Zucker fatty (fa/fa) rat
A homozygous missense mutation (Gln269Pro) in the leptin receptor gene Lepr causes defective leptin receptor signaling, producing obesity, hyperphagia and insulin resistance without leptin ligand deficiency. fa/fa rats do not develop overt diabetes and are regarded as a pre-diabetes model; they show steatosis but do not spontaneously develop NASH.
结局指标
血糖浓度读数。
获得可供光学显微镜观察的 H&E 染色组织切片:细胞核蓝紫色、细胞质与结缔组织纤维粉红色,用于观察组织中细胞的整体结构与形态细节。本方案本身不产出病理评分数值,也不提供特定分子标志物的定量结果。
实验动物
- 组织
- blood
- 组织
- blood
- 年龄
- 22 周龄(PMID:28953222)
- 组织
- liver sections
实验步骤
**Measurement of Blood Glucose ** **Measurement of Blood Glucose ** Blood samples were collected from the tail vein for the determination of blood glucose levels using Accutrend Plus test strips and meter (Roche).
脱蜡(Dewaxing) 石蜡切片置于二甲苯中脱蜡 3 次,每次 00:03:00。 复水(Rehydration) 石蜡切片依次置于 100% 乙醇中 3 次、每次 00:03:00,再置于 95% 乙醇中 1 次、00:03:00,随后用水漂洗。 苏木素核染色(Hematoxylin staining) 石蜡切片以苏木素染液染色 00:00:20,随后在流水下冲洗 00:01:00。 分化与返蓝(Differentiation and bluing) 石蜡切片置于返蓝液中 00:01:00,随后在流水下冲洗 00:01:00;此步使细胞核呈现蓝色。 伊红染色(Eosin staining) 石蜡切片以伊红复染 00:00:10;此步使细胞质成分与结缔组织纤维呈现粉红色。 脱水与透明(Dehydration and Clearing) 石蜡切片依次置于 95% 乙醇中 00:02:00、100% 乙醇中 2 次、每次 00:02:00,随后置于二甲苯中 2 次、每次 00:02:00。 封片(Mounting) 石蜡切片以 Permount 封片介质加盖玻片封固。
模型身份
- 模型名称
- Zucker fatty (fa/fa) rat
- 物种
- rat
- 品系
- Zucker fatty
- 基因型/合子状态
- Lepr fa/fa
- 等位基因
- Lepr fa (Gln269Pro missense)
生物学特性
- 适用研究问题
- Whether leptin-receptor-deficient genetic obesity models human metabolic syndrome and prediabetes, and which stimuli are needed to progress steatosis to NASH.
- 模型局限
- An additional stimulus is required for NASH, and the model is not considered suitable for studying macrovascular complications.
实验方法
实验方案
- 大鼠尾静脉血糖测定
使用Roche Accutrend Plus血糖试纸和读数仪测定大鼠尾静脉血糖。
- 大鼠尾静脉血糖测定
使用Roche Accutrend Plus血糖试纸和读数仪测定大鼠尾静脉血糖。
- Hematoxylin and Eosin (H&E) staining for histopathology
本方案用于固定、石蜡包埋组织切片(4 μm)的苏木素-伊红(H&E)染色:经脱蜡、复水、苏木素核染、返蓝、伊红复染、脱水透明与封片七个步骤,使细胞核呈蓝紫色、细胞质与结缔组织纤维呈粉红色,供光学显微镜下观察组织中细胞与整体结构形态。
配套试剂与材料
- Accutrend Plus 血糖试纸与读数仪
血糖检测耗材与读数平台
- Accutrend Plus 血糖试纸与读数仪
血糖检测耗材与读数平台
- 组织学二甲苯(混合异构体)
脱蜡剂
- Ethanol
复水梯度溶剂(去除二甲苯)
- 95% 乙醇溶液
复水梯度溶剂
- 自来水(组织学染色用)
复水后漂洗用水(后续各流水冲洗共用)
- Epredia Signature Series 苏木素染液(7221)
细胞核染色
- Epredia Signature Series 返蓝液(7301)
返蓝(使细胞核转蓝)
- Epredia Signature Series 伊红 Y 染液(7111)
细胞质与结缔组织纤维复染
- Permount 组织学封片剂(Fisher SP15)
封片介质
参考文献
- Rat Models of Metabolic Syndrome. Methods in Molecular Biology 2019 (PMID:31228162)
- Leptin- and Leptin Receptor-Deficient Rodent Models: Relevance for Human Type 2 Diabetes (PMID:24809394)
- Animal Models of Nonalcoholic Fatty Liver Disease - A Starter's Guide (PMID:28953222)
- Alternatives to the Streptozotocin-Diabetic Rodent. Int Rev Neurobiology 2016 (PMID:27133146)
- protocols.io · vv2 · Michael Hill · This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
- Nathalia L. Oliveira; Samantha Gruenheid. Hematoxylin and Eosin (H&E) staining for histopathology. protocols.io, v1, 2026-07-31. DOI: 10.17504/protocols.io.kxygxr9jdg8j
- Epredia Signature Series Hematoxylin (7221), 产品信息页, Fisher Scientific
- Epredia Signature Series Bluing Reagent (7301), 产品信息页, Fisher Scientific
- Epredia Signature Series Eosin Y (7111), 产品信息页, Fisher Scientific
- Xylene, Fisherbrand HistoPrep (HC7001GAL), 产品信息页, Fisher Scientific
- Fisher Chemical Permount Mounting Medium (SP15 产品族), 产品信息页, Fisher Scientific
- Ethanol (CID 702, CAS 64-17-5), PubChem 化合物标准记录
- ARRIVE 2.0 官方规范