Nass Richard, Hamza Iqbal
Vanderbilt University Medical Center, Nashville, Tennessee, USA.
Curr Protoc Toxicol. 2007 Feb;Chapter 1:Unit1.9. doi: 10.1002/0471140856.tx0109s31.
Significant limitations in vertebrate animal model systems include the time involved, the expense, the fact that in vitro results may not reflect live animal pathology, difficulties in transporting the toxin past the blood brain barrier, and the inability to identify the mechanism of action without some a priori knowledge of the toxin's target. The availability of the complete genome sequence of the nematode C. elegans, coupled with the worm's size, growth rate, ease of culturing, and the realization that basic biological mechanisms and disease processes between worms and humans are highly conserved, makes this genetically tractable model a remarkable opportunity to dissect and identify in vivo the cellular processes involved in toxin-induced cell dysregulation and death. This unit includes protocols for culturing worms on solid and axenic media and acute and chronic exposure parameters for Parkinson's disease-associated toxins and hemin chloride. These methods provide the groundwork for using this powerful model system to further elucidate and understand the molecular mechanisms involved in nutrition as well as toxicological responses relevant to human diseases.
脊椎动物模型系统存在显著局限性,包括所需时间、费用、体外实验结果可能无法反映活体动物病理学情况、将毒素运送通过血脑屏障存在困难,以及在对毒素靶点没有一些先验知识的情况下无法确定作用机制。线虫秀丽隐杆线虫完整基因组序列的可得性,再加上其体型、生长速度、易于培养,以及蠕虫与人类之间基本生物学机制和疾病过程高度保守这一认识,使得这个易于进行基因操作的模型成为剖析和在体内识别毒素诱导的细胞失调和死亡所涉及的细胞过程的绝佳机会。本单元包括在固体和无菌培养基上培养蠕虫的方案,以及帕金森病相关毒素和氯化血红素的急性和慢性暴露参数。这些方法为利用这个强大的模型系统进一步阐明和理解营养以及与人类疾病相关的毒理学反应所涉及的分子机制奠定了基础。