Karoumi A, Meiniel R, Belin M F, Meiniel A
Laboratoire de Biochimie médicale, CJF INSERM 8806, Faculté de Médecine, Clermont-Ferrand, France.
J Neural Transm Gen Sect. 1991;86(3):205-16. doi: 10.1007/BF01250706.
To extend our previous immunochemical investigations in the chick embryo (Karoumi et al., 1990 b), we raised antibodies in the rabbit against crude extracts of the subcommissural organ (SCO) of the bovine. The antiserum labeled A99 was absorbed by crude brain extracts and its specificity was tested by different techniques. Comparison of crude SCO and cerebral hemispheres supernatants after immunoblotting allow to identify specific 98, 60, 52, 42, 38, and 32 kDa polypeptides in the SCO profile. Immunoaffinity chromatography on A99 immunoadsorbent of crude SCO, cerebral hemispheres (CH) and classical ependyma (CE) supernatants was followed by electrophoretical analysis and electrotransfer. Concanavalin A (Con A) and wheat germ agglutinin (WGA) labeling procedures demonstrated the presence of numerous glycopeptides specific of crude SCO supernatants and having an apparent molecular weight ranging from 240 to 50 kDa. In the CH-eluted fraction, 50 and 52 kDa glycopeptides were revealed by ConA and WGA, whereas in the CE-immunopurified fraction no band was visualized. The similarity of the chick embryo and bovine electrophoretic pattern corresponding to the SCO eluted fractions speaks in favour of a high degree of conservation of the SCO secretory material and an evolutionary stability of the antigens recognized by A99IgG.
为了扩展我们之前在鸡胚中的免疫化学研究（Karoumi等人，1990年b），我们用兔制备了针对牛连合下器官（SCO）粗提物的抗体。标记为A99的抗血清用脑粗提物进行吸收，并通过不同技术测试其特异性。免疫印迹后对SCO粗提物和大脑半球上清液进行比较，可在SCO图谱中鉴定出特异性的98、60、52、42、38和32 kDa多肽。对SCO粗提物、大脑半球（CH）和经典室管膜（CE）上清液在A99免疫吸附剂上进行免疫亲和层析，然后进行电泳分析和电转移。刀豆球蛋白A（Con A）和麦胚凝集素（WGA）标记程序表明，SCO粗提物上清液中存在大量糖肽，其表观分子量范围为240至50 kDa。在CH洗脱组分中，ConA和WGA显示出50和52 kDa的糖肽，而在CE免疫纯化组分中未观察到条带。与SCO洗脱组分相对应的鸡胚和牛的电泳图谱相似，这表明SCO分泌物质具有高度保守性，且A99IgG识别的抗原具有进化稳定性。
