Identification of Acetylcholine in Fresh Rat Brain by Combined Gas Chromatography–Mass Spectrometry

Hammar, Carl-Gustaf; Hanin, Israel; Holmstedt, Bo; Kitz, Richard J.; Jenden, Donald J.; Karlén, B

doi:10.1038/220915a0

Cited by 70 publications

(12 citation statements)

References 10 publications

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“…In all analyses reagent blanks were run with the unknowns. For purposes of rigorous identification, samples of whole stellar nerve were prepared without the addition of internal standards and the gas chromatographic effluent was simultaneously monitored at the most prominent peaks of the derivatives of acetylcholine and cholinc (HAMMAR et al, 1968). Complete scans of the mass spectrum were also obtained using a Hewlett Packard Model 5980 gas chromatography/mass spectrometer system.…”

Section: Methodsmentioning

confidence: 99%

Acetylcholine Content of the Schwann Cell and Axon in the Giant Nerve Fibre of the Squid

Villegas

Jenden

1979

Journal of Neurochemistry

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Abstract— Acetylcholine and choline were identified and their concentrations measured, by means of gas chromatography/mass spectrometry, in extracts obtained from nerve fibers of the hindmost stellar nerve of the squid Sepioteuthis sepioidea. These compounds were quantitated in samples of stellar nerve devoid of giant fiber, intact giant nerve fiber, extruded axoplasm, and axoplasm‐free giant nerve fiber sheaths. In 11 samples of stellar nerve devoid of giant fiber, weighing an average of 20.8 ± 2.3 mg (s.e.m.), 756 ± 91 pmol ACh and 8.65 ± 0.62 nmol of choline were found. The total ACh content of the largest fibre in this group (10 μm in diameter), for a 5 cm length of nerve, is in the order of 0.16 pmol. The average wet weights of a single giant nerve fiber (270‐420 μm in diameter) and its separate components (s.e.m.; in mg; number of fibers in parentheses) were: intact fiber, 4.58 ± 0.19 (25); extruded axoplasm, 3.38 ± 0.13 (20); sheaths, 1.21 ± 0.11 (16). The average ACh content per unit weight of sample was about 2‐3 times higher in the sheaths (5‐13 pmol‐mg−1) than in the axoplasm (2‐4 pmol mg−1), whereas the ACh concentrations estimated per unit volume of cellular water were about 40 times higher in the Schwann cell (107‐222 μm) than in the axon (2‐5μm). These experimental findings establish the presence of ACh in the giant nerve fiber of S. sepioidea. They also indicate the Schwann cells themselves as the main source for the release of ACh, responsible for their long‐lasting hyperpolarizations following the conduction of nerve impulse trains by the axon.

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Section: Methodsmentioning

confidence: 99%

Acetylcholine Content of the Schwann Cell and Axon in the Giant Nerve Fibre of the Squid

Villegas

Jenden

1979

Journal of Neurochemistry

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“…Analytical methods for measuring ACh in biological samples, each with differing degrees of sensitivity and specificity (see review by Maruyama et al, 1987), have included the bioassay (Murnagam, 1958), radioenzymatic (Goldberg and McCaman, 1973) and chemiluminescence (Israel and Lesbats, 1982) assays, HPLC with electrochemical detection (HPLC-ED) (Potter et al, 1983), gas chromatography (Stavinoha and Ryan, 1965), and gas chromatography-mass spectrometry (GC-MS) (Hammar et al, 1968). Of these, the radioenzymatic (Consolo et al,1 9 8 7~) and HPLC-ED (Ajima and Kato, 1987;Damsma et al, 19870,b;Tyrefors and Gillberg, 1987) assays recently have been EIVDOGEIVOUS ACETYLCHOLINE RELEASE IA ' VIVO 201 7 modified and applied in quantitating low (less than picomole) amounts of ACh in dialysates of the rat striatum and spinal cord.…”

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confidence: 99%

Drug Effects on the Release of Endogenous Acetylcholine In Vivo: Measurement by Intracerebral Dialysis and Gas Chromatography–Mass Spectrometry

Marien

Richard

1990

Journal of Neurochemistry

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Intracerebral microdialysis was combined with a sensitive and specific gas chromatographic-mass spectrometric assay to measure the release of endogenous acetylcholine in the rat striatum in vivo. In rats anesthetized with urethane (1.2 g/kg i.p.), the levels of striatal acetylcholine dialyzed into a Ringer's perfusate were: (a) reliably measurable only in the presence of physostigmine; (b) stable at between 3 and 8 h of perfusion (30-75 pmol/20 min in the presence of 75 microM physostigmine); (c) reduced by calcium-free Ringer's solution, tetrodotoxin (0.1 microM), and vesamicol (1.0 microM); and (d) increased by elevated potassium (100 mM), atropine (3-300 microM), and haloperidol (0.75 mg/kg i.p.). In conscious unrestrained rats, the spontaneous release of striatal acetylcholine was not altered significantly following the administration of urethane. The changes in acetylcholine release observed in this study are consistent with the known actions of some drugs or ionic conditions on striatal cholinergic neurotransmission and are evident under the condition of urethane anesthesia. The present results demonstrate the sensitivity and suitability of this method for monitoring endogenous striatal acetylcholine release in vivo.

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“…In mass fragmentography, the mass spectrometer is used as a gas chromatographic detector which continously monitors l-3 selected mass numbers of compounds eluted from the gas chromatographic column The ions m/e 137 and 138 are brought alternately into focus [6,8] and the intensities recorded as a function of time ( fig. 1A).…”

Section: Resultsmentioning

confidence: 99%

Detection of biochemical intermediates by mass fragmentography: Mescaline and tetrahydroisoquinoline precursors

Lindgren

Agurell²,

Lundström³

et al. 1971

FEBS Letters

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Identification of Acetylcholine in Fresh Rat Brain by Combined Gas Chromatography–Mass Spectrometry

Cited by 70 publications

References 10 publications

Acetylcholine Content of the Schwann Cell and Axon in the Giant Nerve Fibre of the Squid

Acetylcholine Content of the Schwann Cell and Axon in the Giant Nerve Fibre of the Squid

Drug Effects on the Release of Endogenous Acetylcholine In Vivo: Measurement by Intracerebral Dialysis and Gas Chromatography–Mass Spectrometry

Detection of biochemical intermediates by mass fragmentography: Mescaline and tetrahydroisoquinoline precursors

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