Major goals of research into the neurobiology of learning and memory are to identify (1) brain areas/circuitries that subserve different mnemonic functions and (2) chemistries that encode the memory trace. The discovery that activity modulates neuronal gene expression provided techniques attendant to the first goal and candidates for cellular changes pertinent to the second. Studies in our laboratories have exploited activity-regulated changes in c-fos gene expression to map regions engaged in two-odor discrimination learning, with particular interest in neuronal groups in hippocampus and amygdala. The results of these studies demonstrate that the subdivisions of hippocampus and amygdala do not act in concert across behaviors but are differentially activated depending on task demands. In hippocampus, preferential activation of field CA3 was uniquely associated with initial learning of an odor pair, whereas predominant activation of CA1 occurred with exploration of a novel field and with overtrained responding to odors. The reappearance of precisely the same balance of subfield activation within disparate behavioral contexts was taken to suggest that the hippocampus has basic modes of function that recur in different circumstances and make rather generalized contributions to behavior. Within the amygdala, the basolateral division was most prominently active during task acquisition but not during performance of the well-learned discrimination. Indeed, the amygdala appeared to play the dominant role relative to hippocampus in the early stages of associating positive and negative valences with discriminative cues. These results demonstrate that the balance of neuronal activity both within and between limbic structures changes across sequential stages of odor learning in a fashion that is likely to define behavioral output.

Amygdala↗

PubMedDate not supplied

Memory and the brain: unexpected chemistries and a new pharmacology.

Efforts to characterize long-term potentiation (LTP) and to identify its substrates have led to the discovery of novel synaptic chemistries, computational algorithms, and, most recently, pharmacologies. Progress has also been made in using LTP to develop a "standard model" of how unusual, but physiologically plausible, levels of afferent activity create lasting changes in the operating characteristics of synapses in the cortical telencephalon. Hypotheses of this type typically distinguish induction, expression, and consolidation stages in the formation of LTP. Induction involves a sequence consisting of theta-type rhythmic activity, suppression of inhibitory currents, intense synaptic depolarization, NMDA receptor activation, and calcium influx into dendritic spines. Calcium-dependent lipases, kinases, and proteases have been implicated in LTP induction. Regarding the last group, it has been recently reported that theta pattern stimulation activates calpain and that translational suppression of the protease blocks potentiation. It is thus likely that proteolysis is readily driven by synaptic activity and contributes to structural reorganization. LTP does not interact with treatments that affect transmitter release, has a markedly differential effect on the currents mediated by colocalized AMPA vs NMDA synaptic receptors, changes the waveform of the synaptic current, modifies the effects of drugs that modulate AMPA receptors, and is sensitive to the subunit composition of those receptors. These results indicate that LTP is expressed by changes in AMPA receptor operations. LTP is accompanied by modifications in the anatomy of synapses and spines, something which accounts for its extreme duration (weeks). As with various types of memory, LTP requires about 30 min to consolidate (become resistant to disruption). Consolidation involves adhesion chemistries and, in particular, activation of integrins, a class of transmembrane receptors that control morphology in numerous cell types. Platelet activating factor and adenosine may contribute to consolidation by regulating the engagement of latent integrins. How consolidation stabilizes LTP expression is a topic of intense investigation but probably involves modifications to one or more of the following: membrane environment of AMPA receptors; access of regulatory proteins (e.g., kinases, proteases) to the receptors; receptor clustering; and space available for receptor insertion. Attempts to enhance LTP have focused on the induction phase and resulted in a class of centrally active drugs ("ampakines") that positively modulate AMPA receptors. These compounds promote LTP in vivo and improve the encoding of variety of memory types in animals. Positive results have also been obtained in preliminary studies with humans.

Age Factors↗

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Low levels of calpain activity in Chiroptera brain: implications for mechanisms of aging.

