Thalamocortical Brain Slices Research Articles

Brain-derived neurotrophic factor-dependent unmasking of "silent" synapses in the developing mouse barrel cortex.

Brain-derived neurotrophic factor (BDNF) is a critical modulator of central synaptic functions such as long-term potentiation in the hippocampal and visual cortex. Little is known, however, about its role in the development of excitatory glutamatergic synapses in vivo. We investigated the development of N-methyl-D-aspartate (NMDA) receptor (NMDAR)-only synapses (silent synapses) and found that silent synapses were prominent in acute thalamocortical brain slices from BDNF knockout mice even after the critical period. These synapses could be partially converted to alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR)-containing ones by adding back BDNF alone to the slice or fully converted to together with electric stimulation without affecting NMDAR transmission. Electric stimulation alone was ineffective under the BDNF knockout background. Postsynaptically applied TrkB kinase inhibitor or calcium-chelating reagent blocked this conversion. Furthermore, the AMPAR C-terminal peptides essential for interaction with PDZ proteins postsynaptically prevented the unmasking of silent synapses. These results suggest that endogenous BDNF and neuronal activity synergistically activate AMPAR trafficking into synaptic sites.

Anticonvulsant actions of lamotrigine on spontaneous thalamocortical rhythms.

This study examined the actions of lamotrigine (LTG) on epileptiform discharges resembling generalized absence (GA) and primary generalized tonic-clonic (GTC) seizures in rat thalamocortical (TC) brain slices and attempted to characterize further the cellular mechanisms of action of LTG on neuronal ionic conductances. Rat TC slices generated spontaneous generalized epileptiform discharges after perfusion with a medium containing no added Mg(2+). Using multiple channel extracellular field-potential recordings in thalamus and cortex, the effects of LTG were characterized on two principal variants of activity that are similar to spike-wave discharges (SWDs) of GA epilepsy and GTC seizure discharges. These were termed simple TC burst complexes (sTBCs) and complex TC burst complexes (cTBCs), respectively. With whole-cell patch-clamp recording techniques in acutely dissociated TC neurons, the effects of LTG on GABA (gamma-aminobutyric acid)(A)-receptor-mediated currents and the low-threshold calcium current (I(T)) were examined. In field-potential recording studies in TC slices, both sTBCs and cTBCs were blocked by clinically relevant concentrations of LTG. In patch-clamp recording studies, LTG was found to be ineffective in the modulation of both GABA(A) receptors (GABARs) and I(T) in TC neurons. The efficacy of LTG on both variants of epileptiform discharges in TC slices clearly parallels its broad human clinical spectrum of action. This demonstrates that neurons within the TC system constitute one probable therapeutic target of LTG. However, LTG did not block either GABAR-mediated responses or I(T) in TC neurons. Modulation of these conductances represent likely cellular mechanisms of action of other antiepileptic drugs effective in the control of GA epilepsy. This suggests that LTG may have as yet uncharacterized effects that could combine with its previously defined sodium channel-blocking actions to explain its clinical utility in the control GA seizures.

1809 Neuronal connectivity among barrels in the rat barrel cortex

We studied auditory thalamocortical interactions in vitro, using an auditory thalamocortical brain slice preparation. Cortical activity evoked by electrical stimulation of the medial geniculate nucleus (MGN) was investigated through field potential recordings and voltage sensitive dyes. Experiments were performed in slices obtained from adult mice (9–14 weeks). Stimulus evoked activity was detected in the granular and supragranular layers after a short latency (5–6 ms). In 9–14 weeks old mice infragranular activity was detected in 10 of 24 preparations and was found to be increased in younger mice (p 31–64). In 14 of 24 slices a prominent horizontal spread was observed, which extended into cortical areas lateral to A1. In these experiments, the shortest onset latencies and largest signal amplitudes were located in the supragranular layers of A1. In areas lateral to A1, shortest onset latencies were located in the granular layer, while largest signal amplitudes were found in the supragranular layers. Evoked cortical activity was sensitive to removal of extracellular Ca2+ or application of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 μM). Short repetitive stimulation, resembling thalamic burst activity (three pulses at 100 Hz), resulted in an increase of signal amplitude and excited area by ∼25%, without changing the overall spatiotemporal activity profile. Blockade of N-methyl-d-aspartate receptors by 2-amino-5-phosphonopentanoate (AP5, 50 μM) reduced amplitudes and excited area by ∼15–30%, irrespective of stimulation frequency. Application of bicuculline (10 μM) greatly increased cortical responses to thalamic stimulation. Under these conditions, evoked activity displayed a pronounced horizontal spread in combination with a 2–3-fold increase in amplitude. In conclusion, afferent thalamic inputs primarily activate supragranular and granular layers in the auditory cortex of adult mice. This activation is predominantly mediated by non-NMDA receptors, while GABAA receptor-mediated inhibition limits the horizontal and vertical spread of activity.

