Active zones and receptor surfaces of freeze-fractured crayfish phasic and tonic motor synapses

Deep and superficial flexor muscles in the crayfish abdomen are innervated respectively by small populations of physiologically distinct phasic and tonic motoneurons. Phasic motoneurons typically produce large EPSP's, releasing 100 to 1000 times more transmitter per synapse than their tonic counterparts, and exhibiting more rapid synaptic depression with maintained stimulation. Freeze-fracturing the abdominal flexor muscles yielded images of phasic and tonic synapse-bearing terminals. The two types of synapse are qualitatively similar in ultrastructure, displaying on the presynaptic membrane's P-face synaptic contacts recognized by relatively particle-free oval plaques which are often framed by the muscle fiber's E-face leaflet with its associated receptor particles. Situated within these presynaptic plaques are discrete clusters of large intramembrane particles, forming active zone (AZ) sites specialized for transmitter release. AZs of phasic and tonic synapses are similar: 80% had a range of 15–40 large particles distributed in either paired spherical clusters or in linear form, with a few depressions denoting sites of synaptic vesicle fusion or retrieval around their perimeters. The packing density of particles is similar for phasic and tonic AZs. The E-face of the muscle membrane displays oval-shaped receptor-containing sites made up of tightly packed intramembranous particles. Phasic and tonic receptor particles are packed at similar densities and the measured values resemble those of several other crustacean and insect neuromuscular junctions. Overall, the similarity between phasic and tonic synapses in the packing density of particles at their presynaptic AZs and postsynaptic receptor surfaces suggests similar regulatory mechanisms for channel insertion and spacing. Furthermore, the findings suggest that morphological differences in active zones or receptor surfaces cannot account for large differences in transmitter release per synapse.     

Intramembrane organization of synapses in the lobster stretch receptor organ

Summary The intramembrane organization of axodendritic and neuromuscular synapses in the lobster stretch receptor organ was investigated by freeze-fracturing. Based on ultrastructural criteria which are known to be correlated with physiological properties, we identified three types of synapse: the inhibitory axodendritic, the inhibitory neuromuscular, and the excitatory neuromuscular synapse. Although these synapses have some features in common, each has a characteristic arrangement of intramembrane particles in both the presynaptic and postsynaptic membranes. All three have, in their presynaptic membranes, aggregates of P-face particles and associated depressions representing sites of synaptic vesicle exocytosis, features which together define active zones. However, in the inhibitory axodendritic synapse the P-face contains short ridges in this region. These ridges may occur singly or in pairs oriented in V-shaped configurations. The ridges are decorated with particles along their entire length. In the inhibitory neuromuscular synapse, no ridges are present. Clusters of particles are present, but they are scattered randomly over a large expanse of presynaptic membrane. In the excitatory neuromuscular synapse, isolated clusters of particles are associated with the P-face and are occasionally located on circular elevations of the membrane. The postsynaptic membrane also shows structural diversity in the three types of synapse. In the inhibitory axodendritic synapse, there is no apparent specialization. However, in the inhibitory neuromuscular synapse, P-face particles are arranged in double rows which are separated by particle-free strips of membrane. In the excitatory neuromuscular synapse, particles are confined to a narrow band that borders the synaptic cleft. This band is demarcated by a single intermittent strand of particles arranged in the direction of the long axis of the muscle fibre. Therefore, intramembrane specializations of both the presynaptic and postsynaptic membranes are sufficiently distinctive that three different types of synapse can be recognized.     

A freeze-fracture study of synaptic junction development in the superficial layers of the chick optic tectum

Summary Synaptogenesis in the superficial layers of the rostral pole of the chick optic tectum has been studied using freeze-fracture techniques. The developmental sequence of intramembrane organization at synaptic junctions involves the accumulation and assembly of intramembrane particles into aggregates characteristic of the mature junctions.By embryonic day seven, areas of loosely-arranged clusters of medium-sized particles are observed on the cytoplasmic membrane leaflets (P-faces) of developing neurites. These clusters are characteristic of the intramembrane organization at presynaptic active zones. At later stages, small pits, characteristic of vesicle fusion sites, are observed interspersed among such P-face particle clusters. Complementary intramembrane specializations are also present on the external leaflets (E-faces) of presynaptic membranes at the active zones.Small solitary aggregates of large-sized particles on the E-faces of neurite plasma membranes are also seen at early embryonic stages. As development progresses, these aggregates increase in size and packing density and occupy large oval domains in postsynaptic membranes. These intramembrane specializations may represent the postsynaptic active zones of asymmetric synapses. Another type of intramembrane specialization, observed during the third week of incubation, is characterized by aggregates of small- and medium-sized particles on the P-face of postsynaptic membranes and is often seen directly apposed to the E-face of a presynaptic terminal. This type of intramembrane specialization may represent the postsynaptic active zone region at symmetrical synaptic contacts.     

