Capacitated mammalian spermatozoa undergo a fusion response of their head plasma membrane and the outer acrosomal membrane leading to vesiculation classically known as acrosome reaction. Acrosome reaction occurs in response to various acrosome reaction inducers including zona pellucida proteins, calcium ionophore, dibutyryl cAMP, progesterone, etc. All the acrosome reaction inducers cause a transient of calcium influx into the sperm through voltage-dependent cation channels. Efflux of chloride, stimulation of activity of phospholipases, and phosphorylation of proteins are other known changes introduced by acrosome reaction inducers. Macromolecular organization and dynamics of sperm membranes during the progression of this vesiculation are largely unexplored. In this study, we report that progesterone induced the formation of horizontal microdomains within the exofacial surfaces of sperm membranes, which showed progressive and independent alterations in molecular dynamics. In the light of this observation, we propose that sperm membrane rafts may contain both horizontal and vertical microdomains. (C) 2004 Elsevier Inc. All rights reserved.