Acute Kidney Injury Model Induced by Cisplatin in Adult Zebrafish

斑马鱼 顺铂 急性肾损伤 癌症研究 炎症 肾毒性 医学 标记法 流式细胞术 程序性细胞死亡 免疫系统 药理学 细胞凋亡 生物 病理 免疫学 化疗 内科学 免疫组织化学 生物化学 基因
作者
Camila Morales,Bárbara Nunes Padovani,Mariana Abrantes do Amaral,Guilherme José Bottura de Barros,Izabella Karina Xavier de Oliveira,Meire Ioshie Hiyane,Niels Olsen Saraiva Câmara
出处
期刊:Journal of Visualized Experiments [MyJOVE]
卷期号: (171) 被引量:6
标识
DOI:10.3791/61575
摘要

Cisplatin is commonly used as chemotherapy. Although it has positive effects in cancer-treated individuals, cisplatin can easily accumulate in the kidney due to its low molecular weight. Such accumulation causes the death of tubular cells and can induce the development of Acute Kidney Injury (AKI), which is characterized by a quick decrease in kidney function, tissue damage, and immune cells infiltration. If administered in specific doses cisplatin can be a useful tool as an AKI inducer in animal models. The zebrafish has appeared as an interesting model to study renal function, kidney regeneration, and injury, as renal structures conserve functional similarities with mammals. Adult zebrafish injected with cisplatin shows decreased survival, kidney cell death, and increased inflammation markers after 24 h post-injection (hpi). In this model, immune cells infiltration and cell death can be assessed by flow cytometry and TUNEL assay. This protocol describes the procedures to induce AKI in adult zebrafish by intraperitoneal cisplatin injection and subsequently demonstrates how to collect the renal tissue for flow cytometry processing and cell death TUNEL assay. These techniques will be useful to understand the effects of cisplatin as a nephrotoxic agent and will contribute to the expansion of AKI models in adult zebrafish. This model can also be used to study kidney regeneration, in the search for compounds that treat or prevent kidney damage and to study inflammation in AKI. Moreover, the methods used in this protocol will improve the characterization of tissue damage and inflammation, which are therapeutic targets in kidney-associated comorbidities.
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