Moccia Francesco, Lim Dmitry, Kyozuka Keiichiro, Santella Luigia
Laboratory of Cell Biology, Stazione Zoologica Anton Dohrn, Villa Comunale I-80121, Naples, Italy.
Cell Calcium. 2004 Dec;36(6):515-24. doi: 10.1016/j.ceca.2004.05.004.
In invertebrates oocytes or eggs, the fertilization or activation potential establishes the fast electrical block to polyspermy and, in some species, provides the Ca2+ influx which contributes to the following intracellular Ca2+ wave. In echinoderms, the molecule triggering the activation potential is still unknown. The aim of this study was to assess whether nicotinic acid-adenine dinucleotide phosphate (NAADP) elicited the fertilization potential in starfish oocytes. The changes in membrane potential induced by the sperm were measured in oocytes held at a low resting potential, so that the Ca2+-action potential was inactivated and only the initial slower depolarization caused by the sperm could be studied. Decreasing extracellular Na+ concentration did not prevent the onset of the fertilization potential, while removal of external Ca2+ abolished it. The pre-incubation with SK&F 96365 and verapamil and the pre-injection of BAPTA inhibited the fertilization potential, while the injection of heparin only reduced its duration. The biophysical and pharmacological properties of the sperm-elicited depolarization were similar to those displayed by the NAADP-activated Ca2+-mediated current recently described in starfish oocytes. Indeed, the desensitization of NAADP-receptors prevented the onset of the fertilization potential. Taken together, these data suggest that NAADP could trigger the fertilization potential in starfish oocytes.
在无脊椎动物的卵母细胞或卵子中,受精或激活电位建立了对多精入卵的快速电阻滞,并且在某些物种中,提供了Ca2+内流,这有助于随后的细胞内Ca2+波。在棘皮动物中,触发激活电位的分子仍然未知。本研究的目的是评估烟酰胺腺嘌呤二核苷酸磷酸(NAADP)是否能引发海星卵母细胞的受精电位。在保持低静息电位的卵母细胞中测量精子诱导的膜电位变化,这样Ca2+动作电位就被灭活了,从而只能研究由精子引起的最初较慢的去极化。降低细胞外Na+浓度并不能阻止受精电位的产生,而去除细胞外Ca2+则使其消失。用SK&F 96365和维拉帕米预孵育以及预先注射BAPTA可抑制受精电位,而注射肝素仅缩短其持续时间。精子诱导的去极化的生物物理和药理学特性与最近在海星卵母细胞中描述的NAADP激活的Ca2+介导电流所显示的特性相似。事实上,NAADP受体的脱敏阻止了受精电位的产生。综上所述,这些数据表明NAADP可能触发海星卵母细胞的受精电位。