Evaluation of Fluoro-Jade C Staining: Specificity and Application to Damaged Immature Neuronal Cells in the Normal and Injured Mouse Brain
Evaluation of Fluoro-Jade C Staining: Specificity and Application to Damaged Immature Neuronal Cells in the Normal and Injured Mouse Brain
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DOI：
10.1016/j.neuroscience.2019.11.029
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发表时间：
2020-01-15
期刊：
影响因子：
3.3
通讯作者：
Mori, Tetsuji
中科院分区：
文献类型：
--
作者：
Ikenari, Takuya;Kurata, Hirofumi;Mori, Tetsuji
荧光素C(FJC)染色被广泛应用于所有变性成熟神经元的特异性检测，包括凋亡、坏死和自噬细胞。然而，FJC染色是否能检测到退化的未成熟神经元和神经干细胞/前体细胞仍不清楚。此外，一些相互矛盾的研究表明，FJC及其祖先染料FJ和FJB可以标记静息/激活的星形胶质细胞和小胶质细胞。在本研究中，我们检验了FJC染色在正常和损伤条件下检测成年和胚胎小鼠脑内神经细胞的有效性。在成年啮齿动物脑室下区(SVZ)-吻侧迁移流(RMS)-嗅球(OB)系统中，正常情况下会发生与神经发生相关的细胞凋亡。使用该系统，我们检测到FCJ阳性(+)细胞，其中一些是双皮质激素(DCX)(+)神经母细胞，以及神经元核(Neun)(+)成熟神经元。FJC阴性(-)细胞表达活化的Caspase3，正常脑内可见少量FJC(+)电离钙结合适配器分子1(Iba1)(+)小胶质细胞和FJC(+)/胶质纤维酸性蛋白(GFAP)(+)星形胶质细胞。接下来，我们分析了胚胎脑中神经干细胞/前体细胞的凋亡，分别在胚胎第14天或第10天给予N-乙基-N-亚硝脲(ENU)或乙醇。FJC(+)神经干细胞/前体细胞和表达SRY相关HMG-box 2(SOX2)的神经上皮细胞。令人惊讶的是，退化的间质细胞也是FJC(+)。本研究表明，FJC是各分化阶段变性神经细胞的可靠标记物。但在某些条件下，FJC也能标记变性的非神经细胞。(C)2019 IBRO。爱思唯尔有限公司出版。保留所有权利。
Fluoro-Jade C (FJC) staining is widely used for the specific detection of all degenerating mature neurons, including apoptotic, necrotic, and autophagic cells. However, whether FJC staining can detect degenerating immature neurons and neural stem/precursor cells remains unclear. In addition, some conflicting studies have shown that FJC and its ancestral dyes, Fluoro-Jade (FJ) and FJB, can label resting/activated astrocytes and microglia. In the present study, we examined the validity of FJC staining for the detection of neuronal cells in adult and embryonic mouse brains under normal and injured conditions. In the adult rodent subventricular zone (SVZ)-rostral migratory stream (RMS)-olfactory bulb (OB) system, apoptosis associated with neurogenesis occurs under normal conditions. Using this system, we detected FCJ positive (+) cells, some of which were doublecortin (DCX)(+) neuroblasts, in addition to neuronal nuclei (NeuN)(+) mature neurons. FJC negative (-) apoptotic cells expressing activated Caspase 3 were also observed, and a small number of FJC(+)ionized calcium-binding adaptor molecule 1 (Iba1)(+) microglia and FJC(+)/glial fibrillary acidic protein (GFAP)(+) astrocytes were observed in the normal brain. Next, we analyzed embryonic brains, in which the apoptosis of neural stem/precursor cells was induced by the administration of N-ethyl-N-nitrosourea (ENU) or ethanol at embryonic day 14 or 10, respectively. In those brains, FJC(+) neural stem/precursor cells and neuroepithelial cells expressing SRY-related HMG-box 2 (Sox2) were observed. Surprisingly degenerating mesenchymal cells were also FJC(+). The present study indicates that FJC is a reliable marker for degenerating neuronal cells during all differentiation stages. However, FJC could also label degenerating non-neuronal cells under some conditions. (C) 2019 IBRO. Published by Elsevier Ltd. All rights reserved.