Yuri Geinisman

Induction of long-term potentiation is associated with an increase in the number of axospinous synapses with segmented postsynaptic densities

Long-term potentiation (LTP) is characterized by a long-lasting enhancement of synaptic efficacy which may be due to an increase in synaptic numbers. The present study was designed to verify the validity of this suggestion using recently developed unbiased methods for synapse quantitation. LTP was elicited in young adult rats by high-frequency stimulation of the medial perforant path carried out on each of 4 consecutive days. Potentiated animals were sacrificed 1 h after the fourth stimulation. Stimulated but not potentiated and implanted but not stimulated rats served as controls. Synapses were examined in the middle (MML) and inner (IML) molecular layer of the hippocampal dentate gyrus. Using the stereological disector technique, unbiased estimates of the number of synapses per neuron were differentially obtained for the following morphological synaptic types: axodendritic synapses involving dendritic shafts, non-perforated axospinous synapses exhibiting a continuous postsynaptic density (PSD) and perforated axospinous synapses distinguished by a fenestrated, horseshoe-shaped or segmented PSD. A major finding of this study is that the induction of LTP is accompanied by a selective increase in the number of synapses with segmented PSDs. This change was detected only in the potentiated synaptic field (MML), but not in an immediately adjacent one (IML) which was not directly stimulated during the induction of LTP. It is strongly suggested by the latter finding that the increase in the number of axospinous synapses exhibiting segmented PSDs is associated with LTP. Such a highly selective modification of connectivity, which involves only one particular subtype of synapses in the potentiated synaptic field, is likely to represent a structural substrate of the enduring augmentation of synaptic efficacy typical of LTP.

Age-dependent alterations in hippocampal synaptic plasticity: Relation to memory disorders

In this paper, we review the evidence indicating that the common disturbance in recent memory associated with aging is a consequence of functional and structural impairment in the hippocampal formation. In the Fischer 344 rat, an experimental model of the human age-related memory disorder was developed. The majority of aged rats of this strain show impaired performance in the 8-arm radial maze in a manner typical of young rats with bilateral hippocampal lesions. Aged animals also exhibit rapid decay of LTP and slower kindling of the perforant path-dentate synapse. Furthermore, quantitative morphometric analysis of the hippocampal synaptic architecture revealed that aged, memory-impaired rats had a specific loss of perforated axospinous synapses in the middle third of the dentate gyrus molecular layer; the extent of loss was directly related to the degree of memory dysfunction. Most important was the fact that the electrophysiological and morphological abnormalities did not appear in equally old animals with good memory.

Reduction in Size of Perforated Postsynaptic Densities in Hippocampal Axospinous Synapses and Age-Related Spatial Learning Impairments

A central problem in the neurobiology of normal aging is why learning is preserved in some aged individuals yet impaired in others. To investigate this issue, we examined whether age-related deficits in spatial learning are associated with a reduction in postsynaptic density (PSD) area in hippocampal excitatory synapses (i.e., with a structural modification that is likely to have a deleterious effect on synaptic function). A hippocampus-dependent version of the Morris water maze task was used to separate Long-Evans male rats into young adult, aged learning-unimpaired, and equally aged learning-impaired groups. Axospinous synapses from the CA1 stratum radiatum were analyzed using systematic random sampling and serial section analyses. We report that aged learning-impaired rats exhibit a marked (∼30%) and significant reduction in PSD area, whereas aged learning-unimpaired rats do not. The observed structural alteration involves a substantial proportion of perforated synapses but is not observed in nonperforated synapses. These findings support the notion that many hippocampal perforated synapses become less efficient in aged learning-impaired rats, which may contribute to cognitive decline during normal aging.

