Effects of 4-(2-Hydroxyethyl)-1-piperazine-ethanesulfonic acid (AH5183) on rat cortical synaptosome choline uptake, acetylcholine storage and release

Otero, D. H.; Wilbekin, Finlandia; Meyer, Edwin M.

doi:10.1016/0006-8993(85)91430-1

Cited by 37 publications

(24 citation statements)

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“…AH5183 is the most potent inhibitor of this transport system, half inhibiting AcCho uptake at 40 nM in a noncompetitive manner (7,8). AH5183 also has been found to inhibit the uptake of newly synthesized AcCho into synaptic storage vesicles in PC12 cells (9,10) and, presumably as a secondary effect, to inhibit the release of AcCho from the cat superior cervical ganglion in situ (11) and from rat and mouse brain preparations in vitro (12)(13)(14). Remarkably, the above studies could not demonstrate significant direct effects of AH5183 on high-affinity choline uptake, choline acetyltransferase (ChoAcTase) activity, Ca2l influx, or the vesicular content of previously loaded AcCho.…”

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confidence: 99%

Quantitative autoradiography of brain binding sites for the vesicular acetylcholine transport blocker 2-(4-phenylpiperidino)cyclohexanol (AH5183).

Marien¹,

Parsons²,

Altar³

1987

Proc. Natl. Acad. Sci. U.S.A.

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ABSTRACT2-(4-Phenylpiperidino)cyclohexanol (AH-5183) is a noncompetitive and potent inhibitor of high-affinity acetylcholine transport into cholinergic vesicles. It is reported here that [3H]AH5183 binds specifically and saturably to slide-mounted sections of the rat forebrain (Kd = 1.1 to 2.2 x 10-8 M; Bm.. = 286 to 399 fmol/mg of protein). The association and dissociation rate constants for [3HJAH5183 binding are 8.6x 106 M-1*min-1 and 0.18 mini1, respectively. Bound[31H]AH5183 can be displaced by nonradioactive AH5183 and by the structural analog (2a,3,f,4A,8,8Aa)-decahydro-3-(4-phenyl-l-piperidinyl)-2-naphthalenol but not by 10 ,uM concentrations of the cholinergic drugs acetylcholine, choline, atropine, hexamethonium, eserine, or hemicholinium-3 or by the structurally related compounds 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, 1-methyl-4-phenylpyridine, (±)-N-allylnormetazocine (SKF 10,047), levoxadrol, or dexoxadrol. Brittain and coworkers (1, 2) reported that 2-(4-phenylpiperidino)cyclohexanol (AH5183), a structural isomer of hydroxylated phencyclidine, inhibits the coaxially stimulated contractions of the isolated guinea pig ileum. Because AH5183 did not affect the contractions elicited by the addition of acetylcholine (AcCho) to the bath, it was proposed that AH5183 produces a "selective prejunctional inhibition" of cholinergic neurotransmission (1, 2). Based on the pharmacological characteristics of the neuromuscular blockade produced by AH5183 in vivo (1, 2), in vitro, and in situ (3-5), Marshall hypothesized that AH5183 inhibits the loading of AcCho into synaptic vesicles. In support of Marshall's hypothesis, synaptic vesicles isolated from the electric organ of Torpedo californica, a model system for the mammalian cholinergic synapse (6), have been shown to acquire AcCho by an active transport process that is pharmacologically distinct from other cholinergic systems (7). AH5183 is the most potent inhibitor of this transport system, half inhibiting AcCho uptake at 40 nM in a noncompetitive manner (7, 8). AH5183 also has been found to inhibit the uptake of newly synthesized AcCho into synaptic storage vesicles in PC12 cells (9, 10) and, presumably as a secondary effect, to inhibit the release of AcCho from the cat superior cervical ganglion in situ (11) MATERIALS AND METHODS Materials. The following chemicals were used in this study: AcCho chloride (Fluka); atropine sulfate, choline chloride, eserine sulfate, hemicholinium-3, and hexamethonium bromide (Sigma); (+)-N-allylnormetazocine (SKF 10,047) (Smith Kline & Beckman); and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MePhH4Py) and 1-methyl-4-phenylpyridine (MePhPy+) (Research Biochemicals, Wayland, MA).[3H]AH5183 (15) MePhPy+, 1-methyl-4-phenylpyridine; MePhH4Py, 1-methyl-4-phnenyl-1,2,3,6-tetrahydropyridine . XTo whom reprint requests should be addressed.The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely ...

