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We have examined the trophic effects of conditioned media obtained from purified murine Müller glia cells on chick purified sympathetic or dorsal root ganglia (DRG) neurons and on Retinal Ganglion Cells (RGC) from postnatal mice. Purified murine Müller glia cultures stained positively for vimentin, GFAP or S-100, but were negative for neuronal markers. Murine Müller glial conditioned medium (MMG) was concentrated and at 1:1 dilution supported 100% survival of chick or rat sympathetic neurons after 48 h compared to <5% in controls. Partial purification of the MMG using centriprep concentrators showed that trophic activity is from molecules above 10 kDa. MMG stimulated AKT, ERK and pStat3 in sympathetic neurons. Sympathetic or DRG neuronal survival induced by MMG was blocked by anti-human NGF, but not by anti-human CNTF (sympathetic) or by anti-BDNF (DRGs) neutralizing antibodies. MMG also induced neurite outgrowth in P4 mice retinal explants and on isolated RGC. RGCs plated on top of Müller glia cells had a much better survival rate (>80%, 96 h) compared to laminin+poly-L-lysine substrates. In conclusion, we show that purified mice Müller glia cultures secrete NGF that support peripheral neuronal survival and other unidentified trophic molecules that induce RGC survival and neuritogenesis.

Original publication

DOI

10.1016/j.brainres.2008.02.035

Type

Journal article

Journal

Brain Res

Publication Date

18/04/2008

Volume

1205

Pages

1 - 11

Keywords

Animals, Antibodies, Blocking, Blotting, Western, Brain-Derived Neurotrophic Factor, Cell Count, Cell Survival, Central Nervous System, Ciliary Neurotrophic Factor, Colforsin, Culture Media, Conditioned, Extracellular Signal-Regulated MAP Kinases, Humans, Mice, Nerve Growth Factors, Neurites, Neuroglia, Neurons, Oncogene Protein v-akt, Peripheral Nerves, Rats, STAT3 Transcription Factor, Superior Cervical Ganglion, Sympathetic Nervous System