Active bleb formation is abated in Lytechinus variegatus red spherule coelomocytes after disruption of acto-myosin contractility.

Red spherule coelomocytes are immune cells in the sea urchin Lytechinus variegatus that have been characterized as motile O(2) transport cells. Video microscopy of living red spherule coelomocytes reveals a constitutive, dynamic array of cellular morphologies and movements. Cells continuously send out and retract membrane blebs all over the cell surface as part of their normal cellular physiology. Disruption of microtubules by perfusion with either nocodazole or taxol had no effect on bleb formation or motility. Perfusion with cytochalasin B abated bleb formation and revealed cells that exhibited multiple small spheres attached by short membrane extensions. Attenuation of blebbing and intracellular organelle motility were restored by washing out with cytochalasin B. Treatment with phalloidin also abated bleb formation and revealed a smooth, spherical cellular morphology. The effects of phalloidin were completely reversible after washout. Red spherule coelomocytes treated with blebbistatin rounded up with an irreversible retraction of blebs into surface blebs that were greatly reduced in size, number and motility. Normal cell surface bleb formation and intracellular organelle motility were not restored after washout of the drug. These results indicate that the acto-myosin contractile mechanism contributes to the dynamics of constitutive cell surface membrane blebbing in invertebrate immune cells.