Optimised methods to image hepatic lipid droplets in zebrafish larvae.

IF 4 3区 医学 Q2 CELL BIOLOGY Disease Models & Mechanisms Pub Date : 2024-10-07 DOI:10.1242/dmm.050786
Nouf Khan, Talhah M Salmi, Anthony P Karamalakis, Anjana Ramdas Nair, Kirsten C Sadler, Andrew G Cox
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Abstract

The optical transparency of zebrafish larvae enables visualization of subcellular structures in intact organs, and these vertebrates are widely used to study lipid biology and liver disease. Lipid droplet (LD) presence is a prevalent feature of healthy cells but under conditions such as nutrient excess, toxicant exposure or metabolic imbalance, LD accumulation in hepatocytes can be a harbinger of more severe forms of liver disease. We undertook a comprehensive analysis of approaches useful to investigate LD distribution and dynamics in physiological and pathological conditions in the liver of zebrafish larvae. This comparative analysis of the lipid dyes oil red O (ORO), Nile Red (NR), LipidTox and LipidSpot as well as transgenic LD reporters that rely on EGFP fusions of the LD decorating protein perilipin 2 (PLIN2) demonstrate the strengths and limitations of each approach. These protocols are amenable to detection methods ranging from low resolution stereomicroscopy to confocal imaging, which enables measurements of hepatic LD size, number and dynamics at cellular resolution in live and fixed animals. This resource will benefit investigators studying LD biology in zebrafish disease models.

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斑马鱼幼体肝脏脂滴成像的优化方法。
斑马鱼幼体的光学透明度可使完整器官的亚细胞结构可视化,这些脊椎动物被广泛用于研究脂质生物学和肝脏疾病。脂滴(LD)的存在是健康细胞的普遍特征,但在营养过剩、有毒物质暴露或代谢失衡等条件下,肝细胞中脂滴的积累可能是更严重肝病的先兆。我们对斑马鱼幼体肝脏中生理和病理条件下低密度脂蛋白分布和动态的研究方法进行了全面分析。我们对油红 O(ORO)、尼罗河红(NR)、LipidTox 和 LipidSpot 等脂质染料,以及依赖 EGFP 融合 LD 修饰蛋白周脂素 2(PLIN2)的转基因 LD 报告器进行了比较分析,证明了每种方法的优势和局限性。这些方案适用于从低分辨率体视显微镜到共聚焦成像等各种检测方法,可在活体和固定动物体内以细胞分辨率测量肝脏 LD 的大小、数量和动态。该资源将使研究斑马鱼疾病模型中 LD 生物学的研究人员受益匪浅。
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来源期刊
Disease Models & Mechanisms
Disease Models & Mechanisms 医学-病理学
CiteScore
6.60
自引率
7.00%
发文量
203
审稿时长
6-12 weeks
期刊介绍: Disease Models & Mechanisms (DMM) is an online Open Access journal focusing on the use of model systems to better understand, diagnose and treat human disease.
期刊最新文献
The role of mesenchymal cells in cholangiocarcinoma. High-fat and high-sucrose diets induce an experimental rabbit model for age-related macular degeneration (AMD). Early life cisplatin exposure induces nerve growth factor mediated neuroinflammation and chemotherapy induced neuropathic pain. Hippo signaling cooperates with p53 to regulate lung airway mucous cell metaplasia. Generation of a zebrafish neurofibromatosis model via inducible knockout of nf2.
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