Identification of basic fibroblast growth factor as a cholinergic growth factor from human muscle.
Dissociated embryonic chick ciliary ganglion cells in culture were used as a bioassay to isolate a cholinergic growth-promoting protein from extracts of autopsied adult human muscle. An active protein was purified after acid and salt precipitation of extract, cation exchange, molecular sieving, heparin affinity chromatography, and in some cases, SDS-PAGE. This protein increased levels of choline acetyltransferase activity and ACh synthesis with time in culture. The protein was identified as basic FGF by several criteria. It shared the high affinity for heparin and was the same approximate molecular weight, 18 kD, as basic FGF. Activity was removed from solution by antibodies specific for basic FGF. Recombinant human basic FGF was equally effective in stimulating CAT activity, but was not additive with our purified protein at saturating concentrations. Basic FGF was also found in extracellular matrix and conditioned medium from cultured embryonic chick muscle. The activity could be released from extracellular matrix by treatment with heparinase or high salt extraction. Basic FGF stimulates neurite outgrowth as well as the capacity for transmitter synthesis. Thus, basic FGF is present in embryonic and adult muscle and capable of acting as a growth regulator for cholinergic neurons.
Vaca
K
K
Department of Neurology, Baylor College of Medicine, Houston, TX 77030.
Stewart
S S
SS
Appel
S H
SH
D000109:Acetylcholine / Q000096:biosynthesis*; D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D002795:Choline O-Acetyltransferase / Q000378:metabolism*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005346:Fibroblast Growth Factors / Q000302:isolation & purification* / Q000494:pharmacology; D005726:Ganglia, Parasympathetic / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006133:Growth Substances / Q000302:isolation & purification* / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D009132:Muscles / Q000502:physiology*; D011994:Recombinant Proteins / Q000494:pharmacology
D006133:Growth Substances; D011994:Recombinant Proteins; D005346:Fibroblast Growth Factors; D002795:Choline O-Acetyltransferase; D000109:Acetylcholine