[Retinal ganglion cell neuroprotection in culture].
OBJECTIVE
To study the pig retinal ganglion cell (RGC) survival in culture, analysing the possible neuroprotective effect of retinal Müller glia (RMG) and brain-derived neurotrophic factor (BDNF).


METHODS
Adult pig retina were dissociated and cultured under different conditions: 1) on laminin/poly-D-lysine-coated coverslips in chemically defined medium (CDM); 2) on laminin/poly-D-lysine-coated coverslips in CMD supplemented with BDNF; 3) on confluent monolayer cultures of RMG in CDM; 4) on laminin/poly-D-lysine substrate in conditioned medium obtained from RMG. RGCs were identified by immunocytochemistry using antibody against 68 kDa neurofilament and observed under an fluorescent microscope. RGCs were classified on the basis of the size, number and length of neurites, and their survival was assayed for each treatment.


RESULTS
Confluent RMG substrates and RMG conditioned medium significantly increased the survival of cultured pig RGC. Moreover these two conditions increased the mean area of RGCs and enhanced neurite growth and elongation. Addition of BDNF to culture medium did not modify survival but increased RGC size, neurite number and neurite length.


CONCLUSIONS
These findings demonstrate that factor(s) secreted by RMG exert beneficial effects on adult RGC survival and neurite regeneration in vitro, and might constitute important agent(s) for RGC neuroprotection. BDNF also increases the mean area of RGCs and enhances neurite growth but it does not increase the survival of RGCs.
García
M
M
Departamento Biología Celular, Facultad de Medicina, Universidad del País Vasco, Lejona, Vizcaya, España. gcpvecoe@lg.ehu.es
Ruiz Ederra
J
J
Hernández-Barbáchano
E
E
Urcola
J A
JA
Bilbao
J
J
Araiz
J
J
Durán
J A
JA
Vecino
E
E
D000818:Animals; D015415:Biomarkers; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology*; D048429:Cell Size; D002478:Cells, Cultured / Q000187:drug effects / Q000648:ultrastructure; D018920:Coculture Techniques; D003470:Culture Media / Q000494:pharmacology; D017077:Culture Media, Conditioned / Q000494:pharmacology; D005136:Eye Proteins / Q000032:analysis; D007797:Laminin; D016501:Neurites / Q000648:ultrastructure; D016900:Neurofilament Proteins / Q000032:analysis; D009457:Neuroglia / Q000378:metabolism*; D018696:Neuroprotective Agents / Q000494:pharmacology*; D011107:Polylysine; D012160:Retina / Q000166:cytology; D012165:Retinal Ganglion Cells / Q000145:classification / Q000187:drug effects* / Q000648:ultrastructure; D013552:Swine
D015415:Biomarkers; D019208:Brain-Derived Neurotrophic Factor; D003470:Culture Media; D017077:Culture Media, Conditioned; D005136:Eye Proteins; D007797:Laminin; D016900:Neurofilament Proteins; D018696:Neuroprotective Agents; C066858:neurofilament protein NF 68; D011107:Polylysine