Developmentally regulated release of intraretinal neurotrophic factors in vitro.
The effects of conditioned media either from aggregates or from explants of embryonic chick retinae and of recombinant neurotrophins were tested upon the survival in vitro of ganglion cells in dissociated cell cultures from the retina of newborn rats. Ganglion cells were identified by the detection of retrogradely transported horseradish peroxidase injected bilaterally into the superior colliculus. Conditioned media increased significantly the survival of ganglion cells after 2 days in culture, at a wide range of plating densities, and had no effect upon adhesion of rat retinal cells. Media conditioned by cell ensembles from chick retinae from embryonic day 8 (E8) to E16 had neurotrophic effects. Release of neurotrophic activity peaked at E10 E12, irrespective of the numbers of cells or total concentration of protein in the conditioned media. The active molecules were non-dialyzable and were released either in the presence or in the absence of fetal calf serum. The neurotrophic activity was abolished by trypsinization, and recovered by salting-out with 25 75% ammonium sulfate. NT-4, BDNF and, to a lesser extent, NT-3, increased the survival of ganglion cells in our assay, while NGF had no effect. The data show that chick retinal cells release soluble trophic proteins according to a developmentally regulated pattern. These neurotrophic factors may be involved in local competitive interactions that help control naturally occurring neuron death among ganglion cells of the vertebrate retina.
Ary-Pires
R
R
Instituto de Biofisica Carlos Chagas Filho, UFRJ, Rio de Janeiro, Brazil.
Nakatani
M
M
Rehen
S K
SK
Linden
R
R
D000818:Animals; D000831:Animals, Newborn / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects; D002642:Chick Embryo; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004622:Embryo, Mammalian / Q000502:physiology; D005314:Embryonic and Fetal Development; D066298:In Vitro Techniques; D009414:Nerve Growth Factors / Q000378:metabolism* / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D011994:Recombinant Proteins; D012160:Retina / Q000196:embryology / Q000254:growth & development* / Q000378:metabolism*; D012165:Retinal Ganglion Cells / Q000187:drug effects / Q000502:physiology
D017077:Culture Media, Conditioned; D009414:Nerve Growth Factors; D011994:Recombinant Proteins