Membrane ultrastructure of the giant synapse of the squidLoligo pealei

The ultrastructure of the ‘giant synapse’ of the stellate ganglion of the squid was studied with freeze-fracture and thin-sectioning techniques. A sheath of glial cells separates the pre- and post-synaptic axons. At intervals, round-topped processes of the postsynaptic axon pierce the sheath to contact the presynaptic axon. This area of synaptic contact is marked by a widened intercellular cleft containing electron-dense material and by a cluster of synaptic vesicles within the presynaptic cytoplasm. The number of synaptic vesicles in such clusters was greatly reduced by electrical stimulation of the synapse during fixation. Freeze-fracture reveals a roughly circular patch (0.3 μm diameter) of 10 nm particles on the cytoplasmic leaflet of the presynaptic membrane. A similar patch of particles lies on the external leaflet of the apposed postsynaptic membrane.The squid giant synapse thus consists of multiple small pre- and postsynaptic active zones where neurotransmitter is released from the presynaptic terminal and sensed by postsynaptic receptors. Comparison of the structure of these postsynaptic active zones with those at synapses where the transmitter or transmitter action is known suggests that the excitatory transmitter at this synapse is an amino acid.Presumptive gap junctions, marked by particles in the cytoplasmic leaflet, are found between small-diameter axons in the stellate ganglion but not at the giant synapse. Glial-cell membranes contain aggregates of particles and pits suggestive of gap junctions. The aggregates of pits are embedded within linear arrays of particles which somewhat resemble tight junctions.     

Morphological studies of synapses and varicosities in dissociated cell cultures of sympathetic neurons

Neurons dissociated from the superior cervical ganglia of newborn rats can be grown under conditions which support either adrenergic or cholinergic differentiation. In both cases, the neurons form numerous morphologically specialized synaptic terminals or synapses as well as relatively unspecialized varicosities. The ultrastructure of both types of terminal was compared in mature neuronal cultures and the effects of growth conditions on terminal morphology examined. After aldehyde-osmium fixation, synapses in cultures grown under adrenergic or cholinergic conditions were characterized by asymmetrical membrane specializations comparable to type I or asymmetric synapses; bismuth iodide and ethanolic phosphotungstic acid impregnation of neuronal cultures revealed the presence of characteristic synaptic membrane specializations: a presynaptic grid of dense projections and a wide postsynaptic dense band of uniform thickness. No membrane specializations were apparent in varicosities after aldehyde-osmium fixations or with these stains. Intramembranous particle distributions were examined in freeze-fracture replicas of neurons. Aggregates of large, 10-12 nm particles were found on P-face membrane leaflets of cell bodies and large diameter processes; this distribution is the same as that of synapses in thin-sectioned preparations. These particle aggregates may represent postsynaptic membrane specializations or acetylcholine receptors. The cytoplasmic leaflet of boutons contained large, 12-14 nm particles, which appeared to be concentrated at the region of synaptic contact at putative synapses, but were diffusely distributed in varicosity membranes. Similar large particles were also seen at a much lower density in the membrane E-face. None of these ultrastructural characteristics appeared to vary with transmitter identity or growth conditions. Synaptic vesicle shape, however, did vary in glutaraldehyde-fixed cultures. At all ages examined, neurons grown on monolayers of heart cells contained predominantly round vesicles, whereas neurons grown in the virtual absence of non-neuronal cells possessed pleiomorphic synaptic vesicles. This difference in vesicle shape appeared to be correlated more closely with growth in the presence of non-neuronal cells than with the transmitter present at the time of fixation.     

Freeze-fracture morphology of the vestibular hair cell-primary afferent synapse in the chick

Summary The intramembrane specializations at vestibular hair cell-primary afferent synapses have been identified and characterized in complementary freeze-fracrure replicas from prehatch and hatchling chick cristae and maculae. Hair cell protoplasmic (P) faces at sites where presynaptic bodies are present exhibit small, tightly packed arrays of 9 nm particles. Hair cell external (E) faces have corresponding arrays of pits. Multiple arrays are often observed in contiguity. Opposite the presynaptic bodies, postsynaptic afferent boutons and calyces exhibit a more extensive array of scattered, irregular E-face particles. Corresponding P-fracture faces of afferent boutons and calyces display little topographical specialization opposite these E-face arrays, which are presumed to be the intramembrane correlate of the postsynaptic density. Examination of complementary replicas has allowed identification of the intramembrane synaptic specializations for all membrane faces at the synaptic apposition.     

