التفاصيل البيبلوغرافية
| العنوان: |
On-line monitoring of apoptosis in insulin-secreting cells. |
| المؤلفون: |
Köhler, Martin, Zaitsev, Sergei V., Zaitseva, Irina I., Leibiger, Barbara, Leibiger, Ingo B., Turunen, Mikael, Kapelioukh, Iouri L., Bakkman, Linda, Appelskog, Ioulia B., de Monvel, Jacques Boutet, Imreh, Gabriela, Berggren, Per-Olof, Köhler, Martin |
| المصدر: |
Diabetes; Dec2003, Vol. 52 Issue 12, p2943-2950, 8p, 2 Black and White Photographs, 2 Diagrams, 2 Graphs |
| مصطلحات موضوعية: |
APOPTOSIS, INSULIN, SECRETION, PROTEINS |
| مستخلص: |
Apoptosis was monitored in intact insulin-producing cells both with microfluorometry and with two-photon laser scanning microscopy (TPLSM), using a fluorescent protein based on fluorescence resonance energy transfer (FRET). TPLSM offers three-dimensional spatial information that can be obtained relatively deep in tissues. This provides a potential for future in vivo studies of apoptosis. The cells expressed a fluorescent protein (C-DEVD-Y) consisting of two fluorophores, enhanced cyan fluorescent protein (ECFP) and enhanced yellow fluorescent protein (EYFP), linked by the amino acid sequence DEVD selectively cleaved by caspase-3-1ike proteases. FRET between ECFP and EYFP in C-DEVD-Y could therefore be monitored on-line as a sensor of caspase-3 activation. The relevance of using caspase-3 activation to indicate β-cell apoptosis was demonstrated by inhibiting caspase-3-1ike proteases with Z-DEVD-fmk and thereby showing that caspase-3 activation was needed for high-glucose- and cytokine-induced apoptosis in the β-cell and for staurosporine-induced apoptosis in RINm5F cells. In intact RINm5F cells expressing C-DEVD-Y and in MIN6 cells expressing the variant C-DEVD-Y2, FRET was lost at 155 ± 23 min (n = 9) and 257 ± 59 min (n = 4; mean ± SE) after activation of apoptosis with staurosporine (6 µmol/l), showing that this method worked in insulinproducing cells. [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
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