滇蓝尾蝾螈前肢再生的形态学研究

发布时间：2018-03-14 00:30

本文选题：滇蓝尾蝾螈　切入点：肢体　出处：《昆明医学院》2009年硕士论文　论文类型：学位论文

【摘要】： [目的]:建立再生生物学研究的理想模型,观察滇蓝尾蝾螈前肢肢体的割处再生能力,通过组织形态学观察,了解局部细胞的去分化现象及其细胞来源。 [方法]:取野生成体雌性及雄性滇蓝尾蝾螈共15条,麻醉下自肘部稍下切除右侧前肢,术后3、7、12、15、20、30、45天切取创部新生组织,通过大体形态、组织切片观察肢体的再生以及再生过程中局部组织细胞的形态学变化和再生组织的细胞来源。 [结果]:所有实验动物均健康成活,被切除的右前肢均能再生出缺损的肢体,其过程如下:术后3天,形成创面表皮封闭,也称为顶帽或顶外胚层帽(apicalectodermal cap),顶帽下方存在一群圆形的单个核细胞沿残肢近远中轴生长,同时可见狭长的骨骼肌细胞及成纤维细胞向远端移行,有些细胞胞核可见特异肌源性蛋白Myogenin阳性表达;术后7～15天天,顶帽下方形成再生芽基(blastema),细胞呈现幼稚化,核仁明显,细胞体小,核质比变大,Myogenin阳性表达增多;大约20～30天天,芽基细胞转分化形成成骨细胞,骨骼肌细胞等,部分成骨细胞胞核有Myogenin阳性表达,最终约45天细胞再分化构成一个新的肢体,只是新生肢体较原切除前肢在色泽大小上略有差异。 [结论]:(1)滇蓝尾蝾螈前肢具有完全的“割处再生”能力,可作为再生生物学研究的理想动物模型;(2)滇蓝尾蝾螈前肢的割处再生是邻近组织如骨骼肌细胞迁移、去分化、增殖并再分化的结果;(3)滇蓝尾蝾螈前肢再生芽基中,骨骼肌源性特异性蛋白Myogenin的表达为强阳性,提示骨骼肌细胞是再生芽基的细胞来源之一。
[Abstract]:[objective] to establish an ideal model of regenerative biology, to observe the regeneration ability of the forelimb of salamander, and to understand the dedifferentiation of local cells and the origin of the cells by histomorphological observation. [methods] Fifteen wild adult female and male salamanders were removed from the right forelimb slightly below the elbow under anesthesia. After 3 days of operation, the neovascularization tissues of the trauma region were cut out at 30 and 45 days after operation, and the gross morphology was adopted. Tissue sections were used to observe the regeneration of limbs, the morphological changes of local histocytes and the origin of regenerated tissues. [results]: all the experimental animals survived healthily, and all the right forelimbs resected could regenerate the defective limbs. The process was as follows: 3 days after operation, the wound epidermis was closed. Also known as the top cap or apical ectodermal cap, a group of circular mononuclear cells grow along the distal axis of the remnant limb beneath the cap, and long narrow skeletal muscle cells and fibroblasts can be seen migrating to the distal end. Positive expression of specific myogenic protein Myogenin was found in some cell nuclei. After 7 days, 15 days after operation, regenerated blastocyst was formed under the top cap. The cells were immature, nucleoli were obvious, the cell body was small, and the positive expression of Myogenin was increased with the increase of the ratio of nucleus to cytoplasm; about 20 to 30 days after operation, the positive expression of Myogenin was found in the nucleus of some cells. Budding cells were transformed into osteoblasts, skeletal muscle cells and other osteoblasts. Some osteoblasts showed Myogenin positive expression in their nuclei. Finally, the cells were redifferentiated for about 45 days to form a new limb. However, the color of the new limb was slightly different from that of the original forelimb. [conclusion] the forelimb of the salamander has the ability to regenerate completely and can be used as an ideal animal model for the study of regenerative biology. The regeneration of the forelimb of the salamander is the migration and dedifferentiation of adjacent tissues such as skeletal muscle cells. The expression of skeletal myogenic specific protein (Myogenin) was strongly positive in the regenerated bud of the forelimb of Salamander, suggesting that skeletal muscle cells are one of the cell sources of regenerated bud.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R-332

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