Lysosomal storage diseases are the most common cause of neurodegeneration in children. They are characterised at the cellular level by the accumulation of storage material within lysosomes. There are very limited therapeutic options, and the search for novel therapies has been hampered as few good small animal models are available. Here, we describe the use of light sheet microscopy to assess lipid storage in drug and morpholino induced zebrafish models of two diseases of cholesterol homeostasis with lysosomal dysfunction: First, Niemann–Pick type C disease (NPC), caused by mutations in the lysosomal transmembrane protein NPC1, characterised by intralysosomal accumulation of cholesterol and several other lipids. Second, Smith–Lemli–Opitz syndrome (SLOS), caused by mutations in 7-dehydrocholesterol reductase, which catalyses the last step of cholesterol biosynthesis and is characterised by intralysosomal accumulation of dietary cholesterol. This is the first description of a zebrafish SLOS model. We find that zebrafish accurately model lysosomal storage and disease-specific phenotypes in both diseases. Increased cholesterol and ganglioside GM1 were observed in sections taken from NPC model fish, and decreased cholesterol in SLOS model fish, but these are of limited value as resolution is poor, and accurate anatomical comparisons difficult. Using light sheet microscopy, we were able to observe lipid changes in much greater detail and identified an unexpected accumulation of ganglioside GM1 in SLOS model fish. Our data demonstrate, for the first time in zebrafish, the immense potential that light sheet microscopy has in aiding the resolution of studies involving lysosomal and lipid disorders.

溶酶体贮积病是儿童神经退行性病变的最常见原因。它们在细胞水平上的特征在于溶酶体内的储存材料的积累。治疗选择非常有限，并且由于很少有好的小型动物模型，因此寻找新疗法的工作受到了限制。在这里，我们描述了使用光片显微镜来评估药物和吗啉代诱发的斑马鱼模型中两种溶酶体功能异常的胆固醇稳态的疾病中的脂质存储：首先，由溶酶体跨膜突变引起的尼曼-匹克C型疾病（NPC） NPC1蛋白，其特征为胆固醇和其他几种脂质的溶酶体内积累。其次，由7-脱氢胆固醇还原酶突变引起的史密斯-莱姆利-奥皮兹综合症（SLOS），它催化胆固醇生物合成的最后一步，其特征是膳食胆固醇的溶酶体内积累。这是对斑马鱼SLOS模型的首次描述。我们发现斑马鱼在两种疾病中都能准确地模拟溶酶体贮藏和疾病特异性表型。在取自NPC模型鱼的切片中观察到胆固醇和神经节苷脂GM1升高，而SLOS模型鱼中的胆固醇降低，但由于分辨率差，因此它们的价值有限，并且难以进行精确的解剖比较。使用光片显微镜，我们能够更详细地观察脂质变化，并确定了SLOS模型鱼中神经节苷脂GM1的意外积累。我们的数据首次在斑马鱼中证明，