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V Adam-Vizi

Publications and source records attributed to V Adam-Vizi.

36 records · Page 2Linked to original sources
PubMed1995

Transmitter release by non-receptor activation of the alpha-subunit of guanine nucleotide regulatory protein in rat striatal slices.

The effects of 5 mM NaF + 10 microM AlCl3, a direct activator of guanine nucleotide-binding proteins (G proteins), on the release of [3H]dopamine ([3H]DA), [3H]gamma-aminobutyric acid ([3H]GABA), and [3H]acethylcholine ([3H]ACh) were investigated in slices of rat striatum. When the tissue was exposed to NaF + AlCl3 the release of [3H]DA, [3H]GABA, and [3H]ACh was enhanced significantly. In a calcium-free solution the release of [3H]GABA and [3H]DA was increased by NaF+AlCl3 much more than in the presence of [Ca2+]o. In slice preparations taken from reserpinized animals, in which the vesicular storage of [3H]DA was therefore prevented, NaF + AlCl3 had no effect on [3H]DA release. HPLC analysis of the radioactivity of the perfusate showed that, in the presence of NaF + AlCl3, the content of dihydroxyphenylacetic acid (DOPAC) in perfusate samples increased significantly, while in pargyline-treated animals only the DA content was increased. Inhibition of DA carriers by nomifensine or low temperature prevented the effect of NaF + AlCl3. N-ethylmaleimide (NEM) preincubation did not modify the effect of NaF + AlCl3 on [3H]DA release Neomycin (0.1 mM), a phospholipase C (PLC) inhibitor, significantly decreased the effect of NaF + AlCl3 on [3H]DA and [3H]GABA release. The internal concentration of Ca2+ in synaptosomes was enhanced by NaF + AlCl3 in normal solution. However, [Ca2+]i was not influenced by NaF + AlCl3 in Ca(2+)-free medium. It is concluded that a non-receptor-mediated activation, by NaF + AlCl3, of the alpha-subunit of a G protein, results in a [Ca2+]o-independent release of DA and GABA, but not that of ACh.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1995

Relation of [Ca2+]i to dopamine release in striatal synaptosomes: role of Ca2+ channels.

We compared the effects of KCl and 4-aminopyridine (4-AP) stimulation on the coupling of Ca2+ channel activation to [3H]dopamine ([3H]DA) release in rat striatal synaptosomes and used specific Ca2+ channel blockers to discriminate between the different VSCC's activated by the two stimulatory agents. We found that whereas [3H]DA release is strictly Ca(2+)-dependent in the case of KCl depolarization, 4-AP, at concentrations above 100 microM, progressively causes a large Ca(2+)-independent release of [3H]DA. Thus, at 1 to 3 mM 4-AP, as much as 80-95% of the [3H]DA release is Ca(2+)-independent and can be partially blocked by nomifensine, indicating that some [3H]DA release is occurring through reversal of the DA carrier. Therefore, in the studies relating [Ca2+]i to [3H]DA release we selected 4-AP concentrations lower than 100 microM and corrected for the Ca(2+)-independent release. Under these conditions, we determined that: (1) Ca2+ entry through N-type VSCC's is involved in [3H]DA release both in the case of KCl depolarization (35% inhibition by omega-CgTx) and in 4-AP stimulation (23% inhibition by omega-CgTx); (2) Ca2+ entering through P-type and/or Q-type VSCC's is also involved in [3H]DA release due to 4-AP stimulation (26% inhibition by 200 nM omega-Aga IVA); (3) Neomycin (0.35 mM) inhibited the [3H]DA release due to 4-AP stimulation by about 20% and decreased the KCl induced [3H]DA release by 55%; the effects of neomycin (0.35 mM) and omega-CgTx were additive in both cases, indicating that, at this concentration, the antibiotic does not affect significantly N-type Ca2+ channels; (4) When applied together, omega-CgTx and omega-Aga IVA inhibited the 4-AP stimulated [3H]DA release by about 40-50%, suggesting that the remaining large fraction of the VSCC's activated by 4-AP stimulation are non-N, non-P VSCC's and are coupled to Ca(2+)-dependent [3H]DA release; (5) The contribution of L-type VSCC's is uncertain, since there seemed to be a small contribution in the case of KCl depolarization, but not in the case of 4-AP stimulation. On the whole, the results suggest that the release of [3H]DA in the rat striatal nerve terminals depends on Ca2+ entry through N-, P-, possibly Q-, and other non-N-, non-P-type VSCC's when either KCl or 4-AP stimulation is utilized.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1993