Calcium-dependent neutral proteases ("calpains") have been implicated in degenerative processes in muscles and neurons, suggesting that they might also play a role in age-related brain pathologies and perhaps in brain aging itself. Because Chiroptera exhibit an unusual maximum life span, relative to other mammalian orders, we investigated the activity of these enzymes in the brain of two species of bats. As in other mammals, brain calpain degrades many proteins associated with the cell cytoskeleton. However, enzyme activity is 5-7 fold lower in bat's brain than in a similar-sized mammal, such as the mouse. Moreover, the maximal life span of bats predicted from the equation relating calpain activity and maximal life span across a wide range of mammals is close to the observed values. These results strengthen the hypothesis that calpain activity is somehow linked to the rate at which brains age.

Aging↗

PubMedDate not supplied

Long lasting changes in the spontaneous activity of hippocampal neurons following stimulation of the entorhinal cortex.

The spontaneous activity of hippocampal and denate units was investigated in acute and chronic experiments on adult rats. Spontaneous unitary discharges from both regions were measured before and after brief tetanic stimulation of the entorhinal cortex. An increase in the frequency of spontaneous unit activity was obtained in units recorded from the dentate gyrus and CA3 field of the hippocampus for up to 20 min following stimulation. In chronic recordings of physiologically identified dentate units from freely moving rats similar results were obtained. The findings are discussed with reference to long lasting potentiation of synaptic activity in the perforant path-dentate granule cell connection.

Action Potentials↗

PubMedDate not supplied

Hypertrophy and redistribution of astrocytes in the deafferented dentate gyrus.

The response of the astroglial population of the dentate gyrus molecular layer to removal of that region's primary afferent was investigated using Cajal's gold sublimate method. Deafferentation caused the astrocytes to hypertrophy, an effect which was detectable at 24 hr and maximal at 72-96 hr post-lesion. Following this, the astroglia entered a lengthy period of gradual atrophy. Counts of the astrocytes in the various sublayers of the molecular layer led to the conclusion that these cells migrate into denervated dendritic areas from neighboring, nondeafferented zones.

Animals↗

PubMedDate not supplied

Olfactory tests as possible probes for detecting and monitoring Alzheimer's disease.

One of the characteristics of Alzheimer's disease is the early loss of neurons in pathways involved in processing olfactory information. Olfactory function was assessed in subjects with Alzheimer's disease using a conventional Smell Identification Test and a simple three odor match-to-sample problem. The patients exhibited a diminished capacity to identify common odors but were severely impaired in their ability to use novel odors in a match-to-sample task. Subjects with Parkinson's disease had a severe deficit for identifying common odors with the majority scoring as anosmic. Multiple sclerosis was not accompanied by detectable changes in olfactory functioning. The results of the Alzheimer's group are similar to recent animal studies that have shown lesions of the piriform-entorhinal cortex produce a variety of memory deficits that are particularly acute in tasks involving novel odors.

Aged↗

PubMedDate not supplied

Intracellular injections of EGTA block induction of hippocampal long-term potentiation.

Hippocampal long-term potentiation (LTP) is a remarkably stable facilitation of synaptic responses resulting from very brief trains of high-frequency stimulation. Because of its persistence and modest induction conditions, LTP represents a promising candidate for a substrate of memory. Some progress has been made in localizing the changes responsible for the effect; for example, it has been shown that LTP is not accompanied by changes in the fibre volleys of the test afferents or by generalized alterations of the dendrites of their target cells. However, it is unknown whether the potentiation is due to pre- or postsynaptic changes and there is evidence in favour of each (for example, see refs 5, 6). We now report that intracellular injections of the calcium chelator EGTA block the development of LTP. These results strongly suggest that LTP is caused by a modification of the postsynaptic neurone and that its induction depends on the level of free calcium.

Animals↗

PubMedDate not supplied

Regulation of glutamate receptor binding by the cytoskeletal protein fodrin.

