Cell‑based immunofluorescence assay for screening the neurogenesis potential of new drugs in adult hippocampal neural progenitor cells

• Autorzy: Zhang K. , Li B. , Li P. , Yang X. , Cui H. , Liu X.
• Języki publikacji: EN

Abstrakty

EN

Preclinical studies have suggested that increased adult neurogenesis in the hippocampus might have potential therapeutic effects for Alzheimer’s disease and depression; therefore, it is a target for the treatment of some brain diseases. In this technical communication, we propose a cell‑based fluorescence assay to study the neurogenesis of adult hippocampal progenitor cells that can be used for high‑throughput screening of drugs promoting neurogenesis. Three fluorescent dyes (DAPI, Alexa Fluor 488, and Alexa Fluor 594) and a fluorescence spectrophotometry reader were used, which confirmed that the mutual interference of the three fluorescent dyes is very low. We used this cell‑based fluorescence assay to evaluate the effects of three neurotrophic factors, ciliary neurotrophic factor (CNTF), insulin‑like growth factor 1 (IGF‑1), and IGF‑2 on the promotion of neurogenesis in adult hippocampal neural progenitor cells. The fluorescence intensity ratio of the neuronal marker, class III β‑tubulin, to the housekeeping protein, glyceraldehyde 3‑phosphate dehydrogenase, or nuclear staining dye, DAPI, in CNTF‑treated cells was significantly higher than in control cells. The ratios in IGF‑1‑ and IGF‑2‑treated cells were slightly higher under higher cell density conditions. These results are consistent with those in previous reports; therefore, this report proved the efficacy of this method. Taken together, the results showed that this simple, rapid, and economical cell‑based immunofluorescence assay could be a powerful tool for the rapid screening of drugs that promote adult neurogenesis.

• Wydawca: -
• Czasopismo: Acta Neurobiologiae Experimentalis
• Rocznik: 2019
• Tom: 79
• Numer: 3
• Opis fizyczny: p.302-308,fig.,ref.

Twórcy

autor

Zhang K.

• Department of Neurology, The Second Hospital of Hebei Medical University, Hebei, China

autor

Li B.

• Department of Neurosciences, Georgetown University Medical Center, Washington, USA
• Washington Institute for Health Sciences, Arlington, USA

autor

Li P.

• Department of Neurology, The Second Hospital of Hebei Medical University, Hebei, China

autor

Yang X.

• Department of Neurology, The Second Hospital of Hebei Medical University, Hebei, China

autor

Cui H.

• Department of Human Anatomy, Hebei Medical University, Hebei, China
• Neuroscience Research Center, Medicine and Health Institute, Hebei Medical University, Hebei, China

autor

Liu X.

• Department of Neurology, The Second Hospital of Hebei Medical University, Hebei, China
• Neuroscience Research Center, Medicine and Health Institute, Hebei Medical University, Hebei, China

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Identyfikatory

DOI

10.21307/ane‑2019‑028