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Simultaneous visualization of both signaling cascade activity and end-point gene expression in single cells.

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  • المؤلفون: Weibrecht I;Weibrecht I; Grundberg I; Nilsson M; Söderberg O
  • المصدر:
    PloS one [PLoS One] 2011; Vol. 6 (5), pp. e20148. Date of Electronic Publication: 2011 May 25.
  • نوع النشر :
    Journal Article; Research Support, Non-U.S. Gov't
  • اللغة:
    English
  • معلومة اضافية
    • المصدر:
      Publisher: Public Library of Science Country of Publication: United States NLM ID: 101285081 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1932-6203 (Electronic) Linking ISSN: 19326203 NLM ISO Abbreviation: PLoS One Subsets: MEDLINE
    • بيانات النشر:
      Original Publication: San Francisco, CA : Public Library of Science
    • الموضوع:
      Gene Expression Regulation*/drug effects ; Signal Transduction*/drug effects; Molecular Imaging/*methods ; Single-Cell Analysis/*methods; Benzamides ; Cell Line ; Dual Specificity Phosphatase 6/genetics ; Humans ; Imatinib Mesylate ; Kinetics ; Ligands ; Phosphorylation/drug effects ; Piperazines/pharmacology ; Pyrimidines/pharmacology ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Receptor, Platelet-Derived Growth Factor beta/metabolism ; Time Factors
    • نبذة مختصرة :
      We have developed an approach for simultaneous detection of individual endogenous protein modifications and mRNA molecules in single cells in situ. For this purpose we combined two methods previously developed in our lab: in situ proximity ligation assay for the detection of individual protein interactions and -modifications and in situ detection of single mRNA molecules using padlock probes. As proof-of-principle, we demonstrated the utility of the method for simultaneous detection of phosphorylated PDGFRβ and DUSP6/MKP-3 mRNA molecules in individual human fibroblasts upon PDGF-BB stimulation. Further we applied drugs disrupting the PDGFRβ signaling pathway at various sites to show that this combined method can concurrently monitor the molecular effect of the drugs, i.e. inhibition of downstream signaling from the targeted node in the signaling pathway. Due to its ability to detect different types of molecules in single cells in situ the method presented here can contribute to a deeper understanding of cell-to-cell variations and can be applied to e.g. pinpoint effector sites of drugs in a signaling pathway.
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    • الرقم المعرف:
      0 (Benzamides)
      0 (Ligands)
      0 (Piperazines)
      0 (Pyrimidines)
      0 (RNA, Messenger)
      8A1O1M485B (Imatinib Mesylate)
      EC 2.7.10.1 (Receptor, Platelet-Derived Growth Factor beta)
      EC 3.1.3.48 (DUSP6 protein, human)
      EC 3.1.3.48 (Dual Specificity Phosphatase 6)
    • الموضوع:
      Date Created: 20110608 Date Completed: 20110922 Latest Revision: 20211020
    • الموضوع:
      20221213
    • الرقم المعرف:
      PMC3102075
    • الرقم المعرف:
      10.1371/journal.pone.0020148
    • الرقم المعرف:
      21647446