Lack of involvement of [Ca2+]i in the external Ca(2+)-independent release of acetylcholine evoked by veratridine, ouabain and alpha-latrotoxin: possible role of [Na+]i.

Synaptosomes were challenged by veratridine, ouabain and alpha-latrotoxin, and the release of 14C-acetylcholine (ACh) was measured in the absence of external Ca2+. We wished to test whether Ca2+ mobilized from internal stores triggered the ACh release that was independent of external Ca2+. We found that none of the agents altered the [Ca2+]i in a Ca(2+)-free medium. Buffering the intracellular Ca2+ concentration with BAPTA did not prevent the increase in release of 14C-ACh by veratridine or ouabain in the absence of Ca2+, however, it greatly reduced the release evoked in a Ca(2+)-containing medium. In parallel samples the release of ACh and the change in the internal Na+ concentration ([Na+]i) were measured. It was found that veratridine, ouabain and alpha-latrotoxin all enhanced [Na+]i in a concentration-dependent manner and a good quantitative relationship existed between the increase in [Na+]i and the release of ACh.

Acetylcholine↗
PubMed1993

Effect of alpha-latrotoxin on acetylcholine release and intracellular Ca2+ concentration in synaptosomes: Na(+)-dependent and Na(+)-independent components.

We studied the effect of alpha-latrotoxin (alpha LTX) on [14C]acetylcholine ([14C]ACh) release, intracellular Ca2+ concentration ([Ca2+]i). plasma membrane potential, and high-affinity choline uptake of synaptosomes isolated from guinea pig cortex. alpha LTX (10(-10)-10(-8) M) caused an elevation of the [Ca2+]i as detected by Fura 2 fluorescence and evoked [14C]ACh efflux. Two components in the action of the toxin were distinguished: one that required the presence of Na+ in the external medium and another that did not Displacement of Na+ by sucrose or N-methylglucamine in the medium considerably decreased the elevation of [Ca2+]i and [14C]ACh release by alpha LTX. The Na(+)-dependent component of the alpha LTX action was obvious in the inhibition of the high-affinity choline uptake of synaptosomes. Some of the toxin action on both [Ca2+]i and [14C]ACh release remained in the absence of Na+. Both the Na(+)-dependent and the Na(+)-independent components of the alpha LTX-evoked [14C]ACh release partly required the presence of either Mg2+ or Ca2+. The nonneurotransmitter [14C]choline was released along with [14C]ACh, but this release did not depend on the presence of either Na+ or Ca2+, indicating nonspecific leakage through the plasma membrane. We conclude that there are two factors in the release of ACh from synaptosomes caused by the toxin: (1) cation-dependent ACh release, which is related to (a) Na(+)-dependent divalent cation entry and (b) Na(+)-independent divalent cation entry, and (2) non-specific Na(+)- and divalent cation-independent leakage.

Acetylcholine↗
PubMed1993

Detection of intracellular free Na+ concentration of synaptosomes by a fluorescent indicator, Na(+)-binding benzofuran isophthalate: the effect of veratridine, ouabain, and alpha-latrotoxin.