The erythrocyte cytoskeleton, which consists primarily of a meshwork of spectrin and actin, controls cell shape and the disposition of proteins within the membrane. Proteins similar to spectrin have recently been found in diverse cells and tissues, and it is possible that they mediate the capping of cell-surface receptors, although this has not been demonstrated directly. In neurones, the spectrin-like protein fodrin lines the cortical cytoplasm and may link actin filaments to the membrane. Fodrin has been hypothesized to regulate the number of receptor binding sites on neuronal membranes for the putative neurotransmitter L-glutamate. Micromolar calcium concentrations activate the thiol protease calpain I, induce fodrin degradation and more than double the density of glutamate binding sites; these effects are all blocked by thiol protease inhibitors. We have now used specific antibodies to examine further the role of fodrin proteolysis in regulating glutamate receptors. We report that fodrin antibodies block the fodrin degradation and increase in glutamate binding normally induced by calcium, and so provide direct evidence for control of membrane receptors by a non-erythroid spectrin.

Animals↗

PubMedDate not supplied

Excitatory amino acids inhibit stimulation of phosphatidylinositol metabolism by aminergic agonists in hippocampus.

Since the initial observations in the 1950s a large number of neurotransmitters and hormones have been shown to influence phosphatidylinositol (PI) metabolism in brain and peripheral ganglia (see ref. 3 for review). This has led to the suggestion that PI is part of an intracellular second messenger system for some types of diffusible chemical factors. Consistent with this are recent reports that one of the products of PI turnover (diacylglycerol) stimulates the Ca-dependent phospholipid-dependent protein kinase (kinase C) while a second (inositol trisphosphate) causes the release of calcium from intracellular stores. Thus it is possible that at least some brain neurotransmitters utilize the PI system to produce functional effects that are in addition to and which outlast the very brief physiological responses they elicit. Although it had been anticipated that another class of receptors might inhibit receptor-mediated stimulation of PI breakdown, no clear examples of such effects have been described. We now report that acidic amino acids, which are that acidic amino acids, which are thought to be excitatory neurotransmitters at the majority of brain synapses, strongly inhibit the stimulation of PI metabolism elicited by carbachol, histamine, or by potassium-induced depolarization, without changing the response to noradrenaline. As well as indicating a novel function for the excitatory amino acids, these results suggest that the central nervous system possesses cell-cell interactions of a previously unsuspected type.

Amino Acids↗

PubMedDate not supplied

Translational suppression of calpain blocks long-term potentiation.

Transfection with antisense oligonucleotides was used to reduce calpain 1 activity to approximately 50% of normal values in cultured hippocampal slices. This had no detectable effects on baseline synaptic responses but greatly reduced the incidence and magnitude of long-term potentiation induced with a theta-burst stimulation paradigm. These results suggest that activation of calpain by repetitive bursts of afferent activity, as shown to occur in prior studies, is an essential step in the production of stable increases in synaptic strength.

Animals↗

PubMedDate not supplied

Differential patterns of c-fos mRNA expression in amygdala during successive stages of odor discrimination learning.

Expression of the activity-dependent gene c-fos was used to assess relative levels of neuronal activation in the amygdala and related structures of rats at different stages of odor discrimination learning. In situ hybridization was used to evaluate c-fos mRNA content within the amygdalar subdivisions, the bed nucleus of the stria terminalis, and the hippocampus. After initial exploration of the test apparatus, c-fos mRNA levels were increased in the medial and, to lesser extent, basolateral subdivisions and remained low in the central division. The balance of amygdala to hippocampal labeling favored hippocampus. Rats engaged in familiar nose-poke responses had comparably elevated labeling in the medial and basolateral divisions and low labeling densities in the central division. The ratio of hippocampal to amygdala labeling was at control levels. Rats required to switch from ad libitum responding to cued responding to odors had high basolateral to medial labeling ratios. This was in marked contrast to the medial dominance found in control and exploration rats. Hybridization was substantially more dense in basolateral amygdala than in hippocampal CA1; this imbalance was unique to the group required to form first associations between odors and rewards. Rats performing an overtrained odor discrimination had the least differentiation between amygdalar subdivisions of any behavioral group. The hippocampus-to-amygdala labeling ratio favored hippocampus and was nearly identical to the ratio in exploration rats. These results demonstrate that the balance of activity within the between limbic structures shifts according to behavioral demands. It is suggested that the balances reflect the availability of pertinent afferent cues, interactions between hippocampus and the extended amygdala, and relative levels of activity in the diffuse projections to the limbic system.

Amygdala↗