Eng L A, Metz C B
J Exp Zool. 1980 May;212(2):159-67. doi: 10.1002/jez.1402120202.
Sperm nucleus decondensing activity (NDA) of eggs has been investigated using the gametes of the sea urchin, Lytechinus variegatus. To assay for NDA, isolated sperm heads were exposed to homogenate from mature unfertilized ova and then examined with phase-contrast optics. NDA is retained by dialysis and requires Ca++ at the time of homogenization as well as at the time of assay. Other divalent cations (Mg++, Sr++, Ba++, Mn++) can substitute for Ca++ but are much less effective. NDA is stable to cold (-20 degrees C, -76 degrees C) but not to heat (56 degrees C). The pH activity optimum is in the range 7-9. NDA is present in the supernatant after centrifugation at 150,000 g for 105 minutes. It elutes shortly following the void volume on a Sephacryl S-200 column, with an approximate M.W. of 100,000 daltons.
已利用多斑刺海胆的配子对卵子的精子核解聚活性(NDA)进行了研究。为了测定NDA,将分离的精子头部暴露于成熟未受精卵的匀浆中,然后用相差显微镜进行检查。NDA可通过透析保留,在匀浆时以及测定时都需要Ca++。其他二价阳离子(Mg++、Sr++、Ba++、Mn++)可以替代Ca++,但效果要差得多。NDA对低温(-20℃、-76℃)稳定,但对高温(56℃)不稳定。pH活性最适范围在7-9之间。在150,000 g下离心105分钟后,NDA存在于上清液中。它在Sephacryl S-200柱上的空体积后不久洗脱,分子量约为100,000道尔顿。