A freeze-fracture study of the skate electro receptor

Summary The sensory epithelium lining the ampulla of Lorenzini in the skate was examined by the freeze-fracture technique. Anastomosing tight junctions (zonula occludens) completely encircle the apex of each receptor cell joining it to neighbouring support cells. The tight junctions separate two distinctly different regions of the receptor-cell surface. The apical P-face has numerous large particles while just below the tight junctions of the lateral surface have many smaller particles. On its basal surface each receptor cell makes several evaginating ribbon synapses with an afferent nerve. Three regions of the synaptic evagination can be distinguished on the basis of membrane specializations: 1. At the tip of the evagination a regular array of large particles is found on the P-face of the receptor cell directly opposite a similar regular array of large particles on the P-face of the afferent nerve; 2. just above the tip at a narrow constriction, below which vesicles are not found, a population of large particles on the P-face of the receptor cell opposes a well-defined strip of large particles that cleaves with the E-face of the nerve fibre; 3. at the arch of the synaptic evagination randomly occurring dimples are found on the P-face and protrusions on the E-face of the receptor cell. The density of these protrusions increased in skates that were electrically stimulated. We suggest that the co-extensive arrays of particles at the tip of the ribbon synapse is an intercellular junction; that the active zone of the synapse is at or above the constriction; and that membrane retrieval occurs in the synaptic arch region.     

Freeze-fracture studies on the giant axon and ensheathing Schwann cells of the squid

Summary The giant axons and encompassing sheaths from the stellar nerves of the squidsSepioteuthis sepioidea andLoligo forbesi have been analysed by freeze-fracture. The axolemma exhibits many intramembranous particles (IMPs) that fracture onto the cytoplasmic membrane half-leaflet (P-face); the larger IMPs may be aggregated into clusters. Axoplasmic subsurface cisternae are found beneath this membrane. Clustered or aligned arrays of P-face IMPs are also found on the membranes of the Schwann cells that intimately encapsulate the giant axons as well as ‘capitate’ projections of Schwann cells into the axons. When adjacent Schwann cells abut directly against one another, aligned E-face IMPs are found along the fracture plane of the upturning membranes. These E-face alignments of IMPs have complementary furrows on the Schwann cell membranes which exhibit no complementary structure on the axolemma as they represent the clefts between adjacent glial cells. The other Schwann cell membranes exhibit P-face dimples and E-face (extracellular membrane half-leaflet) protuberances which may reflect endo- or exocytotic activity; alternatively they may represent caveolae. Comparable structures are occasionally observed at axo-glial interfaces. However, those in the Schwann cell membrane could be part of the transverse tubular lattice system which also exists in adaxonal glia. Beyond the Schwann cells, layers of endoneurial cells (fibrocytes) are interleaved by collagen-filled spaces. These cells exhibit extensive cross-fractured intracellular invaginations as well as inpushings of the extracellular matrix material. Their membranes exhibit a large number of IMPs.     

Reciprocal axo-axonal synapses between the common inhibitor and excitor motoneurons in crustacean limb muscles

Summary Nerve terminals of the common inhibitor motoneuron in a crab (Eriphia spinifrons) limb closer muscle and in a crayfish (Procambarus clarkii) limb accessory flexor muscle make neuromuscular synapses with the muscle membrane (postsynaptic inhibition) as well as axo-axonal synapses with the terminals of the excitatory axon (presynaptic inhibition). That transmission is from the inhibitor to the excitor terminals at these axo-axonal synapses is indicated by the occurrence on the inhibitor membrane of presynaptic dense bars denoting sites of transmitter release. Axo-axonal synapses with the opposite polarity, in which transmission is from an excitatory onto an inhibitory terminal, were occasionally seen either adjacent to or separate from the inhibitory axo-axonal synapse. Nerve terminals of the specific inhibitor in the crayfish opener muscle were seen to make numerous axo-axonal output synapses upon excitatory nerve terminals but excitor nerve terminals were not seen to make output synapses onto inhibitor terminals. Thus reciprocal axo-axonal synapses appear to be a feature of the common inhibitor but not of the specific inhibitor. The excitor-to-inhibitor component of these reciprocal synapses may serve to limit transmitter output in the common inhibitor axon by activating glutamate_B receptors which facilitate efflux of K⁺ and hyperpolarization of the membrane.     

