Carroll E J, Cohen J S
Department of Biology, University of California, Riverside 92521.
Mol Reprod Dev. 1990 Jan;25(1):77-86. doi: 10.1002/mrd.1080250114.
The ultrastructure of fertilization envelope (FE) development and the polypeptide spectra of Strongylocentrotus franciscanus and S. droebachiensis envelopes were compared to S. purpuratus. In S. franciscanus, the FE reached its maximum thickness of 67 nm by 3 minutes postinsemination (PI), and final structuralization was observed by 40 minutes PI. The fully formed FE did not have microvillar impressions (casts) and was symmetrical, with outer double laminar elements surrounding an amorphous central region. Isolated S. franciscanus FEs were soluble in reducing and denaturing solvents and the same set of 33 polypeptides ranging from 18.5 to 260 kD was detected in FEs isolated from 10 to 180 minutes PI. The S. droebachiensis FE retained microvillar casts, assumed its definitive form by 3 minutes PI, and was 70 nm thick between microvillar impressions. Isolated S. droebachiensis FEs were partially soluble in reducing and denaturing solvents, and the polypeptide spectra of FEs isolated between 10 and 60 minutes PI were identical and showed 14 polypeptides from 18.5 to 265 kD. Antisera against extracted FEs and the FE extract from S. purpuratus were immunologically cross-reactive (using an enzyme-linked immunosorbent assay) with S. franciscanus and S. droebachiensis FE preparations; immunoblots identified 13 and 5 cross-reactive polypeptides, respectively. Most of the cross-reactive polypeptides were of slightly different molecular weight. Based on comparative ultrastructural, solubility, and electrophoretic data, we suggest that S. droebachiensis FE development is most like that observed in S. purpuratus.
将加州紫海胆、大西洋紫海胆和球海胆受精膜(FE)发育的超微结构以及其多肽谱与紫球海胆进行了比较。在加州紫海胆中,受精膜在授精后3分钟时达到其最大厚度67纳米,并在授精后40分钟时观察到最终的结构化。完全形成的受精膜没有微绒毛印记(铸型),并且是对称的,其外部双层元素围绕着一个无定形的中心区域。分离得到的加州紫海胆受精膜可溶于还原和变性溶剂,并且在从授精后10至180分钟分离得到的受精膜中检测到了相同的一组分子量在18.5至260千道尔顿之间的33种多肽。球海胆的受精膜保留了微绒毛铸型,在授精后3分钟时呈现其最终形态,并且在微绒毛印记之间的厚度为70纳米。分离得到的球海胆受精膜部分可溶于还原和变性溶剂,并且在授精后10至60分钟分离得到的受精膜的多肽谱是相同的,显示出14种分子量在18.5至265千道尔顿之间的多肽。针对提取的受精膜和来自紫球海胆的受精膜提取物制备的抗血清与加州紫海胆和球海胆的受精膜制剂具有免疫交叉反应性(使用酶联免疫吸附测定法);免疫印迹分别鉴定出13种和5种交叉反应性多肽。大多数交叉反应性多肽的分子量略有不同。基于比较超微结构、溶解性和电泳数据,我们认为球海胆受精膜的发育最类似于在紫球海胆中观察到的情况。