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Yuhao Li, Xudan Lei, Yan Sun, Yangwu Fang; The Role of Tumor Necrosis Factor-alpha on the Neurogenesis in Zebrafish Retina. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):1494.
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To investigate the role of tumor necrosis factor-alpha (TNF-α) in the developing zebrafish retina.To investigate the role of tumor necrosis factor-alpha (TNF-α) in the developing zebrafish retina.
Wild-type embryonic zebrafish were used in this study. Morpholino oligonucleotides, complementary to the translation start site of zebrafish TNF-α mRNA sequence, were synthesized and injected into embryos at 1-4-cell stage. The specificity of the translation-blocking was verified by western blot with an anti-TNF-α antibody and whole mount in situ hybridization with a hepatocyte-specific mRNA probe ceruloplasmin. The retinal neurodifferentiation was analyzed by immunohistochemistry with antibodies Zn12, Zpr1 and Zpr-3 which labelled ganglion cells, cones and rods, respectively. An fms mRNA probe and the 4C4 antibody were used to examine the distribution of microglia in the brain and retina.
Targeted knockdown of TNF-α resulted in specific suppression of TNF-α expression and caused a reduction in liver size. In TNF-α morphants, the neurogenesis in was initiated on time, and ganglion cells, cones and rods were well differentiated at 72 hours postfertilization (hpf).
TNF-α is not a key regulator for normal retinal growth and neurogenesis.
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