Abstract
The expression of a cholinergic system during embryonic development is a widespread phenomenon. However, no precise function could be assigned to it during early pre-neural stages and there are only few studies that document when it precisely starts to be expressed. Here, we examined the expression of cholinergic components in a murine embryonic stem cell line by RT-PCR, histochemistry, and enzyme activity measurements; the acetylcholine (ACh) content was measured by HPLC. We have demonstrated that embryonic stem cells express ACh, acetylcholine receptors, choline acetyltransferase (ChAT), acetyl- and butyryl-cholinesterase (AChE and BChE). Butyryl-cholinesterase (BChE) expression was higher than AChE. The cholinesterase activity was down-regulated by adding specific inhibitors to culture medium. Inhibition of BChE led to a reduction of proliferation. This is the first demonstration that mouse embryonic stem cells express the full molecular equipment of a cholinergic system. Locally produced ACh might function as an intercellular signal, modulating the proliferation of stem cells.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Acetylcholine / metabolism
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Acetylcholinesterase / genetics
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Acetylcholinesterase / metabolism
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Animals
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Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide / pharmacology
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Butyrylcholinesterase / genetics
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Butyrylcholinesterase / metabolism
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Cell Line
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Cell Proliferation / drug effects
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Choline O-Acetyltransferase / genetics*
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Choline O-Acetyltransferase / metabolism
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Cholinesterase Inhibitors
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Cholinesterases / genetics*
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Cholinesterases / metabolism
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Embryonic Stem Cells / cytology
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Embryonic Stem Cells / drug effects
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Embryonic Stem Cells / metabolism*
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Gene Expression Profiling*
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Mice
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Receptors, Cholinergic / genetics*
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Receptors, Cholinergic / metabolism
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Reverse Transcriptase Polymerase Chain Reaction
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Tetraisopropylpyrophosphamide / pharmacology
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Time Factors
Substances
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Cholinesterase Inhibitors
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Receptors, Cholinergic
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Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide
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Tetraisopropylpyrophosphamide
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Choline O-Acetyltransferase
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Acetylcholinesterase
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Butyrylcholinesterase
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Cholinesterases
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Acetylcholine