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无尾目光滑爪蟾卵及胚胎胶膜的结构与大分子组成:(蛙胶膜/受精/糖蛋白)

Structure and Macromolecular Composition of the Egg and Embryo Jelly Coats of the Anuran Lepidobatrachus laevis: (frog jelly coat/fertilization/glycoprotein).

作者信息

Carroll Edward J, Wei Susan H, Nagel Glenn M, Ruibal Rodolfo

机构信息

Department of Biology, University of California, Riverside, California 92521.

Department of Chemistry and Biochemistry and Institute for Molecular Biology and Nutrition, California State University, Fullerton, California 92634.

出版信息

Dev Growth Differ. 1991 Feb;33(1):37-43. doi: 10.1111/j.1440-169X.1991.00037.x.

Abstract

Eggs and cleavage-stage embryos of the frog Lepidobatrachus laevis are encased by 3 μm thick vitelline/fertilization envelope and two jelly layers, termed J (innermost) and J (outermost). Based on light and transmission electron microscopy, J had a dense reticular appearance whereas J had a laminar structure. Direct dissolution of the jelly coats was accomplished by reduction of disulfide bonds with 0.08 M 2-mercaptoethanol at pH 10. Soluble jelly preparations were uncontaminated with nucleic acid (A /A =1.44) and yielded an average of 150 μg protein/egg or embryo (n=5). The biochemical composition of the jelly coats in unfertilized eggs was different from that in embryos. When examined via gel permeation chromatography, soluble jelly from unfertilized eggs contained macromolecules which were markedly larger and more heterogeneous (earlier eluting and broader peaks) than jelly from embryos. Differences in the components of jelly from unfertilized eggs and embryos were also observed by electrophoresis, however, a 29,700 molecular weight glycoprotein chain was common to both jelly preparations. The electrophoretic pattern of jelly obtained from parthenogenetically activated eggs was identical to that of unfertilized eggs, therefore the fertilization-associated changes are not due to the exclusive action of cortical granule products.

摘要

光滑爪蟾的卵和卵裂期胚胎被一层3μm厚的卵黄膜/受精膜以及两层胶膜包裹,这两层胶膜分别称为J(最内层)和J(最外层)。基于光学显微镜和透射电子显微镜观察,J呈现出致密的网状外观,而J具有层状结构。通过在pH 10的条件下用0.08 M的2-巯基乙醇还原二硫键,可直接溶解胶膜。可溶性胶膜制剂未被核酸污染(A260/A280 = 1.44),每个卵或胚胎平均可产生150μg蛋白质(n = 5)。未受精卵中胶膜的生化组成与胚胎中的不同。通过凝胶渗透色谱法检测时,未受精卵的可溶性胶膜中含有的大分子明显更大且更具异质性(洗脱更早且峰更宽),比胚胎的胶膜更明显。通过电泳也观察到未受精卵和胚胎的胶膜成分存在差异,然而,两种胶膜制剂中都存在一条分子量为29,700的糖蛋白链。孤雌激活卵的胶膜电泳图谱与未受精卵的相同,因此与受精相关的变化并非仅由皮质颗粒产物的作用引起。

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