BACKGROUND
Vesicular Acetylcholine Transporter (VAChT) is an ~70 kD protein that belongs to a family of transporter proteins that includes two vesicular monoamine transporters (VMAT).(1) Members of this family are thought to concentrate neuro-tramsmitters in synaptic vesicles through exchange of protons for neuro-transmitter. The neurotransmitter is then stored in vesicles until release. VAChT is expressed selectively in all known cholinergic neurons, and double labelling techniques indicate that VAChT and ChAT colocalize.(2) Antibodies to VAChT are thus useful in visualizing acetylcholine-containing neurons. After release of ACh from the synaptic vesicles, transmission is stopped when ACh is broken down in the synaptic clefts by acetylcholinesterase. The choline formed from this breakdown is recycled by the high-affinity choline transporter (HAChT).
SPECIFICITY AND PREPARATION
The antibodies recognize rat and mouse VAChT. They do not cross-react with guinea-pig, rabbit or cat VAChT. The antibodies do not recognize vesicular monoamine transporter. A synthetic peptide corresponding to the carboxy-terminus of rat VAChT was used as immunogen. The antiserum is routinely tested by immunohistochemistry of frozen rat brain tissue sections.
USAGE
The VAChT antiserum is useful as a marker for cholinergic neurons in both the central and peripheral nervous systems where it stains fibers and cell bodies. Applications include immunohistochemistry of frozen tissue sections. We find that a dilution of 1:10,000 is optimal. However, the optimal dilution may vary between 1:1000-10,000 depending on the experimental system. Rat brain is suggested as a positive tissue control. The antiserum has also been shown to recognize VAChT in tissue-cultured cells, including PC12 rat adrenal pheochromocytoma cells (ATCC CRL 1721) and primary cultures of spinal motorneurons.
REFERENCES
1. Bejanin, S., R. Cervini, J. Mallet, and S. Berrard. 1994. A unique gene organization for two cholinergic markers, choline acetyltransferase and a putative vesicular transporter of acetylcholine. J. Biol. Chem. 269:21944-21947.
2. Gilmor, M.L., N.R. Nash, A. Roghani, R.H. Edwards, H. Yi, S.M. Hersch, and A.I. Levey. 1996. Expression of the putative vesicular acetylcholine transporter in rat brain and localization in cholinergic synaptic vesicles. J. Neurosci. 16:2179-2190.
| Cat. No. | Description | Isotype | Size |
| 60596E | Polyclonal goat anti-VAChT | Goat Igs | 0.1ml |
PharMingen supplies an extensive line of antibodies and reagents for Neuroscience research, including:
PharMingen supplies an extensive line of antibodies and reagents for Neuroscience research, including:
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