A novel fluorescent Na+ indicator, Na(+)-binding benzofuran isophthalate (SBFI), was used to follow changes in the intracellular free Na+ concentration ([Na+]i) of synaptosomes. The dye, when loaded into synaptosomes in the form of its acetoxymethyl ester, was responsive to changes of [Na+]. Calibration was made using the 340/380 nm excitation ratio when the cytoplasmic Na+ concentration was equilibrated with different concentrations of extracellular Na+ in the presence of 2 microM gramicidin D. The basal value of [Na+]i in synaptosomes in the presence of 140 mM extracellular Na+ was found to be 10.9 +/- 1.8 mM. Veratridine, which opens potential-dependent Na+ channels, caused a sudden increase in [Na+]i in a concentration-dependent manner (1-20 microM), whereas the effect of ouabain (20 and 50 microM), the inhibitor of the plasma membrane Na+,K(+)-ATPase, was more gradual. The rise in the fluorescence intensity upon addition of veratridine was prevented completely by 2 microM tetrodotoxin. alpha-Latrotoxin, the black widow spider toxin, caused an increase in the fluorescence intensity, which became evident 1 min after the addition of the toxin. The rate of increase was proportional to the concentration of the toxin (0.19-1.5 nM). This report confirms our earlier finding demonstrating a Na(+)-dependent component in the action of alpha-latrotoxin, and shows that changes in [Na+]i in synaptosomes can be followed by SBFI.

Animals↗
PubMed1992

Parameters not influenced by vesamicol: membrane potential, calcium uptake, and internal calcium concentration of synaptosomes.

In our previous study vesamicol, an inhibitor of the acetylcholine transporter of the cholinergic vesicles, inhibited veratridine-evoked external Ca(2+)-dependent acetylcholine release from striatal slices but did not influence acetylcholine release observed in Ca(2+)-free medium (4). Here we examined if the effect of veratridine on membrane potential, Ca(2+)-uptake, and intracellular Ca2+ concentration of synaptosomes was altered by vesamicol in parallel with the inhibition of acetylcholine release. The depolarizing effect of 10 microM veratridine (from 67 +/- 2.3 mV resting membrane potential to 50.7 +/- 2.5 mV) was not significantly influenced by vesamicol (1-20 microM). Vesamicol (1-20 microM) had no effect on either the overall curve of the veratridine-evoked 45Ca2+ uptake or the amount of Ca2+ taken up by synaptosomes. Veratridine caused a rise in intrasynaptosomal Ca2+ concentration as measured by Fura2 fluorescence, and the same increase both in characteristics and in magnitude was observed in the presence of vesamicol (20 microM). The K(+)-evoked (40 mM) increase of Ca2+ uptake and of intracellular calcium concentration were also unaltered by vesamicol. In high concentration (50 microM) vesamicol inhibited both the fall in membrane potential and the elevated Ca2+ uptake by veratridine, indicating a possible nonspecific effect on potential-dependent Na+ channels at this concentration. Vesamicol, in lower concentration (20 microM) when neither of the above parameters was changed, completely prevented veratridine-evoked increase of [14C]acetylcholine release. This was observed only when vesamicol was present in the media throughout the experiment after loading the preparation with [14C]choline.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗
PubMed1991

Ca2+o-independent veratridine-evoked acetylcholine release from striatal slices is not inhibited by vesamicol (AH5183): mobilization of distinct transmitter pools.

The effect of 2-(4-phenylpiperidino)cyclohexanol (AH5183 or vesamicol), a compound known to block the uptake of acetylcholine (ACh) into cholinergic synaptic vesicles, on the release of endogenous and [14C]ACh from slices of rat striatum was investigated. ACh release was evoked either by electrical stimulation or by veratridine. The effect of electrical stimulation was entirely dependent on external Ca2+. By contrast, veratridine (40 microM) also enhanced ACh release in the absence of Ca2+. Indeed, with veratridine two components were clearly distinguished: one dependent on external Ca2+ and the other not. Vesamicol inhibited [14C]ACh release evoked by both veratridine and electrical stimulation in the presence of external Ca2+, provided it was added to the tissue prior to loading with [14C]choline. With the same treatment vesamicol only slightly affected the release of endogenous ACh. Under the same conditions the Ca2(+)-independent [14C]ACh release evoked by veratridine was not prevented by vesamicol. The differential responsiveness to vesamicol suggests that ACh pools involved in Ca2+o-dependent ACh release are different from those mobilized during Ca2+o-independent ACh release.

