Identification of a potent neurotrophic substance for ciliary ganglionic neurons in fetal calf serum as insulin-like growth factor II
| Autor: | T, Motoike, K, Unsicker |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Neurons
Platelet-Derived Growth Factor Dose-Response Relationship Drug Epidermal Growth Factor Cell Survival Heparin Blotting Western Antibodies Monoclonal Ganglia Parasympathetic Nerve Tissue Proteins Chick Embryo Hydrogen-Ion Concentration Chromatography Affinity Molecular Weight Insulin-Like Growth Factor II Chromatography Gel Animals Cattle Ciliary Neurotrophic Factor Nerve Growth Factors Insulin-Like Growth Factor I |
| Zdroj: | Journal of neuroscience research. 56(4) |
| ISSN: | 0360-4012 |
| Popis: | When fetal calf serum (FCS) alone is used as a trophic support for cultured chicken parasympathetic ciliary ganglionic (cCG) neurons, it does not show any survival-promoting effects on these neurons. When FCS is applied to heparin-affinity chromatography, however, potent survival-promoting activity is obtained in the fraction eluted with 0.5 M NaCl. Using cCG neurons as a bioassay system, this neurotrophic activity was purified by a combination of heparin-affinity chromatography, gel filtration chromatography, and Sep-Pak C18 cartridge. The 40-50-kDa fractions from the gel filtration column with strong survival-promoting activity were shown to contain insulin-like growth factor II (IGF-II) by immunoblot analysis. By acidification, the survival-promoting activity and IGF-II were translocated together from the 40-50-kDa to the 7-10-kDa fractions, and the survival-promoting activity in the 7-10-kDa fractions was blocked by an anti-IGF-II neutralizing monoclonal antibody. These results indicate that the neurotrophic substance in 0.5 M NaCl-eluate from heparin-affinity chromatography is IGF-II and that mechanisms may exist in vivo for the activation of latent IGF-II, whose biological effects may be blocked by its specific binding proteins. |
| Databáze: | OpenAIRE |
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