Structural and molecular biological study of duplication of centrioles during ciliogenesis

在大多数动物细胞中，高尔基体位于核外区，几乎固定在中心体周围。中心体由一对中心粒和中心体基质组成，使微管成核。TGN 38和golgin-97等高尔基体基质蛋白在细胞周期中定位于中心体，而高尔基体58 K蛋白在中心体的定位尚未见报道。在本研究中，我们研究了高尔基体58 K蛋白在MDCK，KD，HeLa细胞和人和实验动物的输卵管柱状上皮中的定位。用结构和细胞生物学方法研究了高尔基体蛋白在MDCK、KD和HeLa细胞中心粒的定位，发现高尔基体58 K抗体阳性。高尔基体58 K在整个细胞周期中始终与中心体结合。高尔基体58 K与中心体的结合不受秋水仙胺或布雷菲德菌素A处理的干扰。冷处理对高尔基体58 K与中心体的结合也没有影响。输卵管上皮主要由纤毛细胞和分泌细胞组成，有或无初级纤毛。位于柱状上皮细胞顶端的中心体和基底体延伸的纤毛被Golgi 58 K染色阳性。这些观察结果表明，中心粒保持在含有高尔基体共同分子的基质内，即使在迁移到顶端细胞质和/或纤毛形成后。高尔基体基质蛋白可能在纤毛发生过程中对中心粒的复制起重要作用。

In most animal cells, the Golgi apparatus is located in the juxtanuclear region and appears almost stationary around the centrosome. The centrosome, consisting of a pair of centrioles and the centrosomal matrix, nucleates microtubules. Several Golgi matrix proteins including TGN 38 and golgin-97 have been reported to be localized to the centrosome throughout the cell cycle, but localization of Golgi 58K protein to the centrosome has not been studied as yet. In the present study, we investigated the localization of Golgi 58K protein in MDCK, KD, and HeLa cells and in the oviductal columnar epithelium of human and experimental animals. Localization of Golgi proteins to centrioles was investigated with structural and cell biological methods.The centrosome was positively stained with anti Golgi 58K antibodies in MDCK, KD and HeLa cells. Golgi 58K remained associated with the centrosome throughout the cell cycle. Association of Golgi 58K to the centrosome was not disturbed by the colcemid or brefeldin A treatment. Cold treatment also had no effect on the association of Golgi 58K to the centrosome. The oviduct epithelium mainly consisted of ciliated cells and secretory cells with or without primary cilia. The centrosome located in the apical region of columnar epithelial cells and basal bodies extending cilia were positively stained with Golgi 58K. These observations indicate that centrioles are maintained within the matrix containing common molecules to the Golgi apparatus, even after the migration to the apical cytoplasm and or formation of cilia. Golgi matrix proteins might have important roles on duplication of centrioles during ciliogenesis.