Hansjürgen Matthies

The effect of dopaminergic D1 receptor blockade during tetanization on the expression of long-term potentiation in the rat CA1 region in vitro

The effect of dopaminergic D1 receptor blockade on the expression of long-term potentiation (LTP) was investigated in the rat hippocampal CA1 region in vitro by extracellular recordings (by measuring the population spike amplitude and the field EPSP). The presence of the very selective D1 receptor blocker SCH 23390 at a concentration of 0.1 microM during tetanization with 3 trains of 100 impulses (100 Hz) resulted in a prevention of late LTP stages (greater than 1-2 h). When SCH 23390 was added to the bath medium immediately after tetanization, an influence on established LTP could not be observed during the first 3 h investigated.

Polymyxin B, an inhibitor of protein kinase C, prevents the maintenance of synaptic long-term potentiation in hippocampal CA1 neurons.

The involvement of protein kinase C (PKC)-mediated processes in mechanisms of long-term potentiation (LTP) was suggested by recent studies which have demonstrated a correlation between PKC activation and LTP. However, it was not possible to tell whether there is a causal relationship between the two events. Therefore, we have examined the induction and maintenance of LTP in rat hippocampal slices in the presence of a relatively selective PKC inhibitor, using extracellular electrophysiological techniques. Bath application of 0.1-100 microM polymyxin B did not influence the occurrence of post-tetanic and long-term potentiation usually seen in test responses 1 and 10 min after a 100-Hz/1 s tetanic stimulation of stratum radiatum fibers. However, 20 microM polymyxin B significantly depressed the increase in population spike amplitude and population excitatory postsynaptic potential (EPSP) slope from 30 to 120 min onwards, following repeated tetanization. Immediately after the drug application only weak and reversible effects were seen by the same parameters in test responses of a non-tetanized control input. A late (greater than 6 h) heterosynaptic potentiation of the population spike in the control input was blocked by polymyxin B treatment. Whereas the EPSP-LTP was fully blocked, some potentiation of the population spike still remained, suggesting the independence of PKC of the additional spike (E/S) potentiation for the first 6 h. These results provide direct evidence that the PKC activation is not essential for the initial phase of LTP, but is a necessary condition for a medium and a late, protein synthesis-dependent phase in this monosynaptic pathway, i.e. for the maintenance of synaptic LTP.

Two sensitive periods for the amnesic effect of anisomycin

Impairment of retention of a brightness discrimination in rats was obtained when anisomycin (80 microgram bilaterally into both hippocampi) was injected 10 min before and 80 min after training or 240 and 360 min after training. No amnesia was observed when anisomycin was injected 45 and 165 min post training. The two separate sensitive periods for the amnesic effect of the inhibitor obviously correspond to the two phases of increased protein synthesis during the consolidation of the same learning procedure. The results support the previous findings of the two independent and qualitatively different macromolecular processes. They also argue for the inhibition of protein synthesis as an important mechanism in the amnesic effect of anisomycin.

Kindling and its consequences on learning in rats

To study the learning performance of pentylenetetrazol- and amygdala-kindled Wistar rats we used the following learning tests: short-term memory was tested in the response-to-change model, brightness discrimination was tested in a Y-chamber, and two-way active avoidance learning was tested in a shuttle-box. Short-term memory was not impaired by both kindling procedures. Considering two-way active avoidance learning the performance of pentylenetetrazol (PTZ)-kindled rats was significantly diminished. This effect persists over a period of 4 weeks. However, amygdala (AMY)-kindled rats acquired this task like the controls. In brightness discrimination reaction (BDR) the learning performance of PTZ-kindled animals was not influenced. Although the acquisition of BDR was nearly identical, the 24-h retention was remarkably diminished in AMY-kindled rats. It was hypothesized that the different kindling procedures interfere in different ways and extent with neuronal circuits resulting in different functional impairments.

Long-term potentiation induced in dendrites separated from rat's CA1 pyramidal somata does not establish a late phase

In 14 transversal hippocampal slices from rats the extracellular field-EPSP (excitatory postsynaptic potential) in the dendritic region of the CA1 subfield was recorded after isolating the apical dendrites from their somata by microsurgical cuts through the proximal stratum radiatum. Contrary to intact slices which, after tetanic stimulation of the Schaffer collaterals, showed long-term potentiation (LTP) until the end of registration after 8-10 h, LTP declined in the isolated dendrites to baseline values after 3 h. It is concluded that de-novo protein synthesis in postsynaptic cell bodies might be necessary for the maintenance of late phases of LTP.

Accumulation of c-fos mRNA in rat hippocampus during acquisition of a brightness discrimination.

