Instituto de Fisiología y Biofísica Prof. Bernardo Houssay (IFIBIO-Houssay) UBA y Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina.
Departamento de Fisiología, Facultad de Medicina, Universidad de Buenos Aires (UBA), Buenos Aires, Argentina.
Eur J Neurosci. 2018 Nov;48(9):3019-3042. doi: 10.1111/ejn.14113. Epub 2018 Sep 30.
The adult zebrafish is considered a useful model for studying mechanisms involved in tissue growth and regeneration. We have characterized cytotoxic damage to the retina of adult zebrafish caused by the injection of cobalt chloride (CoCl ) into the vitreous cavity. The CoCl concentration we used primarily caused injury to photoreceptors. We observed the complete disappearance of cones, followed by rods, across the retina surface from 28 to 96 hr after CoCl injury. The loss of 30% of bipolar cells was also observed by 50 hr after lesion (hpl). CoCl injury provoked a strong induction of the proliferative activity of multipotent Müller glia and derived progenitors. The effect of CoCl on retina cells was significantly reduced by treatment with glutamate ionotropic receptor antagonists. Cone photoreceptor regeneration occurred 25 days after injury. Moreover, a single dose of CoCl induced vascular damage and regeneration, whereas three injections of CoCl administered weekly provoked neovascular-like changes 20 days after injury. CoCl injury also caused microglial reactivity in the optic disc, retina periphery and fibre layer. CoCl -induced damage enhanced pluripotency and proneural transcription factor gene expression in the mature retina 72 hpl. Tumour necrosis factor alpha, vascular endothelial growth factor (VEGF) and VEGF receptor mRNA levels were also significantly enhanced by 72 hpl. The injury paradigm we have described in this work may be useful for the discovery of signalling molecules and pathways that participate in the regenerative response and it may serve as a model to screen for compounds that could potentially treat aberrant angiogenesis.
成年斑马鱼被认为是研究组织生长和再生相关机制的有用模型。我们描述了将氯化钴（CoCl ）注射到玻璃体腔后对成年斑马鱼视网膜造成的细胞毒性损伤。我们使用的 CoCl 浓度主要导致光感受器损伤。我们观察到，在 CoCl 损伤后 28 至 96 小时，从视网膜表面开始，锥体完全消失，随后是杆状细胞。在损伤后 50 小时，还观察到双极细胞损失了 30%。CoCl 损伤引发多潜能的 Müller 胶质细胞及其衍生祖细胞的增殖活性强烈诱导。用谷氨酸离子型受体拮抗剂处理可显著降低 CoCl 对视网膜细胞的作用。在损伤后 25 天发生锥体光感受器再生。此外，单次 CoCl 剂量可引起血管损伤和再生，而每周注射三次 CoCl 可在损伤后 20 天引起类似新生血管的变化。CoCl 损伤还导致视盘、视网膜周边和纤维层中小胶质细胞的反应性。CoCl 诱导的损伤可增强成熟视网膜中多能性和原神经转录因子基因的表达，在损伤后 72 小时达到高峰。肿瘤坏死因子-α、血管内皮生长因子（VEGF）和 VEGF 受体 mRNA 水平在 72 小时也显著增强。在这项工作中描述的损伤模型可能有助于发现参与再生反应的信号分子和途径，并且可以作为筛选可能治疗异常血管生成的化合物的模型。
