Dendritic spines of rat cerebellar Purkinje cells: serial electron microscopy with reference to their biophysical characteristics

We have used serial electron microscopy and 3-dimensional reconstructions of dendritic spines from Purkinje spiny branchlets of normal adult rats to evaluate 2 questions about the relationship of spine geometry to synaptic efficacy. First, do relationships between spine geometry and other anatomical indicators of synaptic activity suggest that spine size and shape might be associated with synaptic efficacy? Reconstructed spines were graphically edited into head and neck compartments; the area of the postsynaptic density (PSD) was measured; the volume of spine smooth endoplasmic reticulum (SER) was computed; and all of the vesicles in the axonal varicosities were counted. Spine head volume and the volume of SER contained in the head are well correlated with the area of the PSD and the number of vesicles in the presynaptic axonal varicosity. Spine neck diameter does not fluctuate with PSD area, head volume, or the vesicle number. These results suggest that the dimensions of the spine head, but not of the spine neck, are likely to reflect differences in synaptic efficacy. Second, does the geometry of cerebellar spine necks reduce the transfer of synaptic charge to the recipient dendrite from the theoretical maximum that could be transferred if the synapse were on a dendritic shaft? Comparison of volume to surface area showed that the spine heads are approximately spherical and the necks are approximately cylindrical. Application of results from a biophysical model that assumed these geometrical shapes for spines (Wilson, 1984) showed that the cerebellar spine necks are unlikely to reduce transfer of synaptic charge by more than 5-20% even if their SER were to completely block passage of current through the portion of the neck that it occupies. We suggest that the constricted spine neck diameter might serve to isolate metabolic events in the vicinity of activated synapses by reducing diffusion to neighboring synapses, without significantly influencing the transfer of synaptic charge to the postsynaptic dendrite.     

Dimensions and density of dendritic spines from rat dentate granule cells based on reconstructions from serial electron micrographs

In the hippocampus, most excitatory synapses are located on dendritic spines. It has been postulated that the geometry of spines and/or the postsynaptic density (PSD) influences synaptic efficiency and may contribute to the expression of plastic processes such as learning or long-term potentiation (LTP). Based on three-dimensional reconstructions of dentate granule cell dendrites from serial electron micrographs, we have measured head dimensions, neck cross-sectional areas, neck length, and PSD area and form of 115 spines of dentate granule cells in the medial perforant path termination zone. All dimensions showed a large variability, with up to 100-fold differences in values. A calculated diffusion index for transport of molecules through the reconstructed neck varied over a 100-fold range. The neck and head dimensions were moderately positively correlated, whereas the PSD area was strongly correlated with head volume. Distribution histograms and scatter plots of various spine dimensions did not reveal any systematic clustering, suggesting that there is a continuum of spine geometries rather than distinct classes for granule cell dendritic spines in the middle molecular layer. Transversely (n = 13) and longitudinally (n = 27) sectioned dendrites had mean spine densities of 2.66 and 1.01 spines/μm, respectively, uncorrected for so-called hidden spines. Bifurcating spines made up 2.1% of the total spine number in transversely and 2.3% in longitudinally sectioned dendrites. The twin spine heads never shared the same presynaptic bouton. Fenestrated or split PSDs shared the same presynaptic element in all but two cases, arguing against PSD division as an intermediate step in synapse formation. J. Comp. Neurol. 377:15-28, 1997. © 1997 Wiley-Liss, Inc.     

Critical assessment of the involvement of perforations,spinules, and spine branching in hippocampal synapse formation

Several studies propose that long-term enhancement of synaptic transmission between neurons results from the enlargement, perforation, and splitting of synapses and dendritic spines. Unbiased analyses through serial electron microscopy were used to assess the morphological basis for synapse splitting in hippocampal area CA1. Few perforated synapses and almost no split (i.e., branched) spines occurred at postnatal day 15, an age of high synaptogenesis; thus, synapse splitting is unlikely to be important during development. The synapse splitting hypothesis predicts an intermediate stage of branched spines with both heads sharing the same presynaptic bouton. Ninety-one branched dendritic spines were traced through serial sections, and the different branches never synapsed with the same presynaptic bouton. Projections from spines, called “spinules,” have been thought to extend from perforations in the postsynaptic density (PSD), thereby dividing the presynaptic bouton. Forty-six spinules were traced, and only 13% emerged from perforations in the PSD. Most spinules emerged from the edges of nonperforated PSDs, or from spine necks, where they extended into boutons that were not presynaptic to the spine. In summary, these morphological characteristics are inconsistent with synapse and spine splitting. An alternative is discussed whereby perforated synapses and spinules are transient components of synaptic activation, and branched spines appear from synapses forming in close proximity to one another. J. Comp. Neurol. 398:225–240, 1998. © 1998 Wiley-Liss, Inc.     

