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1.
The sensorimotor area of rat cerebral cortex was subjected to repeated electrical stimulation at 10-min intervals, with resultant formation and progressive lengthening of self-sustained after-discharges (SSAD). One and 60 min after the third SSAD ended, we carried out an electron microscopy morphometric analysis of the agranular synaptic vesicles in type I synapses (after Gray) in the second cortical layer of the homotopic area of the unstimulated hemisphere. One minute after the seizure ended, 5.8% enlargement of the synaptic vesicles compared with the control was demonstrated in zone II of the synapse (0.1-0.2 micron from the active zone of the synapse). Neither the size nor the shape of the synaptic vesicles in the other parts of the synaptic apparatus altered. Sixty min after the seizure ended, a 5.5% enlargement of the synaptic vesicles in zone I (0.0-0.1 micron) and a 5.4% enlargement of those in zone II was found. The synaptic vesicles in zone I in the experimental animals were more oval than in the controls. Our findings support the vesicular theory and testify that hyperfunction, up to temporary exhaustion of the synaptic apparatuses, produces a change in the transmitter content of the synaptic vesicles. A raised amount of transmitter in the synaptic vesicles near the active zone could be one of the factors responsible for continued hyperexcitability of the tissue one hour after the seizure had ended. The results likewise support the concept of two mechanisms of synaptic vesicle formation, and hence of the existence of two different vesicle populations. 相似文献
2.
The effect of ethanol on the distribution of synaptic vesicles in cortical synaptosomes of the rat 总被引:1,自引:0,他引:1
F Hajós M K?amán A Csillag J Kisgy?rgy 《Acta morphologica Academiae Scientiarum Hungaricae》1979,27(3):193-198
Ethanol was shown to cause a redistribution of synaptic vesicles in incubated synaptosomes. While the number of synaptosomes containing synaptic vesicles attached to the presynaptic membrane decreased markedly, an increase in the number of synaptosomes lacking membrane-vesicle associations was observed. The findings support the possibility of a presynaptic action of ethanol and point to the role of membrane-attached vesicles in synaptic transmission. 相似文献
3.
Löw P Norlin T Risinger C Larhammar D Pieribone VA Shupliakov O Brodin L 《European journal of cell biology》1999,78(11):787-793
Exocytosis - syntaxin - synaptobrevin - SNARE synaptic vesicle The lamprey giant reticulospinal synapse can be used to manipulate the molecular machinery of synaptic vesicle exocytosis by presynaptic microinjection. Here we test the effect of disrupting the function of the SNARE protein SNAP-25. Polyclonal SNAP-25 antibodies were shown in an in vitro assay to inhibit the binding between syntaxin and SNAP-25. When microinjected presynaptically, these antibodies produced a potent inhibition of the synaptic response. Ba2+ spikes recorded in the presynaptic axon were not altered, indicating that the effect was not due to a reduced presynaptic Ca2+ entry. Electron microscopic analysis showed that synaptic vesicle clusters had a similar organization in synapses of antibody-injected axons as in control axons, and the number of synaptic vesicles in apparent contact with the presynaptic plasma membrane was also similar. Clathrin-coated pits, which normally occur at the plasma membrane around stimulated synapses, were not detected after injection of SNAP-25 antibodies, consistent with a blockade of vesicle cycling. Thus, SNAP-25 antibodies, which disrupt the interaction with syntaxin, inhibit neurotransmitter release without affecting the number of synaptic vesicles at the plasma membrane. These results provide further support to the view that the formation of SNARE complexes is critical for membrane fusion, but not for the targeting of synaptic vesicles to the presynaptic membrane. 相似文献
4.
