成年小鼠大脑中髓鞘形成少突胶质细胞的可视化。

IF 4.2 3区 医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Journal of Neurochemistry Pub Date : 2024-09-04 DOI:10.1111/jnc.16218
Kiichi Yokoyama, Yuichi Hiraoka, Yoshifumi Abe, Kenji F Tanaka
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引用次数: 0

摘要

少突胶质细胞(OL)从少突胶质细胞前体细胞(OPCs)分化而来,被认为会产生两个群体:前髓鞘化少突胶质细胞和髓鞘化少突胶质细胞。最近的单细胞 RNA 序列数据将这些群体细分为新形成(NFOLs)、髓鞘形成(MFOLs)和成熟(MOLs)少突胶质细胞。然而,哪个新提出的群体对应于髓鞘化前或髓鞘化的少突胶质细胞尚不清楚。我们重点研究了NFOL特异性长非编码少突胶质细胞1基因(LncOL1),并试图利用四环素可控基因诱导系统对LncOL1启动子控制下的NFOL进行标记。我们证明髓鞘前OLs表达LncOL1,而MFOL特异性基因Ctps不表达,这表明NFOLs对应于髓鞘前OLs,而MFOLs和MOLs对应于髓鞘OLs。随后,我们生成了一种LncOL1-tTA小鼠,其中在LncOL1转录起始位点下游插入了一个四环素转录激活剂(tTA)盒。通过将 LncOL1-tTA 小鼠与 tetO 报告小鼠杂交,我们产生了 LncOL1-tTA::tetO-黄色荧光蛋白(YFP)双转基因(LncOL1-YFP)小鼠。虽然 LncOL1 是非编码,但在 LncOL1-YFP 小鼠中检测到了 YFP,这表明 tTA 翻译成功。意想不到的是,我们发现 LncOL1-tTA 驱动的 YFP+ 细胞的形态与 LncOL1+ 髓鞘化前 OL 的形态不同,标记的细胞显示为髓鞘化 OL。我们通过其 RNA 表达证明,YFP 标记的 OL 是 MFOL,而不是 MOL。利用延迟诱导 YFP 的独特特性,我们试图确定 MFOLs 是否不断从 OPCs 中获得供应并分化为 MOLs,或者 MFOLs 是否暂停分化并在成人大脑中维持这一阶段。为了实现这一目标,我们用 X 射线照射 LncOL1-YFP 成年人大脑,以消耗正在分裂的 OPCs 及其后代。照射后,YFP标记的OL熄灭,表明成年OPC在一个时期内分化为MOL。我们建立了一个新的转基因小鼠品系,该品系可对MFOLs进行基因标记,为研究成体少突发生的动态提供了可靠的工具。
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Visualization of myelin-forming oligodendrocytes in the adult mouse brain.

Oligodendrocyte (OL) differentiation from oligodendrocyte precursor cells (OPCs) is considered to result in two populations: premyelinating and myelinating OLs. Recent single-cell RNA sequence data subdivided these populations into newly formed (NFOLs), myelin-forming (MFOLs), and mature (MOLs) oligodendrocytes. However, which newly proposed population corresponds to premyelinating or myelinating OLs is unknown. We focused on the NFOL-specific long non-coding oligodendrocyte 1 gene (LncOL1) and sought to label NFOLs under the control of the LncOL1 promoter using a tetracycline-controllable gene induction system. We demonstrated that LncOL1 was expressed by premyelinating OLs and that the MFOL-specific gene, Ctps, was not, indicating that NFOLs correspond to premyelinating OLs and that MFOLs and MOLs correspond to myelinating OLs. We then generated a LncOL1-tTA mouse in which a tetracycline transactivator (tTA) cassette was inserted downstream from the LncOL1 transcription initiation site. By crossing the LncOL1-tTA mice with tetO reporter mice, we generated LncOL1-tTA::tetO-yellow fluorescent protein (YFP) double-transgenic (LncOL1-YFP) mice. Although LncOL1 is non-coding, YFP was detected in LncOL1-YFP mice, indicating successful tTA translation. Unexpectedly, we found that the morphology of LncOL1-tTA-driven YFP+ cells was distinct from that of LncOL1+ premyelinating OLs and that the labeled cells instead appeared as myelinating OLs. We demonstrated from their RNA expression that YFP-labeled OLs were MFOLs, but not MOLs. Using the unique property of delayed YFP induction, we sought to determine whether MFOLs are constantly supplied from OPCs and differentiate into MOLs, or whether MFOLs pause their differentiation and sustain this stage in the adult brain. To achieve this objective, we irradiated adult LncOL1-YFP brains with X-rays to deplete dividing OPCs and their progeny. The irradiation extinguished YFP-labeled OLs, indicating that adult OPCs differentiated into MOLs during a single period. We established a new transgenic mouse line that genetically labels MFOLs, providing a reliable tool for investigating the dynamics of adult oligodendrogenesis.

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来源期刊
Journal of Neurochemistry
Journal of Neurochemistry 医学-神经科学
CiteScore
9.30
自引率
2.10%
发文量
181
审稿时长
2.2 months
期刊介绍: Journal of Neurochemistry focuses on molecular, cellular and biochemical aspects of the nervous system, the pathogenesis of neurological disorders and the development of disease specific biomarkers. It is devoted to the prompt publication of original findings of the highest scientific priority and value that provide novel mechanistic insights, represent a clear advance over previous studies and have the potential to generate exciting future research.
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