Synaptic potentiation induces increased glial coverage of excitatory synapses in CA1 hippocampus

Patterns of activity that induce synaptic plasticity at excitatory synapses, such as long‐term potentiation, result in structural remodeling of the postsynaptic spine, comprising an enlargement of the spine head and reorganization of the postsynaptic density (PSD). Furthermore, spine synapses represent complex functional units in which interaction between the presynaptic varicosity and the postsynaptic spine is also modulated by surrounding astroglial processes. To investigate how activity patterns could affect the morphological interplay between these three partners, we used an electron microscopic (EM) approach and 3D reconstructions of excitatory synapses to study the activity‐related morphological changes underlying induction of synaptic potentiation by theta burst stimulation or brief oxygen/glucose deprivation episodes in hippocampal organotypic slice cultures. EM analyses demonstrated that the typical glia‐synapse organization described in in vivo rat hippocampus is perfectly preserved and comparable in organotypic slice cultures. Three‐dimensional reconstructions of synapses, classified as simple or complex depending upon PSD organization, showed significant changes following induction of synaptic potentiation using both protocols. The spine head volume and the area of the PSD significantly enlarged 30 min and 1 h after stimulation, particularly in large synapses with complex PSD, an effect that was associated with a concomitant enlargement of presynaptic terminals. Furthermore, synaptic activity induced a pronounced increase of the glial coverage of both pre‐ and postsynaptic structures, these changes being prevented by application of the NMDA receptor antagonist D‐2‐amino‐5‐phosphonopentanoic acid. These data reveal dynamic, activity‐dependent interactions between glial processes and pre‐ and postsynaptic partners and suggest that glia can participate in activity‐induced structural synapse remodeling. © 2009 Wiley‐Liss, Inc.     

Three-dimensional structure of dendritic spines and synapses in rat hippocampus (CA1) at postnatal day 15 and adult ages: implications for the maturation of synaptic physiology and long-term potentiation.

It has long been hypothesized that changes in dendritic spine structure may modify the physiological properties of synapses located on them. Due to their small size, large number, and highly variable shapes, standard light microscopy of Golgi impregnations and electron microscopy (EM) of single thin sections have not proved adequate to identify most spines in a sample or to quantify their structural dimensions and composition. Here we describe a new approach, the series sample, that was developed to classify by shape and subcellular composition all of the spines and synapses in a sample of neuropil by viewing them through serial EM sections. Spines in each class are then randomly selected for serial reconstruction and measurement in three dimensions. This approach was used to assess whether structural changes in hippocampal CA1 spines could contribute to the enhanced synaptic transmission and the greater endurance of long-term potentiation (LTP) that occur with maturation. Our results show a near doubling in the total density of synapses in the neuropil and along reconstructed dendrites between postnatal day 15 (PND 15) and adult ages. However, this doubling does not occur uniformly across all spine and synapse morphologies. Thin spines, mushroom spines containing perforated postsynaptic densities (PSDs) and spine apparatuses, and branched spines increase by about four-fold in density between PND 15 and adult ages. In contrast, stubby spines decrease by more than half and no change occurs in mushroom spines with macular PSDs or in dendritic shaft synapses. The stubby spines that remain are smaller in adults than at PND 15 and the mushroom spines are larger, while no change occurs in the three-dimensional structure of thin spines. Only a few spine necks at either age are constricted or long enough to attenuate charge transfer; therefore, the doubling in synapses should mediate the enhancement of synaptic transmission that occurs with maturation. In addition, LTP is not likely to be mediated by widening of spine necks at either age. However, the constricted spine necks could serve to concentrate specific molecules at activated synapses, thereby enhancing the specificity and endurance of LTP with maturation. These results demonstrate that the new series sample method combined with three-dimensional reconstruction reveals quantitative changes in the frequency and structure of spines and synapses that are not discernable by other methods and are likely to have dramatic effects on synaptic physiology and plasticity.     