Training rats to attain a foot-shock-motivated brightness discrimination in a Y-maze results in an early and transient increase of hippocampal c-fos mRNA levels. Maximal accumulation was observed immediately after training, returning to basal levels during the following 2 h. A similar increase was obtained when rats were subjected to a pseudotraining with an equal number of runs, but with random pairing of the choice of bright and dark alleys with foot shock. It is suggested that induction of hippocampal c-fos mRNA expression is a necessary, but not sufficient, prerequisite for the formation of long-term memory trace. This early gene expression seems rather to correspond to an initial stage induced by complex stimulus presentation of both the training and the pseudotraining procedure. The subsequent late synthesis or processing of target proteins finally contributing to the formation of a permanent trace requires the action of further convergent signals to principal cells, probably mediating reward or emotional influences.

The duration of long-term potentiation in the CA1 region of the hippocampal slice preparation.

The duration of long-term potentiation (LTP) of the monosynaptic excitatory Schaffer collateral-commissural input to hippocampal neurons of the CA1 region was examined in the in vitro slice. Relatively stable evoked potentials were obtained under conventional perfusion conditions at least for 10 hours. Tetanic stimulation (100 Hz, 1 sec) increased the population spike (pop-spike) amplitude by about 150% and the slope of the field-EPSP by about 30% over the pre-LTP baseline, whereas the latency and peak latency of the pop-spike decreased. In comparison to control experiments (same number of stimuli at 0.2 Hz) the differences were statistically significant for 2 hr (field-EPSP) and for greater than or equal to 10 hr (pop-spike), respectively. Repeated tetanization (3 X 100 Hz/1 sec), however, substantially prolongs EPSP-LTP (greater than or equal to 10 hr) and doubles the approximated half-life of pop-spike LTP. The threshold current intensity to elicit pop-spike responses decreased after the induction of LTP. Furthermore, the smaller field-EPSP values necessary to evoke near-threshold pop-spikes demonstrate an E-S potentiation (left-shift) at least in the low-intensity range. While the total duration of potentiation of the different parameters has not been determined, all the above mentioned effects could be observed at least 10 hr following the repeated tetanization. It is proposed that the slice preparation is suitable for the investigation of mechanisms of a postulated late phase of LTP if appropriate conditions are used.

Effect of serotonin on Y-maze retention and hippocampal protein synthesis in rats

The effect of serotonin (5-HT) on consolidation of a brightness discrimination reaction was investigated in rats. Five microgram 5-HT, injected intrahippocampally immediately after training, impaired retention of the brightness discrimination tested 24 hr later. In biochemical experiments, leucine incorporation into hippocampal proteins in vivo was 32% inhibited by 5 microgram 5-HT. Leucine incorporation into proteins of hippocampal slices in vitro was decreased by 5 x 10(-5)M 5-HT. The results seem to support Essmans assumption that inhibition of brain protein synthesis by 5-HT may be responsible for the memory impairment. But also some other possibilities for a mechanism of 5-HT amnesia must be discussed.

Calcium-induced long-term potentiation in the hippocampal slice: Characterization of the time course and conditions

A transient increase in extracellular calcium concentration causes a long-lasting enhancement of radiatum fibers evoked excitatory postsynaptic potential and population spike responses of CA1 pyramidal neurons which resembles long-term potentiation (LTP). The duration of this potentiation is much longer than described previously and is probably limited by the survival of the preparation itself (greater than 8 hr). Therefore, Ca-induced LTP can be used for the investigation of a postulated late phase of LTP. Ca effects were activity-independent, since the subsequently evoked responses were facilitated even when the presynaptic fibers were not concurrently stimulated during or immediately after superfusion with the high Ca medium. In contrast, if too frequent testing of the synaptic input was done during the high Ca pulse, a short lasting depression instead of potentiation was observed. A lower extracellular magnesium concentration in the standard medium (1.3 instead of 2.0 mM MgSO4) prevents the potentiation of the EPSP at least for the first few hours. Presumably, both tetanus- and Ca-induced LTP share some common mechanisms, since an additional tetanization after Ca induction was not followed by an additional LTP. Compared to the potentiation following tetanization, the Ca-induced LTP was, however, not accompanied by a potentiation of the EPSP/spike ratio within the range of the population spike threshold intensity.

c-fos Protooncogene expression in rat hippocampus and entorhinal cortex following tetanic stimulation of the perforant path

The elevated expression of the c-fos protooncogene has been proposed to be a marker of cell activation leading to a long term cellular response. In this communication we compared the c-fos mRNA accumulation in the hippocampus (i.e. postsynaptic cells) and entorhinal cortex (i.e. presynaptic cells) following high (tetanic) and low frequency electrical stimulation of the perforant path. Using Northern blot analysis we have found that high frequency stimulation elevates c-fos expression in both hippocampus and entorhinal cortex, and the increase of c-fos mRNA levels in the entorhinal cortex is less pronounced, but longer lasting, than in the hippocampus. Slight increase of c-fos mRNA levels has been also observed in low frequency treated animals in the entorhinal cortex, but not in the hippocampus. These findings raise the question about differences in mechanisms involved in c-fos activation in both parts of the brain after stimulation which evokes long term potentiation (LTP) of synaptic efficacy.