Synapse restructuring associated with the maintenance phase of hippocampal long-term potentiation

Synapses in the middle molecular layer of the rat dentate gyrus were analyzed by electron microscopy during the maintenance phase of long‐term potentiation (LTP). LTP was induced by high‐frequency stimulation of the medial perforant path carried out on each of 4 consecutive days. The dentate gyrus was examined electron microscopically 13 days following the fourth stimulation. At this time point, synaptic responses were still significantly enhanced relative to baseline, although the extent of their potentiation was lower than 1 hour after the last high‐frequency stimulation. Stimulated, but not potentiated, rats served as controls. Using the stereological double disector method, estimates of the number of different morphological types of synapses per postsynaptic neuron were obtained. The number of asymmetrical axodendritic synapses increased (by 28%) during LTP maintenance, whereas the number of other synaptic types was not significantly altered. Our previous work demonstrated that the induction of LTP is followed by a selective increase in the number of axospinous perforated synapses with multiple, completely partitioned, transmission zones. Thus, the induction and maintenance phases of LTP are characterized by different structural synaptic alterations. These alterations may be related to each other as indicated by another finding of the present study regarding the existence of perforated synapses that appear to be transitional between axospinous and axodendritic junctions. This suggests a model of structural synaptic plasticity associated with LTP in which some axospinous perforated synapses increase in numbers shortly after the induction of LTP and are then converted into axodendritic ones during LTP maintenance.

Aged rats need a preserved complement of perforated axospinous synapses per hippocampal neuron to maintain good spatial memory

Spatial working memory, which crucially depends on the structural integrity of the hippocampal formation and its afferent connections, is impaired in the most, but not all, of aged rats. This study was designed to verify whether aged animals that do not exhibit the spatial memory deficit are the ones in which the hippocampal synaptic connectivity remains preserved with advancing chronological age. Young adult rats with good spatial memory, aged rats with impaired spatial memory and equally aged rats with intact spatial memory were compared. The number of synapses per neuron was estimated in the hippocampal dentate gyrus. The most important results were obtained when axospinous synapses were divided into perforated and non-perforated ones according to the appearance of their postsynaptic density. A significant decrease in the number of perforated synapses was found in memory-impaired aged rats as compared to either young adults or aged animals without memory deficits. The number of non-perforated synapses per neuron was diminished in memory-deficient aged rats relative to young adults, but not to memory-intact aged rats. However, it was only the loss of perforated synapses which correlated with the degree of spatial memory impairment. Thus, aged rats need a preserved complement of hippocampal perforated synapses to maintain good spatial memory.

Distance-Dependent Differences in Synapse Number and AMPA Receptor Expression in Hippocampal CA1 Pyramidal Neurons

The ability of synapses throughout the dendritic tree to influence neuronal output is crucial for information processing in the brain. Synaptic potentials attenuate dramatically, however, as they propagate along dendrites toward the soma. To examine whether excitatory axospinous synapses on CA1 pyramidal neurons compensate for their distance from the soma to counteract such dendritic filtering, we evaluated axospinous synapse number and receptor expression in three progressively distal regions: proximal and distal stratum radiatum (SR), and stratum lacunosum-moleculare (SLM). We found that the proportion of perforated synapses increases as a function of distance from the soma and that their AMPAR, but not NMDAR, expression is highest in distal SR and lowest in SLM. Computational models of pyramidal neurons derived from these results suggest that they arise from the compartment-specific use of conductance scaling in SR and dendritic spikes in SLM to minimize the influence of distance on synaptic efficacy.

Aging, spatial learning, and total synapse number in the rat CA1 stratum radiatum

The aim of this study was to determine whether spatial learning deficits in aged rats are associated with a loss of hippocampal synapses. The Morris water maze task was used to assess the spatial learning capacity of young and aged rats and to attribute aged animals to learning-impaired and learning-unimpaired groups. The number of axospinous synapses in the entire volume of the CA1 stratum radiatum was estimated with unbiased stereological techniques. The results show that the total number of all axospinous synapses and of their perforated and nonperforated subtypes remains constant in the CA1 stratum radiatum of aged learning-impaired rats as compared to aged learning-unimpaired rats and to young adults. Thus, neither age-related deficits in spatial learning nor advanced chronological age are associated with a loss of axospinous synapses from the rat CA1 stratum radiatum.

