Gandhi S, Santelli J, Mitchell D H, Stiles J W, Sanadi D R
Mech Ageing Dev. 1980 Feb;12(2):137-50. doi: 10.1016/0047-6374(80)90090-1.
This paper describes a technique capable of establishing and maintaining large, age-synchronous populations of the nematode Caenorhabditis elegans. The technique has three essential components: a rich chemical medium; a method for producing and harvesting mass quantities of eggs; and 5-fluorodeoxyuridine (FUdR), an inhibitor of DNA synthesis. A culture of worms is filtered through glass wool or a wire screen to isolate young larvae. Eggs laid by these worms after they mature are collected over a period of 4-6 hours and allowed to hatch. A low level of FUdR (25 microM) is added just before the larvae reach maturity. This timing is important to avoid developmental abnormalities. The adults lay eggs in the presence of FUdR but the eggs do not hatch, which maintains the synchrony of the culture. Many aging characteristics appear to be similar in treated and untreated worms, such as the time of cessation of egg production, the appearance of visible and behavioral age-related changes, and the mean lifespan. This system thus seems suitable for large-scale biochemical analysis of certain aspects of aging in C. elegans.
本文描述了一种能够建立和维持线虫秀丽隐杆线虫大量年龄同步群体的技术。该技术有三个基本组成部分：丰富的化学培养基；一种生产和收获大量虫卵的方法；以及DNA合成抑制剂5-氟脱氧尿苷（FUdR）。通过玻璃棉或金属丝网过滤虫体培养物以分离幼虫。这些虫体成熟后产下的卵在4至6小时内收集并使其孵化。在幼虫接近成熟时加入低水平的FUdR（25微摩尔）。这个时间点很重要，可避免发育异常。成虫在有FUdR的情况下产卵，但卵不会孵化，从而保持培养物的同步性。在经处理和未经处理的虫体中，许多衰老特征似乎相似，如产卵停止时间、与年龄相关的可见和行为变化的出现以及平均寿命。因此，该系统似乎适用于对秀丽隐杆线虫衰老某些方面进行大规模生化分析。