Summary Horseradish peroxidase (HRP) was introduced directly into the cerebral cortex of adult rats, which were allowed to survive for 60 min before perfusion fixation. After the tissue had been incubated to demonstrate HRP at the LM and EM levels, blocks of cortical tissue were taken at varying distances from the injection site. These eight blocks of tissue constituted a time sequence for HRP diffusion.Qualitative examination of the presynaptic terminals showed that the most commonly encountered profiles are the plain synaptic vesicles, many of which accumulate tracer. In some terminals labelled vesicles are lined-up in tubular fashion. Other profiles commonly labelled are coated vesicles, tubular and vacuolar cisternae, and plain and coated pinocytotic vesicles.Quantitative analyses based on the number of terminals containing labelled profiles demonstrate an early rise in the rate of labelling of both plain synaptic vesicles and coated vesicles, after which synaptic vesicle labelling rises slowly towards a plateau. By contrast, there is a late parallel increase in the rate of labelling of coated vesicles and cisternae. A more detailed analysis, based on the actual numbers of labelled and total profiles within each presynaptic terminal, highlight early and late periods of rapid labelling for plain synaptic vesicles, coated vesicles and cisternae. A further aspect of HRP incorporation studied, concerns its uptake into four delineated regions of the presynaptic terminal.Our data indicate that membrane uptake into the presynaptic terminal is accomplished mainly via coated vesicles, although plain synaptic vesicles may also be involved. Coated vesicles, in turn, appear to give rise directly to plain synaptic vesicles, with some coalescing to produce vacuolar cisternae. The latter are involved in a two-way interchange of membrane with tubular cisternae, plain synaptic vesicles and coated vesicles. An additional source of plain synaptic vesicles are the tubular cisternae. Exocytosis of plain synaptic vesicles constitutes the mechanism by which transmitter is released from the presynaptic terminal.Supported by the Nuffield Foundation. We are grateful to Mr. M. Austin for help with the photography 相似文献
5.
The docking of synaptic vesicles at active zones on the presynaptic plasma membrane of axon terminals is essential for their fusion with the membrane and exocytosis of their neurotransmitter to mediate synaptic impulse transmission. Dense networks of macromolecules, called active zone material, (AZM) are attached to the presynaptic membrane next to docked vesicles. Electron tomography has shown that some AZM macromolecules are connected to docked vesicles, leading to the suggestion that AZM is somehow involved in the docking process. We used electron tomography on the simply arranged active zones at frog neuromuscular junctions to characterize the connections of AZM to docked synaptic vesicles and to search for the establishment of such connections during vesicle docking. We show that each docked vesicle is connected to 10-15 AZM macromolecules, which fall into four classes based on several criteria including their position relative to the presynaptic membrane. In activated axon terminals fixed during replacement of docked vesicles by previously undocked vesicles, undocked vesicles near vacated docking sites on the presynaptic membrane have connections to the same classes of AZM macromolecules that are connected to docked vesicles in resting terminals. The number of classes and the total number of macromolecules to which the undocked vesicles are connected are inversely proportional to the vesicles' distance from the presynaptic membrane. We conclude that vesicle movement toward and maintenance at docking sites on the presynaptic membrane are directed by an orderly succession of stable interactions between the vesicles and distinct classes of AZM macromolecules positioned at different distances from the membrane. Establishing the number, arrangement and sequence of association of AZM macromolecules involved in vesicle docking provides an anatomical basis for testing and extending concepts of docking mechanisms provided by biochemistry. 相似文献
6.
CHANGES IN THE FINE STRUCTURE OF THE NEUROMUSCULAR JUNCTION OF THE FROG CAUSED BY BLACK WIDOW SPIDER VENOM 总被引:19,自引:9,他引:10
Application of black widow spider venom to the neuromuscular junction of the frog causes an increase in the frequency of miniature end-plate potentials (min.e.p.p.) and a reduction in the number of synaptic vesicles in the nerve terminal. Shortly after the increase in min.e.p.p. frequency, the presynaptic membrane of the nerve terminal has either infolded or "lifted." Examination of these infoldings or lifts reveals synaptic vesicles in various stages of fusion with the presynaptic membrane. After the supply of synaptic vesicles has been exhausted, the presynaptic membrane returns to its original position directly opposite the end-plate membrane. The terminal contains all of its usual components with the exception of the synaptic vesicles. The only other alteration of the structures making up the neuromuscular junction occurs in the axon leading to the terminal. Instead of completely filling out its Schwann sheath, the axon has pulled away and its axoplasm appears to be denser than the control. The relation of these events to the vesicle hypothesis is discussed. 相似文献
7.
