Yamamoto T
Shikwa Gakuho. 1989 Oct;89(10):1549-603.
The present work used 2 groups of Wistar rats, weighing 100g each. Rats in the first group served as the controls; those in the second group were given 2 mg/kg of vinblastine in single intravenous injections. The animals were then fixed by perfusion with a mixture of 0.1% glutaraldehyde and 4% paraformaldehyde. After having been dissected out of the jaws, upper incisors were demineralized in EDTA and prepared into longitudinal sections (3 microns thick) for immunohistochemical demonstration of alpha-tubulin by means of indirect methods using an anti-alpha-tubulin monoclonal antibody as the primary antibody and peroxidase-labeled anti-mouse sheep IgG as the secondary antibody. 1. Control group. In controls, diffuse alpha-tubulin reaction was observed in the distal cytoplasm of inner enamel epithelial cells, differentiating ameloblasts, and ameloblasts in the stage of matrix formation. In the ameloblasts, the reaction was especially strong in the distal ends and Tomes processes in the matrix-formation stage. It gradually decreased until the transitional stage, in which the ameloblasts regained intense reaction to alpha-tubulin. Reaction to alpha-tubulin was generally low in the enamel-maturation stage in both smooth-ended and ruffle-ended ameloblasts but grew somewhat intense in the stage in which ferritines were precipitated in the cells. Although dental papilla cells reacted only faintly to alpha-tubulin, when they started differentiating into odontoblasts, reaction grew stronger in their distal end regions and in processes extending from the distal ends into the dentin. But this reaction decreased and ceased at a middle dentin level. Fairly high reaction was observed in the cells of the outer enamel epithelium, the stratum intermedium, the stellate reticulum, the papillary layer, the pulp and the dental sac. 2. Experimental group. Reaction to alpha-tubulin described in the preceding sections generally decreased in from 1 to 6 hours after vinblastine administration. Reaction recovery appeared to begin at from 12 to 24 hours and almost reached control degree by 48 hours after administration. Following the decrease of alpha-tubulin reaction in ameloblasts and odontoblasts, their shapes and polarities changed dramatically. In other cells, however, in spite of decreased alpha-tubulin reactions, no noticeable morphological alteration took place.
本研究使用了两组Wistar大鼠，每组大鼠体重均为100克。第一组大鼠作为对照组；第二组大鼠单次静脉注射2毫克/千克长春碱。然后用0.1%戊二醛和4%多聚甲醛的混合液灌注固定动物。从颌骨中取出后，上切牙在乙二胺四乙酸（EDTA）中脱矿，并制成纵向切片（3微米厚），以抗α-微管蛋白单克隆抗体为一抗，过氧化物酶标记的抗小鼠羊IgG为二抗，采用间接法进行α-微管蛋白的免疫组织化学显示。1. 对照组。在对照组中，在内釉上皮细胞、分化中的成釉细胞和基质形成阶段的成釉细胞的远端细胞质中观察到弥漫性的α-微管蛋白反应。在成釉细胞中，在基质形成阶段，远端和托姆斯突处的反应尤为强烈。反应逐渐减弱，直至过渡阶段，此时成釉细胞对α-微管蛋白的反应再次增强。在釉质成熟阶段，平滑端和成皱端的成釉细胞对α-微管蛋白的反应通常较低，但在细胞中铁蛋白沉淀的阶段反应有所增强。尽管牙乳头细胞对α-微管蛋白的反应很微弱，但当它们开始分化为成牙本质细胞时，其远端区域和从远端延伸至牙本质中的突起处的反应会增强。但这种反应在牙本质中层水平会减弱并停止。在外釉上皮细胞、中间层、星网状层、乳头层、牙髓和牙囊的细胞中观察到相当高的反应。2. 实验组。在长春碱给药后1至6小时内，上述各节所述的对α-微管蛋白的反应普遍降低。反应恢复似乎在给药后12至24小时开始，到48小时时几乎达到对照程度。在成釉细胞和成牙本质细胞中α-微管蛋白反应降低后，它们的形状和极性发生了显著变化。然而，在其他细胞中，尽管α-微管蛋白反应降低，但未发生明显的形态改变。
