Grzegorz Hess

Quantal analysis of paired-pulse facilitation in guinea pig hippocampal slices

Intracellular excitatory postsynaptic potentials (EPSPs) were recorded in area CA1 of hippocampal slices from guinea pigs. With paired-pulse stimulation of stratum radiatum, the second stimulus (interval of 40-50 ms) produced an EPSP with enlarged amplitude. Two methods of quantal analysis showed an increase in quantal content of the facilitated EPSP and a smaller increase in quantal size. The correlation between amplitudes of the first and second EPSP was usually insignificant. The results favour a presynaptic location of the mechanisms of the paired-pulse facilitation and suggest increases in the average of transmitter quanta released by presynaptic volley as well as increases in the amount of transmitter in each quantum.

N-methyl-d-aspartate receptor mediated component of field potentials evoked in horizontal pathways of rat motor cortex

To identify potential sites of synaptic modification of intrinsic cortical circuits, the contribution of the N-methyl-D-aspartate type of glutamate receptors to field potentials evoked in horizontal and oblique intracortical pathways was examined in rat motor cortex slice preparations. Presumably monosynaptic, short latency responses with a prominent negativity (-0.4 to -2.0 mV) were recorded in both superficial (across layer III) and deep (across layer V) horizontal pathways at a distance of approximately equal to 500 microns lateral to electrical stimulation sites and in oblique V-III pathway (-0.3 to -1.6 mV). Bath application of the N-methyl-D-aspartate receptor antagonist D,L-2-amino-5-phosphonovaleric acid (100 microM) reversibly decreased field potentials. Although decreases were observed in all components of the waveform, the most pronounced effect was on the late phase of the response. D,L-2-Amino-5-phosphonovaleric acid produced on average a 22% decrease in area, 12% in initial slope and 11% in peak amplitude of responses. Combined application of 100 microM D,L-2-amino-5-phosphonovaleric acid and a non-N-methyl-D-aspartate glutamate receptor antagonist, 6-cyano-7-nitro- or 6,7-dinitro-quinoxaline-2,3- dione (10-20 microM), eliminated all but a small, early and presumably non-synaptic response. In 18 of 23 cases, the relative contribution of the D,L-2-amino-5-phosphonovaleric acid-sensitive component was unrelated to field potential magnitude, suggesting that this component is present in all fiber classes. It is concluded that glutamate is the major transmitter of horizontal connections of layers II/III and layer V, as well as in the oblique V-III pathway. While most glutamatergic transmission is relayed by other glutamate receptor subtypes, N-methyl-D-aspartate receptor activation contributes a small but consistent part of ordinary transmission in each of these pathways in vitro. The results further suggest that a potential for N-methyl-D-aspartate receptor-mediated synaptic modification exists in intrinsic horizontal pathways of both superficial and deep layers of rat motor cortex.

5-HT7 receptors increase the excitability of rat hippocampal CA1 pyramidal neurons

In the CA1 area of rat hippocampal slices, a combined application of 5-CT, a potent 5-HT(1A) and 5-HT(7) receptor agonist, and WAY 100635, a selective 5-HT(1A) receptor antagonist, resulted in a reversible increase of the CA1 extracellular population spike amplitude. In whole-cell recording from identified pyramidal neurons, the effects of 5-CT applied in the presence of WAY 100635 involved a reduction of the slow afterhyperpolarization (sAHP) and the frequency adaptation of action potential firing, which could be blocked by a specific 5-HT(7) receptor antagonist SB 269970. The results indicate that the activation of 5-HT(7) receptors increases the excitability of hippocampal CA1 pyramidal cells.

Repeated administration of imipramine attenuates glutamatergic transmission in rat frontal cortex.