A freeze-fracture study of synaptogenesis in the distal retina of larvalXenopus

Summary Synapse formation between photoreceptor, bipolar and horizontal cells of the larvalXenopus retina was studied by the freeze-fracture technique. Photoreceptors and horizontal cells were joined by ribbon synapses; photoreceptor and bipolar cells by basal junctions. Gap junctions were found between photoreceptors and between horizontal cells. Horizontal cell dendrites invaginated receptor bases before the plasma membrane of either cell showed zones of intramembrane (IMP) particle accumulation. Subsequently the receptor cell began to form a synaptic ridge where P-face IMPs aggregated at a protrusion of the surface membrane. The length of the ridge and the density of its IMPs increased between larval stages 40 and 56. Cross-fractured views of receptor cytoplasm at different larval stages showed that synaptic ribbons and synaptic vesicles developed in conjunction with the ridge. Plasmalemmal deformations suggesting sites of vesicle fusion or uptake were noted adjacent to the apex of the ridge. Horizontal cell dendritic membrane first accumulated P-face IMPs at several small regions; subsequently the IMPs became aligned over a broad membrane area. Both rod- and cone-related horizontal cell dendrites also manifested a loose patch of E-face IMPs which subsequently was transformed into a linear array. Basal junctions were characterized by a P-face IMP aggregate in the photoreceptor membrane and an E-face IMP aggregate in the bipolar cell membrane. Basal junctions appeared suddenly in a mature configuration at larval stage 42.     

Structural-functional differences of a crab motoneuron to four stomach muscles

A motor unit in the stomach of the blue crab, Callinectes sapidus, consists of four separate muscles involved in different aspects of the trituration and filtering of food. Motor nerve terminals to two of the muscles (CPV7a and GM5) release small amounts of transmitter (low-output) while those to the other two muscles (CV2 and CV3) release between three and five-fold greater amounts (high-output). Structural features underlying the disparity in synaptic strength were analysed with thin serial-section electron microscopy. Nerve terminals were similar in their volume percent of mitochondria, clear vesicles and dense core vesicles among the four muscles. This was also the case for the number and size of synaptic contacts. However, presynaptic dense bars representing active zones were longer and occurred more frequently at high-output synapses than at low-output ones. High-output synapses were also characterized by the close spacing of adjacent dense bars. The longer and more closely spaced dense bars at high-output synapses would be factors in the generation of larger synaptic potentials in these terminals compared to their low-output counterparts. Other factors, however, need to be considered to fully account for the physiological differences in synaptic strength among the four muscles.     

Membrane structure of parallel-fibre synaptic terminals in the cerebellum of the jaundiced Gunn rat: freeze-fracture and E-PTA study

Summary Parallel-fibre synaptic membranes were examined by freeze-fracture and ethanolic-phosphotungstic acid methods in the cerebellum of homozygous (j/j) Gunn rats with hereditary jaundice. Parallel-fibre synapses with dendritic spines of Purkinje cell were severely affected since many Purkinje cells degenerated during the early postnatal period. Some parallel-fibre synaptic terminals lacked their postsynaptic partners and faced astrocytic processes from 18 days of age to the adult stage. These parallel-fibre terminals contained clusters of synaptic vesicles adjacent to synaptic membranes, and synaptic membranes and synaptic cleft materials were identical to those of parallel fibres with postsynaptic partners, as visualized by conventional electron microscopy with osmium tetroxide postfixation and staining of sections with uranyl acetate and lead citrate. In freeze-fractured specimens, the presynaptic membrane of parallel fibres had diffusely distributed large particles and tiny pits on the P-face and protuberances on the E-face, together representing synaptic vesicle attachment sites. Such vesicle attachment sites were present on the presynaptic membranes of parallel fibres without postsynaptic partners from day 18 to the adult stage. After ethanolic-phosphotungstic acid staining, parallel-fibre terminals displayed presynaptic dense projections, intercleft materials and postsynaptic thickening, but some parallel fibres lacked postsynaptic thickening. These observations suggest that the presynaptic membrane structure of the parallel fibre is preserved, even in the absence of a postsynaptic partner, in j/j cerebella. A mechanism for persistence of presynaptic membrane structures without postsynaptic partners in j/j cerebella is discussed.     

