TY - JOUR
T1 - Mammalian phospholipase C zeta induces oocyte activation from the sperm perinuclear matrix
AU - Fujimoto, S
AU - Yoshida, N
AU - Fukui, T
AU - Amanai, M
AU - Isobe, T
AU - Itagaki, C
AU - Izumi, T
AU - Perry, Anthony C F
PY - 2004
Y1 - 2004
N2 - Mammalian sperm-borne oocyte activating factor (SOAF) induces oocyte activation from a compartment that engages the oocyte cytoplasm, but it is not known how. A SOAF-containing extract (SE) was solubilized from the submembrane perinuclear matrix, a domain that enters the egg. SE initiated activation sufficient for full development. Microinjection coupled to tandem mass spectrometry enabled functional correlation profiling of fractionated SE without a priori assumptions about its chemical nature. Phospholipase C-zeta (PLCzeta) correlated absolutely with activating ability. Immunoblotting confirmed this and showed that the perinuclear matrix is the major site of 72-kDa PLCzeta. Oocyte activation was efficiently induced by 1.25 fg of sperm PLCzeta, corresponding to a fraction of one sperm equivalent (similar to0.03). Immunofluorescence microscopy localized sperm head PLCzeta to a post-acrosomal region that becomes rapidly exposed to the ooplasm following gamete fusion. This multifaceted approach suggests a mechanism by which PLCzeta originates from an oocyte-penetrating assembly the sperm perinuclear matrix-to induce mammalian oocyte activation at fertilization.
AB - Mammalian sperm-borne oocyte activating factor (SOAF) induces oocyte activation from a compartment that engages the oocyte cytoplasm, but it is not known how. A SOAF-containing extract (SE) was solubilized from the submembrane perinuclear matrix, a domain that enters the egg. SE initiated activation sufficient for full development. Microinjection coupled to tandem mass spectrometry enabled functional correlation profiling of fractionated SE without a priori assumptions about its chemical nature. Phospholipase C-zeta (PLCzeta) correlated absolutely with activating ability. Immunoblotting confirmed this and showed that the perinuclear matrix is the major site of 72-kDa PLCzeta. Oocyte activation was efficiently induced by 1.25 fg of sperm PLCzeta, corresponding to a fraction of one sperm equivalent (similar to0.03). Immunofluorescence microscopy localized sperm head PLCzeta to a post-acrosomal region that becomes rapidly exposed to the ooplasm following gamete fusion. This multifaceted approach suggests a mechanism by which PLCzeta originates from an oocyte-penetrating assembly the sperm perinuclear matrix-to induce mammalian oocyte activation at fertilization.
UR - http://dx.doi.org/10.1016/j.ydbio.2004.07.025
U2 - 10.1016/j.ydbio.2004.07.025
DO - 10.1016/j.ydbio.2004.07.025
M3 - Article
SN - 0012-1606
VL - 274
SP - 370
EP - 383
JO - Developmental Biology
JF - Developmental Biology
IS - 2
ER -