Acetylcholine accumulation and release by hybrid NG108-15, glioma and neuroblastoma cells--role of a 16kDa membrane protein in release.
A procedure is described to fill up cells in culture with ACh and study its calcium dependent release, by-passing the synthesis steps. Whether differentiated or not with dbc-AMP, the NG108-15 cells efficiently released ACh when stimulated with calcium and ionophore A23187. The release was also studied in the parent C6-BU-1 and N18TG2 cells. It was found that C6-BU-1 released ACh much better that N18TG2 in spite of their glial origin. The internalization by NG108-15 cells of an antisense oligonucleotide probe hybridizing the 16 kDa proteolipid messenger common to mediatophore and to the V-ATPase reduced ACh release indicated a role of this proteolipid in ACh translocation. This characteristic protein was found in the membrane extract of NG108-15 cells and also in the C6-BU-1 cells, but its amount was strongly reduced in the N18TG2 cell line and in the NG108-15 cells having internalized the antisense probe.
Israël
M
M
Département de Neurochimie, C.N.R.S., Gif-sur-Yvette, France.
Lesbats
B
B
Synguelakis
M
M
Joliot
A
A
D000109:Acetylcholine / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism; D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D005910:Glioma / Q000378:metabolism*; D006822:Hybrid Cells; D008565:Membrane Proteins / Q000502:physiology*; D051379:Mice; D008970:Molecular Weight; D009447:Neuroblastoma / Q000378:metabolism*; D015345:Oligonucleotide Probes; D016376:Oligonucleotides, Antisense; D051381:Rats; D014162:Transfection; D014407:Tumor Cells, Cultured
D008565:Membrane Proteins; D015345:Oligonucleotide Probes; D016376:Oligonucleotides, Antisense; D000001:Calcimycin; D003994:Bucladesine; D000251:Adenosine Triphosphatases; D000109:Acetylcholine; D002118:Calcium