A freeze-fracture study of photoreceptor presynaptic membranes during ribbon synapse formation

Summary The outer plexiform layer (OPL) of the developing chick retina from 11 embryonic days to 11/2 weeks posthatching has been studied by freeze-fracture to characterize changes in the membrane structure of photoreceptor terminals during synaptogenesis. At early stages, the undifferentiated photoreceptor synaptic base is characterized by a sparse distribution of intramembrane particles on the inner leaflet (P-face). Later, as the synaptic base begins to differentiate by extending filopodia into the OPL, numerous small aggregates of large particles appear between and on filopodial surfaces. Many of the aggregates occupy crater-like depressions, which are seen in cross-fractures through the underlying cytoplasm to be associated with vesicular invaginations of the presynaptic membrane. Corresponding thin sections through these regions at this time reveal immature arciform densities and coated vesicles fusing with the presynaptic membrane adjacent to these densities. At later stages, many of the particle aggregates on the photoreceptor membrane appear to have coalesced into longer arrays overlying ridges surrounded by numerous vesicle fusion sites. These intramembrane changes correlate with the formation of the mature arciform density-synaptic ribbon specialization in the photoreceptor presynaptic terminal and with physiological maturation of the chick retina.     

Orthogonal arrays of particles in plasma membranes of the gastric parietal cell

Freeze-fracture of rat gastric mucosa revealed a specific set of intramembranous particles in the plasma membrane of the parietal cells. The particles were small and of square shape and formed orthogonal arrays in the P-face with corresponding orthogonal arrays of pits in the E-face. Arrays, scattered among usual globular particles, were particularly numerous at the basal pole of the cell and less concentrated on the lateral side. They were not present in the apical microvillar membrane nor in the membranes of intracellular tubulovesicles. As in other cell types in which similar arrays were described previously (e.g., astrocytes, “light” cells of the kidney collecting tubule), their presence in parietal cell membranes suggest some specialized function of these membranes not shared by plasma membranes showing only a population of globular particles. This function has yet to be identified.     

Coordinated multivesicular release at a mammalian ribbon synapse

Traditional models of synaptic transmission hold that release sites within an active zone operate independently. Although the release of multiple vesicles (multivesicular release; MVR) from single active zones occurs at some central synapses, MVR is not thought to require coordination among release sites. Ribbon synapses seem to be optimized to release many vesicles over an extended period, but the dynamics of MVR at ribbon synapses is unknown. We examined MVR at a ribbon synapse in a retinal slice preparation using paired recordings from presynaptic rod bipolar and postsynaptic AII amacrine cells. When evoked release was highly desynchronized, discrete postsynaptic events were larger than quantal miniature excitatory postsynaptic currents (mEPSCs) but had the same time course. The amplitude of these multiquantal mEPSCs, which seem to arise from the essentially simultaneous release of multiple vesicles, was reduced by lowering release probability. The release synchrony reflected in these multivesicular events suggests that release within an active zone is coordinated during MVR.     

Evidence for an ephaptic feedback in cortical synapses: postsynaptic hyperpolarization alters the number of response failures and quantal content.

The amplitude of excitatory postsynaptic potentials and currents increases with membrane potential hyperpolarization. This has been attributed to an increase in the driving force when the membrane potential deviates from the equilibrium potential of the respective ions. Here we report that in a subset of neocortical and hippocampal synapses, postsynaptic hyperpolarization affects traditional measures of transmitter release: the number of failures, coefficient of variation of response amplitudes, and quantal content, suggesting increased presynaptic release. The result is compatible with the hypothesis of Byzov on the existence of electrical (or "ephaptic") linking in purely chemical synapses. The linking, although negligible at neuromuscular junctions, could be functionally significant in influencing transmitter release at synapses with high resistance along the synaptic cleft. Our findings necessitate reconsideration of classical amplitude-voltage relations for such synapses. Thus, synaptic strength may be enhanced by hyperpolarization of the postsynaptic membrane potential. The positive ephaptic feedback could account for "all-or-none" excitatory postsynaptic potentials at some cortical synapses, large evoked and spontaneous multiquantal events and a high efficacy of large "perforated" synapses whose number increases following behavioural learning or the induction of long-term potentiation.     

Morphological correlates of synaptic transmission in lamprey spinal cord.

