Cellular and subcellular localization of Rab10 and phospho-T73 Rab10 in the mouse and human brain.
Cellular and subcellular localization of Rab10 and phospho-T73 Rab10 in the mouse and human brain.
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DOI：
10.1186/s40478-023-01704-9
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发表时间：
2023-12-18
影响因子：
7.1
通讯作者：
中科院分区：
文献类型：
--
作者：
富含亮氨酸重复序列激酶2（LRRK 2）的常染色体显性致病突变导致帕金森病（PD）。最常见的突变G2019 S-LRRK 2增加LRRK 2的激酶活性，导致其底物的过度磷酸化。这些底物之一Rab 10在保守的Thr 73残基（pRab 10）处被磷酸化，并且是在各种组织中表达的最丰富的LRRK 2 Rab GTP酶之一。Rab 10参与神经退行性疾病，包括PD和阿尔茨海默病，这使得精确定位Rab 10和pRab 10在大脑中的细胞和亚细胞定位成为理解其功能作用以及翻译后修饰如何影响功能的重要一步。为了确定抗体对Rab 10磷酸化形式（pRab 10）的特异性，将Rab 10特异性反义寡核苷酸脑室内注射到小鼠脑中。此外，Rab 10敲除诱导的神经元（从人诱导多能干细胞分化）用于测试pRab 10抗体特异性。为了放大pRab 10的弱免疫荧光信号，利用酪胺信号放大。Rab 10和pRab 10表达于皮层、纹状体和黑质背侧部。在G2019 S-LRRK 2敲入小鼠中，pRab 10的免疫荧光增强。神经元、星形胶质细胞、小胶质细胞和少突胶质细胞均表达Rab 10和pRab 10。Rab 10与内质网、溶酶体和trans-Golgi网络标记共定位，而pRab 10不定位于这些细胞器。然而，pRab 10确实与小鼠和人类皮质中突触前末端的标记物重叠，包括α-突触核蛋白。本研究的结果表明Rab 10和pRab 10在本研究中测试的所有脑区和细胞类型中表达，但pRab 10在突触前末端富集。由于Rab 10是LRRK 2激酶底物，PD中G2019 S-LRRK 2的激酶活性增加可能影响Rab 10介导的疾病神经元突触前末端的膜运输。在线版本包含补充材料，可通过10.1186/s40478-023-01704-9获得。
Autosomal dominant pathogenic mutations in Leucine-rich repeat kinase 2 (LRRK2) cause Parkinson’s disease (PD). The most common mutation, G2019S-LRRK2, increases the kinase activity of LRRK2 causing hyper-phosphorylation of its substrates. One of these substrates, Rab10, is phosphorylated at a conserved Thr73 residue (pRab10), and is one of the most abundant LRRK2 Rab GTPases expressed in various tissues. The involvement of Rab10 in neurodegenerative disease, including both PD and Alzheimer’s disease makes pinpointing the cellular and subcellular localization of Rab10 and pRab10 in the brain an important step in understanding its functional role, and how post-translational modifications could impact function. To establish the specificity of antibodies to the phosphorylated form of Rab10 (pRab10), Rab10 specific antisense oligonucleotides were intraventricularly injected into the brains of mice. Further, Rab10 knock out induced neurons, differentiated from human induced pluripotent stem cells were used to test the pRab10 antibody specificity. To amplify the weak immunofluorescence signal of pRab10, tyramide signal amplification was utilized. Rab10 and pRab10 were expressed in the cortex, striatum and the substantia nigra pars compacta. Immunofluorescence for pRab10 was increased in G2019S-LRRK2 knockin mice. Neurons, astrocytes, microglia and oligodendrocytes all showed Rab10 and pRab10 expression. While Rab10 colocalized with endoplasmic reticulum, lysosome and trans-Golgi network markers, pRab10 did not localize to these organelles. However, pRab10, did overlap with markers of the presynaptic terminal in both mouse and human cortex, including α-synuclein. Results from this study suggest Rab10 and pRab10 are expressed in all brain areas and cell types tested in this study, but pRab10 is enriched at the presynaptic terminal. As Rab10 is a LRRK2 kinase substrate, increased kinase activity of G2019S-LRRK2 in PD may affect Rab10 mediated membrane trafficking at the presynaptic terminal in neurons in disease. The online version contains supplementary material available at 10.1186/s40478-023-01704-9.
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影响因子：
15.1
作者：
Belluzzi E;Gonnelli A;Cirnaru MD;Marte A;Plotegher N;Russo I;Civiero L;Cogo S;Carrion MP;Franchin C;Arrigoni G;Beltramini M;Bubacco L;Onofri F;Piccoli G;Greggio E
通讯作者：
Greggio E
影响因子：
12.7
作者：
Fan Y;Nirujogi RS;Garrido A;Ruiz-Martínez J;Bergareche-Yarza A;Mondragón-Rezola E;Vinagre-Aragón A;Croitoru I;Gorostidi Pagola A;Paternain Markinez L;Alcalay R;Hickman RA;Düring J;Gomes S;Pratuseviciute N;Padmanabhan S;Valldeoriola F;Pérez Sisqués L;Malagelada C;Ximelis T;Molina Porcel L;Martí MJ;Tolosa E;Alessi DR;Sammler EM
通讯作者：
Sammler EM
影响因子：
5.3
作者：
Deng, Cai-Yun;Lei, Wen-Liang;Luo, Zhen-Ge
通讯作者：
Luo, Zhen-Ge
影响因子：
5.3
作者：
Beccano-Kelly DA;Kuhlmann N;Tatarnikov I;Volta M;Munsie LN;Chou P;Cao LP;Han H;Tapia L;Farrer MJ;Milnerwood AJ
通讯作者：
Milnerwood AJ
影响因子：
3.3
作者：
Glodowski, Doreen R.;Chen, Carlos Chih-Hsiung;Rongo, Christopher
通讯作者：
Rongo, Christopher