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PubMed1977

Synaptic vesicle fraction devoid of adenosine triphosphatase activity from bovine caudatolenticular nuclei and thalamus.

1. As a part of studies on the mechanism by which catecholamines are released from the nerve terminals, the synaptic vesicle fraction was isolated from bovine caudatolenticular nuclei and thalamus by differential centrifugation essentially according to the method of Kadota and Kadota (17). 2. Further centrifugation on a sucrose density gradient of the synaptic vesicle fraction by the method of Whittaker et al. (1) yielded white materials on the upper portion of 0.4 M sucrose, which consisted of vesicles averaging 600-800 A in diameter, and did not show Mg2+-dependent ATpase activity. On the other hand, the denser materials centering on 0.6 M sucrose, consisting of a mixture of microsomes and synaptic vesicles of 400-500 A diameter, showed an ATpase activity activated by either Mg2+ or Ca2+ but not inhibited by ouabain. 3. The white materials on 0.4 M sucrose were almost free of mitochondria, but they contained a large amount of non-heme iron, as reported elsewhere (2). Furthermore, the protein components analyzed on SDS-polyacrylamide gels were similar to those already reported for purified synaptic vesicles (3). 4. Based on these results, the white materials were assumed to be synaptic vesicles devoid of Mg2+-dependent ATPase activity.

Adenosine Triphosphatases↗
PubMed1978

Electrogenic behavior of synaptic vesicles from Torpedo californica.

Electrical potential changes in pure synaptic vesicles from Torpedo californica were monitored with the fluorescent dye 3,3'-dipropylthiadicarbocyanine iodide. Vesicles resuspended in variable external sodium ion in the presence of gramicidin established sodium ion membrane diffusion potentials. Vesicles resuspended in choline or acetylcholine chloride became hyperpolarized upon addition of gramicidin. Hyperpolarization was subsequently partially reversed spontaneously by choline or acetylcholine influx, which was confirmed by gel filtration, to yield a new, less negative, stable membrane potential. Thus, acetylcholine and choline are taken up electrogenically by synaptic vesicles.

Acetylcholine↗
PubMed1977

Chlorpromazine induces population increase of synaptic vesicles in the rat hypothalamic ventromedial nucleus.

The population of synaptic vesicles per synaptic area in the hypothalamic ventromedial nucleus of rats treated with chlorpromazine (CP) for 3 days was compared with that of controls. In type I synapse, which contained 400-800 A small spherical vesicles (s-vesicles), together with 800-1200 A large spherical dense-cored vesicles (lc-vesicles), the population of s-vesicles increased 57.8% in the CP rats, In type II synapse, which contained only s-vesicles, the rate of increase among CP animals was 15.4%. Findings suggested that CP blocks the release of the vesicles from the presynaptic site or inhibits the transport function for biogenic amines through the presynaptic membrane, and type I synapse may be aminergic.

Animals↗
PubMed1976

Nucleoside triphosphate and cation requirement for dopamine uptake by plain synaptic vesicles isolated from rat cerebrums.

Plain synaptic vesicles were partially purified from rat cerebrums and their [3H]dopamine uptake was investigated. The addition of MgCl2 plus ATP and of CaCl2 plus ATP increased the uptake 16 times and 5.5 times, respectively, higher than the control level, whereas the separate addition of these agents augmented the level at most 2.5 times. GTP stimulated the uptake as well as ATP, whereas UTP was one-third as effective and CTP, ADP, and AMP were all ineffective. Adenylyl imidodiphosphate was not only ineffective, but strongly reduced the ATP-dependent uptake. The half-maximal level of the ATP-dependent uptake was reached within 1.5 min after the start of incubation at 25 degrees C. The incorporation peaked at 5-10 min, then gradually declined to half-maximum at 45 min. This decline was prevented by the further addition of small amounts of CaCl2. There was no uptake at 0 degrees C, and the incorporation rate was at least 2 times faster at 37 degrees C than it was at 25 degrees C. The apparent Michaelis constant for [3H] dopamine was 1.6 muM. The half maximal inhibition of uptake was obtained at 0.1 muM reserpine; neither colchicine nor ouabain showed significant inhibition. The ATP-dependent uptake was not affected by K+, Na+, and Cl-, but was drastically decreased by isotonic phosphate buffer.