SYNAPTIC VESICLE DEPLETION AND RECOVERY IN CAT SYMPATHETIC GANGLIA ELECTRICALLY STIMULATED IN VIVO : Evidence for Transmitter Secretion by Exocytosis 总被引:8,自引:8,他引:0
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This study examined the ultrastructure of presynaptic terminals after short periods of vigorous acetylcholine (ACh) secretion in the cat superior cervical ganglion in vivo. Experimental trunks of cats anesthetized with chloralose-urethane were stimulated supra-maximally for periods of 15–30 min and at several frequencies including the upper physiological range (5–10 Hz). Stimulated and contralateral control ganglia from each animal were fixed by intra-arterial aldehyde perfusion, processed simultaneously, and compared by electron microscopy. Stimulation produced an absolute decrease in the number of synaptic vesicles, an enlargement of axonal surface membrane, and distinct alterations in the shape of presynaptic terminals. Virtually complete recovery occurred within 1 h after stimulation at 10 Hz for 30 min. These results support the hypothesis that ACh release at mammalian axodendritic synapses occurs by exocytosis of synaptic vesicles resulting in the incorporation of vesicle membrane into the presynaptic membrane and that synaptic vesicles subsequently are reformed from plasma membrane. 相似文献
8.
An in vitro model to study the molecular control of binding of highly purified synaptic vesicles to presynaptic plasma membranes has been developed. Presynaptic plasma membranes were immobilized by dotting onto nitrocellulose, and binding of iodinated synaptic vesicle membranes was studied under varying experimental conditions. Synaptic vesicles bind to presynaptic plasma membranes in the presence of Ca2+ and ATP. Binding is reduced in the presence of EGTA and abolished by the calmodulin antagonist trifluoperazine. Vesicle binding is stimulated 5-fold after incubation--prior to dotting--of presynaptic plasma membranes with ATP in the presence of the phorbol-ester 12-O-tetradecanoylphorbol-13-acetate (1 microM) and 2.5-fold after preincubation with Ca2+ (50 microM). Pretreatment of plasma membranes with alkaline phosphatase strongly reduces vesicle binding. Microsomes prepared from bovine liver did not bind to presynaptic plasma membranes. Our results suggest that activation of protein kinase C and Ca2+ stimulate binding of synaptic vesicles to the presynaptic membrane. In the intact nerve terminal this interaction may represent an initial step in synaptic vesicle exocytosis. 相似文献
9.
The organization of cytoplasm at the presynaptic active zone of a central nervous system synapse 总被引:19,自引:0,他引:19
The axoplasm at the presynaptic active zone of excitatory synapses between parallel fibers and Purkinje cell spines contains a meshwork of distinct filaments intermingled with synaptic vesicles, seen most clearly after the rapid freezing, freeze-etch technique of tissue preparation. One set of filaments extends radially from synaptic vesicles and intersects similar filaments associated with vesicles as well as larger filaments arising from the presynaptic membrane. The small, vesicle-associated filaments appear to link synaptic vesicles to one another and to enmesh them in the vicinity of the synaptic junction. The vesicle-associated filaments could be synapsin I because they have the same molecular dimensions and are distributed in the same pattern as synapsin I immunoreactivity. 相似文献
10.
Yu Sun Mytyl Aiga Eileen Yoshida Patrick O. Humbert Shernaz X. Bamji 《Molecular biology of the cell》2009,20(14):3390-3400
An understanding of how synaptic vesicles are recruited to and maintained at presynaptic compartments is required to discern the molecular mechanisms underlying presynaptic assembly and plasticity. We have previously demonstrated that cadherin–β-catenin complexes cluster synaptic vesicles at presynaptic sites. Here we show that scribble interacts with the cadherin–β-catenin complex to coordinate vesicle localization. Scribble and β-catenin are colocalized at synapses and can be coimmunoprecipitated from neuronal lysates, indicating an interaction between scribble and β-catenin at the synapse. Using an RNA interference approach, we demonstrate that scribble is important for the clustering of synaptic vesicles at synapses. Indeed, in scribble knockdown cells, there is a diffuse distribution of synaptic vesicles along the axon, and a deficit in vesicle recycling. Despite this, synapse number and the distribution of the presynaptic active zone protein, bassoon, remain unchanged. These effects largely phenocopy those observed after ablation of β-catenin. In addition, we show that loss of β-catenin disrupts scribble localization in primary neurons but that the localization of β-catenin is not dependent on scribble. Our data supports a model by which scribble functions downstream of β-catenin to cluster synaptic vesicles at developing synapses. 相似文献
11.
