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MitoPark mouse

MitoPark mice delete Tfam in midbrain dopamine neurons and develop progressive nigrostriatal degeneration, reduced striatal dopamine, impaired dopamine release and age-dependent motor deficits. Studies in 20-30 week old animals also detect depression- and anxiety-like behavior and altered cortical physiology.

结局指标

以荧光显微镜在脑切片中观察TH阳性胞体与纤维并采集图像；本方案输出为免疫荧光染色与成像结果，不包含体视学细胞计数、纤维密度阈值或纹状体多巴胺浓度测定。

实验动物

脑切片酪氨酸羟化酶（TH）免疫荧光染色
年龄
20-30 weeks in cited studies
性别
混合
组织
substantia nigra and striatum
脑切片酪氨酸羟化酶（TH）免疫荧光染色
年龄
20-30 weeks in cited studies
性别
混合
组织
substantia nigra and striatum

实验步骤

脑切片洗涤 用4% PFA/PBS洗涤脑切片一次。 去除固定液 以轻柔摇动或旋转5分钟，确保彻底去除固定液。 PBS漂洗 用PBS短暂漂洗，去除残留PFA。 封闭液配制 配制含10%驴血清和0.3% Triton-X的PBS封闭液。 封闭 室温下将切片在封闭液中孵育1小时，以减少非特异性结合。 均匀封闭 将切片置于摇床或旋转器上，使切片均匀接触封闭液。 一抗工作液配制 按下列组成配制一抗工作液。 加入鸡抗TH一抗 鸡抗TH抗体（Abcam，货号ab76442）。 稀释一抗 用含2.5%驴血清和0.3% Triton-X的PBS将一抗按1:200稀释。 一抗孵育 切片与一抗在室温孵育1小时。 湿盒孵育 在湿盒中孵育，防止液体蒸发。 一抗后洗涤 用PBS洗涤切片三次，每次5分钟。 轻柔洗涤 洗涤过程中保持轻柔摇动或旋转。 二抗孵育 与针对鸡IgG的荧光偶联二抗按1:1000在室温孵育1小时。 避光 避光以保护荧光。 终末洗涤 用PBS洗涤切片三次，每次5分钟。 避光洗涤 在避光条件下完成洗涤，以维持荧光完整性。 贴片 将切片贴于包被载玻片上。 Mowiol封片 用Mowiol封片介质覆盖切片并加盖玻片。 干燥 避光干燥至少30分钟后再成像。 荧光显微镜成像 使用Leica DM6000 B显微镜和10×物镜观察切片。 选择滤光片 使用合适的荧光滤光片组检测二抗信号。 调整曝光 调整曝光设置以优化信噪比。

模型身份

模型名称
MitoPark mouse
物种
mouse
品系
MitoPark (DAT-cre; Tfam loxP/loxP)
遗传背景
C57BL/6
基因型／合子状态
Dopamine-neuron-specific deletion of Tfam (mitochondrial transcription factor A) using DAT-cre
等位基因
Tfam loxP/loxP; DAT-cre

生物学特性

表型出现阶段
Progressive; dopamine dysfunction and motor deficits emerge with age. Published endpoints include 20-30 weeks and older, with sex-dependent differences in onset.
适用研究问题
Mitochondrial DNA maintenance in dopamine neuron survival; progression of nigrostriatal degeneration; motor and affective phenotypes; mitochondrial-targeted and GLP-1 receptor interventions.
模型局限
The model uses a conditional Tfam deletion rather than a Parkinson disease-causing mutation. Phenotype depends on Cre/lox timing and background, and it does not reproduce Lewy body pathology. This is a literature reference model, not a statement of company inventory or completed experiments.

