Role of tumor necrosis factor-alpha in zebrafish retinal neurogenesis and myelination

• 作者: Jian-Lin Cui Shi-Jiao Cai Xu-Dan Lei Yan Sun Yang-Wu Fang Yu-Hao Li
• 来源期刊: 国际眼科杂志：英文版
• 学科: 医学
• 关键词: tumor necrosis factor-alpha RETINA NEUROGENESIS MYELINATION ZEBRAFISH
• 年，卷（期）: 2016,（6）
• 页码范围: 831-837
• 页数: 7页
• 分类号: R77

摘要

AIM: To investigate the role of tumor necrosis factoralpha(TNF-α) in zebrafish retinal development and myelination.METHODS: Morpholino oligonucleotides(MO), which are complementary to the translation start site of the wild-type embryonic zebrafish TNF-α m RNA sequence,were synthesized and injected into one- to four-cell embryos. The translation blocking specificity was verified by Western blotting using an anti-TNF-α antibody,whole-mount in situ hybridization using a hepatocyte-specific m RNA probe ceruloplasmin(cp), and co-injection of TNF-α MO and TNF-α m RNA. An atonal homolog 7(atoh7) m RNA probe was used to detect neurogenesis onset. The retinal neurodifferentiation was analyzed by immunohistochemistry using antibodies Zn12, Zpr1, and Zpr3 to label ganglion cells, cones, and rods, respectively. Myelin basic protein(mbp) was used as a marker to track and observe the myelination using whole-mount in situ hybridization.RESULTS: Targeted knockdown of TNF-α resulted in specific suppression of TNF-α expression and a severely underdeveloped liver. The co-injection of TNF-α MO and m RNA rescued the liver development. Retinal neurogenesis in TNF-α morphants was initiated on time.The retina was fully laminated, while ganglion cells,cones, and rods were well differentiated at 72 hours post-fertilization(hpf). mbp was expressed in Schwann cells in the lateral line nerves and cranial nerves from 3days post-fertilization(dpf) as well as in oligodendrocytes linearly along the hindbrain bundles and the spinal cord from 4 dpf, which closely resembled its endogenous profile.CONCLUSION: TNF-α is not an essential regulator for retinal neurogenesis and optic myelination.