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Evaluation of Fluoro-Jade C Staining: Specificity and Application to Damaged Immature Neuronal Cells in the Normal and Injured Mouse Brain.

Neuroscience | 2020

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.

Pubmed ID: 31785351 RIS Download

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CB6-Tg(Gad1-EGFP)G42Zjh/J (tool)

RRID:IMSR_JAX:007677

Mus musculus with name CB6-Tg(Gad1-EGFP)G42Zjh/J from IMSR.

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Anti-Sox2 (antibody)

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Anti Iba1, Rabbit antibody (antibody)

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Sox-2 (Y-17) Antibody (antibody)

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Iba1 antibody (antibody)

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Goat polyclonal antibody against T (Brachyury) (NCBI Gene ID: 6862) (antibody)

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Anti-ACTIVE Caspase-3 Polyclonal Antibody, Unconjugated (antibody)

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Anti-Sox2 (antibody)

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This polyclonal targets SOX2

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Sox-2 (Y-17) Antibody (antibody)

RRID:AB_2286684

This polyclonal targets SOX2

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Anti Iba1, Rabbit antibody (antibody)

RRID:AB_839504

This polyclonal targets Iba1

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Iba1 antibody (antibody)

RRID:AB_2224402

This polyclonal targets AIF1

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Anti-Green Fluorescent Protein (antibody)

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This monoclonal targets Green Fluorescent Protein

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Goat polyclonal antibody against T (Brachyury) (NCBI Gene ID: 6862) (antibody)

RRID:AB_10014654

This polyclonal targets T (Brachyury) (NCBI Gene ID: 6862)

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Anti-ACTIVE Caspase-3 Polyclonal Antibody, Unconjugated (antibody)

RRID:AB_430875

This polyclonal targets ACTIVE& x00AE; Caspase-3

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