Neuronal connectivity among barrels in the rat barrel cortex

We studied auditory thalamocortical interactions in vitro, using an auditory thalamocortical brain slice preparation. Cortical activity evoked by electrical stimulation of the medial geniculate nucleus (MGN) was investigated through field potential recordings and voltage sensitive dyes. Experiments were performed in slices obtained from adult mice (9–14 weeks). Stimulus evoked activity was detected in the granular and supragranular layers after a short latency (5–6 ms). In 9–14 weeks old mice infragranular activity was detected in 10 of 24 preparations and was found to be increased in younger mice (p 31–64). In 14 of 24 slices a prominent horizontal spread was observed, which extended into cortical areas lateral to A1. In these experiments, the shortest onset latencies and largest signal amplitudes were located in the supragranular layers of A1. In areas lateral to A1, shortest onset latencies were located in the granular layer, while largest signal amplitudes were found in the supragranular layers. Evoked cortical activity was sensitive to removal of extracellular Ca2+ or application of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 μM). Short repetitive stimulation, resembling thalamic burst activity (three pulses at 100 Hz), resulted in an increase of signal amplitude and excited area by ∼25%, without changing the overall spatiotemporal activity profile. Blockade of N-methyl-d-aspartate receptors by 2-amino-5-phosphonopentanoate (AP5, 50 μM) reduced amplitudes and excited area by ∼15–30%, irrespective of stimulation frequency. Application of bicuculline (10 μM) greatly increased cortical responses to thalamic stimulation. Under these conditions, evoked activity displayed a pronounced horizontal spread in combination with a 2–3-fold increase in amplitude. In conclusion, afferent thalamic inputs primarily activate supragranular and granular layers in the auditory cortex of adult mice. This activation is predominantly mediated by non-NMDA receptors, while GABAA receptor-mediated inhibition limits the horizontal and vertical spread of activity.

Functional GABAergic Synaptic Connection in Neonatal Mouse Barrel Cortex

Intracortical inhibition is crucial to proper functioning of the mature neocortex, yet, paradoxically, is reported to be rare or absent in the neonatal animal. We reexamined this issue by recording whole-cell postsynaptic currents (PSCs) of barrel cortex neurons in thalamocortical brain slices from neonatal mice. Monosynaptic, excitatory thalamocortical responses were elicited in layers V/VI neurons as early as postnatal day 0 (P0, the first 24 hr after birth) and in presumptive layer IV as early as P2. At very low stimulation frequencies, the monosynaptic response was invariably followed by a prolonged (up to 1 sec) synaptic barrage, which fatigued at stimulus repetition rates of 2/min or higher. This barrage consisted of postsynaptic responses to spiking activity in neighboring cortical cells, because (1) it could also be evoked by intracortical stimulation in coronal slices and (2) it was abolished by antagonists to NMDA receptors (NMDARs), even when NMDARs on the recorded cell were under a voltage-dependent block. Some of the larger polysynaptic events changed polarity at a negative reversal potential and were blocked by GABAA receptor (GABAAR) antagonists, with a concurrent enhancement of the extracellular field potential, indicating that they were GABAAR- mediated, CI-dependent inhibitory PSCs (IPSCs). We conclude that a network of functional intracortical GABAAR-mediated synaptic connections exists from the earliest postnatal ages, although it gives rise to responses that differ from mature IPSCs in reversal potential and latency.

Anticonvulsant drug effects on spontaneous thalamocortical rhythms in vitro: phenytoin, carbamazepine, and phenobarbital.

When perfused with a medium containing no added Mg2+, rodent thalamocortical brain slices generate spontaneous generalized thalamocortical discharges of several types. Two of these discharges, termed simple and complex thalamocortical burst complexes (sTBCs and cTBCs), are physiologically and pharmacologically similar to the spike-wave discharges of generalized absence epilepsy and to the discharges underlying generalized tonic-clonic seizures, respectively. In a further characterization of the pharmacology of generalized thalamocortical discharges recorded in rodent thalamocortical slices, the actions of anticonvulsants effective in control of partial and generalized tonic-clonic seizures, but not generalized absence seizures, were studied on these rhythms. The effects of phenytoin, carbamazepine, and phenobarbital were tested against sTBCs and cTBCs recorded in vitro in rodent thalamocortical slices. When applied in clinically relevant concentrations, phenytoin and carbamazepine were very effective in reducing or blocking cTBCs. These drugs were much less effective in controlling sTBCs. Phenobarbital was effective in controlling both sTBCs and cTBCs, but the level of block was greater for cTBCs. Therefore, it appears that sTBCs and cTBCs are quite distinct in their relative sensitivity to anticonvulsant drugs, and this differential sensitivity parallels the relative effectiveness of these drugs in controlling generalized absence and generalized tonic-clonic seizures.