Kuhns W J, Bramson S, Simpson T L, Burkart W, Jumblatt J, Burger M M
Eur J Cell Biol. 1980 Dec;23(1):73-9.
Specific rabbit antisera were prepared against purified aggregation factor and its membrane-associated receptor, baseplate, derived from the marine sponge. Microciona prolifera. They were utilized in conjunction with fluorescent-labeled goat anti-rabbit IgG in an assay to demonstrate the surface localizations of both components. The specificity of antibody preparations for AF and BP was demonstrated through inhibition of the rotation-mediated assay by homotypic antibody. This study confirms the presence of aggregation factor on the surface of disaggregated sponge cells maintained in the presence of the divalent cations, Ca++ and Mg++, and its absence when cells are maintained in Ca++ and Mg++-free seawater. The location of BP could also be demonstrated on the cell surface. Aggregation factors and baseplate appear to be heavily distributed on archeocytes and choanocytes, but are localized less intensely on gray cells. Gray cells are typified by yellowish autofluorescence of their intracellular granules in stained and control preparations. The reaction of anti-Microciona aggregation factor with its homotypic factor appeared to be species specificity judged by immunofluorescence assays and by inhibition of rotation-mediated assay by anti-homotypic AF since antibodies prepared against heterotypic AF preparations were unreactive.
制备了针对从海洋海绵微拟球藻中纯化的聚集因子及其膜相关受体基板的特异性兔抗血清。它们与荧光标记的山羊抗兔IgG一起用于测定，以证明这两种成分的表面定位。通过同型抗体对旋转介导测定的抑制，证明了抗体制备物对聚集因子（AF）和基板（BP）的特异性。本研究证实，在存在二价阳离子Ca++和Mg++的情况下维持的分散海绵细胞表面存在聚集因子，而在无Ca++和Mg++的海水中维持细胞时则不存在。基板的位置也可以在细胞表面得到证明。聚集因子和基板似乎大量分布在原细胞和领细胞上，但在灰色细胞上的定位不太强烈。灰色细胞的典型特征是在染色和对照制剂中其细胞内颗粒呈现淡黄色自发荧光。通过免疫荧光测定以及抗同型AF对旋转介导测定的抑制判断，抗微拟球藻聚集因子与其同型因子的反应似乎具有种属特异性，因为针对异型AF制剂制备的抗体没有反应。
