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English words and Examples of Usage
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Sentences for "choline "
Before the availability of antibodies against nicotinic acetylcholine receptors of dipteran flies, cholinergic pathways have been analyzed by more indirect methods, mainly by immunocytochemistry against choline acetyltransferase (ChAT), an enzyme of the acetylcholine metabolism [ 38, 39] or autoradiography of 125I-?
-bungarotoxin binding [ 40, 41].
Some of the choline used for ACh synthesis is stored in membrane phosphatidylcholine (PC).
Previous studies have shown that this choline is liberated from PC in a one-step reaction, thus indicating that it is produced by a phospholipase D (PLD, EC 3.
4) -an enzyme that hydrolyzes the phospholipid into choline and phosphatidic acid (PA) [ 4 ] .
Previous studies have shown that choline derived from PC is indeed used for ACh synthesis [ 4 14 15 16 ] and that this choline is generated by a PLD [ 4 14 ] .
This pathway may become particularly important when extracellular circulating choline concentrations are low (e.
during dietary choline deficiency) or when ACh synthesis and release are accelerated by high neuronal activity, thus generating a high demand for choline [ 16 ] .
In the present study, we used a cholinergic cell line, SN56 [ 17 ] , which is derived from the fusion of septal neurons of postnatal day 21 mice with N18TG2 murine neuroblastoma cells, as a model for cholinergic neurons, in order to determine the possible role of PLD 1 and PLD2 in generating choline for ACh synthesis.
We found that the cells express exclusively PLD2 and that downregulating PLD2 expression reduces ACh synthesis, while overexpressing and/or stimulating PLD2 activity increases the synthesis of the neurotransmitter, indicating that PLD2-catalyzed hydrolysis of PC generates choline for ACh synthesis in cholinergic cells.
These data show that SN56 cells express exclusively PLD2 and thus experimental manipulations of the activity of this PLD isoform were subsequently employed to determine if this enzyme is capable of producing choline for ACh synthesis in cholinergic cells.
Under these conditions the choline precursor used for the synthesis of ACh is derived entirely from the intracellular pool, which includes the choline in PC [ 4 15 ] .
Free choline accumulation was also significantly increased by PMA.
The magnitude of this effect tended to be small (approximately 15%), presumably because free choline is a rapidly turning over metabolic intermediate that does not accumulate in cells (Fig.
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