NAADP and InsP3 play distinct roles at fertilization in starfish oocytes.
NAADP participates in the response of starfish oocytes to sperm by triggering the fertilization potential (FP) through the activation of a Ca2+ current which depolarizes the membrane to the threshold of activation of the voltage-gated Ca2+ channels. The aim of this study was to investigate whether this Ca2+ influx is linked to the onset of the concomitant InsP3-mediated Ca2+ wave by simultaneously employing Ca2+ imaging and single-electrode intracellular recording techniques. In control oocytes, the sperm-induced membrane depolarization always preceded by a few seconds the onset of the Ca2+ wave. Strikingly, the self-desensitization of NAADP receptors either abolished the Ca2+ response or resulted in abnormal oocyte activation, i.e., the membrane depolarization followed the Ca2+ wave and the oocyte was polyspermic. The inhibition of InsP3 signaling only impaired the propagation of the Ca2+ wave and shortened the FP. The duration of FP was also reduced in low-Na+ sea water. Finally, uncaged InsP3 produced a Ca2+ increase, which depolarized the membrane upon the activation of a Ca2+-sensitive cation current. These results support the hypothesis that Ca2+ entry during the NAADP-triggered FP is required for the onset of the Ca2+ wave at fertilization. The InsP3-mediated Ca2+ wave, in turn, may interact with the NAADP-evoked depolarization by activating a Ca2+-dependent Na+ entry.
Moccia
Francesco
F
Laboratory of Cell Signaling, Stazione Zoologica Anton Dohrn, Villa Comunale, 80121 Naples, Italy. moccia@szn.it
Nusco
Gilda A
GA
Lim
Dmitry
D
Kyozuka
Keiichiro
K
Santella
Luigia
L
D000818:Animals; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling; D004594:Electrophysiology; D005260:Female; D005306:Fertilization; D008564:Membrane Potentials; D009249:NADP / Q000031:analogs & derivatives* / Q000502:physiology; D009865:Oocytes / Q000378:metabolism* / Q000502:physiology; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000502:physiology*; D012964:Sodium / Q000378:metabolism; D013215:Starfish
D009249:NADP; C024376:NAADP; D012964:Sodium; D017853:Phosphotransferases (Alcohol Group Acceptor); C051038:Inositol 1,4,5-trisphosphate 3-kinase; D002118:Calcium