Analysis of synaptic ultrastructure without fixative using high-pressure freezing and tomography

Electron microscopy allows the analysis of synaptic ultrastructure and its modifications during learning or in pathological conditions. However, conventional electron microscopy uses aldehyde fixatives that alter the morphology of the synapse by changing osmolarity and collapsing its molecular components. We have used high-pressure freezing (HPF) to capture within a few milliseconds structural features without aldehyde fixative, and thus to provide a snapshot of living synapses. CA1 hippocampal area slices from P21 rats were frozen at -173 degrees C under high pressure to reduce crystal formation, and synapses on dendritic spines were analysed after cryosubstitution and embedding. Synaptic terminals were larger than after aldehyde fixation, and synaptic vesicles in these terminals were less densely packed. Small filaments linked the vesicles in subgroups. The postsynaptic densities (PSDs) exhibited filamentous projections extending into the spine cytoplasm. Tomographic analysis showed that these projections were connected with the spine cytoskeletal meshwork. Using immunocytochemistry, we found as expected GluR1 at the synaptic cleft and CaMKII in the PSD. Actin immunoreactivity (IR) labelled the cytoskeletal meshwork beneath the filamentous projections, but was very scarce within the PSD itself. ProSAP2/Shank3, cortactin and Ena/VASP-IRs were concentrated on the cytoplasmic face of the PSD, at the level of the PSD projections. Synaptic ultrastructure after HPF was different from that observed after aldehyde fixative. The boutons were larger, and filamentous components were preserved. Particularly, filamentous projections were observed linking the PSD to the actin cytoskeleton. Thus, synaptic ultrastructure can be analysed under more realistic conditions following HPF.     

Slices have more synapses than perfusion-fixed hippocampus from both young and mature rats.

Hippocampal slices have long been used to investigate properties of synaptic transmission and plasticity. Here, for the first time, synapses in slices have been compared quantitatively with synapses occurring in perfusion-fixed hippocampus, which is presumed to represent the natural in vivo state. Relative to perfusion-fixed hippocampus, a remarkable 40-50% increase in spine number occurs in adult hippocampal slices, and a 90% increase occurs in slices from postnatal day 21 rats. Serial EM shows that all of the dendritic spines have normal synapses with presynaptic and postsynaptic elements; however, not all spine types are affected uniformly. Stubby and mushroom spines increase in the adult slices, and thin, mushroom, and branched spines increase in the immature slices. More axonal boutons with multiple synapses occur in the slices, suggesting that the new synapses form on preexisting axonal boutons. The increase in spine and synapse number is evident within a couple of hours after preparing the slices. Once the initial spine induction has occurred, no further change occurs for up to 13 hr in vitro, the longest time investigated. Thus, the spine increase is occurring during a period when there is little or no synaptic activity during the first hour, and the subsequent stabilization in spine synapse numbers is occurring after synaptic activity returns in the slice. These findings suggest that spines form in response to the loss of synaptic activity when slices are removed from the rest of the brain and during the subsequent 1 hr recovery period.     

The effects of amphetamine on synaptic plasticity in rat's medial prefrontal cortex

Morphometric analysis of medial prefrontal cortex (layer VI) of rats treated daily with amphetamine in a dose of 2.5 mg/kg during 3 weeks was performed on the electron microscopic level. The efficacy of the amphetamine dosage was tested on behavioral observation. Synapses on dendritic shafts and spines were studied. The density of axo-dendritic synapses increase on 74%, while the density of synapses on spine's neck decreased on 53%. Most synaptic parameters measured in axo-dendritic (1) and axo-spinous (2) synapses increased significantly under the influence of 2.5 mg/kg dose of AMPH: area of presynaptic terminal increased on 35% (1) and 21% (2), length of postsynaptic density increased on 13% (1) and 12% (2), area of spine increase on 25%. But the density of synaptic vesicles near the active zone decrease (1-on 16.5%, 2-on 20%).     

