Soluble and Membrane-bound Forms of Choline-o-Acetyltransferase Activity in the Purely Cholinergic Neurons of Torpedo

Eder-Colli, Lorenza; Amato, Stefano; Froment, Yvette

doi:10.1007/978-1-349-08639-9_10

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“…Effect of antibody 8/38A on AChE and on ACh release induced in Torpedo synaptosomes We were interested in knowing whether our monoclonal antibody, besides inhibiting ChAT activity (Eder-Colli et al, 1987 was able also to affect other cholinergic functions. We first tested if the an-tibody was able to recognize the AChE that is present in purified SPMs.…”

Section: Immunoblotting Analysismentioning

confidence: 99%

A Monoclonal Antibody Raised Against Plasma Membrane of Cholinergic Nerve Terminals of the Torpedo Inhibits Choline Acetyltransferase Activity and Acetylcholine Release

Eder-Colli¹,

Briand²,

Pellegrinelli³

et al. 1989

Journal of Neurochemistry

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Monoclonal antibodies were raised against the synaptosomal plasma membranes (SPMs) purified from the electric organ of the Torpedo. One antibody that reacts preferentially with SPMs rather than with other membrane fractions isolated from this tissue was previously found to inhibit hydrophilic and amphiphilic choline-O-acetyltransferase (ChAT) activity. On immunoblots of SPMs, this antibody recognizes two polypeptides of 135 and 66 kilodaltons that are related; the 66-kilodalton polypeptide appears to exist as a monomer and as a dimer in SPMs. The antibody was also able to inhibit the calcium-dependent release of acetylcholine in Torpedo synaptosomes without affecting the total neurotransmitter content. This inhibition was dependent on the antibody concentration and was observed when the release was elicited by either KCl depolarization or the calcium ionophore A23187; this suggests that inhibition was not mediated by a blockage of the depolarization-activated calcium influx. The inhibition could not be prevented by atropine, a result indicating that the antibody does not block release by mimicking the action of acetylcholine on presynaptic muscarinic autoreceptors. Thus, the antigen recognized by this antibody appeared to be involved in acetylcholine release; this antigen could be membrane-bound ChAT, another protein of the SPMs, or both.

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Section: Immunoblotting Analysismentioning

confidence: 99%