Choline acetyltransferase activity and mass fragmentographic measurement of acetylcholine in specific nuclei and tracts of rat brain

Cheney, D. L.; LeFevre, Henry F.; Racagni, Giorgio

doi:10.1016/0028-3908(75)90107-0

Cited by 278 publications

(25 citation statements)

References 24 publications

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“…Finally, "-'I-rhNGF bound to virtually all sites labeled by 125I-muNGF (medial septum, diagonal band, substantia innominata; refs. [16][17][18] (36,37); the ACh synthetic enzyme, choline acetyltransferase (ChAT) (38,39); the ACh degradative enzyme, acetylcholinesterase (38); and ACh itself (50). NGF NGFR antibodies (7, 11-14, 34, 40) and '25I-muNGF (16)(17)(18) and localized to cholinergic cell bodies and dendrites.…”

Section: Discussionmentioning

confidence: 99%

Recombinant human nerve growth factor is biologically active and labels novel high-affinity binding sites in rat brain.

Altar¹,

Burton²,

Bennett³

et al. 1991

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

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MATERIALS AND METHODSProduction, Purification, and lodination of rhNGF. Purified (>98%) rhNGF was produced by the Research Collaborations group at Genentech, using transfected Chinese hamster ovary cell cultures and anion-exchange chromatography (25) as described (26,27). rhNGF stimulated PC12 (rat pheochromocytoma cell line) neurite outgrowth (51) with an EC50 value of 39 and 52 pg/ml in two experiments conducted by different personnel in separate laboratories (Fig. 1). rhNGF was iodinated (28) with modifications (26, 27) and used within 1-6 days in all but one study, since decreases in the proportion of total 1251-rhNGF binding that was displaced by 100 nM rhNGF were noted after this time. Two-and 3-week-old uCi/,ug; 3580 Ci/mmol of rhNGF dimer; 1 Ci = 37 GBq) stimulated neurite outgrowth in the PC12 bioassay with an EC50 of 49 ± 19 pg/ml (2-week-old tracer; n = 4) and 49 ± 0.8 pg/ml (3-week-old tracer; n = 3). The 1-day-old batch of tracer produced a half-maximal stimulation of PC12 neurite outgrowth at 35 pg/ml. Without a biochemical separation of labeled and unlabeled species, we cannot exclude the possibility that biological activity is due to unlabeled rhNGF. However, the high degree of rhNGF labeling (1.65:1 ratio of 1251 to rhNGF dimer), the retrograde transport of 125I-rhNGF, which can be blocked by rhNGF (26), and the indistinguishable potencies of rhNGF and 125I-rhNGF in the PC12 assay (Fig. 1) indicate that 1251_ rhNGF is active in bioassays of NGF function.Brain Sections. Brains from male Sprague-Dawley rats were frozen in isopentane at -15°C either separately, for anatomical mapping, or as a group of four to five brain hemispheres (22), for association, dissociation, and equilibrium saturation analyses. Sections (12 ,um) were thawmounted onto gelatin-coated glass microscope slides and stored frozen for up to 1 month at -70°C.

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Section: Discussionmentioning

confidence: 99%

Recombinant human nerve growth factor is biologically active and labels novel high-affinity binding sites in rat brain.

Altar¹,

Burton²,

Bennett³

et al. 1991

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

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“…We recently determined (34) that concentrations of [3H]AH5183 binding in 14-18 brain areas correlated highly (r = 0.88 -0.97) and significantly (P < 0.01) with regional levels of ChoAcTase (35,36), AcCho esterase (16,37), and AcCho itself (35,36). The [3H]AH5183 binding site in the forebrain was also strikingly similar to the autoradiographic distribution (38,39) and pH optimum and binding affinity (38,40) …”

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confidence: 95%

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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“…On the basis of the blocking action of atropine the post-synaptic cholinoceptors may be classified as of the muscarinic type. Cholinergic involvement in the functioning of the raphe nuclei has been indicated previously by a number of histological and histochemical studies (Lewis & Shute, 1967;Palkovits & Jacobwitz, 1974;Cheney, LeFevre & Racagni, 1975). Recently it has also been shown that 13H1-quinillidinyl-benzilate (QNB), a potent cholinoceptor antagonist, bind specificially to muscarinic postsynaptic receptors (Yamamura, Kuhar & Snyder, 1974).…”

Section: Discussionmentioning

confidence: 88%

Electrocortical Changes Induced by the Perfusion of Noradrenaline, Acetylcholine and Their Antagonists Directly Into the Dorsal Raphé Nucleus of the Cat

Key

Krzywoskinski

1977

British J Pharmacology

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I The electrocortical changes induced by the perfusion of drugs directly into the dorsal raphe nucleus of the cat encephale isole' preparation have been studied. 2 (-)-Noradrenaline (NA), (-)-adrenaline, or (-)isoprenaline (Isop) produced intermittent or sustained electrocortical desynchronization during the perfusion period. 3 These changes were markedly attenuated or completely abolished by the prior perfusion of (+)-sotalol or (-)-propranolol, but not by equimolecular concentrations of (+)-propranolol. 4 The effects of NA or Isop were also blocked by phentolamine, whereas phenoxybenzamine either potentiated the responses to NA and Isop or mimicked the effects of these catecholamines. 5The effect of dopamine was similar to that induced by NA, but could not be elicited at all of the perfusion sites where NA was effective. It could be blocked by (±)-sotalol or (-)-propranolol and also by the prior perfusion of fusaric acid. 6 Acetylcholine (ACh) increased, or initiated, electrocortical synchronization. These effects could be antagonized by sensory stimulation, the prior perfusion of atropine, or the simultaneous perfusion of NA at the same site. 7 Lignocaine, induced prolonged electrocortical desynchronization, behavioural alerting and an increased responsiveness to sensory stimulation. 8 The results have been discussed in relation to the possible involvement of inhibitory,-adrenoceptors and facilitatory cholinoceptors (muscarinic) in the functioning of the dorsal raphe nucleus.

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Choline acetyltransferase activity and mass fragmentographic measurement of acetylcholine in specific nuclei and tracts of rat brain

Cited by 278 publications

References 24 publications

Recombinant human nerve growth factor is biologically active and labels novel high-affinity binding sites in rat brain.

Recombinant human nerve growth factor is biologically active and labels novel high-affinity binding sites in rat brain.

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

Electrocortical Changes Induced by the Perfusion of Noradrenaline, Acetylcholine and Their Antagonists Directly Into the Dorsal Raphé Nucleus of the Cat

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