Acetylcholine↗
PubMed1988

Botulinum toxin-induced ADP-ribosylation and inhibition of exocytosis are unrelated events.

The hypothesis that inhibition of secretion by botulinum neurotoxin type D occurs by an intracellular process involving ADP-ribosylation has been directly tested by measuring both the extent of inhibition of secretion and of ADP-ribosylation in the same cells. Although the inhibitory effect of unpurified toxin closely parallels intracellular ribosylation, the two events are clearly unrelated, as using purified D and C3 toxins together with their antibodies, each of these events can be either stimulated or inhibited independently of each other.

Adenosine Diphosphate Ribose↗
PubMed1988

The effects of nitroprusside and putative agonists on guanylate cyclase activity in squid giant axons.

cGMP content of axoplasm from the giant axon of Loligo forbesi was investigated after subjecting the axon to various treatments. Repetitive electrical stimulation or depolarisation by high K+ caused no change in cGMP content. Glutamate and serotonin were also without effect. The nicotinic agonist carbachol (100 microM) increased cGMP levels by 90% (n = 5). A large transient elevation of cGMP content was evoked by external nitroprusside (10 nM-20 microM in intact axons. Nitroprusside injected into both extruded axoplasm and intact axons also increased cGMP content, the stimulation being considerably higher in intact axons where the axolemma was also present. Nitroprusside was also active in axons where the soluble cytoplasmic components were washed out by internal perfusion.

Animals↗
PubMed1987

Depolarization of brain cortex slices and synaptosomes by lithium. Determination of K+-equilibrium potential in cortex slices.

K+-equilibrium potential was determined in brain cortex slices of rat by measuring 86Rb+ distribution between the extra- and intracellular space. The ratio of internal to external Rb+ concentration was 39 +/- 1.8, corresponding to a resting membrane potential of 93.8 mV. Li+ (1-126 mM) decreased the membrane potential in both cortex slices and synaptosomes in a concentration-dependent manner. The presence of 1 mM Li+ was enough to cause a slight but distinct depolarization. During incubation in Li+-containing medium slices took up K+; however, for depolarization the presence of extracellular Li+ seemed to be necessary.

Animals↗
PubMed1987

Relation of acetylcholine release to Ca2+ uptake and intraterminal Ca2+ concentration in guinea-pig cortex synaptosomes.

[14C]Acetylcholine (ACh) release and parallel alterations in 45Ca2+ uptake and intrasynaptosomal free CA2+ concentration ([Ca2+]i) were measured in guinea-pig brain cortex synaptosomes. Depolarization by high K+ concentrations caused a rapid transient increase in Ca2+ uptake, terminating within 60 s (rate constant = 0.060 s-1; t1/2 = 11.6 s). This resulted in a rapid increase (within 1 s) in [Ca2+1]i, which then fell to a maintained but still-elevated plateau level (t1/2 for the decline was 15 s). Peaks of [Ca2+]i showed a sigmoidal dependence on depolarization, contrasting with the simple linear dependence of plateau levels of [Ca2+]i. The K+-evoked ACh release also had two phases: a fast initial increase (t1/2 = 11.3 s), which terminated within 60 s, was followed by a slow additional increase during sustained depolarizations of up to 10 min. Depolarization by veratridine led to a slow gradual increase in Ca2+ uptake (t1/2 = 130 s) over a 10-min incubation period, whereas an elevated plateau level of [Ca2+]i was achieved within 2 min (without a rapid peak elevation). The Ca2+-dependent fraction of the veratridine-evoked ACh release correlated with the increase in [Ca2+]i rather than with Ca2+ uptake. Using two different methods of depolarization partially circumvented the time limitations imposed by a buffering Ca2+ indicator and we suggest that, in the main, ACh is released in bursts associated with [Ca2+]i transients.

