John Larson

Alterations in synaptic transmission and long-term potentiation in hippocampal slices from young and aged PDAPP mice

Synaptic transmission and plasticity were studied in the CA1 field of hippocampal slices from young and aged transgenic mice over-expressing a mutant form of the human amyloid precursor protein (PDAPP mice). The transgenic mice at 4-5 months of age, prior to the formation of amyloid-beta peptide deposits in these animals, differed from non-transgenic control mice in three respects: (1) paired-pulse facilitation (PPF) was enhanced; (2) responses to high frequency stimulation bursts were distorted; (3) long-term potentiation (LTP) decayed more rapidly. More striking was the profound reduction in the size of synaptic responses and frequent loss of field potentials that were found in the transgenic mice at 27-29 months, an age at which they exhibit numerous amyloid plaques, neuritic dystrophy, and gliosis. Control mice at these ages did not show such dramatic effects. PPF was reduced in aged transgenic mice, compared to aged controls; however, LTP was still in evidence, although direct comparisons of its induction conditions in aged transgenic and control mice were compromised by the profound differences in field potentials between the two groups. These results point to two conclusions: (1) altered synaptic communication appears in PDAPP mice in advance of amyloid plaque formation and probably involves changes in presynaptic calcium kinetics; (2) the disturbances in synaptic transmission that appear when abundant plaques and Alzheimers-like neuropathology are present in the transgenic mice are not necessarily accompanied by a disproportionate loss of long-term synaptic plasticity.

Reversal of LTP by theta frequency stimulation

Reversal of long-term potentiation (LTP) by physiological stimulation was tested in the CA1 field of hippocampal slices. In control medium, a one minute episode of 5 Hz (theta frequency) stimulation beginning 1-3 min after LTP had no effect on the degree of potentiation measured 30 min later. However, in the presence of norepinephrine (200 microM), 5 Hz stimulation reduced LTP by about 30%. Theta frequency stimulation was only effective when administered within 10 min of LTP induction and had no lasting effects on non-potentiated synapses. Stimulation at 1 Hz did not reverse LTP and stimulation at 10 Hz was no more effective than 5 Hz stimulation. LTP could be nearly completely reversed by theta frequency stimulation when potentiation was induced by milder and more naturalistic stimulation patterns. Under these conditions, LTP reversal was blocked by an antagonist of adenosine A1 receptors. These results suggest that the hippocampal theta rhythm promotes both the induction of LTP and its subsequent reversal with the latter process involving activation of adenosine receptors. Reversal of LTP may function to refine or sharpen recently encoded representations.

Stimulation of NMDA receptors induces proteolysis of spectrin in hippocampus

Stimulation of N-methyl-D-aspartate (NMDA) receptors was found to induce proteolysis of brain spectrin in hippocampal slices. The effect was dependent upon extracellular calcium, blocked by the antagonist 2-amino-5-phosphonovalerate (AP5), and was not reproduced by potassium-induced depolarization. These results are consistent with the hypothesis that the involvement of NMDA receptors in plasticity and excitotoxicity is at least partially mediated by calcium-activated proteolysis of cytoskeletal proteins.

Theta pattern stimulation and the induction of LTP: the sequence in which synapses are stimulated determines the degree to which they potentiate.

Induction of long-term potentiation (LTP) by asynchronous stimulation of converging afferents was studied in hippocampal slices. Three stimulation electrodes were positioned to activate separate groups of Schaffer-commissural inputs to a population of CA1 pyramidal cells. Patterned stimulation consisted of a single coincident priming pulse to all 3 electrodes followed by a burst of 4 pulses (100 Hz) to the first input (S1) at a delay of 180 ms, to the second (S2) at a delay of 200 ms, and to the third (S3) at a delay of 220 ms. This pattern was repeated 10 times at 5-s intervals. The magnitude of LTP induced (measured 20 min after stimulation) was greatest for the first stimulated input, intermediate for the second, and least for the third. Intracellular recordings indicated that the greatest postsynaptic depolarization occurred during the period of S2 stimulation; thus the magnitude of LTP induced was not simply dependent on the degree of depolarization during afferent activation. Rather, sustained depolarization after synaptic activation could contribute to LTP induction by prolonging the activity of N-methyl-D-aspartate receptor-gated channels. Earlier-arriving bursts may also trigger an inhibitory process that reduces the effectiveness of later bursts for inducing LTP.

Development of hippocampal long-term potentiation is reduced by recently introduced calpain inhibitors

The effects of two recently synthesized inhibitors of calpains, calpain inhibitor I (CiI) and calpain inhibitor II (CiII) were tested on the development of long-term potentiation (LTP) in region CA1 of rat hippocampus. Slices maintained in 100 microM of CiI or CiII showed an initial degree of potentiation after theta burst stimulation that, in contrast to controls, slowly decayed across time. The effects of CiI and CiII appeared to be independent of possible actions on the physiological mechanisms that take place during the induction stage of LTP. Since these inhibitors are more potent and specific than leupeptin in blocking calpain activity, their effects on LTP can be more convincingly ascribed to a selective blockade of the calcium-sensitive protease. Accordingly, the results favor the idea that a proteolytic event of the kind found after N-methyl-D-aspartate receptor activation is an intermediary step in the development of LTP.