The dye Procion brown was used to identify in the light and electron microscope, synaptic contacts made between monosynaptically coupled neurons in the lamprey spinal cord whose synaptic interaction had been recorded. Synaptic contacts were made on different dendrites of the postsynaptic cell at different distances from the soma. Some of the contacts were made on dentritic spines and some on the smooth shaft of the dentrites. Serial sections through synaptic contacts made on dendritic processess of the postsynaptic cells were used for three-dimensional reconstruction of the synapses using computer graphics techniques. The computer reconstructions and detailed examination of the serial EM micrographs revealed the large proliferation of membrane involved in making these en passant synapses as well as the morphological changes due to stimulation of the presynaptic axon. These changes include depletion of synaptic vesicles and formation of complex vesicles and synaptic cisternae. Besides chemical synaptic contacts, four electrotonic contacts were located, confirming the mixed electrochemical synaptic response recorded from the postsynaptic cell. The mean quantum content was estimated and compared with the estimate of the available transmitter pool, assuming the quantal release hypothesis applies at these synapses. The total transmitter pool was estimated by counting all synaptic vesicles in all synaptic contacts. It was estimated that about 6% of the total transmitter pool is available for release at these synapses. This compares with less than 1% at the neuromuscular junction and about 20% at sympathetic synapses. These results support the hypothesis that synaptic vesicles may be recycled as described by Heuser and Reese (22) at the neuromuscular junction. Ongoing studies are investigating the effect on a variety of synaptic junctions to stimulation for different periods of time of presynaptic axons. The methods described in this study can also be used to test the models of synaptic interaction on dendritic trees described by Rall (39) and Jack and Redman (24).     

Vesicle number does not predict postsynaptic measures of miniature synaptic activity frequency in cultured cortical neurons

We tested the hypothesis that heterogeneity in the frequency of miniature synaptic activity reflects differences in the number of vesicles present in presynaptic terminals. Using imaging techniques, we measured dendritic miniature synaptic calcium transients attributed to the spontaneous release of single transmitter quanta. Following imaging, the identified neurons were processed for serial transmission electron microscopy. At sites of quantal Ca(2+) transients mediated by N-methyl-D-aspartate receptors, we confirmed the presence of excitatory synapses and measured the total number of vesicles and the number of docked vesicles.We observed no correlation between the frequency of spontaneous miniature activity and either the total vesicle number or the number of docked vesicles. We conclude that the presynaptic vesicle complement as measured by ultrastructural analysis does not necessarily determine the frequency of spontaneous activity at synapses mediated by N-methyl-D-aspartate receptors.     

Ultrastructural correlates of experimentally altered transmitter release efficacy in frog motor nerve terminals

After experimentally inducing long term changes in transmitter release, a series of frog neuromuscular junctions were studied with intracellular recording and then semi-serially sectioned and examined in the electron microscope. Transmitter release per unit length of motor nerve terminal was well correlated with several measures of the length of individual presynaptic active zones and with the number of mitochondria per terminal. Total release from each terminal correlated with estimates of the total amount of active zone. This study of neuromuscular junctions in sartorius muscles of the frog Rana pipiens was undertaken to search for ultrastructural correlates of the increase in transmitter release efficacy that follows denervation of the contralateral sartorius. This treatment typically results in greatly enhanced release at some synapses while others appear unaffected. In the present study, nine identified junctions with known physiological properties were sectioned every 6 micron throughout much of their length to yield 40-105 cross-sectional profiles per junction. Overall, these 9 synapses showed a 33-fold range in quantal transmitter release and an 18-fold range in release per unit nerve terminal. Release correlated with estimates of active zone size. No correlations were found between release and the density of synaptic vesicles adjacent to active zones. Our results suggest that active zones in motor nerve terminals are plastic structures, and that changes in active zone size may be the structural basis of long term changes in transmitter release and synaptic efficacy.     

Retrograde modulation of presynaptic release probability through signaling mediated by PSD-95-neuroligin

The structure and function of presynaptic and postsynaptic components of the synapse are highly coordinated. How such coordination is achieved and the molecules involved in this process have not been clarified. Several lines of evidence suggest that presynaptic functionalities are regulated by retrograde mechanisms from the postsynaptic side. We therefore sought postsynaptic mechanisms responsible for trans-synaptic regulation of presynaptic function at excitatory synapses in rat hippocampal CA1 pyramidal neurons. We show here that the postsynaptic complex of scaffolding protein PSD-95 and neuroligin can modulate the release probability of transmitter vesicles at synapse in a retrograde way, resulting in altered presynaptic short-term plasticity. Presynaptic beta-neurexin serves as a likely presynaptic mediator of this effect. Our results indicate that trans-synaptic protein-protein interactions can link postsynaptic and presynaptic function.