Presynaptic Ultrastructural Plasticity Along CA3→CA1 Axons During Long‐Term Potentiation in Mature Hippocampus

In area CA1 of the mature hippocampus, synaptogenesis occurs within 30 minutes after the induction of long‐term potentiation (LTP); however, by 2 hours many small dendritic spines are lost, and those remaining have larger synapses. Little is known, however, about associated changes in presynaptic vesicles and axonal boutons. Axons in CA1 stratum radiatum were evaluated with 3D reconstructions from serial section electron microscopy at 30 minutes and 2 hours after induction of LTP by theta‐burst stimulation (TBS). The frequency of axonal boutons with a single postsynaptic partner was decreased by 33% at 2 hours, corresponding perfectly to the 33% loss specifically of small dendritic spines (head diameters <0.45 μm). Docked vesicles were reduced at 30 minutes and then returned to control levels by 2 hours following induction of LTP. By 2 hours there were fewer small synaptic vesicles overall in the presynaptic vesicle pool. Clathrin‐mediated endocytosis was used as a marker of local activity, and axonal boutons containing clathrin‐coated pits showed a more pronounced decrease in presynaptic vesicles at both 30 minutes and 2 hours after induction of LTP relative to control values. Putative transport packets, identified as a cluster of less than 10 axonal vesicles occurring between synaptic boutons, were stable at 30 minutes but markedly reduced by 2 hours after the induction of LTP. APV blocked these effects, suggesting that the loss of axonal boutons and presynaptic vesicles was dependent on N‐methyl‐D‐aspartic acid (NMDA) receptor activation during LTP. These findings show that specific presynaptic ultrastructural changes complement postsynaptic ultrastructural plasticity during LTP. J. Comp. Neurol. 521:3898–3912, 2013. © 2013 Wiley Periodicals, Inc.     

Vezatin is essential for dendritic spine morphogenesis and functional synaptic maturation

Vezatin is an integral membrane protein associated with cell-cell adhesion complex and actin cytoskeleton. It is expressed in the developing and mature mammalian brain, but its neuronal function is unknown. Here, we show that Vezatin localizes in spines in mature mouse hippocampal neurons and codistributes with PSD95, a major scaffolding protein of the excitatory postsynaptic density. Forebrain-specific conditional ablation of Vezatin induced anxiety-like behavior and impaired cued fear-conditioning memory response. Vezatin knock-down in cultured hippocampal neurons and Vezatin conditional knock-out in mice led to a significantly increased proportion of stubby spines and a reduced proportion of mature dendritic spines. PSD95 remained tethered to presynaptic terminals in Vezatin-deficient hippocampal neurons, suggesting that the reduced expression of Vezatin does not compromise the maintenance of synaptic connections. Accordingly, neither the amplitude nor the frequency of miniature EPSCs was affected in Vezatin-deficient hippocampal neurons. However, the AMPA/NMDA ratio of evoked EPSCs was reduced, suggesting impaired functional maturation of excitatory synapses. These results suggest a role of Vezatin in dendritic spine morphogenesis and functional synaptic maturation.     

Diversity of thalamorecipient spine morphology in cat visual cortex and its implication for synaptic plasticity

A feature of spine synapses is the existence of a neck connecting the synapse on the spine head to the dendritic shaft. As with a cable, spine neck resistance (R_neck) increases with increasing neck length and is inversely proportional to the cross‐sectional area of the neck. A synaptic current entering a spine with a high R_neck will lead to greater local depolarization in the spine head than would a similar input applied to a spine with a lower R_neck. This could make spines with high R_neck more sensitive to plastic changes since voltage sensitive conductances, such as N‐methyl‐D‐aspartic acid (NMDA) channels can be more easily activated. This hypothesis was tested using serial section electron microscopic reconstructions of thalamocortical spine synapses and spine necks located on spiny stellate cells and corticothalamic cells from area 17 of cats. Thalamic axons and corticothalamic neurons were labeled by injections of the tracer biotinylated dextran amine (BDA) in the dorsal lateral geniculate nucleus (dLGN) of anesthetized cats and spiny stellates were filled intracellularly in vivo with horseradish peroxidase. Twenty‐eight labeled spines that formed synapses with dLGN boutons were collected from three spiny stellate and four corticothalamic cells and reconstructed in 3D from serial electron micrographs. Spine length, spine diameter, and the area of the postsynaptic density were measured from the 3D reconstructions and R_neck of the spine was estimated. No correlation was found between the postsynaptic density size and the estimated spine R_neck. This suggests that forms of plasticity that lead to larger synapses are independent of spine neck resistance. J. Comp. Neurol. 521:2058–2066, 2013. © 2012 Wiley Periodicals, Inc.     

