Retroviral-mediated gene transfer of the porcine choline acetyltransferase: a model to study the synthesis and secretion of acetylcholine in mammalian cells.
We have constructed a recombinant retrovirus that expresses choline acetyltransferase (ChAT) by placing the porcine enzyme cDNA under the control of the 5' long terminal repeat of the retroviral vector pMMuLV. Using retrovirus-mediated gene transfer, we have expressed ChAT in astroglial (STR-SVLT) and neuroendocrine (RIN) cell lines. Both genetically modified cell types synthesize acetylcholine (ACh). ACh is also present in the culture medium at a low concentration relative to that found in the modified cells. This result suggests that the synthesized ACh is retained within the cells and released by these two cell types. Release of ACh is not increased in the presence of the calcium ionophore A23187 or by depolarizing concentrations of potassium in either STR-SVLT or in RIN cells. The implications of these studies for understanding ACh release mechanisms are discussed.
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Laboratoire de Genetique Moléculaire de la Neurotransmission et des Processus Neurodegeneratifs, Centre National de la Recherche Scientifique, Gif-sur-Yvette, France.
Horellou
P
P
Leroy
C
C
Mallet
J
J
D000109:Acetylcholine / Q000096:biosynthesis* / Q000378:metabolism*; D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D002795:Choline O-Acetyltransferase / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors; D008954:Models, Biological; D012190:Retroviridae / Q000235:genetics*; D013552:Swine; D014162:Transfection
D000001:Calcimycin; D002795:Choline O-Acetyltransferase; D000109:Acetylcholine; D002118:Calcium