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2019 | 79 | 3 |
Tytuł artykułu

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

Autorzy
Warianty tytułu
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.
Słowa kluczowe
EN
Wydawca
-
Rocznik
Tom
79
Numer
3
Opis fizyczny
p.302-308,fig.,ref.
Twórcy
autor
  • Department of Neurology, The Second Hospital of Hebei Medical University, Hebei, China
autor
  • Department of Neurosciences, Georgetown University Medical Center, Washington, USA
  • Washington Institute for Health Sciences, Arlington, USA
autor
  • Department of Neurology, The Second Hospital of Hebei Medical University, Hebei, China
autor
  • Department of Neurology, The Second Hospital of Hebei Medical University, Hebei, China
autor
  • Department of Human Anatomy, Hebei Medical University, Hebei, China
  • Neuroscience Research Center, Medicine and Health Institute, Hebei Medical University, Hebei, China
autor
  • Department of Neurology, The Second Hospital of Hebei Medical University, Hebei, China
  • Neuroscience Research Center, Medicine and Health Institute, Hebei Medical University, Hebei, China
Bibliografia
  • An WF, Tolliday N (2010) Cell‑based assays for high‑throughput screening. Mol Biotechnol 45: 180–186.
  • Bergmann O, Spalding KL, Frisén J (2015) Adult neurogenesis in humans. Cold Spring Harb Perspect Biol 7: a018994.
  • Boldrini  M, Fulmore CA, Tartt AN, Simeon LR, Pavlova I, Poposka  V, Rosoklija GB, Stankov A, Arango V, Dwork AJ, Hen R, Mann JJ (2018) Hu‑ man hippocampal neurogenesis persists throughout aging. Cell Stem Cell 22: 589–599.
  • Chen H, Tung YC, Li B, Iqbal K, Grundke‑Iqbal I (2007) Trophic factors coun‑ teract elevated FGF‑2‑induced inhibition of adult neurogenesis. Neuro‑ biol Aging 28: 1148–1162.
  • Guo W, Patzlaff NE, Jobe EM, Zhao X (2012) Isolation of multipotent neural stem or progenitor cells from both the dentate gyrus and subventricular zone of a single adult mouse. Nat Protoc 7: 2005–2012.
  • Han MH, Lee EH, Koh SH (2016) Current opinion on the role of neurogen‑ esis in the therapeutic strategies for Alzheimer disease, Parkinson dis‑ ease, and ischemic stroke; considering neuronal voiding function. Int Neurourol J 20: 276–287.
  • Iqbal K, Grundke‑Iqbal I (2011) Opportunities and challenges in developing Alzheimer disease therapeutics. Acta Neuropathol 122: 543–549.
  • Kohl TO, Ascoli CA (2017) Direct and indirect cell‑based enzyme‑linked im‑ munosorbent assay. Cold Spring Harb Protoc 2017(5). doi: 10.1101.
  • Kuhn HG, Eisch AJ, Spalding K, Peterson DA (2016) Detection and pheno‑ typic characterization of adult neurogenesis. Cold Spring Harb Perspect Biol 8: a025981.
  • Liu Q, Wu C, Cai H, Hu N, Zhou J, Wang P (2014) Cell‑based biosensors and their application in biomedicine. Chem Rev 114: 6423–6461.
  • Miller BR, Hen R (2015) The current state of the neurogenic theory of de‑ pression and anxiety. Curr Opin Neurobiol 30: 51–58.
  • Nierode G, Kwon PS, Dordick JS, Kwon SJ (2016) Cell‑based assay design for high‑content screening of drug candidates. J Microbiol Biotechnol 26: 213–225.
  • Sun T, Wang XJ, Xie SS, Zhang DL, Wang XP, Li BQ, Ma  W, Xin H (2011) A comparison of proliferative capacity and passaging potential between neural stem and progenitor cells in adherent and neurosphere cultures. Int J Dev Neurosci 29: 723–731.
  • Toda T, Parylak SL, Linker SB, Gage FH (2019) The role of adult hippo‑ campal neurogenesis in brain health and disease. Mol Psychiatry 24: 67–87.
  • Urbán N, Guillemot F (2014) Neurogenesis in the embryonic and adult brain: same regulators, different roles. Front Cell Neurosci 8: 396.
  • Winner B, Winkler J (2015) Adult neurogenesis in neurodegenerative dis‑ eases. Cold Spring Harb Perspect Biol 7: a021287.
  • Zang R, Li D, Tang IC, Wang J, Yang ST (2012) Cell‑based assays in high‑throughput screening for drug discovery. Int J Biotechnol Wellness Ind 1: 31–51.
  • Zhang J, Jiao J (2015) Molecular biomarkers for embryonic and adult neural stem cell and neurogenesis. Biomed Res Int 2015: 727542.
Typ dokumentu
Bibliografia
Identyfikatory
Identyfikator YADDA
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