N N Bogolepov A S Pushkin V A Chernosvitova V A Dereviagin G N Kryzhanovski? 《Arkhiv anatomii, gistologii i émbriologii》1977,73(11):30-38
Changes in synaptic ultrastructure of the external geniculate body (EGB) were investigated in rats when a generator of pathologically intensified excitation (GPIE) was produced in this nucleus under the influence of tetanus toxin (TT). At the period of pronounced convulsive activity (24 h after TT injection), synaptic changes were estimated electronmicroscopically and with quantitative comparison of the materials from three groups. The first group included EGB synapses where TT was injected, the second group included contralateral EGB synapses and the third included EGB from the rats injected with inactivated toxin. By means of electron optic computer "Klassimat" average amount of round, flat, anomalous and adjacent to the presynaptic membrane vesicles was measured, average relative length of the active zone, average area of the presynaptic terminal, average relative section areas of pre- and postsynaptic cytoplasm condensation were estimated. In the area of GPIE formation, under the influence of TT, the increased amount of the vesicles related to the presynaptic membrane and that of flat vesicles were statistically significant. At the same time, the synaptic terminals, by the number of vesicles, have bimodal, while the control groups have unimodal distribution. 相似文献
12.
Immunoisolation of two synaptic vesicle pools from synaptosomes: a proteomics analysis 总被引:1,自引:0,他引:1
Morciano M Burré J Corvey C Karas M Zimmermann H Volknandt W 《Journal of neurochemistry》2005,95(6):1732-1745
The nerve terminal proteome governs neurotransmitter release as well as the structural and functional dynamics of the presynaptic compartment. In order to further define specific presynaptic subproteomes we used subcellular fractionation and a monoclonal antibody against the synaptic vesicle protein SV2 for immunoaffinity purification of two major synaptosome-derived synaptic vesicle-containing fractions: one sedimenting at lower and one sedimenting at higher sucrose density. The less dense fraction contains free synaptic vesicles, the denser fraction synaptic vesicles as well as components of the presynaptic membrane compartment. These immunoisolated fractions were analyzed using the cationic benzyldimethyl-n-hexadecylammonium chloride (BAC) polyacrylamide gel system in the first and sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the second dimension. Protein spots were subjected to analysis by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI TOF MS). We identified 72 proteins in the free vesicle fraction and 81 proteins in the plasma membrane-containing denser fraction. Synaptic vesicles contain a considerably larger number of protein constituents than previously anticipated. The plasma membrane-containing fraction contains synaptic vesicle proteins, components of the presynaptic fusion and retrieval machinery and numerous other proteins potentially involved in regulating the functional and structural dynamics of the nerve terminal. 相似文献
13.
Andreas Grabrucker Bianca Vaida Jürgen Bockmann Tobias M. Boeckers 《Cell and tissue research》2009,338(3):333-341
Hippocampal neurons in dissociated cell culture are one of the most extensively used model systems in the field of molecular
and cellular neurobiology. Only limited data are however available on the normal time frame of synaptogenesis, synapse number
and ultrastructure of excitatory synapses during early development in culture. Therefore, we analyzed the synaptic ultrastructure
and morphology and the localization of presynaptic (Bassoon) and postsynaptic (ProSAP1/Shank2) marker proteins in cultures
established from rat embryos at embryonic day 19, after 3, 7, 10, 14, and 21 days in culture. First excitatory synapses were
identified at day 7 with a clearly defined postsynaptic density and presynaptically localized synaptic vesicles. Mature synapses
on dendritic spines were seen from day 10 onward, and the number of synapses steeply increased in the third week. Fenestrated
or multiple synapses were found after 14 or 21 days, respectively. So-called dense-core vesicles, responsible for the transport
of proteins to the active zone of the presynaptic specialization, were seen on cultivation day 3 and 7 and could be detected
in axons and especially in the presynaptic subcompartments. The expression and localization of the presynaptic protein Bassoon
and of the postsynaptic molecule ProSAP1/Shank2 was found to correlate nicely with the ultrastructural results. This regular
pattern of development and maturation of excitatory synapses in hippocampal culture starting from day 7 in culture should
ease the comparison of synapse number and morphology of synaptic contacts in this widely used model system. 相似文献
14.