Adenosine Triphosphate↗
PubMedDate not supplied

Quantification of synaptic vesicles in hippocampus of aging rats and initial studies of possible relations to neurophysiology.

Synaptic vesicle populations were quantified in Schaffer-commissural synapses which terminate on CA1 pyramidal cell apical dendrites, in aging and young-mature rats. Vesicles were found to be reduced with age, and this effect was most pronounced in the oldest animals (e.g., 28 months) within the aged group. Numerical density of synaptic vesicles in aged rat hippocampus was reduced whether expressed as vesicles per terminal, vesicles per mu 2 of terminal, or vesicles per mu2 of terminal corrected for shrinkage or swelling as assessed by mitochondrial cross-sectional diameters. Counted synaptic terminal areas were not significantly different in the aged animals, although a trend toward reduced terminal size with aging was seen. The latter observation apparently rules out increased terminal swelling in aged rat synapses as a basis for reduced vesicle density. In some rats, neurophysiological studies were concomitantly performed. A significant correlation was found between the amplitude of the monosynaptic population spike after 20 min of 4 Hz stimulation of the Schaffer-commissural fibers and synaptic vesicle populations in terminals of these stimulated pathways. However, because of a low n this result must be viewed as preliminary.

Aging↗
PubMed1975

The involvement of lysophosphoglycerides in neurotransmitter release; the composition and turnover of phospholipids of synaptic vesicles of guinea-pig cerebral cortex and Torpedo electric organ and the effect of stimulation.

(1) Crude synaptosomal fractions (P2) derived from guinea-pig cerebral cortex were incubated in the presence of 50 mM KCl in a Krebs-glucose medium. Torpedo marmorata electric organs were stimulated electrically in vivo at 5 pulses/sec for 30 min by electrodes placed on the electric lobe. Synaptic vesicles were isolated from each source and the phospholipid compositions analysed and compared with vesicles from unstimulated controls. (2) Lysophosphatidylcholine was the only lysophosphoglyceride demonstrable in the synaptic vesicles from either source and its low levels did not increase as a result of chemical or electircal stimulation. In each case there was a close similarity of the phospholipid distributions in the vesicles taken from control and stimulated samples. (3) Control experiments indicated extensive decreases in the acetylcholine content of the vesicles from the stimulated electric organ and smaller decreases in the acetylcholine content of the synaptic vesicles from stimulated crude synaptosomal fractions. These fractions were found to respire linearly in the presence of 10 mM glucose and the vesicle fractions were shown to have low levels of contaiminating membranes as judged by marker enzyme analyses. (4) Crude synaptosomal fractions from guinea-pig cerebral cortex were incubated in a Krebs-glucose medium with labelled fatty acids and [3H]glucose in the presence or absence of 50 mM KCl. Subsynaptosomal fractionation was carried out and specific radioactivities of phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine and phosphatidylinositol were determined in fractions D (synaptic vesicles), E (microsomes) and H (disrupted synaptosomes). The release of neurotransmitter did not significantly enhance the labelling of phospholipids in any of the fractions studied as compared with phospholipids from unstimulated fractions. This was found after two incubation times and using [14C]oleate, [14C]arachidonate, [3H]palmitate and [3H]glucose.

Acetylcholine↗
PubMed1976

A critical evaluation of the relationship between the presynaptic network, synaptic vesicles and dense projections in central synapses.

Synapses of the oculomotor nucleus of Echidna have been examined ultrastructurally with the aim of integrating data obtained from osmicated and nonosmicated PTA stained material. Particular emphasis has been laid on the relationship between the synaptic vesicles of the osmicated material and the presynaptic network and vesicular grid of the PTA material. This relationship has been explored qualitatively by examining osmicated material of varying qualities of fixation. Such material contains dense projections in addition to synaptic vesicles, and various vesicular network appearances. A variety of measurement techniques have shown that the PTA network is characterised by reticular strands, spaces, and regular hexagonal units smaller than vesicles, these observations prompting the formulation of a "vesicle-network coincidence" model of the presynaptic terminal. This model has been tested by tracing the profiles of vesicles within the PTA network and comparing their size and shape frequency distributions with those of osmicated synaptic vesicles. The distributions have been found to be essentially similar, suggesting that vesicles can be located within the network, and that the hexagonal network units are formed only in the presence of an underlying vesicular matrix. Additionally, the following points have emerged: 1) the dense projections in the two types of material appear to be quivalent; 2) a loose correlation exists between dense projections and vesicles in osmicated terminals, increase in the area of the dense projections being associated with a decrease in the area of the vesicles; 3) network and dense projection units are similar. In view of the similarity between network and dense projection units, the demonstrated vesicular basis of the network raises the question of whether dense projections are entirely independent structures, or whether they depend in part for their existence on the nearby presence of synaptic vesicles.

