Gonatas J O, Gonatas N K, Stieber A, Louvard D
Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia 19104-6079.
J Neurochem. 1987 Nov;49(5):1498-506. doi: 10.1111/j.1471-4159.1987.tb01020.x.
An antiserum was raised against fractions of the Golgi apparatus of neurons from rat brain. Immunoblots of these fractions with the antiserum showed two principal bands of 185 and 150 kilodaltons (kd) in apparent molecular mass. The antiserum reacted with five or six bands of 200, 150, 130, 100-110, 64, and 40 kd in apparent molecular mass in immunoblots of several crude brain membrane fractions. Affinity-purified antibodies from the different gel bands transferred to nitrocellulose paper were used in immunoblot and immunocytochemical studies. Antibodies eluted from the 200-, 150-, 100-110-, and 64-kd bands reacted not only with the corresponding band but also with the other three bands. Antibodies eluted from the 40-kd band stained only the corresponding band. On light and/or electron microscopic immunocytochemistry, the antiserum stained the Golgi apparatus of rat neurons, glia, liver, and kidney tubule cells. Weaker, segmented, and less consistent staining was observed in nuclear envelopes, rough endoplasmic reticulum, and plasma membranes of neurons. Antibodies eluted from the bands at 200, 150, 100-110, and 64 kd stained intermediate cisterns of the Golgi apparatus of neurons. These findings suggest that a group of related polypeptides of brain membranes is preferentially expressed or enriched in the Golgi apparatus of neurons. Polypeptides with apparent molecular masses of 185 and 150 kd probably represent moieties endogenous to membranes of the neuronal Golgi apparatus.
制备了针对大鼠脑神经元高尔基体组分的抗血清。用该抗血清对这些组分进行免疫印迹分析，结果显示出两条主要条带，表观分子量分别为185和150千道尔顿（kd）。在几种粗制脑膜组分的免疫印迹中，该抗血清与表观分子量为200、150、130、100 - 110、64和40 kd的五条或六条条带发生反应。将从不同凝胶条带转移至硝酸纤维素膜上的亲和纯化抗体用于免疫印迹和免疫细胞化学研究。从200 - kd、150 - kd、100 - 110 - kd和64 - kd条带洗脱的抗体不仅与相应条带反应，还与其他三条条带反应。从40 - kd条带洗脱的抗体仅对相应条带染色。在光镜和/或电镜免疫细胞化学实验中，该抗血清对大鼠神经元、神经胶质细胞、肝细胞和肾小管细胞的高尔基体进行了染色。在神经元的核膜、粗面内质网和质膜中观察到较弱、呈节段性且不太一致的染色。从200、150、100 - 110和64 kd条带洗脱的抗体对神经元高尔基体的中间扁平囊进行了染色。这些发现表明，一组相关的脑膜多肽在神经元高尔基体中优先表达或富集。表观分子量为185和150 kd的多肽可能代表神经元高尔基体膜的内源性部分。