D. J. Stelzner 《Cell and tissue research》1971,120(3):332-345
Summary Routine electron microscopy and a zinc iodide-osmium tetroxide technique (ZIO), recently found to be specific for synaptic vesicles, were used to study the origin of synaptic vesicles during postnatal development in the lumbosacral enlargement of the albino rat. In immature nervous tissue, a large number of vesicles, indistinguishable from synaptic vesicles (S vesicles), were found in the Golgi apparatus and in different portions of the axon where they were often intermingled with elements of the smooth endoplasmic reticulum (SER). Ten to twenty percent of these S vesicles within the Golgi apparatus as well as the majority of these vesicles in all parts of the axon were positive to ZIO. Much of the SER in axons was also positive. The number of vesicles and elements of the SER showed some decrease in the non-terminal portion of axons on day 21 and even more of a decrease in adult neurons. These data suggest that synaptic vesicles are produced in the Golgi apparatus and SER in immature neurons. The decrease in S vesicles and SER in adult neurons suggests a drop in synaptic vesicle production after synaptogenesis has ended. In addition, the material that has been studied shows that ZIO staining is not limited to synaptic vesicles during development since oligodendroglia and endothelial cells are also stained during this period. 相似文献
15.
The actin cytoskeleton and neurotransmitter release: an overview 总被引:12,自引:0,他引:12
Here we review evidence that actin and its binding partners are involved in the release of neurotransmitters at synapses. The spatial and temporal characteristics of neurotransmitter release are determined by the distribution of synaptic vesicles at the active zones, presynaptic sites of secretion. Synaptic vesicles accumulate near active zones in a readily releasable pool that is docked at the plasma membrane and ready to fuse in response to calcium entry and a secondary, reserve pool that is in the interior of the presynaptic terminal. A network of actin filaments associated with synaptic vesicles might play an important role in maintaining synaptic vesicles within the reserve pool. Actin and myosin also have been implicated in the translocation of vesicles from the reserve pool to the presynaptic plasma membrane. Refilling of the readily releasable vesicle pool during intense stimulation of neurotransmitter release also implicates synapsins as reversible links between synaptic vesicles and actin filaments. The diversity of actin binding partners in nerve terminals suggests that actin might have presynaptic functions beyond synaptic vesicle tethering or movement. Because most of these actin-binding proteins are regulated by calcium, actin might be a pivotal participant in calcium signaling inside presynaptic nerve terminals. However, there is no evidence that actin participates in fusion of synaptic vesicles. 相似文献
16.
The formation and refinement of synaptic connections are key steps of neural development to establish elaborate brain networks. To investigate the functional role of protein tyrosine phosphatase (PTP) σ, we employed an olfactory sensory neuron (OSN)-specific gene manipulation system in combination with in vivo imaging of transparent zebrafish embryos. Knockdown of PTPσ enhanced the accumulation of synaptic vesicles in the axon terminals of OSNs. The exaggerated accumulation of synaptic vesicles was restored to the normal level by the OSN-specific expression of PTPσ, indicating that presynaptic PTPσ is responsible for the regulation of synaptic vesicle accumulation. Consistently, transient expression of a dominant-negative form of PTPσ in OSNs enhanced the accumulation of synaptic vesicles. The exaggerated accumulation of synaptic vesicles was reproduced in transgenic zebrafish lines carrying an OSN-specific expression vector of the dominant-negative PTPσ. By electron microscopic analysis of the transgenic line, we found the significant increase of the number of OSN-mitral cell synapses in the central zone of the olfactory bulb. The density of docked vesicles at the active zone was also increased significantly. Our results suggest that presynaptic PTPσ controls the number of OSN-mitral cell synapses by suppressing their excessive increase. 相似文献
17.