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confidence: 99%

Quantitative autoradiography of brain binding sites for the vesicular acetylcholine transport blocker 2-(4-phenylpiperidino)cyclohexanol (AH5183).

Marien¹,

Parsons²,

Altar³

1987

Proc. Natl. Acad. Sci. U.S.A.

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“…and Collier, 1986). Because AH 5 183 does not block extracellular choline transport appreciably at the low concentration (75 nM) used in our study (Otero et al, 1985;Jope and Johnson, 1986), it would appear likely that it blocks synthesis at the vesicular surface by reducing an accelerated transport of ACh into synaptic vesicle release sites, by inhibiting the veratridine-induced activation of that ChAT fraction associated with these sites, or by suppressing both processes. For example, as described already, AH 5 183 reduced the ACh content of the "monodisperse" vesicle fraction during veratridine depolarization of tissue at a time when repletion of this fraction with newly synthesized ACh appeared important to the maintenance of evoked release.…”

Section: Subcsllulor Fractionsmentioning

confidence: 77%

“…Because a water-soluble fraction of ChAT believed to be cytosolic is also activated during depolarization of brain tissue (Carroll, 1987), it could also potentially contribute to the depolarization-induced increase in brain ACh synthesis. We predicted that the drug AH 5 183 might be helpful in discriminating between the water-and detergent-soluble ChAT fractions because of its known inhibitory actions at synaptic vesicles on ACh transport (Anderson et al, 1983) and release (CarroJ, 1985;Otero et al, 1985;Jope and Johnson, 1986;R%n$ and Collier, 1986) as well as its ability to obviate the increase in ACh synthesis that occurs when brain tissue is depo-l_arized with high K+ levels (Jope and Johnson, 1986;NEn? and Collier, 1986).…”

Section: Discussionmentioning

confidence: 99%

“…For example, the water-soluble ChAT fraction, which differs physically from the detergent-soluble ChAT fraction , may float freely in the cytosol and form that ACh released under resting conditions. Also, it reduces the release of ACh from brain tissue during either high-K+-level (Carro!l, 1985; Otero et al, 1985;Jope and Johnson, 1986;RiEn? This latter suggestion can now be tested because of the existence of the cholinergic neurotoxin 2-(4-phenylpiperidino)cyclohexanol (AH 5 183 or vesamicol), a drug that acts at synaptic vesicles, rather than in the cytosol, to inhibit the transport, release, and synthesis of ACh.…”

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Effect of 2‐(4‐Phenylpiperidino)cyclohexanol (AH 5183) on the Veratridine‐Induced Increase in Acetylcholine Synthesis by Rat Hippocampal Tissue

