Choline acetyltransferase activity of spinal cord cell cultures increased by co-culture with muscle and by muscle-conditioned medium.
Activity of the enzyme choline acetyltransferase (CAT), which mediates the synthesis of the neurotransmitter, acetylcholine, was increased up to 20- fold in spinal cord (SC) cells grown in culture with muscle cells for 2 wk. This increase was directly related to the duration of co-culture as well as to the cell density of both the SC and muscle involved and was not affected by the presence of the acetylcholine receptor blocking agent, alpha-bungarotoxin. Glutamic acid decarboxylase (GAD) activity was often markedly decreased in SC-muscle cultures while the activities of acetylcholinesterase and several other enzymes were little changed. Increased CAT activity was also observed when SC cultures were maintained in medium which had been conditioned by muscle cells or by undifferentiated cells from embryonic muscle. Muscle-conditioned medium (CM) did not affect the activities of SC cell GAD or acetylcholinesterase. Dilution or concentration of the CM directly affected its ability to increase SC CAT activity , as did the duration and timing of exposure of the SC cells to the CM. The medium could be conditioned by muscle cells in the presence or absence of serum, and remained effective after dialysis or heating to 58 degrees C. Membrane filtration data were consistent with the conclusion that the active material(s) in CM had a molecular weight in excess of 50,000 daltons. We conclude that large molecular weight material that is released by muscle cells is capable of producing a specific increase in CAT activity of SC cells.
D000123:Acetyltransferases / Q000378:metabolism*; D001769:Blood; D002038:Bungarotoxins / Q000494:pharmacology; D002478:Cells, Cultured; D002795:Choline O-Acetyltransferase / Q000378:metabolism*; D003470:Culture Media; D003956:Dialysis; D005968:Glutamate Decarboxylase / Q000378:metabolism; D006358:Hot Temperature; D008970:Molecular Weight; D009132:Muscles / Q000166:cytology*; D009474:Neurons / Q000201:enzymology; D013116:Spinal Cord / Q000166:cytology / Q000201:enzymology*; D013997:Time Factors
D002038:Bungarotoxins; D003470:Culture Media; D000123:Acetyltransferases; D002795:Choline O-Acetyltransferase; D005968:Glutamate Decarboxylase