Large, second-order neurons of locust ocelli, or L-neurons, make some output connections that transmit small changes in membrane potential and can sustain transmission tonically. The synaptic connections are made from the axons of L-neurons in the lateral ocellar tracts, and are characterized by bar-shaped presynaptic densities and densely packed clouds of vesicles near to the cell membrane. A cloud of vesicles can extend much of the length of this synaptic zone, and there is no border between the vesicles that are associated with neighboring presynaptic densities. In some axons, presynaptic densities are associated with discrete small clusters of vesicles. Up to 6% of the volume of a length of axon in a synaptic zone can be occupied with a vesicle cloud, packed with 4.5 to 5.5 thousand vesicles per microm(3). Presynaptic densities vary in length, from less than 70 nm to 1.5 microm, with shorter presynaptic densities being most frequent. The distribution of vesicles around short presynaptic densities was indistinguishable from that around long presynaptic densities, and vesicles were distributed in a similar way right along the length of a presynaptic density. Within the cytoplasm, vesicles are homogeneously distributed within a cloud. We found no differences in the distribution of vesicles in clouds between locusts that had been dark-adapted and locusts that had been light-adapted before fixation. 相似文献
18.
Zefirov AL 《Rossi?skii fiziologicheski? zhurnal imeni I.M. Sechenova / Rossi?skaia akademiia nauk》2007,93(5):544-562
Presynaptic nerve terminals contain a great number ofsynaptic vesicles filled with neurotransmitter. The transmission of information in synapses is mediated by release of transmitter from vesicles: exocytosis, after their fusion with presynaptic membrane. At the functioning synapses, the continuous recycling of synaptic vesicles occurs (vesicle cycle), which provides multiple reuse of vesicular membrane material during synaptic activity. Vesicle cycle consists of large number of steps, including vesicle fusion--exocytosis, formation of new vesicles--endocytosis, vesicle sorting, filling of vesicles with transmitter, intraterminal vesicle transport driving the vesicles to different vesicle pools and preparing to next exocytic event. At this paper, I presented the latest literature and our data regarding the steps and mechanisms of vesicle cycle at synapses. Special attention was paid to neuromuscular synapse as the most thoroughly investigated and as my favorite preparation. 相似文献
19.
In response to calcium influx, some of the synaptic vesicles in presynaptic terminals fuse rapidly with the presynaptic membrane, allowing fast synaptic transmission. The regulated recycling of synaptic vesicles at the terminals is required for a sustained release of neurotransmitters. Localization of 'ready to be released' vesicles in close vicinities to voltage-gated calcium channels enables the rapid release of neurotransmitters. Thus, recycling vesicles must translocate from the sites of endocytosis to these release sites. However, the sub-cellular organization that supports this local vesicular traffic remains poorly understood. We will review the results of various electron microscopy studies, which have begun to unveil the structure of presynaptic terminals. 相似文献
20.
W Kummer 《Regulatory peptides》1992,37(2):135-142
Calcitonin gene-related peptide-immunoreactive (CGRP-IR) nerves within guinea-pig peribronchial ganglia were studied at ultrastructural level using pre-embedding immunohistochemistry. Preterminal CGRP-IR axons were unmyelinated and contained singular immunoreactive dense core vesicles. CGRP-IR axon terminals were filled with numerous non-reactive small clear vesicles and few immunoreactive dense core vesicles. Some of these terminals were presynaptic to large neuronal processes emerging from local ganglion cells. Another population of presynaptic varicosities lack CGRP-IR. Within CGRP-IR terminals, non-reactive clear vesicles were clustered at the presynaptic membrane whereas CGRP-IR large vesicles remained in some distance from the synaptic cleft. The present observations indicate that: (1) at least two neurochemically different types of synaptic input exist to guinea-pig peribronchial ganglia. (2) CGRP-IR presynaptic terminals probably utilize a non-peptide transmitter for fast synaptic transmission, whilst the peptides are likely to be released parasynaptically and may act in a modulatory fashion. 相似文献