Ikenari, Takuya Department of Biological Regulation, School of Health Science, Faculty of Medicine, Tottori University
Kurata, Hirofumi Department of Biological Regulation, School of Health Science, Faculty of Medicine, Tottori University / Division of Child Neurology, Department of Brain and Neurosciences, Faculty of Medicine, Tottori University / Department of Pediatrics, National Hospital Organization, Kumamoto Saishunso National Hospital Researchers DB KAKEN
Satoh, Takemasa Division of Neurobiology, School of Life Sciences, Faculty of Medicine, Tottori University Researchers DB KAKEN
Hata, Yoshio Division of Neurobiology, School of Life Sciences, Faculty of Medicine, Tottori University / Division of Integrative Bioscience, Institute of Regenerative Medicine and Biofunction, Tottori University Graduate School of Medical Sciences Researchers DB KAKEN
neural stem/precursor cells
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.
Copyright © 2019 IBRO. Published by Elsevier Ltd. All rights reserved. This manuscript version is made available under the CC-BY-NC-ND 4.0 license https://creativecommons.org/licenses/by-nc-nd/4.0/
Ikenari, T., Kurata, H., Satoh, T., Hata, Y., & Mori, T. (2020). Evaluation of Fluoro-Jade C Staining: Specificity and Application to Damaged Immature Neuronal Cells in the Normal and Injured Mouse Brain. Neuroscience, 425, 146-156.
Faculty of Medicine/Graduate School of Medical Sciences/University Hospital