The effects of repeated administration of a tricyclic antidepressant, imipramine, lasting 14 days (10 mg/kg p.o., twice daily), were studied ex vivo in rat frontal cortex slices prepared 48 h after last dose of the drug. In slices prepared from imipramine-treated animals the mean frequency, and to a lesser degree the mean amplitude, of spontaneous excitatory postsynaptic currents recorded from layer II/III pyramidal neurons, were decreased. These effects were accompanied by a reduction of the initial slope ratio of pharmacologically isolated N-methyl-D-aspartate to AMPA/kainate receptor-mediated stimulation-evoked excitatory postsynaptic currents. Imipramine treatment also resulted in a decrease of extracellular field potentials evoked in layer II/III by stimulation of underlying sites in layer V. These results indicate that chronic treatment with imipramine results in an attenuation of the release of glutamate and an alteration in the postsynaptic reactivity of ionotropic glutamate receptors in rat cerebral cortex.

Facilitation of long-term potentiation in layer II/III horizontal connections of rat motor cortex following layer I stimulation: route of effect and cholinergic contributions

Abstract The ability of layer I activation to facilitate the induction of long-term potentiation (LTP) in layer II/III horizontal connections of motor cortex (MI) was examined in rat brain slice preparations. Field potentials evoked in layer I and layer II/III horizontal pathways were recorded from radially aligned MI sites. While theta burst stimulation (TBS) of layer II/III pathways alone failed to induce LTP, simultaneous TBS of layer I and layer II/III inputs on alternate sides of the recording electrodes induced LTP in the layer II/III input in 8 out of 13 slices (mean change +20±6%; N=13). In the same cases, the layer I connections showed mixed effects: LTP in three cases, LTD in five cases, and no modification in five slices. Despite the facilitatory effect of layer I activation on layer II/III LTP induction, we found that the critical circuitry for this effect was outside layer I. Cutting the layer I fibers selectively in the slice did not prevent layer II/III LTP induction, while cuts preserving only layer I blocked layer II/III LTP after conjoint I+II/III TBS. Cholinergic fibers were evaluated as candidates for the facilitatory effect because they branch widely in both layers and they are thought to participate in synaptic modification. The cholinergic contribution to layer II/III LTP facilitation was investigated using bath application of muscarinic antagonists. Muscarinic blockade prevented facilitation of layer II/III LTP by layer I coactivation. Instead, conjoint stimulation in 10 µM atropine produced long-term depression (LTD) of layer II/III (–18±9%; N=11) as well as of layer I (–21±6%; N=11) horizontal responses. These results indicate that connections formed within layer I are ineffective in promoting LTP in the deeper-lying horizontal connections; the critical route by which layer I stimulation influenced LTP induction required the circuitry in the deeper layers, particularly the cholinergic system. Thus, it appears that diffuse cholinergic afferents provide an additional route to regulate activity-dependent synaptic modificaton in horizontal cortical connections.

Repeated administration of antidepressants decreases field potentials in rat frontal cortex.

The effects of repeated administration of a tricyclic antidepressant, imipramine, and a selective serotonin reuptake blocker, citalopram, for 14 days (10 mg/kg p.o., twice daily), were studied ex vivo in rat frontal cortex slices prepared 48 h after last dose of the drug. Treatment with both antidepressants resulted in a decrease in the amplitude of field potentials evoked in layer II/III by stimulation of underlying sites in layer V. The amplitude ratio of pharmacologically isolated N-methyl-D-aspartic acid (NMDA) to alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)/kainate receptor-mediated components of the field potential was reduced. These results indicate that chronic treatment with imipramine or citalopram results in an attenuation of glutamatergic synaptic transmission in the cerebral cortex.

Sensory learning-induced enhancement of inhibitory synaptic transmission in the barrel cortex of the mouse

In adult mice, repetitive pairing of stimulation of mystacial vibrissae with an electrical shock to the tail induces expansion of the cortical representation of stimulated vibrissae accompanied by elevation of the GABAergic markers. Here, we show that this associative learning paradigm results in a selective increase in the frequency of spontaneous inhibitory postsynaptic currents in layer IV excitatory neurons located within the barrel representing stimulated vibrissae, evident 24 h after the end of training. The mean amplitude of spontaneous inhibitory postsynaptic potentials recorded from excitatory neurons was unchanged. Recordings from layer IV excitatory and fast spiking neurons showed that the training induced changes neither in the mean frequency nor it the mean amplitude of spontaneous excitatory postsynaptic currents. On the other hand, the mean amplitude of field potentials evoked by the stimulation of layer VI and recorded in layer IV was significantly reduced. These data indicate that aversive training results in a selective and long‐lasting enhancement of GABAergic transmission within the cortical representation of stimulated vibrissae, which may result in a decrease in layer VI‐evoked field responses.

