&lt;b&gt;SPECIFICITY OF IONIC FIXATION WITH SILICOTUNGSTIC ACID FOR CYTOCHEMICAL LOCALIZATION AND IDENTIFICATION OF ACETYLCHOLINE IN SYNAPTIC &lt;/b&gt;&lt;b&gt;VESICLES &lt;/b&gt;

Ohoka, Tadakazu; Tsuji, Shigeru

doi:10.2220/biomedres.9.335

Cited by 9 publications

(4 citation statements)

References 18 publications

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“…By using this method, a plasticity of the nerve afferents to nigrostriatal dopaminergic neurons was evidenced in Parkinson's disease, although the cholinergic nature of all the stained terminals could not be assessed (5). Moreover, ionic fixation seemed more specific for ACh in peripheral nervous system (84,105) than in the brain where the concentrations of neurotransmitters are the highest. Since, until now, ionic fixation by STA is the one and only method allowing in situ insolubilization of ACh, further works must be performed in order to unequivocally characterize the compounds of the precipitates observed under electron microscope.…”

Section: Perspectivesmentioning

confidence: 96%

Historical landmarks in the histochemistry of the cholinergic synapse: perspectives for future researches

Anglade¹,

Larabi-Godinot

2010

Biomed. Res.

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Nearly one hundred years ago, acetylcholine (ACh) was proposed as a chemical agent responsible for nerve transmission at the synapse, the junction area between one neuron and its target cell. Since it has been proved that ACh played, indeed, a major role in the functioning of the nerve system in the vertebrates, cholinergic nerve transmission became a basic field of study in neuroscience. The birth of histochemistry and its ulterior developments allowed in situ localization of the molecular agents related to the functioning of the cholinergic synapse. This report presents historical landmarks in the histochemistry of major cholinergic agents (acetylcholinesterase, nicotinic acetylcholine receptor, choline acetyltransferase, and ACh), a domain which has greatly contributed to the knowledge of the nerve system. It is emphasized that despite extraordinary progresses made in this field, basic problems, such as in situ localization of ACh, still remain to be solved.

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Section: Perspectivesmentioning

confidence: 96%

Historical landmarks in the histochemistry of the cholinergic synapse: perspectives for future researches

Anglade¹,

Larabi-Godinot

2010

Biomed. Res.

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“…Moreover, ionic fixation seemed more specific for acetylcholine in the peripheral nerve system [37,41] than in the brain where the concentrations of neurotransmitters are the highest. Since, until now, ionic fixation by STA is the unique method allowing in situ precipitation of acetylcholine, further works must be performed in order to unequivocally characterize the compounds of the precipitates observed under an electron microscope.…”

Section: Ionic Fixation Of Acetylcholine-like Cations: Sun and Rain On Original Researchmentioning

confidence: 98%

“…In vitro, experimental models and ultrastructural observation of cholinergic and non-cholinergic nerve terminals in the frog heart enabled to partly answer to the criticisms. The results suggested a relative specificity of ionic fixation by STA for acetylcholine compared to the other neurotransmitters[41].…”

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confidence: 89%

Shigeru Tsuji (1936-2008): A Master in the Histochemistry of the Cholinergic Synapse – Insight for Future Researches

2019

Journal of Neurophysiology and Neurological Disorders

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The present paper is a contribution to the memory of a scientist and, thereby, a report about an outstanding research achievement that may help future investigators in the field of cholinergic nerve transmission. Indeed, Shigeru Tsuji realized a basic analysis of the histochemical reactions used to detect acetylcholinesterase activity. Moreover, he remains, until now, a pioneer in the in situ localization of the quaternary ammonium compounds, such as choline and acetylcholine.Always open-minded, Tsuji remarkably understood, at the end of his life, the relevance of initiating research in the newly emerging field of quantal neurobiology. Here, his significant findings will be put together to emphasize that they provide both enlightening data and very useful tools for further works in the domain of the cholinergic synapse.

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“…The chemical reaction between STA and these ACh-like cations gave point-like precipitates in the synaptic vesicles. Preferential precipitation of ACh and choline by STA and its homologous compounds compared to biogenic amines was proved in vitro (13).…”

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confidence: 98%

Further Studies on Electron Microscopic Localization of Acetylcholine-Like Cations in Frog Neuromuscular Junctions.

Tsuji

Motelica-Heino

1993

Acta Histochem. Cytochem

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In the present work we have localized at the electron microscope level acetylcholine (ACh) in frog neuromuscular junctions, simultaneously in storage (synaptic vesicles) and releasing space (synaptic cleft) by a cytochemical method, previously described (19), which involved the precipitation of ACh-like cations with silicotungstic acid. The fixation took place in different physiological states (resting, during electrical stimulation and recovery). At rest, ACh was localized in the synaptic vesicles as point-like precipitates and in the synaptic cleft beneath some but not all the "active zones" as laminar precipitates. We think that the precipitates seen in the synaptic vesicles correspond to stored ACh, while those in the synaptic cleft to ACh released through the "active zones". After a prolonged tetanic electrical stimulation, laminar precipitates covered the whole axoplasm of the nerve terminals, and it appeared that they extend beyond the nerve terminal membrane in contact with Schwann cells. The laminar precipitates found in the nerve terminals might correspond to a newly synthetized axoplasmic ACh (or leaked into the axoplasm from the synaptic vesicles) while the precipitates which filled whole synaptic cleft are presumed to be ACh released by electrical stimulation of the motor nerve. During the recovery period, the distributions of both types of precipitates became similar to those observed at rest. This cytochemical fixation method of ACh-like cations appears to be a useful "tool" for the quick capture of the neurotransmitter ACh in the synaptic vesicles, axoplasm and synaptic cleft, and therefore helpful for identification of cholinergic nerve terminals.

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<b>SPECIFICITY OF IONIC FIXATION WITH SILICOTUNGSTIC ACID FOR CYTOCHEMICAL LOCALIZATION AND IDENTIFICATION OF ACETYLCHOLINE IN SYNAPTIC </b><b>VESICLES </b>

Cited by 9 publications

References 18 publications

Historical landmarks in the histochemistry of the cholinergic synapse: perspectives for future researches

Historical landmarks in the histochemistry of the cholinergic synapse: perspectives for future researches

Shigeru Tsuji (1936-2008): A Master in the Histochemistry of the Cholinergic Synapse – Insight for Future Researches

Further Studies on Electron Microscopic Localization of Acetylcholine-Like Cations in Frog Neuromuscular Junctions.

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