Drebrin a content correlates with spine head size in the adult mouse cerebral cortex

Synaptic activities alter synaptic strengths at the axospinous junctions, and such changes are often accompanied by changes in the size of the postsynaptic spines. We have been exploring the idea that drebrin A, a neuron-specific actin-binding protein localized on the postsynaptic side of excitatory synapses, may be a molecule that links synaptic activity to the shape and content of spines. Here, we performed electron microscopic immunocytochemistry with the nondiffusible gold label to explore the relationship among levels of drebrin A, the NR2A subunit of N-methyl-D-aspartate receptors, and the size of spines in the perirhinal cortex of adult mouse brains. In contrast to the membranous localization within neonatal spines, most immunogold particles for drebrin A were localized to the cytoplasmic core region of spines in mature spines. This distribution suggests that drebrin within adult spines may reorganize the F-actin network at the spine core, in addition to its known neonatal role in spine formation. Drebrin A-immunopositive (DIP) spines exhibited larger spine head areas and longer postsynaptic densities (PSDs) than drebrin A-immunonegative (DIN) spines (P < 0.001). Furthermore, spine head area and PSD lengths correlated positively with drebrin A levels (r = 0.47 and 0.40). The number of synaptic NR2A immunolabels was also higher in DIP spines than in DIN spines, whereas their densities per unit lengths of PSD were not significantly different. These differences between the DIP and the DIN spines indicate that spine sizes and synaptic protein composition of mature brains are regulated, at least in part, by drebrin A levels.     

Three-dimensional analysis of the structure and composition of CA3 branched dendritic spines and their synaptic relationships with mossy fiber boutons in the rat hippocampus.

This paper is the third in a series to quantify differences in the composition of subcellular organelles and three-dimensional structure of dendritic spines that could contribute to their specific biological properties. Proximal apical dendritic spines of the CA3 pyramidal cells receiving synaptic input from mossy fiber (MF) boutons in the adult rat hippocampus were evaluated in three sets of serial electron micrographs. These CA3 spines are unusual in that they have from 1 to 16 branches emerging from a single dendritic origin. The branched spines usually contain subcellular organelles that are rarely found in adult spines of other brain regions including ribosomes, multivesicular bodies (MVB), mitochondria, and microtubules. MVBs occur most often in the spine heads that also contain smooth endoplasmic reticulum, and ribosomes occur most often in spines that have spinules, which are small nonsynaptic protuberances emerging from the spine head. Most of the branched spines are surrounded by a single MF bouton, which establishes synapses with multiple spine heads. The postsynaptic densities (PSDs) occupy about 10-15% of the spine head membrane, a value that is consistent with spines from other brain regions, with spines of different geometries, and with immature spines. Individual MF boutons usually synapse with several different branched spines, all of which originate from the same parent dendrite. Larger branched spines and MF boutons are more likely to synapse with multiple MF boutons and spines, respectively, than smaller spines and boutons. Complete three-dimensional reconstructions of representative spines with 1, 6, or 12 heads were measured to obtain the volumes, total surface areas, and PSD surface areas. Overall, these dimensions were larger for the complete branched spines than for unbranched or branched spines in other brain regions. However, individual branches were of comparable size to the large mushroom spines in hippocampal area CA1 and in the visual cortex, though the CA3 branches were more irregular in shape. The diameters of each spine branch were measured along the cytoplasmic path from the PSD to the origin with the dendrite, and the lengths of branch segments over which the diameters remained approximately uniform were computed for subsequent use in biophysical models. No constrictions in the segments of the branched spines were thin enough to reduce charge transfer along their lengths.(ABSTRACT TRUNCATED AT 400 WORDS)     

Chemically induced long-term potentiation increases the number of perforated and complex postsynaptic densities but does not alter dendritic spine volume in CA1 of adult mouse hippocampal slices