Acetylcholine↗
PubMed1986

Calcium uptake of rat brain synaptosomes as a function of membrane potential under different depolarizing conditions.

The uptake of 45Ca2+ was measured in brain synaptosomes under conditions designed to depolarize the membranes. Membrane potential was estimated from the distribution of 86Rb+ between the intra- and extracellular compartments. K+ depolarization (8-60 mM) only increased "a2+ uptake beyond a threshold depolarization of about 10 mV, whereas veratridine (5-40 microM) induced an increased Ca2+ uptake at a concentration which depolarized the membrane less than this threshold. Ouabain did not enhance Ca2+ uptake, but the depolarization it produced did not reach threshold. Ca2+ influx already stimulated by K+ depolarization can be further enhanced by veratridine without any parallel change in membrane potential. Only the pathway mediating Ca2+ uptake during K+ depolarization can be inactivated: Ca2+ uptake evoked by K+ depolarization is decreased in synaptosomes pre-depolarized in the presence of a high concentration of K+. In contrast, pre-depolarization does not change Ca2+ uptake evoked by veratridine. Ca2+ channel blockers, such as verapamil and diltiazem but not nifedipine, in concentrations of 10-100 microM, decrease the stimulation of Ca2+ uptake by high K+ concentration without influencing depolarization, whereas the effect of veratridine on Ca2+ uptake is only inhibited when its effect on Na+ channels is also prevented. It is concluded that Ca2+ uptake during K+ depolarization proceeds through voltage-dependent Ca2+ channels similar to those of squid axon, whereas veratridine activates an additional Ca2+ entry, possibly via influx through open Na+ channels. Different quantitative relationships are found between acetylcholine release of synaptosomes and the amount of Ca2+ taken up by different mechanisms: the same amount of Ca2+ uptake is accompanied by a greater increase of acetylcholine release if the uptake is induced by K+ depolarization rather than veratridine.

Acetylcholine↗
PubMed1984

Release of acetylcholine from rat brain synaptosomes by various agents in the absence of external calcium ions.

The relationship between 86Rb+ distribution across synaptosomal membrane and [14C]acetylcholine (ACh) release have been studied in a rat brain cortex synaptosomal preparation using K+, ouabain and veratridine depolarization. Decrease in membrane potential, approximated from the 86Rb+ distribution, is accompanied by an increase in [14C]ACh release, but the extent of the increase at a certain depolarization is dependent on how the depolarization is induced. A substantial depolarization by K+ is necessary to enhance ACh release, as compared to ouabain and veratridine where only a slight depolarization is accompanied by an increase in ACh release. In Ca2+-free, EGTA-containing medium ouabain and veratridine can also increase [14C]ACh release. The relationship between membrane potential and ACh release is very similar in the presence of ouabain and veratridine both in Ca2+-containing and Ca2+-free medium. The effect of ouabain and veratridine on the Na-K exchange pump is different; ouabain can completely abolish Na-K-ATPase activity and 86Rb+ uptake of synaptosomes, whereas veratridine does not seem to influence the activity of the pump. m-Chloro-carbonylcianid phenyl hydrazon (50-500 nM) increases [14C]ACh release in a concentration-dependent manner without a considerable change of membrane potential or Na-K pump activity. The Ca2+ ionophore A 23187 induces a substantial increase in [14C]ACh release in the absence of external Ca2+. In this case neither Na-K pump activity nor membrane potential of synaptosomes is changed. A possible role of intracellular Ca2+ mobilization as a consequence of increased intracellular Na+ concentration in some depolarization-induced transmitter release is discussed.

Acetylcholine↗
PubMed1984

Presynaptic inhibitory effect of Met-enkephalin on [14C] acetylcholine release from the myenteric plexus and its interaction with muscarinic negative feedback inhibition.

