Ca2+ oscillation-inducing phospholipase C zeta expressed in mouse eggs is accumulated to the pronucleus during egg activation

Abstract Sperm-specific phospholipase C zeta (PLCζ) is known to induce intracellular Ca 2+ oscillations and egg activation when expressed in mouse eggs by injection of RNA encoding PLCζ. We investigated the expression level and spatial distribution of PLCζ in the egg in real time and in relation to the initiation and termination of Ca 2+ oscillations by monitoring fluorescence of a yellow fluorescent protein ‘Venus’ fused with PLCζ. Ca 2+ oscillations similar to those at fertilization were induced at 40–50 min after RNA injection, when expressed PLCζ reached 10–40 × 10 −15 g in the egg. PLCζ–Venus increased up to 3 h and attained a steady level at 4–5 h. Interestingly, PLCζ–Venus is accumulated to the pronucleus (PN) formed at 5–6 h and continuously increased there. Ca 2+ oscillations stopped in most eggs before initiation of the accumulation. A variant of PLCζ that lacks three EF hand domains was much less effective in induction of Ca 2+ oscillations and little accumulated in the pronucleus, indicating a critical role of those domains. The ability of the accumulation to the pronucleus qualifies PLCζ for a strong candidate of the Ca 2+ oscillation-inducing sperm factor, which is introduced into the ooplasm upon sperm–egg fusion and concentrated to the pronucleus after inducing egg activation.