Animals↗
PubMed1978

Visualization of proteinaceous granules in the clear synaptic vesicles.

A method for demonstration of electron-dense particles within clear synaptic vesicles from various areas of the CNS as well as from neuromuscular junctions of rat is described. Electron-dense granules of 70-250 A were visible in the center of the synaptic vesicles, or in some cases excentrically situated and bound to the vesicular membrane. Digestion with proteolytic enzymes lead to a negative reaction, whereas treatment with hyaluronidase and neuraminidase, as well as the lipid extraction had no effect. Based on the obtained data, it may be assumed that this method manifests the proteinaceous structures.

Animals↗
PubMed1976

Interrelationships between Golgi, GERL and synaptic vesicles in the nerve cells of insect and gastropod ganglia.

In addition to demonstrating synaptic vesicles, staining with the zinc-iodide-osmium tetroxide (ZIO) method reveals the presence of positively reacting GERL membranes in association with the Golgi complex and lysosomes in the nerve cell bodies within ganglia from the locust Schistocerca gregaria and the gastropod molluscs, Limnaea stagnalis and Helix aspersa. A positive response to ZIO occurs in certain Golgi vesicles and saccules, in GERL (Golgi-endoplasmic-reticulum-lysosomes), in multivesicular bodies as well as residual bodies and in small vesicles and cisternae of axonal smooth endoplasmic reticllum (ER). The interrelationships between these organelles are considered in view of the similarity of the ZIO localization to phosphatase-rich sites in the neuronal perikarya and with respect to the possibility that components of the synaptic vesicles are formed in the Golgi region of the cell and migrate via the axonal smooth ER to the synaptic regions.

Animals↗
PubMed1975

[Effect of carbidine on the content and storage of adrenergic neurotransmitter in the synaptic vesicles].

The influence of carbidine, an original psychotropic drug, on the adrenergic neurotransmitter content and storage in the sympathetic nerves was studied with the use of cytochemical electron microscopy. The influence of carbidine on the uptake of the exogenous noradrenaline (NA) in the synaptic vesicles was also studied. Carbidine was found to be capable to decreasing the NA storage in the synaptic vesicles and failed to block the accumulation of the exogenous NA in the synaptic vesicles.

Amines↗
PubMed1976

Characterization of ATPases of plain synaptic vesicle and coated vesicle fractions isolated from rat brains.

The plain synaptic vesicle and the ocated vesicle fractions were isolated from rat brains, and the ATPase [EC 3.6.1.3] activities were characterized in terms of ionic effects, drug effects, and protein components. Coated vesicle fraction contained three times as much actomysin-like proteins as plain vesicle fraction, although both fractions had an identical ratio of actin-like protein to myosin-like protein. The ATPases of these two fractions were activated by both Mg2+ and Ca2+, and, in the presence of either of the cations, were inhibited by KCl. Reserpine activated plain vesicle ATPase only in the presence of Cl-. Colchicine and vinblastine inhibited coated vesicle ATPase only. The results are consistent with the view that actomyosin-like proteins are involved in the synaptic retrieval process.

Actins↗
PubMed1978

The effect of pentobarbital, chloralhydrate, ether and protoveratrine on the distribution of synaptic vesicles in rat cortical synaptosomes.

Rat cortical synaptosomes were incubated in a saline medium. On the addition of pentobarbital, chloralhydrate, ether and protoveratrine a redistribution of synaptic vesicles was revealed by electron microscopy. The anaesthetics decreased the number of synaptic vesicles attached to the presynaptic membrane while protoveratrine increased them. It is assumed that these agents affect the mechanism of vesicle reformation from the synaptosomal membrane and/or vesicle migration towards the synaptic cleft.