Chronic administration of a thiol-proteinase inhibitor blocks long-term potentiation of synaptic responses

It has been proposed that activation of a calcium-sensitive protease (calpain) is a crucial step in the induction of long-term potentiation (LTP). To test this hypothesis, we used chronic recording techniques to measure the effects of intraventricular infusion of leupeptin, a calpain inhibitor, on LTP in the hippocampus. Rats implanted bilaterally with stimulating electrodes in the Schaffer-commissural system and one recording electrode in the apical dendrites of field CA1 were fitted with osmotic mini-pumps delivering either leupeptin (20 mg/ml) or saline at a rate of 0.5 microliter/h into the lateral ventricle. Short bursts of high-frequency stimulation with the bursts delivered at 5/s were used to induce LTP in those animals which had stable responses for several days. Rats in the saline group (n = 11) exhibited an immediate LTP effect that remained in place over successive days of testing, while only 3 of 13 leupeptin treated animals showed evidence of LTP 24 h after high-frequency stimulation, and in only one of those was a sizeable effect recorded over several days. The average change in responses at the 24-h test point was +33% for the controls and +4% for the leupeptin group (P less than 0.01). The block of LTP induction was reversible, since high-frequency stimulation applied after disconnecting the pumps led to a robust LTP effect that lasted for several days in 6 of 7 animals tested. There were no detectable differences in baseline responses in the presence and absence of leupeptin.

Anoxia reveals a vulnerable period in the development of long-term potentiation

Transient anoxia occurring 1-2 min after high-frequency stimulation selectively prevented the stable expression of long-term potentiation (LTP). Anoxia occurring after this brief vulnerable period did not reverse LTP. Experiments on the duration of anoxia necessary to block LTP expression indicated that simply reducing synaptic transmission was insufficient but that membrane depolarization was not required. The effects of anoxia on LTP were blocked by antagonists of A1 adenosine receptors. It is concluded that LTP develops in about one minute and that the chemistries operating in this period are easily disrupted by an event triggered by adenosine receptors.

Lesions of entorhinal cortex produce a calpain-mediated degradation of brain spectrin in dentate gyrus. I. Biochemical studies

Lesions of the rat entorhinal cortex cause extensive synaptic restructuring and perturbation of calcium regulation in the dentate gyrus of hippocampus. Calpain is a calcium-activated protease which has been implicated in degenerative phenomena in muscles and in peripheral nerves. In addition, calpain degrades several major structural neuronal proteins and has been proposed to play a critical role in the morphological changes observed following deafferentation. In this report we present evidence that lesions of the entorhinal cortex produce a marked increase in the breakdown of brain spectrin, a substrate for calpain, in the dentate gyrus. Two lines of evidence indicate that this effect is due to calpain activation: (i) the spectrin breakdown products observed following the lesion are indistinguishable from calpain-generated spectrin fragments in vitro; and (ii) their appearance can be reduced by prior intraventricular in fusion of leupeptin, a calpain inhibitor. Levels of spectrin breakdown products are increased as early as 4 h post-lesion, reach maximal values at 2 days, and remain above normal to some degree for at least 27 days. In addition, a small but significant increase in spectrin proteolysis is also observed in the hippocampus contralateral to the lesioned side in the first week postlesion. At 2 days postlesion the total spectrin immunoreactivity (native polypeptide plus breakdown products) increases by 40%, suggesting that denervation of the dentate gyrus produces not only an increased rate of spectrin degradation but also an increased rate of spectrin synthesis. These results indicate that calpain activation and spectrin degradation are early biochemical events following deafferentation and might well participate in the remodelling of postsynaptic structures. Finally, the magnitude of the observed effects as well as the stable nature of the breakdown products provide a sensitive assay for neuronal pathology.

Long-term potentiation: Persisting problems and recent results

In this paper we discuss recent experimental results pertinent to three unresolved issues regarding the long-term potentiation (LTP) effect: the nature of its enduring substrates, the biochemical mechanisms that produce it, and its potential role in memory. LTP appears to be triggered by a postsynaptic influx of calcium and is associated with alterations in the shape of dendritic spines and probably the formation of new synapses. We discuss the possibility that morphological reorganization also modifies membrane surface chemistry of synaptic elements. Evidence is presented that LTP is not associated with changes in presynaptic calcium currents. Activation of protein kinase C is shown to be insufficient for the induction of LTP, although it may play a modulatory role. The hypothesis that activation of a calcium-sensitive protease (calpain) is pivotal to the establishment of LTP is supported by experiments showing that a calpain inhibitor, leupeptin, blocks LTP. Furthermore, activation of NMDA receptors, an event implicated in LTP induction, is accompanied by calcium-sensitive proteolysis of spectrin, a major dendritic cytoskeletal protein. The finding that stimulation patterns designed to mimic naturally-occurring cell discharge patterns are highly effective for LTP induction greatly strengthens the hypothesis that LTP actually occurs during the encoding of information in cortical systems. Potential contributions of LTP to learning are explored using computer simulations of a simple cortical network.

Theta burst stimulation is optimal for induction of LTP at both apical and basal dendritic synapses on hippocampal CA1 neurons

The efficacy of stimulation patterns consisting of brief high frequency bursts repeated at various intervals to induce long-term potentiation (LTP) at synapses on apical and basal dendrites of CA1 hippocampal neurons was tested in vitro. Both apical and basal dendritic synapses exhibited maximal LTP after bursts repeated at 5-10 Hz, i.e. close to the frequency of the endogenous hippocampal theta rhythm. As at apical dendritic synapses, LTP at basal dendritic synapses was blocked by an antagonist of NMDA receptors. Basal dendritic LTP was significantly greater in magnitude than apical dendritic LTP, although the reason for this is unknown.