Real-Time Dynamics of Ca2+, Caspase-3/7, and Morphological Changes in Retinal Ganglion Cell Apoptosis under Elevated Pressure

摘要: Quantitative information on the dynamics of multiple molecular processes in individual live cells under controlled stress is central to understanding cell behavior interest and establishment reliable models. Here, apoptosis regulator intracellular Ca(2+), effector caspase-3/7, morphological changes, as well temporal correlation between them at single level, are examined retinal gangling line (differentiated RGC-5 cells) undergoing elevated hydrostatic pressure using a custom-designed imaging platform that allows long-term real-time simultaneous molecular-level physiological changes large numbers (beyond field-of-view typical microscopy) pressure. This examination revealed Ca(2+) elevation with transient or peaks less than 0.5 hour duration appearing early stages (typically 5 hours after onset 100 mmHg pressure) followed by gradual caspase-3/7 activation late later hours). The data reveal strong peak occurrence neurite retraction body shrinkage. suggests elevation, through its impact ion channel activity water efflux, likely responsible for apoptotic changes. Moreover, show significant cell-to-cell variation activation, an inevitable consequence stochastic nature underlying biochemical reactions not captured conventional assays based population-averaged cellular responses. study provides, first time, statistically level enable refinements testing models mitochondria-mediated apoptosis. Further, developed approach has direct significance variety signaling pathway phenomena.