实验方法

实验方案

  • 脑切片酪氨酸羟化酶（TH）免疫荧光染色

    脑切片经4% PFA/PBS洗涤、10%驴血清/0.3% Triton-X封闭后，以鸡抗TH抗体ab76442（1:200）和抗鸡IgG荧光二抗（1:1000）孵育，Mowiol封片并以荧光显微镜成像，用于TH阳性胞体与纤维的定位观察。

  • 脑切片酪氨酸羟化酶（TH）免疫荧光染色

    脑切片经4% PFA/PBS洗涤、10%驴血清/0.3% Triton-X封闭后，以鸡抗TH抗体ab76442（1:200）和抗鸡IgG荧光二抗（1:1000）孵育，Mowiol封片并以荧光显微镜成像，用于TH阳性胞体与纤维的定位观察。

配套试剂与材料

参考文献

  1. Cong L; Muir ER; Chen C; Qian Y; Liu J; Biju KC. Multimodal MRI Evaluation of the MitoPark Mouse Model of Parkinson's Disease.. PloS one. 2016. PMID:27003179.
  2. Chen YH; Wang V; Huang EY; Chou YC; Kuo TT; Olson L. Delayed Dopamine Dysfunction and Motor Deficits in Female Parkinson Model Mice.. International journal of molecular sciences. 2019. PMID:31835787.
  3. Extraction identity record 06371e2919da93954789eaf39c8d615a5a42b0c841f70275941692bb903fd5e3
  4. Tseng KY; Kuo TT; Chang PK; Chou JL; Huang EY; Greig NH. PT320, a GLP-1 receptor agonist, mitigates nucleus accumbens-associated depression- and anxiety-like behaviors in MitoPark mice of Parkinson's disease.. Journal of Parkinson's disease. 2026. PMID:42585299.
  5. Marta Graziano, Ioannis Mantas, Konstantinos Meletis. Immunohistochemistry for Tyrosine Hydroxylase (TH) Detection in Brain Sections. protocols.io, v1, 2025-02-20. DOI:10.17504/protocols.io.6qpvr9jwpvmk/v1. Protocol status: Working; License: Creative Commons Attribution License.
  6. Abcam. Tyrosine Hydroxylase antibody (neuronal marker) ab76442 product page, accessed 2026-10-19: Key facts species field lists Mouse; product details list rat samples; applications listed as IHC-P/IHC-FrFl.
  7. Sundström E, Strömberg I, Tsutsumi T, Olson L, Jonsson G. Studies on the effect of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) on central catecholamine neurons in C57BL/6 mice. Comparison with three other strains of mice. Brain Res. 1987;405(1):26-38. DOI:10.1016/0006-8993(87)90986-3; PMID:2882814. 原文以TH免疫组化显示MPTP处理后C57BL/6小鼠黑质致密部多巴胺能胞体减少70%、纹状体DA纤维近乎完全丢失。
  8. Sauer H, Oertel WH. Progressive degeneration of nigrostriatal dopamine neurons following intrastriatal terminal lesions with 6-hydroxydopamine: a combined retrograde tracing and immunocytochemical study in the rat. Neuroscience. 1994;59(2):401-415. DOI:10.1016/0306-4522(94)90605-X; PMID:7516500. 原文在大鼠纹状体6-OHDA损伤后以TH免疫反应细胞计数显示黑质TH阳性细胞进行性减少。
  9. Oiwa Y, Sanchez-Pernaute R, Harvey-White J, Bankiewicz KS. Progressive and extensive dopaminergic degeneration induced by convection-enhanced delivery of 6-hydroxydopamine into the rat striatum: a novel rodent model of Parkinson disease. J Neurosurg. 2003;98(1):136-144. DOI:10.3171/jns.2003.98.1.0136; PMID:12546362. 原文在大鼠6-OHDA模型以形态学标准定义黑质细胞丢失>80%，并报告纹状体多巴胺下降>95%。
  10. Huang et al. Therapeutic Effects of Noninvasive Electrical Stimulation in Combination Transplantation of Human Adipose-Derived Stem Cells-Derived Dopaminergic Neuron on the Monkey Model of Parkinson's Disease. MedComm. 2026. DOI:10.1002/mco2.70595. Sections 4.1 and 4.13 document cynomolgus monkeys and Abcam ab76442 with anti-chicken fluorescent secondary in brain sections.
  11. ARRIVE 2.0 官方规范
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