Qingming Luo — Page 3 | Search PubMed

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Real-time detection of caspase-2 activation in a single living HeLa cell during cisplatin-induced apoptosis.

Caspase-2 is important for the mitochondrial apoptotic pathway, however, the mechanism by which caspase-2 executes apoptosis remains obscure. We carry out the first measurements of the dynamics of caspase-2 activation in a single living cell by a FRET (fluorescence resonance energy transfer) probe. Two FRET probes are constructed that each encoded a CRS (caspase-2 or caspase-3 recognition site) fused with a cyan fluorescent protein (CFP) and a red fluorescent protein (DsRed) (CFP-CRS-DsRed). Using these probes, we found that during cisplatin-induced apoptosis, caspase-2 activation occurred more slowly than did activation of caspase-3; additionally, caspase-2 activation was initiated much earlier than that of caspase-3.

Antineoplastic Agents↗

PubMedDate not supplied

In vivo optical reflectance imaging of spreading depression waves in rat brain with and without focal cerebral ischemia.

Spreading depression (SD) waves occur in focal cerebral ischemia of the brain. Optical reflectance imaging at 550 +/- 10-nm wavelength using a charge-coupled device (CCD) camera, called optical intrinsic signal imaging (OISI) in the neuroscience community, provides high resolution imaging of SD waves based on changes in blood perfusion. We present optical images of SD waves in normal rat brain induced by a pinprick, and the spontaneous SD waves that follow middle cerebral artery occlusion (MCAO). The images of change in reflectance are calculated as A = (I-I(o))I(o), where I is pixel intensity as some timepoint and I(o) is the initial intensity just prior to an SD wave. Difference images B = [I(i)-I(i-1)]I(o), where I(i) is the image at time i and I(i-1) is the previous image at time i-1 (a 6.4-s interval), significantly sharpen the boundaries between leading and trailing edges of the SD wave. Maximum rate-of-change images C = max(B) display the maximum pixel value of B within the duration of a single SD wave, and provide an image that visualizes the entire penumbra. The penumbra appear bright due to a rapid drop in perfusion, while the normal brain and infarct area appear dark.

Animals↗