for ∼25% of total EPSP size, and current–voltage (I–V) relationships of the underlying EPSCs showed inward rectification. In contrast, in similar experiments after the light period, EPSPs were PhTx insensitive with linear I–V characteristics, indicating that CP-AMPARs were less abundant. Combined EEG and EMG recordings confirmed that slow-wave sleep-associated delta wave power peaked at the onset of the more quiescent, lights-on phase of the cycle. Subsequently, we show that burst firing, a characteristic action potential discharge mode of layer V pyramidal neurons during slow-wave sleep has a dual impact on synaptic AMPA receptor composition: repetitive burst firing without synaptic stimulation eliminated CP-AMPARs by activating serine/threonine phosphatases. Additionally, repetitive burst-firing paired with EPSPs led to input-specific long-term depression (LTD), affecting Ca2+ impermeable AMPARs via protein kinase C signaling. In agreement with two parallel mechanisms, simple bursts were ineffective after the light period but paired bursts induced robust LTD. In contrast, incremental LTD was generated by both conditioning protocols after the dark cycle. Together, our results demonstrate qualitative changes at neocortical glutamatergic synapses that can be induced by discharge patterns characteristic of non-rapid eye movement sleep.

Dendritic calcium currents in thalamic relay cells

Techniques of in vitro whole-cell recording and compartmental modeling were combined to investigate dendritic structure and calcium currents in individual thalamocortical neurons. Voltage-clamp recordings of I T obtained in intact and dissociated ventrobasal neurons were used to constrain I T conductances and passive parameters incorporated in morphorealistic models. High dendritic I T densities were found necessary to establish congruent model behavior. Several methodologies based on axial resistance conservation were developed to algorithmically reduce thalamocortical morphology into a behaviorally congruent low-order compartmental model. This type of simplified model is suitable for investigating the functional role played by distal T- current localization at the network level in sleep oscillations and epilepsy.

GABAB receptor subtypes differentially regulate thalamic spindle oscillations

ABSTRACT Following the discovery of GABAB receptors by Norman Bowery and colleagues, cloning and biochemical efforts revealed that GABAB receptors assemble multi‐subunit complexes composed of principal and auxiliary subunits. The principal receptor subunits GABAB1a, GABAB1b and GABAB2 form two heterodimeric GABAB(1a,2) and GABAB(1b,2) receptors that can associate with tetramers of auxiliary KCTD (K+ channel tetramerization domain) subunits. Experiments with subunit knock‐out mice revealed that GABAB(1b,2) receptors activate slow inhibitory postsynaptic currents (sIPSCs) while GABAB(1a,2) receptors function as heteroreceptors and inhibit glutamate release. Both GABAB(1a,2) and GABAB(1b,2) receptors can serve as autoreceptors and inhibit GABA release. Auxiliary KCTD subunits regulate the duration of sIPSCs and scaffold effector channels at the receptor. GABAB receptors are well known to contribute to thalamic spindle oscillations. Spindles are generated through alternating burst‐firing in reciprocally connected glutamatergic thalamocortical relay (TCR) and GABAergic thalamic reticular nucleus (TRN) neurons. The available data implicate postsynaptic GABAB receptors in TCR cells in the regulation of spindle frequency. We now used electrical or optogenetic activation of thalamic spindles and pharmacological experiments in acute slices of knock‐out mice to study the impact of GABAB(1a,2) and GABAB(1b,2) receptors on spindle oscillations. We found that selectively GABAB(1a,2) heteroreceptors at TCR to TRN cell synapses regulate oscillation strength, while GABAB(1b,2) receptors control oscillation frequency. The auxiliary subunit KCTD16 influences both oscillation strength and frequency, supporting that KCTD16 regulates network activity through GABAB(1a,2) and GABAB(1b,2) receptors. This article is part of the “Special Issue Dedicated to Norman G. Bowery”. HIGHLIGHTSOptogenetic or electrical activation of TRN neurons generates thalamic spindles with similar frequencies and strength.GABAB receptors regulate both oscillation frequency and strength of thalamic spindles.GABAB(1a,2) heteroreceptors regulate oscillation strength at glutamatergic inputs to TRN cells.GABAB(1b,2) receptors regulate oscillation frequency, mainly at postsynaptic sites in TCR cells.The auxiliary GABAB receptor subunit KCTD16 regulates oscillation strength and frequency.