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生成 Armcx1 条件性敲除小鼠。

Generation of an Armcx1 Conditional Knockout Mouse.

机构信息

University Program in Genetics and Genomics, Duke University Graduate School, Durham, North Carolina, USA.

Department of Ophthalmology, Duke University School of Medicine, Durham, North Carolina, USA.

出版信息

Genesis. 2024 Aug;62(4):e23615. doi: 10.1002/dvg.23615.

DOI:10.1002/dvg.23615
PMID:39139090
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11364276/
Abstract

Armadillo repeat-containing X-linked protein-1 (Armcx1) is a poorly characterized transmembrane protein that regulates mitochondrial transport in neurons. Its overexpression has been shown to induce neurite outgrowth in embryonic neurons and to promote retinal ganglion cell (RGC) survival and axonal regrowth in a mouse optic nerve crush model. In order to evaluate the functions of endogenous Armcx1 in vivo, we have created a conditional Armcx1 knockout mouse line in which the entire coding region of the Armcx1 gene is flanked by loxP sites. This Armcx1 line was crossed with mouse strains in which Cre recombinase expression is driven by the promoters for β-actin and Six3, in order to achieve deletion of Armcx1 globally and in retinal neurons, respectively. Having confirmed deletion of the gene, we proceeded to characterize the abundance and morphology of RGCs in Armcx1 knockout mice aged to 15 months. Under normal physiological conditions, no evidence of aberrant retinal or optic nerve development or RGC degeneration was observed in these mice. The Armcx1 mouse should be valuable for future studies investigating mitochondrial morphology and transport in the absence of Armcx1 and in determining the susceptibility of Armcx1-deficient neurons to degeneration in the setting of additional heritable or environmental stressors.

摘要

Armcx1 蛋白是一种跨膜蛋白,其包含多个 Armadillo 重复序列,但目前其功能仍不清楚。该蛋白在神经元中能够调控线粒体的运输,其过表达能够诱导胚胎神经元的轴突生长,并能促进视神经挤压模型中小鼠视网膜神经节细胞(RGC)的存活和轴突再生。为了研究 Armcx1 蛋白在体内的功能,我们构建了条件性 Armcx1 基因敲除小鼠,该小鼠的 Armcx1 基因编码区被loxP 位点所包围。利用 Armcx1 条件性敲除小鼠分别与表达 Cre 重组酶的β-actin 和 Six3 启动子小鼠杂交,实现了 Armcx1 基因在小鼠体内的敲除。通过基因敲除确认后,我们进一步研究了 15 月龄的 Armcx1 基因敲除小鼠中 RGC 的数量和形态。在正常生理条件下,这些小鼠的视网膜或视神经发育以及 RGC 变性均无异常。该 Armcx1 基因敲除小鼠将有助于进一步研究 Armcx1 缺失对线粒体形态和运输的影响,以及确定 Armcx1 缺失神经元在其它遗传或环境应激因素下易变性的机制。

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本文引用的文献

1
Comprehensive single-cell atlas of the mouse retina.小鼠视网膜的综合单细胞图谱。
iScience. 2024 May 8;27(6):109916. doi: 10.1016/j.isci.2024.109916. eCollection 2024 Jun 21.
2
Armcx1 Reduces Neurological Damage Via a Mitochondrial Transport Pathway Involving Miro1 After Traumatic Brain Injury.Armcx1 通过创伤性脑损伤后涉及 Miro1 的线粒体转运途径减少神经损伤。
Neuroscience. 2024 May 3;545:111-124. doi: 10.1016/j.neuroscience.2024.03.009. Epub 2024 Mar 16.
3
Armcx1 attenuates secondary brain injury in an experimental traumatic brain injury model in male mice by alleviating mitochondrial dysfunction and neuronal cell death.
Armcx1 通过减轻线粒体功能障碍和神经元细胞死亡来减轻雄性小鼠实验性颅脑损伤模型中的二次脑损伤。
Neurobiol Dis. 2023 Aug;184:106228. doi: 10.1016/j.nbd.2023.106228. Epub 2023 Jul 15.
4
Roles of syntaphilin and armadillo repeat-containing X-linked protein 1 in brain injury after experimental intracerebral hemorrhage.突触结合蛋白和富含装甲蛋白重复序列的 X 连锁蛋白 1 在实验性脑出血后脑损伤中的作用。
Neurosci Lett. 2023 Jul 13;809:137300. doi: 10.1016/j.neulet.2023.137300. Epub 2023 May 13.
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Progressive optic atrophy in a retinal ganglion cell-specific mouse model of complex I deficiency.一种复合体 I 缺陷的视网膜神经节细胞特异性小鼠模型中的进行性视神经萎缩。
Sci Rep. 2020 Oct 1;10(1):16326. doi: 10.1038/s41598-020-73353-0.
6
Miro: A molecular switch at the center of mitochondrial regulation.Miro:线粒体调控中心的分子开关。
Protein Sci. 2020 Jun;29(6):1269-1284. doi: 10.1002/pro.3839. Epub 2020 Feb 24.
7
Formation of retinal direction-selective circuitry initiated by starburst amacrine cell homotypic contact.由星状双极细胞同源接触引发的视网膜方向选择性电路的形成。
Elife. 2018 Apr 3;7:e34241. doi: 10.7554/eLife.34241.
8
Mitochondrial dynamics, transport, and quality control: A bottleneck for retinal ganglion cell viability in optic neuropathies.线粒体动态、运输和质量控制:视神经病变中视网膜神经节细胞存活的瓶颈。
Mitochondrion. 2017 Sep;36:186-192. doi: 10.1016/j.mito.2017.08.014. Epub 2017 Sep 1.
9
The Mammalian-Specific Protein Armcx1 Regulates Mitochondrial Transport during Axon Regeneration.哺乳动物特有的蛋白质Armcx1在轴突再生过程中调节线粒体运输。
Neuron. 2016 Dec 21;92(6):1294-1307. doi: 10.1016/j.neuron.2016.10.060.
10
Number of People Blind or Visually Impaired by Glaucoma Worldwide and in World Regions 1990 - 2010: A Meta-Analysis.1990 - 2010年全球及世界各地区因青光眼致盲或视力受损的人数:一项荟萃分析。
PLoS One. 2016 Oct 20;11(10):e0162229. doi: 10.1371/journal.pone.0162229. eCollection 2016.