Decrease in the number of synapses in the senescent brain: A quantitative electron microscopic analysis of the dentate gyrus molecular layer in the rat

Axo-dendritic synapses were counted in electron micrographs taken from the middle third of the dentate gyrus molecular layer of young adult and senescent Fischer 344 rats. A significant decrease in the number of synapses was found in senescent animals relative to young ones. This loss of synapses, which involved all the morphological varieties of axo-dendritic synaptic contacts in the dentate gyrus molecular layer, appeared to be unrelated to changes in dimensions of synapses, tissue volume or number of postsynaptic granule cells. It is proposed that the age-related loss of synaptic contacts might be attributed to a reduced capacity of senescent brains for synaptic regeneration and remodelling.

Differences in the expression of AMPA and NMDA receptors between axospinous perforated and nonperforated synapses are related to the configuration and size of postsynaptic densities

Axospinous synapses are traditionally divided according to postsynaptic density (PSD) configuration into a perforated subtype characterized by a complex‐shaped PSD and nonperforated subtype exhibiting a simple‐shaped, disc‐like PSD. It has been hypothesized that perforated synapses are especially important for synaptic plasticity because they have a higher efficacy of impulse transmission. The aim of the present study was to test this hypothesis. The number of postsynaptic AMPA receptors (AMPARs) is widely regarded as the major determinant of synaptic efficacy. Therefore, the expression of AMPARs was evaluated in the two synaptic subtypes and compared with that of NMDA receptors (NMDARs). Postembedding immunogold electron microscopy was used to quantify the immunoreactivity following single labeling of AMPARs or NMDARs in serial sections through the CA1 stratum radiatum of adult rats. The results showed that all perforated synapses examined were immunopositive for AMPARs. In contrast, only a proportion of nonperforated synapses (64% on average) contained immunogold particles for AMPARs. The number of immunogold particles for AMPARs was markedly and significantly higher in perforated synapses than in immunopositive nonperforated synapses. Although all synapses of both subtypes were NMDAR immunopositive perforated synapses contained significantly more immunogold particles for NMDARs than nonperforated ones. Multivariate analysis of variance revealed that the mode of AMPAR and NMDAR expression is related to the complexity of PSD configuration, not only to PSD size. These findings support the notion that perforated synapses may evoke larger postsynaptic responses relative to nonperforated synapses and, hence, contribute to an enhancement of synaptic transmission associated with some forms of synaptic plasticity. J. Comp. Neurol. 468:86–95, 2004. Published 2003 Wiley‐Liss, Inc.

Anatomical evidence for convergence of olfactory, gustatory, and visceral afferent pathways in mouse cerebral cortex

Flavor perception requires the neural integration of olfactory, gustatory and, possibly, visceral afferent information. Presently, it is not known where, or how this integration takes place in the brain. Neuroanatomical data presented here suggest that pathways subserving these sensory modalities converge in mouse insular cortex after surprisingly few synaptic relays. Orthograde transport of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP) was used to label main olfactory bulb (MOB) efferents. A projection into layer I of insular cortex was present in every case. Bulb transections were made to provoke anterograde degeneration and EM analysis confirmed that the olfactory projection to insular cortex was a terminal pathway. WGA-HRP injections in the MOB-recipient zone of insular cortex resulted in ortho and retrograde labeling of ascending and descending gustatory-visceral afferent pathways. It is concluded that in the mouse, there is a remarkably direct convergence of olfactory and gustatory-visceral sensory pathways in insular cortex. Together with the descending connections from insular cortex to the amygdala and to brainstem autonomic structures, it is possible that the cortical integration of olfactory and gustatory-visceral information could modulate mechanisms involved in food selection and autonomic reactions relating to the chemical senses. Basic mechanisms subserving flavor perception might be usefully modelled in mouse insular cortex.