The effect of Met-enkephalin on the release of radioactivity (14C) from a myenteric plexus-longitudinal muscle preparation loaded with [14C]choline has been investigated under different conditions, when the muscarinic receptor mediated negative feedback inhibition was operative or when it was completely excluded by atropine. Separation of the [14C]acetylcholine (ACh) and [14C] choline components of the released radioactivity revealed that during 45-min incubation periods about 3.2% of the ACh store became labeled and that during stimulation only the release of [14C]ACh increased above resting level. The fractional release at rest measured in the 5-min collection period was 1.07 +/- 0.09 X 10(-2) in the absence and 1.56 +/- 0.07 X 10(-2) in the presence of physostigmine. Met-enkephalin had no effect on the release of ACh evoked by a 2-Hz stimulation when cholinesterase was inhibited by physostigmine. However, in the presence of atropine or in the absence of cholinesterase inhibition, the release by stimulation was significantly higher and subject to inhibition by Met-enkephalin. The present results indicate that Met-enkephalin is able to reduce ACh release only under those conditions in which the negative feedback modulation is negligible and the release is not yet reduced completely. These findings also suggest that in the myenteric plexus there is no independent population of cholinergic neurons exclusively sensitive to either ACh or to Met-enkephalin; cholinergic varicosities are equipped with at least both types of receptors we studied. When the effect of Met-enkephalin on twitch tension and on the release of radioactivity was studied simultaneously, a positive correlation was found.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗
PubMed1983

Studies on the osmotic disruption and resealing of synaptosomes.

The release of lactate dehydrogenase and K+ when synaptosomes are exposed to resuspension in media of various osmolarity has been investigated in order to measure their disruption. Even when resuspended in distilled water a significant percentage (10-20%) of lactate dehydrogenase and K+ remains unreleased. The particles containing these substances sediment to the same density as synaptosomes. Synaptosomes retaining their internal organelles after hypoosmotic treatment can be seen in electron micrographs. Resealing of disrupted synaptosomes was measured by the inclusion of [14C]sucrose. The resealing is spontaneous, essentially complete (80-90%) within 20 min and not noticeably affected by temperature, pH, or the addition of fusogen. The synaptosome preparation after hypoosmotic disruption will therefore contain some undisrupted synaptosomes with some or all of their complement of cytoplasmic constituents, as well as resealed synaptosomes. The retention of the ability of the hypoosmotically treated preparation to convert [14C]choline to [14C]acetylcholine is demonstrated as an example of the disproportionate effect these undisrupted particles have on its properties.

Acetylcholine↗
PubMed1982

Na-K activated ATPase and the release of acetylcholine and noradrenaline.

1. It has been shown that different experimental conditions known to inhibit Na-K-activated ATPase, and enzyme present in the neuronal membranes, are able to promote transmitter release (ACh, NA, etc.) from different tissues, simply by making the membrane leaky. 2. Under physiological conditions, Ca entering the cell transiently inhibits membrane ATPase, resulting in a transient change in membrane permeability and a subsequent release of transmitter. 3. When membrane ATPase inhibitor was used one part of the release proved to be Ca-independent. This finding indicates that the voltage and Ca-dependent link of transmitter release can be by-passed by direct membrane ATPase inhibitors (ouabain). 4. Neurochemical and electrophysiological evidence was obtained on mouse diaphragm that most of the released ACh is cytoplasmic and Na-K ATPase inhibition is responsible for its release. 5. The stimulation of membrane ATPase (by switching off K and its readmission) results in an inhibition of both ACh and noradrenaline release evoked by axonal stimulation. 6. It is suggested that, in those cases where the varicose axon terminals do not make synaptic contact, the transmitter released from the cytoplasmic pool contributes to the transmission, since during diffusion (sometimes few thousand nm) transmitter of different origins becomes mixed up.

Acetylcholine↗