Ca2+ oscillation-inducing phospholipase C zeta expressed in mouse eggs is accumulated to the pronucleus during egg activation.
Sperm-specific phospholipase C zeta (PLC zeta) is known to induce intracellular Ca(2+) oscillations and egg activation when expressed in mouse eggs by injection of RNA encoding PLC zeta. We investigated the expression level and spatial distribution of PLC zeta 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 PLC zeta. Ca(2+) oscillations similar to those at fertilization were induced at 40-50 min after RNA injection, when expressed PLC zeta reached 10-40 x 10(-15) g in the egg. PLC zeta-Venus increased up to 3 h and attained a steady level at 4-5 h. Interestingly, PLC zeta-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 PLC zeta 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 PLC zeta 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.
Yoda
Ayako
A
Department of Physiology, Tokyo Women's Medical University School of Medicine, Shinjuku-ku, Tokyo 162-8666, Japan.
Oda
Shoji
S
Shikano
Tomohide
T
Kouchi
Zen
Z
Awaji
Takeo
T
Shirakawa
Hideki
H
Kinoshita
Katsuyuki
K
Miyazaki
Shunichi
S
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D017930:Genes, Reporter; D007527:Isoenzymes / Q000378:metabolism; D051379:Mice; D010063:Ovum / Q000201:enzymology* / Q000378:metabolism; D054801:Phosphoinositide Phospholipase C; D054803:Phospholipase C delta; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D010738:Type C Phospholipases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism
D007527:Isoenzymes; D011993:Recombinant Fusion Proteins; D010738:Type C Phospholipases; D054801:Phosphoinositide Phospholipase C; D054803:Phospholipase C delta; C499234:Plcz1 protein, mouse; D002118:Calcium