Examination of the morphological correlates of long-term potentiation (LTP) in the hippocampus requires the analysis of both the presynaptic and postsynaptic elements. However, ultrastructural measurements of synapses and dendritic spines following LTP induced via tetanic stimulation presents the difficulty that not all synapses examined are necessarily activated. To overcome this limitation, and to ensure that a very large proportion of the synapses and spines examined have been potentiated, we induced LTP in acute hippocampal slices of adult mice by addition of tetraethylammonium (TEA) to a modified CSF containing an elevated concentration of Ca(2+) and no Mg(+). Quantitative electron microscope morphometric analyses and three-dimensional (3-D) reconstructions of both dendritic spines and postsynaptic densities (PSDs) in CA1 stratum radiatum were made on serial ultrathin sections. One hour after chemical LTP induction the proportion of macular (unperforated) synapses decreased (50%) whilst the number of synapses with simple perforated and complex PSDs (nonmacular) increased significantly (17%), without significant changes in volume and surface area of the PSD. In addition, the surface area of mushroom spines increased significantly (13%) whilst there were no volume differences in either mushroom or thin spines, or in surface area of thin spines. CA1 stratum radiatum contained multiple-synapse en passant axons as well as multiple-synapse spines, which were unaffected by chemical LTP. Our results suggest that chemical LTP induces active dendritic spine remodelling and correlates with a change in the weight and strength of synaptic transmission as shown by the increase in the proportion of nonmacular synapses.     

Organization of mitochondria in olfactory bulb granule cell dendritic spines.

In contrast to dendritic spines with only postsynaptic functions, the spines of olfactory bulb granule cells subserve both pre- and postsynaptic roles. In single sections these spines were previously seen to contain mitochondria, most likely needed to provide energy for presynaptic functions, but their frequency and distribution were unknown. In order to understand the organization of mitochondria in these specialized dendritic appendages, we have studied the geometry and cytoplasmic organization of granule cell spines with computer-assisted reconstructions of serial electron micrographs. The spine heads were seen to be elliptical in shape with a single pair of reciprocal synapses on the concave face apposed to the mitral/tufted cell dendrite. Mitochondria were found localized in the spine neck as well as the spine head and often extended between the two compartments. Based on their variable distribution it seems reasonable to suggest that these mitochondria are motile and move in and out of spine compartments from the parent dendrite. Spine apparatus was apparent in most of the spines as membrane bound cisterns of smooth endoplasmic reticulum located close to mitochondria. The possible role of spine apparatus in facilitating the movement of mitochondria in the necks and heads of granule cell spines in the absence of microtubules is discussed.     

Axospinous synapses with segmented postsynaptic densities: a morphologically distinct synaptic subtype contributing to the number of profiles of ‘perforated’ synapses visualized in random sections

Axospinous synapses were examined in the molecular layer of the rat dentate gyrus. Serial section analysis of synapses, which exhibited a discontinuity of the postsynaptic density (PSD) in at least one consecutive section, was performed. Reconstruction of each discontinuous PSD was made in a plane perpendicular to that of serial sections. The results obtained confirm earlier observations that profiles of 'perforated' synapses visualized in random sections of osmicated material are produced by sectioning of synapses with perforated and horseshoe-shaped PSDs. Additionally, it has been found that two other synaptic subtypes, namely synapses with notched and segmented PSD, contribute to the number of profiles of 'perforated' synapses. Synaptic contacts with notched PSD are characterized by an indentation of an otherwise continuous PSD, relatively small dimensions and simple shape. They appear to be unrelated to the category of synapses with discontinuous PSD. Synaptic contacts with segmented PSD are distinguished by the presence of 2-5 discrete PSD segments at the interface between a presynaptic axon terminal and a postsynaptic dendritic spine. Some PSD segments exhibit 1-3 perforations, while others are horseshoe-shaped. It is postulated that the segmented PSD may evolve through the stages of perforated and horseshoe-shaped PSD to form a specialized synaptic contact of an unusually high efficacy. Every PSD segment is a component of a separate synaptic complex, each one comparable to that of a small, simple-shaped synapse. A concerted activation of several synaptic complexes belonging to a single synaptic junction may provide a mechanism for an amplification of synaptic transmission.     