Long-term depression and long-term potentiation in horizontal connections of the barrel cortex

Synaptic plasticity of horizontally orientated connections between barrels, in the barrel cortex of adult mice, was studied in slice preparations cut across rows of barrels. Field potentials were evoked in the middle of one barrel column (in layer IV or V) and recorded in the neighbouring barrel (in layer IV and V). In layer IV, long‐term depression (LTD) by 26.5 ± 5% was first induced by a low‐frequency stimulation (2 Hz) applied for 10 min. After 30 min, theta‐burst stimulation was delivered to previously depressed connections, resulting in long‐term potentiation (LTP) by 28.8 ± 11.8%. When theta‐burst stimulation was delivered without an earlier low‐frequency stimulation, no LTP was induced. Similar results were obtained in layer V connections (LTD: 40.6 ± 12.5%; LTP: 26.9 ± 12.5%). In layer IV, the application of 100 µm d,l‐2‐amino‐5‐phosphonovaleric acid (APV), an antagonist of NMDA receptors, blocked the induction of both LTD and LTP. These experiments show that a potential for synaptic plasticity is retained in granular and infragranular layers of adult mice.

Brief neck restraint stress enhances long-term potentiation and suppresses long-term depression in the dentate gyrus of the mouse.

We studied the effects of brief (10 min) neck restraint on long-term potentiation (LTP) and long-term depression (LTD) in mouse dentate gyrus (DG) slices. Brain slices were prepared immediately after neck restraint and LTP/LTD induction was attempted 3.5h later. LTP enhancement and LTD suppression was observed in slices prepared from stressed animals. The corticosterone plasma concentration was elevated approx. fourfold just after the neck restraint session. To examine the role of glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) activation in mediating the effects of neck restraint on LTP and LTD, the animals were pretreated with either the specific GR antagonist RU38486, the specific MR antagonist spironolactone, the corticosterone synthesis inhibitor metyrapone, or vehicle Tween 80. Injection with Tween 80 and subsequent neck restraint did not influence LTP enhancement or LTD suppression, but the effects of neck restraint were prevented by RU38486, spironolactone and metyrapone pretreatment. These data indicate that acute stress caused by brief neck restraint enhances LTP and impairs LTD in the dentate gyrus and that these effects are mediated by activation of GRs and MRs.

Increased excitability of cortical neurons induced by associative learning: an ex vivo study.

In adult mice, classical conditioning in which whisker stimulation is paired with an electric shock to the tail results in a decrease in the frequency of head movements, induces expansion of the cortical representation of stimulated vibrissae and enhances inhibitory synaptic interactions within the ‘trained’ barrels. We investigated whether such a simple associative learning paradigm also induced changes in neuronal excitability. Using whole‐cell recordings from ex vivo slices of the barrel cortex we found that layer IV excitatory cells located in the cortical representation of the ‘trained’ row of vibrissae had a higher frequency of spikes recorded at threshold potential than neurons from the ‘untrained’ row and than cells from control animals. Additionally, excitatory cells within the ‘trained’ barrels were characterized by increased gain of the input–output function, lower amplitudes of fast after‐hyperpolarization and decreased effect of blocking of BK channels by iberiotoxin. These findings provide new insight into the possible mechanism for enhanced intrinsic excitability of layer IV excitatory neurons. In contrast, the fast spiking inhibitory cells recorded in the same barrels did not change their intrinsic excitability after the conditioning procedure. The increased excitability of excitatory neurons within the ‘trained’ barrels may represent the counterpart of homeostatic plasticity, which parallels enhanced synaptic inhibition described previously. Together, the two mechanisms would contribute to increase the input selectivity within the conditioned cortical network.