Cyclic AMP elevation is sufficient to promote the survival of spinal motor neurons in vitro.
The short-term survival of highly purified embryonic spinal motor neurons (SMNs) in culture can be promoted by many peptide trophic factors, including brain-derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF), fibroblast growth factor (FGF), glial-derived neurotrophic factor (GDNF), and hepatocyte growth factor (HGF). We have asked whether these peptides are sufficient to promote the long-term survival of purified E15 SMNs. Contrary to previous reports, we find that when SMNs are cultured in serum-free medium containing a single peptide trophic factor only approximately one-third of the cells survive for 3 d in culture. When multiple factors are combined, additive effects on survival are observed transiently, but by 7 d of culture the majority of SMNs has died. Surprisingly, when cAMP levels are elevated, the majority of SMNs extend processes and survive for 1 week in culture in the absence of peptide trophic factors, even in low-density cultures. A combination of five peptide trophic factors, together with cAMP elevation, promotes the long-term survival of most of the SMNs in serum-free culture for 3 weeks. These findings provide useful culture conditions for studying the properties of SMNs and have implications for the treatment of motor neuron diseases.
Hanson
M G
MG
Stanford University School of Medicine, Department of Neurobiology, Stanford, California 94305-5125, USA.
Shen
S
S
Wiemelt
A P
AP
McMorris
F A
FA
Barres
B A
BA
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000818:Animals; D019485:Bone Morphogenetic Proteins / Q000378:metabolism; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology; D002470:Cell Survival / Q000502:physiology; D020934:Ciliary Neurotrophic Factor; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism*; D004958:Estradiol / Q000494:pharmacology; D051100:Glial Cell Line-Derived Neurotrophic Factor; D017228:Hepatocyte Growth Factor / Q000494:pharmacology; D006854:Hydrocortisone / Q000494:pharmacology; D007004:Hypoglycemic Agents / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D009046:Motor Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D009410:Nerve Degeneration / Q000378:metabolism; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins / Q000494:pharmacology; D010100:Oxygen / Q000494:pharmacology; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D011374:Progesterone / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013116:Spinal Cord / Q000166:cytology*; D013974:Thyroxine / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology
D019485:Bone Morphogenetic Proteins; D019208:Brain-Derived Neurotrophic Factor; D020934:Ciliary Neurotrophic Factor; C495041:Gdnf protein, rat; D051100:Glial Cell Line-Derived Neurotrophic Factor; D007004:Hypoglycemic Agents; D007328:Insulin; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D010726:Phosphodiesterase Inhibitors; D005576:Colforsin; D011374:Progesterone; D004958:Estradiol; D014212:Tretinoin; D017228:Hepatocyte Growth Factor; D000242:Cyclic AMP; D013974:Thyroxine; D010100:Oxygen; D015056:1-Methyl-3-isobutylxanthine; D006854:Hydrocortisone