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氚标记胸腺嘧啶核苷([3H]-TdR)与无性繁殖绦虫沃氏中殖孔绦虫(同物异名:柯氏中殖孔绦虫)细胞命运的免疫细胞化学追踪

Tritiated thymidine ([3H]-TdR) and immunocytochemical tracing of cellular fate within the asexually dividing cestode Mesocestoides vogae (syn. M. corti).

作者信息

Smith A G, McKerr G

机构信息

Cancer and Ageing Research Group, University of Ulster at Coleraine, Co. Londonderry, Northern Ireland.

出版信息

Parasitology. 2000 Jul;121 ( Pt 1):105-10. doi: 10.1017/s0031182099006010.

DOI:10.1017/s0031182099006010
PMID:11085229
Abstract

This report documents the presence of an active thymidine kinase (TK) system within Mesocestoides vogae tetrathyridia as quantified by tritiated thymidine ([3H]-TdR) incorporation using liquid scintillation counting. A 100-fold increase in [3H]-TdR incorporation was observed at 37 degrees C when compared with its incorporation at 0 degrees C. Thymidine's competitive analogue, BrdU, competed for sites within newly replicated DNA. Immunohistochemical trials performed here using antibodies against BrdU identified cells that have entered and passed through S-phase. Positively stained nuclei were most numerous at the anterior tip of tetrathyridia especially within the ganglia, lesser numbers of these cells occurred along the growing commissure and amongst surface tegumental cytons suggesting that stem cells do not exist in one region but are found throughout the entire body. As M. vogae has no internal organ systems the major sites for cell proliferation are those exhibiting maximal cell recruitment and undergoing tissue repair. These results show that it is possible to monitor changes in the cell recruitment pattern within this cestode. Thus use of BrdU and immunohistochemistry demonstrates how spatial arrangement and cellular reorganization can be successfully traced within M. vogae.

摘要

本报告记录了通过使用液体闪烁计数法测定氚标记胸腺嘧啶核苷([3H]-TdR)掺入量来量化的沃氏中殖孔绦虫四膜幼虫体内活跃的胸苷激酶(TK)系统的存在情况。与0℃时的掺入量相比,在37℃时观察到[3H]-TdR掺入量增加了100倍。胸腺嘧啶核苷的竞争性类似物溴脱氧尿苷(BrdU)竞争新复制DNA中的位点。在此进行的免疫组织化学试验使用抗BrdU抗体鉴定出已进入并通过S期的细胞。阳性染色的细胞核在四膜幼虫的前端尤其是神经节内最多,沿着生长中的连合部和表面皮层细胞体中这些细胞的数量较少,这表明干细胞并非存在于一个区域,而是遍布整个身体。由于沃氏中殖孔绦虫没有内部器官系统,细胞增殖的主要部位是那些表现出最大细胞募集并正在进行组织修复的部位。这些结果表明,可以监测该绦虫体内细胞募集模式的变化。因此,使用BrdU和免疫组织化学证明了如何在沃氏中殖孔绦虫体内成功追踪空间排列和细胞重组情况。

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