Abstract
In cholinergic neurons choline is directed to three main pathways; (1) conversion to phosphorylcholine (PCh) and cytidine diphosphate choline (CDP-choline) for the synthesis of phosphatidylcholine, (2) acylation to the neurotransmitter acetylcholine and (3) oxidation to betaine for the formation of methionine. Thus, the distribution of choline among the different metabolites is important for a better understanding of the regulation of these pathways in neurons. A non-HPLC method for the simultaneous separation of five choline metabolites found in neurons is described. High voltage electrophoresis (HVE) was combined with thin layer chromatography (TLC) to separate choline, PCh, CDP-choline, acetylcholine and betaine. This method is useful in studying the distribution of choline among its different metabolites in radiotracer experiments. Aqueous metabolites from leukemia inhibitory factor treated LA-N-2 cells labeled with [methyl-3H]choline were separated by HVE followed by TLC in the same dimension. Although the separation appeared to be complete, some 'tailing' by PCh significantly elevated the radioactivity measured in CDP-choline. This tailing of PCh was confirmed by subjecting radiolabeled PCh alone to this multiple separation method. Contamination of CDP-choline by PCh was eliminated by subjecting the samples to HVE followed by TLC in the second dimension. This two-dimensional approach was consistently reproducible and achieved excellent resolution of all five metabolites. In addition, this technique also resolved a sixth choline-containing metabolite, glycerophosphorylcholine (GPC), a breakdown product of phosphatidylcholine. Copyright (C) 1999 Elsevier Science B.V.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 13-21 |
| Number of pages | 9 |
| Journal | Journal of Neuroscience Methods |
| Volume | 90 |
| Issue number | 1 |
| DOIs | |
| State | Published - Aug 1 1999 |
| Externally published | Yes |
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Keywords
- Acetylcholine
- Betaine
- CDP-choline
- Glycerophosphorylcholine
- Phosphorylcholine
ASJC Scopus subject areas
- Neuroscience(all)
Cite this
Non-HPLC separation of water-soluble choline metabolites by two-dimensional high voltage electrophoresis and thin layer chromatography. / Utal, A. K.; Coleman, Paul.
In: Journal of Neuroscience Methods, Vol. 90, No. 1, 01.08.1999, p. 13-21.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Non-HPLC separation of water-soluble choline metabolites by two-dimensional high voltage electrophoresis and thin layer chromatography
AU - Utal, A. K.
AU - Coleman, Paul
PY - 1999/8/1
Y1 - 1999/8/1
N2 - In cholinergic neurons choline is directed to three main pathways; (1) conversion to phosphorylcholine (PCh) and cytidine diphosphate choline (CDP-choline) for the synthesis of phosphatidylcholine, (2) acylation to the neurotransmitter acetylcholine and (3) oxidation to betaine for the formation of methionine. Thus, the distribution of choline among the different metabolites is important for a better understanding of the regulation of these pathways in neurons. A non-HPLC method for the simultaneous separation of five choline metabolites found in neurons is described. High voltage electrophoresis (HVE) was combined with thin layer chromatography (TLC) to separate choline, PCh, CDP-choline, acetylcholine and betaine. This method is useful in studying the distribution of choline among its different metabolites in radiotracer experiments. Aqueous metabolites from leukemia inhibitory factor treated LA-N-2 cells labeled with [methyl-3H]choline were separated by HVE followed by TLC in the same dimension. Although the separation appeared to be complete, some 'tailing' by PCh significantly elevated the radioactivity measured in CDP-choline. This tailing of PCh was confirmed by subjecting radiolabeled PCh alone to this multiple separation method. Contamination of CDP-choline by PCh was eliminated by subjecting the samples to HVE followed by TLC in the second dimension. This two-dimensional approach was consistently reproducible and achieved excellent resolution of all five metabolites. In addition, this technique also resolved a sixth choline-containing metabolite, glycerophosphorylcholine (GPC), a breakdown product of phosphatidylcholine. Copyright (C) 1999 Elsevier Science B.V.
AB - In cholinergic neurons choline is directed to three main pathways; (1) conversion to phosphorylcholine (PCh) and cytidine diphosphate choline (CDP-choline) for the synthesis of phosphatidylcholine, (2) acylation to the neurotransmitter acetylcholine and (3) oxidation to betaine for the formation of methionine. Thus, the distribution of choline among the different metabolites is important for a better understanding of the regulation of these pathways in neurons. A non-HPLC method for the simultaneous separation of five choline metabolites found in neurons is described. High voltage electrophoresis (HVE) was combined with thin layer chromatography (TLC) to separate choline, PCh, CDP-choline, acetylcholine and betaine. This method is useful in studying the distribution of choline among its different metabolites in radiotracer experiments. Aqueous metabolites from leukemia inhibitory factor treated LA-N-2 cells labeled with [methyl-3H]choline were separated by HVE followed by TLC in the same dimension. Although the separation appeared to be complete, some 'tailing' by PCh significantly elevated the radioactivity measured in CDP-choline. This tailing of PCh was confirmed by subjecting radiolabeled PCh alone to this multiple separation method. Contamination of CDP-choline by PCh was eliminated by subjecting the samples to HVE followed by TLC in the second dimension. This two-dimensional approach was consistently reproducible and achieved excellent resolution of all five metabolites. In addition, this technique also resolved a sixth choline-containing metabolite, glycerophosphorylcholine (GPC), a breakdown product of phosphatidylcholine. Copyright (C) 1999 Elsevier Science B.V.
KW - Acetylcholine
KW - Betaine
KW - CDP-choline
KW - Glycerophosphorylcholine
KW - Phosphorylcholine
UR - http://www.scopus.com/inward/record.url?scp=0032788951&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032788951&partnerID=8YFLogxK
U2 - 10.1016/S0165-0270(99)00059-X
DO - 10.1016/S0165-0270(99)00059-X
M3 - Article
C2 - 10517269
AN - SCOPUS:0032788951
VL - 90
SP - 13
EP - 21
JO - Journal of Neuroscience Methods
JF - Journal of Neuroscience Methods
SN - 0165-0270
IS - 1
ER -