Degeneration of dopaminergic neurons and impaired intracellular trafficking in Atp13a2 deficient zebrafish
Degeneration of dopaminergic neurons and impaired intracellular trafficking in Atp13a2 deficient zebrafish
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Atp13a2 缺陷斑马鱼的多巴胺能神经元退化和细胞内运输受损
DOI：
10.1016/j.ibror.2020.05.002
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发表时间：
2020
期刊：
影响因子：
2.6
通讯作者：
Matsui Hideaki
中科院分区：
文献类型：
--
作者：
Nyuzuki Hiromi;Ito Shinji;Nagasaki Keisuke;Nitta Yohei;Matsui Noriko;Saitoh Akihiko;Matsui Hideaki
ATP 13 A2是青少年型帕金森病（PARK 9，Parkinson's disease 9）的常染色体隐性遗传致病基因，也称为Kufor-Rakeb综合征。该疾病的特征是左旋多巴反应性帕金森综合征、核上性凝视麻痹、痉挛和痴呆。在此之前，我们已经报道了Atp 13 a2缺陷的青鳉鱼表现出多巴胺能神经变性和溶酶体功能障碍，表明溶酶体自噬损伤可能是帕金森病的关键发病机制之一。在这里，我们使用CRISPR/Cas9基因编辑建立了Atp 13 a2缺陷斑马鱼。我们发现后结节和蓝斑的TH +神经元数量明显减少（多巴胺能神经元，4个月时为64%，12个月时为37%，分别为P< 0.001和P < 0.05;去甲肾上腺素神经元，4个月时为52%，12个月时为40%，分别为p< 0.001和p < 0.05），证明了多巴胺能神经元的变性。此外，我们发现在Atp 13 a2缺陷的斑马鱼中，组织蛋白酶D蛋白表达降低（60%，p< 0.05）。透射电子显微镜分析使用中间间脑样品Atp 13 a2缺陷斑马鱼显示溶酶体样小体与囊泡积累和指纹样结构，表明溶酶体功能障碍。此外，在Atp 13 a2缺陷的斑马鱼中，通过液相色谱串联质谱，用囊泡与高尔基体融合注释的蛋白质表达显著降低（p< 0.001），表明细胞内运输受损。因此，我们得出结论，Atp 13 a2缺陷斑马鱼表现出多巴胺能神经元变性，溶酶体功能障碍和细胞内运输障碍的可能性，这可能是帕金森病的关键致病机制。
ATP13A2is the autosomal recessive causative gene for juvenile-onset Parkinson’s disease (PARK9, Parkinson’s disease 9), also known as Kufor-Rakeb syndrome. The disease is characterized by levodopa-responsive Parkinsonism, supranuclear gaze palsy, spasticity, and dementia. Previously, we have reported that Atp13a2 deficient medaka fish showed dopaminergic neurodegeneration and lysosomal dysfunction, indicating that lysosome-autophagy impairment might be one of the key pathogeneses of Parkinson’s disease. Here, we established Atp13a2 deficient zebrafish using CRISPR/Cas9 gene editing. We found that the number of TH + neurons in the posterior tuberculum and the locus coeruleus significantly reduced (dopaminergic neurons, 64 % at 4 months and 37 % at 12 months,p< 0.001 andp< 0.05, respectively; norepinephrine neurons, 52 % at 4 months and 40 % at 12 months,p< 0.001 andp< 0.05, respectively) in Atp13a2 deficient zebrafish, proving the degeneration of dopaminergic neurons. In addition, we found the reduction (60 %,p< 0.05) of cathepsin D protein expression in Atp13a2 deficient zebrafish using immunoblot. Transmission electron microscopy analysis using middle diencephalon samples from Atp13a2 deficient zebrafish showed lysosome-like bodies with vesicle accumulation and fingerprint-like structures, suggesting lysosomal dysfunction. Furthermore, a significant reduction (p< 0.001) in protein expression annotated with vesicle fusion with Golgi apparatus in Atp13a2 deficient zebrafish by liquid-chromatography tandem mass spectrometry suggested intracellular trafficking impairment. Therefore, we concluded that Atp13a2 deficient zebrafish exhibited degeneration of dopaminergic neurons, lysosomal dysfunction and the possibility of intracellular trafficking impairment, which would be the key pathogenic mechanism underlying Parkinson’s disease.
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影响因子：
56.9
作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle
影响因子：
46.9
作者：
通讯作者：
--
DOI：
10.1016/s0168-0102(00)81020-2
发表时间：
2000
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
M. Koike;H. Nakanishi;P. Saftig;J. Ezaki;K. Isahara;Y. Ohsawa;W. Schulz-Schaeffer;T. Watanabe-T.-Wat
通讯作者：
M. Koike;H. Nakanishi;P. Saftig;J. Ezaki;K. Isahara;Y. Ohsawa;W. Schulz-Schaeffer;T. Watanabe-T.-Wat
影响因子：
16.6
作者：
Bento CF;Ashkenazi A;Jimenez-Sanchez M;Rubinsztein DC
通讯作者：
Rubinsztein DC