Carroll

Ivy

1988

Journal of Neurochemistry

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The intent of this study was to determine whether the drug 2-(4-phenylpiperidino)cyclohexanol (AH 5183 or vesamicol) might inhibit the veratridine-induced increase in acetylcholine (ACh) synthesis by reducing the veratridine-induced activation of a detergent-soluble choline-O-acetyltransferase (EC 2.3.1.6; ChAT) fraction associated with a vesicle-bound store of ACh. When minces of rat hippocampal tissue were loaded with [14C]choline and subsequently depolarized with veratridine, an increase in the synthesis of [14C]ACh occurred that could be abolished by L-AH 5183 (75 nM). When minces were depolarized with veratridine in the presence of L-AH 5183 (75 nM), the depolarization-induced activation of a detergent-soluble ChAT fraction associated with a vesicle-bound store of ACh was blocked. Conversely, the veratridine-induced activation of a water-soluble ChAT fraction believed to be cytosolic was not. AH 5183 also blocked the repletion of the vesicle-bound store with newly synthesized ACh following veratridine-induced depletion of ACh, a result that appeared to be mediated by an effect on the synthesis of ACh at the vesicular surface. These results suggest that veratridine depolarization of rat hippocampal nerve terminals stimulates the synthesis of ACh by activating a detergent-soluble fraction of ChAT closely associated with synaptic vesicle release sites. ACh synthesis and transport at the vesicular surface may be influenced by a common AH 5183-sensitive regulatory protein.

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“…Physostigmine was not present in these experiments. Vesamicol blocks vesicular refilling of cholinergic neurons [Otero et al, 1985;Suskiw and Toth, 19861. By incubating striatal slices in 100 p M vesamicol30 min prior to and during exposure to [3H]choline, cholinergic vesicles should be initially emptied and remain depleted of tritium for the duration of the experiment.…”

Section: T3h1ach Releasementioning

confidence: 99%

N‐(n‐propyl)‐n‐(3‐fluoro‐4‐pyridinyl)‐1h‐3‐methylindol‐1‐amine hydrochloride (HP 184): In vitro spontaneous release of acetylcholine and norepinephrine

Smith¹,

Brougham²,

Huger³

et al. 1993

Drug Development Research

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It has been shown that a single injection of N-(n-propyl)-N-(3-fluoro-4-pyridinyl)-l H-3-methylindol-l-amine hydrochloride) (HP 184) (0.64.8 mglkg, or 2-15 p,moles/kg, sc) reversed passive avoidance deficits in rats with combined cholinergic and noradrenergic lesions. This report describes the effects of HP 164 on NE and ACh release from rat brain slices. In contrast to 4-aminopyridine (CAP), tyramine, or veratridine, HP 184 only enhanced spontaneous release and had no effect on electrical stimulation (ES). Chromatographic analysis showed that the spontaneous release from [3H]choline-loadecl striatal slices correlated with increased release of ACh, not choline efflux. HP 184 also enhanced P\Ch spontaneous release in the absence of both extracellular calcium and functional vesicles (as defined by ES and vesamicol). In frontal cortical slices, HP 184 caused [3H]NE release in a calcium independent fashion different from that induced by veratridine, and was not affected by uptake blockers. In contrast to the cholinergic profile, the [3H]NE release induced by HP 184 required intact storage vesicles, since release was blocked by reserpine pretreatment. The results show tlhat HP 184 can release both NE and ACh in vitro, and, coupled with the dual lesion results, suggest it may have use in diseases involving cholinergic and noradrenergic deterioration. o 1993 Wiiey-Liss, Inc.

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Effects of 4-(2-Hydroxyethyl)-1-piperazine-ethanesulfonic acid (AH5183) on rat cortical synaptosome choline uptake, acetylcholine storage and release

Cited by 37 publications

References 11 publications

Quantitative autoradiography of brain binding sites for the vesicular acetylcholine transport blocker 2-(4-phenylpiperidino)cyclohexanol (AH5183).

Quantitative autoradiography of brain binding sites for the vesicular acetylcholine transport blocker 2-(4-phenylpiperidino)cyclohexanol (AH5183).

Effect of 2‐(4‐Phenylpiperidino)cyclohexanol (AH 5183) on the Veratridine‐Induced Increase in Acetylcholine Synthesis by Rat Hippocampal Tissue

N‐(n‐propyl)‐n‐(3‐fluoro‐4‐pyridinyl)‐1h‐3‐methylindol‐1‐amine hydrochloride (HP 184): In vitro spontaneous release of acetylcholine and norepinephrine

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