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Immunofluorescence/fluorescence assessment of brain-derived neurotrophic factor, c-Fos activation, and apoptosis in the brain of zebrafish (Danio rerio) larvae exposed to glufosinate
NeuroToxicology ( IF 3.4 ) Pub Date : 2018-09-14 , DOI: 10.1016/j.neuro.2018.09.003
Selim Çomakli , Mine Köktürk , Ahmet Topal , Mustafa Özkaraca , Saltuk Buğrahan Ceyhun

In this study, we investigated the potential neuro-toxicological mechanism of the glufosinate in the brain of zebrafish larvae in terms of BDNF and c-Fos proteins by evaluating apoptosis, immunofluorescence BDNF, and c-FOS activation. We also measured survival rate, hatching rate, and body malformations during 96 h exposure time. For this purpose, zebrafish embryos were treated with graded concentrations of dosing solutions (0.5, 1, 3, and 5 ppm) of glufosinate. End of the treatment, acridine orange staining was used to detect apoptotic cells in the brain of zebrafish larvae at 96 hpf. Texas Red and FITC/GFP labeled protein-specific antibodies were used in immunofluorescence assay for BDNF and c-FOS, respectively. The results have indicated that exposure to glufosinate caused to embryonic death, hatching delay, induction of apoptosis, increasing of c-FOS activity and the level of BDNF in a dose-dependent manner. As a conclusion, we suggested that c-Fos might play a role in the regulation of BDNF which responses to prevent the cell from apoptosis even in case of unsuccessful in zebrafish larvae exposed to glufosinate.



中文翻译:

暴露于草铵膦的斑马鱼(Danio rerio)幼虫脑源性神经营养因子,c-Fos活化和脑细胞凋亡的免疫荧光/荧光评估

在这项研究中,我们通过评估细胞凋亡,免疫荧光BDNF和c-FOS活化作用,以BDNF和c-Fos蛋白的形式研究了斑马鱼幼虫大脑中草铵膦的潜在神经毒理学机制。我们还测量了96小时暴露时间下的存活率,孵化率和身体畸形。为此,用梯度浓度的草铵膦剂量溶液(0.5、1、3和5 ppm)处理斑马鱼胚胎。治疗结束时,a啶橙染色用于检测斑马鱼幼虫大脑在96 hpf时的凋亡细胞。Texas Red和FITC / GFP标记的蛋白特异性抗体分别用于BDNF和c-FOS的免疫荧光测定。结果表明,暴露于草铵膦会导致胚胎死亡,孵化延迟,诱导细胞凋亡,剂量依赖性增加c-FOS活性和BDNF水平。结论是,我们建议c-Fos可能在BDNF的调节中起作用,即使在未成功暴露于草铵膦的斑马鱼幼虫中,c-Fos仍可响应以阻止细胞凋亡。

更新日期:2018-09-14
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