POWER SPECTRAL ANALYSIS OF THE SHAPE OF fMLP-STIMULATED GRANULOCYTES. A TOOL FOR THE STUDY OF CYTOSKELETAL ORGANIZATION UNDER NORMAL AND PATHOLOGICAL CONDITIONS

Abstract

ABSTRACTfMLP (N-formyl-methionyl-leucyl-phenylalanine) is a powerful activator of granulocytes, eliciting different metabolic responses, such as generation of reactive oxygen species, production-dates printpubdate="05/03/02" of arachidonic acid metabolites, and release of lysosomal enzymes. fMLP determines also a dramatic rearrangement of the actin cytoskeleton; under non-gradient conditions this entails characteristic alterations in cell shape (chemokinesis), while under gradient conditions it is instrumental in promoting cell migration up the gradient (chemotaxis). Here we analyze mathematically the cell contour of fMLP-stimulated human granulocytes stimulated with fMLP under non-gradient conditions, using the methods for study of stochastic series. The cell contours were drawn and divided into 200 segments of equal linear length and the angles between consecutive segments were computed. The derived series of angles were examined for autocorrelations and from the autocorrelation function the power spectrum was calculated. Our results show that the pattern of lamellipodial extensions of the cell membrane is not entirely randomly-designed, but it is partly regulated by deterministic components, as revealed by the presence of statistically significant periodicities. Soon after fMLP stimulation, the power spectrum of the cell contours exhibits a single distinct peak at frequency 0.07, indicating a prevalence of prominent lamellipodia, each one covering in the average 1/15 of the linearized cell contour. Some 30 min after fMLP stimulation the power spectrum becomes flatter (indicating a general decrement of the deterministic component), but still presents one single peak; the latter is shifted to the right (frequency 0.13), indicating the prevalence of less prominent and regular, but more numerous, protrusions, each one covering 1/20 to 1/30 of the cell contour.