Use of suppression subtractive hybridization to identify genes regulated by ciliary neurotrophic factor in postnatal retinal explants.

摘要: Purpose: The retinal progenitors are multipotential, and the decision taken by a progenitor to differentiate along particular path depends on both cell-intrinsic cell-extrinsic factors. Ciliary neurotrophic factor (CNTF), member of interleukin-6 (IL-6) family, added rat postnatal inhibits rod photoreceptor cell differentiation, promotes Muller glia genesis enhances expression bipolar neuron markers. We hypothesized that those transcripts regulated during CNTF-influenced differentiation may be involved in choice fate. Our aim was isolate these genes, characterize their retina, subsequently focus candidates promote differentiation. Methods: Retinas were cultured vitro as explants at day 0 (P0) absence or presence CNTF for six days. Transcripts after days (DIV) selected subtraction suppressive hybridization (SSH) cloned two libraries. UC6 DC6 libraries contained genes upregulated downregulated, respectively, 6DIV. Results: In first library, UC6, eight clones representing seven different isolated up-regulated CNTF. 21 clones, 17 appeared down-regulated Genes classified categories, such protein modification, signal transduction, regulation transcription according Gene Ontology Annotation. Conclusions: Among 24 our study revealed 11 (two nine downregulated) potentially biological effects.