Complement C5a Alters the Membrane Potential of Neutrophils during Hemorrhagic Shock

Figure 3

(a) Change in the shape and granularity of PMN after C5a stimulation. Influence of 1 minute C5a 100 ng/ml exposure on SSC-A (indicator for granularity) and FSC-A (indicator for changes in cell shape) on human PMN. (b) C5a-induced change in cell shape cannot be achieved by forced depolarization. Change in FSC-A for human PMN (n = 10–12) after incubation with 10 minutes in buffers plus 1 minute of C5a 100 ng/ml. (c) Independency of cell shape from membrane potential. Difference of membrane potential in human PMN subgroups (n = 10–12). Shown is the relation between the 25% upper and lower part of cells in relation to the average fluorescence. (d) C5a-induced change in cell granularity cannot be achieved by forced depolarization. Change in SSC-A for human PMN (–12) after incubation with 10 minutes in buffer plus 1 minute of C5a 100 ng/ml minutes in total. (e) Independency of cell granularity from membrane potential. Difference of membrane potential in human PMN subgroups (–12). Shown is the relation between the 25% upper and lower part of cells in relation to the average fluorescence. (f) Representative illustration of determination of membrane potential. DiBAC₄(3)’s fluorescence was measured in the FITC channel. Each sample consists of Ctrl (PMN in K4.5 = light gray), PMN exposed to C5a 100 ng/ml in K4.5 for one minute (gray), and PMN incubated in K15 (dark gray). , , and .

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