Effects of brain-derived neurotrophic factor on cell survival, differentiation and patterning of neuronal connections and Müller glia cells in the developing retina.
The aim of the present study was to determine the influence of brain-derived neurotrophic factor (BDNF) on survival, phenotype differentiation and network formation of retinal neurons and glia cells. To achieve a defined concentration and constant level of BDNF over several days, experiments were performed in an organotypic culture of the developing rat retina. After 6 days in vitro, apoptosis in the different cell layers was determined by TUNEL staining and cell-type-specific antibodies were used to identify distinct neuronal cell types and Müller cells. Cultured retinas treated with BDNF (100 ng BDNF/mL medium) were compared with untreated as well as with age-matched in vivo retinas. Quantitative morphometry was carried out using confocal microscopy. BDNF promoted the in vitro development and differentiation of the retina in general, i.e. the number of cells in the nuclear layers and the thickness of the plexiform layers were increased. For all neurons, the number of cells and the complexity of arborizations in the synaptic layers were clearly up-regulated by BDNF. In control cultures, the synaptic stratification of cone bipolar cells within the On- and Off-layer of the inner plexiform layer was disturbed and a strong reactivity of Müller cell glia was observed. These effects were not present in BDNF-treated cultures. Our data show that BDNF promotes the survival of retinal interneurons and plays an important role in establishing the phenotypes and the synaptic connections of a large number of neuronal types in the developing retina. Moreover, we show an effect of BDNF on Müller glia cells.
Pinzón-Duarte
Germán
G
Experimental Ophthalmology, University Eye Hospital, Röntgenweg 11, 72076 Tübingen, Germany.
Arango-González
Blanca
B
Guenther
Elke
E
Kohler
Konrad
K
D000367:Age Factors; D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000502:physiology; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology*; D064032:Calbindin 2; D064026:Calbindins; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002452:Cell Count; D002454:Cell Differentiation / Q000187:drug effects* / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D005136:Eye Proteins; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D005974:Glutamate-Ammonia Ligase / Q000378:metabolism; D051597:Hippocalcin; D007150:Immunohistochemistry / Q000379:methods; D020287:In Situ Nick-End Labeling / Q000379:methods; D008074:Lipoproteins; D018613:Microscopy, Confocal / Q000379:methods; D009419:Nerve Tissue Proteins; D009457:Neuroglia / Q000166:cytology / Q000187:drug effects*; D009474:Neurons / Q000187:drug effects* / Q000502:physiology; D009924:Organ Culture Techniques; D010320:Parvalbumins / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism; D051381:Rats; D051603:Recoverin; D012160:Retina / Q000166:cytology*; D064030:S100 Calcium Binding Protein G / Q000378:metabolism; D013997:Time Factors
D019208:Brain-Derived Neurotrophic Factor; D064032:Calbindin 2; D064026:Calbindins; D002135:Calcium-Binding Proteins; D005136:Eye Proteins; D005904:Glial Fibrillary Acidic Protein; D008074:Lipoproteins; D009419:Nerve Tissue Proteins; D010320:Parvalbumins; C496733:Rcvrn protein, rat; D064030:S100 Calcium Binding Protein G; D051603:Recoverin; D051597:Hippocalcin; D011493:Protein Kinase C; C081173:glutamine synthetase I; D005974:Glutamate-Ammonia Ligase