Animals↗
PubMed1979

Changes in the distribution and size of synaptic vesicles in neuromuscular junctions of Locusta migratoria after stimulation and rest.

The distribution and size of synaptic vesicles in excitatory terminals of the extensor tibiae muscle were determined after stimulation at frequencies varying from 0.5 to 100 Hz and after subsequent rest. Only in preparations stimulated at the higher frequencies do these parameters differ from those in the controls. The synaptic vesicles in the nonsynaptic areas of these terminals are depleted in number, and the remaining vesicles are reduced in size. These effects are reversed after a 1 h rest.

Animals↗
PubMed1978

Purification of synaptic vesicles from elasmobranch electric organ and the use of biophysical criteria to demonstrate purity.

We have purified cholinergic synaptic vesicles from the electric organs of two related marine elasmobranchs, Torpedo californica and Narcine brasiliensis, to a specific activity higher than had previously been obtained. We have demonstrated the homogeneity of the vesicles by biophysical criteria. The purification scheme consisted of differential centrifugation, flotation equilibrium in sucrose density gradients, and permeation chromatography on glass bead columns of average pore size 3000 A. Our criteria for purity were that bound acetylcholine, bound nucleotide triphosphate, protein, and lipid--phosphorus behave identically when vesicles were analyzed by procedures which depend on vesicle size, density, and charge. Contaminants were not detected when vesicles were fractionated by preparative and analytical sedimentation, by preparative equilibrium sedimentation using glycerol density gradients, or by electrophoresis in Ficoll density gradients. Pure synaptic vesicles, which have been purified 290-fold from the initial homogenate, contain per mg of protein: 8 mumol of acetylcholine, 3 mumol of ATP, and 7 mumol of lipid phosphorus. These procedures may be of general value in the purification of membrane vesicles.

Animals↗
PubMed1975

Influence of castration on incorporation of exogenous peroxidase into "synaptic" vesicles of the median eminence. II. The perivascular part of the palisade zone in male rats.

In the median eminence of male rats, nerve profiles in the immediate vicinity of portal capillaries have been divided into 4 categories on the basis of their vesicular content: profiles a with agranular "synaptic" vesicles of about 50 nm, b with similar agranular vesicles and also with granular vesicles of mainly 60-140 nm, c with granular vesicles alone and d without vesicles. Twenty-four hours after castration, the percentage of profiles of category a was significantly increased when compared with sham-operated animals, whereas the percentage of profiles of category b was significantly decreased. After intravenous injection of exogenous peroxidase, especially the nerve profiles located in direct contact with the outer basement membrane of the portal capillaries contained peroxidase positive "synaptic" vesicles. Injection off peroxidase after castration resulted in a significant increase in the percentage of nerve profiles containing both peroxidase positive and peroxidase negative "synaptic" vesicles (category a). It is suggested that, in certain nerve terminals, castration may lead to release off the content of granular vesicles, which may contain gonadotropin releasing factor. This release then may cause that nerve terminals with agranular vesicles and granular vesicles (category b) change in their vesicular content and are therefore classified as terminals with only agranular vesicles (category a). An increased turnover rate in the ultrastructurally affected terminals may be reflected in increased uptake of exogenous peroxidase. The observations imply that certain neurones projecting to portal capillaries in the median eminence are, directly or indirectly, sensitive to changes in the level of gonadal steroids.

Animals↗
PubMed1979

5'-triphosphate recycles independently of acetylcholine in cholinergic synaptic vesicles.

The effect of hemicholinium-3 (HC-3) on vesicular contents in acetylcholine (ACh) and 5-triphosphate (ATP) and the vesicular incorporation of 14C-label derived from [14C]choline ,nd 3H-label derived from [3H]adenosine was investigated after low frequency stimulation (with a subsequent rest period) of the Torpedo electric organ. HC-3 (100 microM) caused an increased depletion of vesicular ACh and blocked the incorporation of 14C-label whereas contents in vesicular ATP and 3H-incorporation were identical with and without HC-3. HC-3 also blocked the recovery of electrical response of the tissue after stimulation but did not cause a change in vesicle numbers. The result suggest that synaptic vesicles continue to recycle ATP in the absence of recycling of ACh and that vesicular uptake and storage of the two components are not coupled to each other.

Acetylcholine↗