Green J D, Glas P S, Cheng S D, Lynn J W
Department of Anatomy, Louisiana State University Medical Center, New Orleans 70112.
Mol Reprod Dev. 1990 Feb;25(2):177-85. doi: 10.1002/mrd.1080250211.
Eggs of the sea urchin Strongylocentrotus purpuratus were fertilized in normal and in several chloride-deficient sea waters ([ Cl-]: normal greater than isethionate greater than methyl sulfonate greater than bromide). The fertilization envelopes (FE) were thinner and failed to harden, and the characteristic I-T transition did not occur. The permeability of the experimental FEs, as determined by release of protein from the perivitelline space, increased in the order of decreasing [Cl-]. Release of the enzymes beta-1,3-glucanase and cortical granule protease were not significantly altered. On the other hand, release of ovoperoxidase was increased three to four times in bromide sea water. Furthermore, a dose-response was observed in varying concentrations of bromide-normal sea water. With decreasing chloride (increasing bromide) concentration, more ovoperoxidase activity was observed. Cytochemical localization of ovoperoxidase activity with diaminobenzidine revealed almost a total lack of staining of FEs from bromide-substituted sea water. The results suggest that in chloride-deficient sea waters protein incorporation into the nascent FE is impaired. At least in the case of bromide, the incorporation of ovoperoxidase into the nascent FE was also inhibited.
将紫海胆的卵在正常海水以及几种缺氯海水中受精([Cl-]:正常海水>羟乙基磺酸盐海水>甲磺酸盐海水>溴化物海水)。受精膜(FE)更薄且无法硬化,并且未出现特征性的I-T转变。通过卵周隙中蛋白质的释放来测定,实验受精膜的通透性按照[Cl-]降低的顺序增加。β-1,3-葡聚糖酶和皮质颗粒蛋白酶的释放没有显著变化。另一方面,在溴化物海水中,卵过氧化物酶的释放增加了三到四倍。此外,在不同浓度的溴化物-正常海水中观察到了剂量反应。随着氯化物浓度降低(溴化物浓度增加),观察到更多的卵过氧化物酶活性。用二氨基联苯胺对卵过氧化物酶活性进行细胞化学定位显示,来自溴化物替代海水的受精膜几乎完全没有染色。结果表明,在缺氯海水中,新生受精膜中蛋白质的掺入受到损害。至少在溴化物的情况下,卵过氧化物酶掺入新生受精膜也受到抑制。