Intracellular injections of a soluble sperm factor trigger calcium oscillations and meiotic maturation in unfertilized oocytes of a marine worm

How sperm trigger activating calcium transients in eggs remains a central, unresolved question in fertilization biology. To determine if a soluble sperm factor can generate a fertilization-like calcium response in the absence of sperm-egg binding, aqueous extracts of sperm from the nemertean worm Cerebratulus lacteus were mixed with Ca2+-sensitive fluorescent dyes and injected into unfertilized, metaphase-I-arrested oocytes. Based on confocal imaging analyses, unfertilized oocytes that had been injected with sperm extract routinely produced oscillating Ca2+ waves and resumed meiotic maturation in a manner that closely resembled normal fertilization. Calcium oscillations and maturation were typically lacking in control oocytes that had been (i) injected with buffer alone or with buffer containing added calcium, (ii) given external treatments of the sperm factor, or (iii) injected with extracts made from cells other than sperm. Boiling or protease treatment essentially abolished the potency of the sperm extract, and nonboiled extracts retained full activity in >10-kDa fractions, but not in <10-kDa fractions. Collectively, such findings suggest that the sperm of C. lacteus possess a soluble protein that can bypass oolemmal surface receptors to act within the ooplasm as a trigger of repetitive Ca2+ waves and meiotic maturation. Results obtained in this study are discussed with respect to the minimum amount of extract needed for egg activation and whether the oscillogenic substance is sufficiently concentrated in a single sperm to play a biological role during fertilization. (C) 1997 Academic Press.