Quantitative ultrastructural changes in rat cortical synapses during early-, mid- and late-adulthood

Quantitative ultrastructural analysis of rat parietal cortex was undertaken to determine the nature of the synaptic changes occurring in the molecular layer over a series of ages in early- (3 months), mid- (6 and 10 months) and late- (17 months) adulthood.The total number of synapses remained constant until 10 months of age, but decreased significantly by 17 months. Asymmetrical synapses on dendritic shafts were lost earlier (by 6 months) than asymmetrical synapses on dendritic spines (by 17 months). Symmetrical axodendritic synapses remained constant throughout adulthood.Analysis of synaptic terminal parameters revealed the following. Both individual and total presynaptic terminal areas decreased over the age range studied. Individual and total postsynaptic terminal areas, however, remained constant over the 3–17-month period. Positive correlations were obtained for the relationships between presynaptic terminal area and both age and synaptic vesicle number. The presynaptic terminal area was largest and contained the greatest number of vesicles at 3 months of age. This age was, in addition, characterized by the least numbers of mitochondria in the presynaptic terminal and spine apparatus in the postsynaptic terminal. The vacuolar and tubular cisternae of the presynaptic terminal were considerably reduced at 17 months.These data suggest that in the molecular layer of the cerebral cortex the period of adulthood is characterized by a diversity of synaptic changes. The 3-months age may reflect the end of the developmental phase and may be marked by changes in synaptic functional activity. The asymmetrical axodendritic synapses may constitute an intermediate form of synapse, capable of being transformed into axospinous synapses as dendritic spines continue to be formed in the adult.     

Three-dimensional organization of cell adhesion junctions at synapses and dendritic spines in area CA1 of the rat hippocampus

Recent work has emphasized the role of adhesion molecules in synaptic plasticity, including long-term potentiation in the hippocampus. Such adhesion molecules are concentrated in junctions that are characterized by dense thickenings on both sides of the junction and are called puncta adhaerentia (PA). Reconstruction from serial electron microscopy was used to determine the location and size of PA in the stratum radiatum of hippocampal area CA1, where many of the previous functional studies have been performed. PAs were found at the edges of synapses on 33% of dendritic spines. The areas occupied by PA were variable across different types of synapses, occupying 0.010 ± 0.005 μm² at macular synapses and 0.034 ± 0.031 μm² at perforated synapses. Another zone, called a vesicle-free transition zone (VFTZ), was identified. Like the PA, this zone also had no presynaptic vesicles and was located at the edges of synapses; however, unlike the PA, the presynaptic thickening was less than the postsynaptic thickening. Together, 45% of spine synapses had PA and/or VFTZ occupying 23 ± 11% of the total junctional area between axons and spines. PA also occurred at nonsynaptic sites involving neuronal as well as glial elements. Most (64%) of these PAs occurred between nonsynaptic portions of dendritic spines and neighboring astrocytic processes. Smooth endoplasmic reticulum was often apposed to one or both sides of the synaptic and the nonsynaptic PA. These findings provide further data as a structural basis for understanding the roles of cell adhesion junctions in hippocampal synaptic function and plasticity. J. Comp. Neurol. 393:58–68, 1998. © 1998 Wiley-Liss, Inc.     

The effects of haloperidol on synaptic plasticity in rat's medial prefrontal cortex

Morphometric analysis of Medial prefrontal cortex (layer VI) of rats treated daily with haloperidol in a dose of 0.1 mg/kg during 3 weeks was performed on the electron microscopic level. The efficacy of the haloperidol dosage was tested on the amphetamine psychosis model. Synapses on dendritic shafts and dendritic spines were studied. The density of synapses on dendritic shafts increased on 51%, while on spine's neck it decreased on 19%. There were significant changes of some synaptic parameters only in axo-dendritic synapses: area of presynaptic terminal decreased on 13% (p less than 0.05), length of postsynaptic density decreased on 15% (p less than 0.05), but the density of synaptic vesicles near the active zone increased on 10% (p less than 0.05).     

Synaptic plasticity in rat medial prefrontal cortex under chronic haloperidol treatment produced behavioral sensitization

Morphometric analysis of the population of synapses in medial prefrontal cortex (layer VI) of rat treated daily with haloperidol in a dose of 1.0 mg/kg during 3 weeks was performed on the electronmicroscopic level. The density of synapses on dendritic shafts increased on 29% and on the head of spine it increased on 20%. Nearly all the parameters measured in axo-spinous synapses were significantly decreased (area of presynaptic terminal, summarized area of mitochondriae per terminal, number of synaptic vesicles near the active zone, area of postsynaptic spine). There were no differences between the parameters of axo-dendritic synapses in control and haloperidol-treated rats. The data suggest that haloperidol might have induced the formation of the new synapses and may have changed the efficacy of the synaptic transmission in axo-spinous synapses on the head of spine.