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Parkin knockout rat

Parkin knockout rats show altered dopamine handling and mitochondrial function with mild degeneration.

Outcome measures

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。

Experimental animals

脑切片酪氨酸羟化酶（TH）免疫荧光染色
Age
age as reported in the cited study
Tissue
substantia nigra and striatum
脑切片酪氨酸羟化酶（TH）免疫荧光染色
Age
age as reported in the cited study
Tissue
substantia nigra and striatum

Experimental procedures

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。

Model identity

Model name
Parkin knockout rat
Species
rat
Genotype / zygosity
Parkin knockout
Allele
Parkin knockout

Biological characteristics

Phenotype stage
Adult; stress-dependent.
Research questions
Parkin mitophagy and dopamine neuron maintenance.
Limitations
Review-supported rat knockout with mild baseline phenotype. Literature reference only.

Experimental methods

Experimental protocols

  • 脑切片酪氨酸羟化酶（TH）免疫荧光染色

    Brain sections are washed with 4% PFA in PBS, blocked with 10% donkey serum/0.3% Triton-X, incubated with chicken anti-TH ab76442 (1:200) and a chicken-IgG fluorescent secondary antibody (1:1000), mounted in Mowiol and imaged by fluorescence microscopy to localize TH-positive cell bodies and fibers.

  • 脑切片酪氨酸羟化酶（TH）免疫荧光染色

    Brain sections are washed with 4% PFA in PBS, blocked with 10% donkey serum/0.3% Triton-X, incubated with chicken anti-TH ab76442 (1:200) and a chicken-IgG fluorescent secondary antibody (1:1000), mounted in Mowiol and imaged by fluorescence microscopy to localize TH-positive cell bodies and fibers.

Reagents and materials

References

  1. Kin K; Yasuhara T; Kameda M; Date I. Animal Models for Parkinson's Disease Research: Trends in the 2000s.. International journal of molecular sciences. 2019. PMID:31671557.
  2. Shen Z; Ma L; Mellick GD. Strategies for Optimizing Genetic Mouse Models to Enhance the Understanding of Parkinson's Disease.. Biomedicines. 2026. PMID:42193484.
  3. 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.
  4. 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.
  5. 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。
  6. 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。
  7. 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%。
  8. 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.
  9. ARRIVE 2.0 guidelines
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