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BCL-x(L)-dependent light scattering by apoptotic cells

  1. Author:
    Boustany, N. N.
    Tsai, Y. C.
    Pfister, B.
    Joiner, W. M.
    Oyler, G. A.
    Thakor, N. V.
  2. Author Address

    Rutgers State Univ, Dept Biomed Engn, Piscataway, NJ 08854 USA. NCI, Lab Prot Dynam & Signaling, Frederick, MD 21701 USA. Univ Penn, Sch Med, Dept Neurosurg, Philadelphia, PA 19104 USA. Johns Hopkins Univ, Dept Biomed Engn, Baltimore, MD USA. Univ Maryland, Sch Med, Dept Neurol, Baltimore, MD 21201 USA. Baltimore VA Med Ctr, Res Serv, Baltimore, MD USA Boustany, NN, Rutgers State Univ, Dept Biomed Engn, 617 Bowser Rd, Piscataway, NJ 08854 USA
    1. Year: 2004
    2. Date: DEC
  1. Journal: Biophysical Journal
    1. 87
    2. 6
    3. Pages: 4163-4171
  2. Type of Article: Article
  1. Abstract:

    We measured the intensity ratio of wide-to-narrow angle scatter, optical scatter image ratio (OSIR), in single cells during apoptosis and after overexpression of the mitochondria-bound antiapoptotic protein lBCL-x(L). OSIR is sensitive to particle size/shape for objects with wavelength-scale dimensions, and was used as a morphometric measure of cellular response. Three cell variants were treated with staurosporine (STS): nontransfected parental CSM14.1, CSM14.1 stably expressing yellow fluorescent protein (YFP) with diffuse YFP fluorescence, and apoptosis-resistant CSM14.1 stably expressing the fusion protein construct YFP-BCL-x(L) with YFP fluorescence localized on the mitochondria. After treatment with 1 or 2 muM STS, the measured OSIR decreased monotonically by similar to25% in the nontransfected and YFP variants, and reached a steady-state value 40 - 60 min after STS treatment. The decrease in OSIR at the onset of apoptosis preceded phosphatidyl serine exposure by 5 h. In the YFP-BCL- x(L) cell variant, the initial OSIR was already similar to24% lower than the initial OSIR in YFP and nontransfected cells, and only decreased by <10% after STS treatment. Alterations in light scattering by cells overexpressing BCL-x(L) even before apoptosis induction raise interesting questions as to the role of BCL-x(L) in conferring apoptosis resistance by preconditioning the cells and possibly altering mitochondrial morphology

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External Sources

  1. WOS: 000225426700051

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