测量线虫固体介质的寿命
Summary
在这篇文章中，我们提出了一个测量寿命固体培养基上保持线虫用紫外线灭活的细菌性食物的一般协议。
Abstract
老化是一种退化性的过程，特点是逐渐恶化的细胞成分和细胞器造成的死亡率。线虫<em>线虫</em用来研究老化生物学的一个主要模式出现。因为随着年龄的增加，几乎每一个生物子系统进行功能下降，寿命是利益的考虑老化率时的主要终点。线虫的寿命通常定义为一种动物的天数仍然对外界刺激的的反应。线虫可以在液体介质中或板固体培养基上，并测量两个条件下的寿命的开发技术已。这里我们提出一个广义的协议和测量寿命固体线虫的生长介质保持线虫喂食的紫外线杀死细菌的饮食。这些程序可以很容易地适应各种常见的条件下测定的寿命，包括饮食活的细菌，限制饮食，和RNA干扰组成。
Protocol
第1部分：准备线虫生长介质（NGM）板本节介绍了在寿命实验中使用的固体NGM板的准备。一个基本的寿命实验需要两种类型的板块：NGM的标准板，它含有任何添加剂，并安培/ FUDR板，氨苄青霉素（AMP）和fluorodeoxyuridine（FUDR）添加到了NGM。氨苄青霉素是用来防止外来细菌污染。 FUDR抑制细胞分裂，降低产蛋量，防止种蛋。 FUDR为长寿分析使用，不影响成人的寿命，不再需要转移蠕?…
Discussion
长寿的遗传控制已被广泛研究， 在 C 线虫 ，主要是由于与寿命可确定的易用性和快速性。在这篇文章中讨论的协议描述为获得重现生命跨度的数据， 在 C的基本框架也可以应用于线虫和relatednematode物种。做一些简单的改动，这些程序可适应各种条件下的寿命来衡量。在这里，我们将讨论几种常见的变化，包括活的细菌，细菌剥夺限制饮食，RNA干扰（RNAi），和无毒品NGM?…
Acknowledgements
这项工作是支持老年学奖和美国国立卫生研究院资助1R01AG031108 – 01 MKGS是由美国国立卫生研究院培训资助P30AG013280支持格伦/ AFAR突破。 Mk为中老年埃利森医学基金会新学者。
Materials
| Material Name | Type | Company | Catalogue Number | Comment |
|---|---|---|---|---|
| Agar | Reagent | Fisher Scientific (BD Diagnostic Systems) | DF0145-17-0 (214530) | |
| Ampicillin | Reagent | MidSci | 0339 | |
| BactoTryptone | Reagent | Fisher Scientific (BD Diagnostic Systems) | DF0123-17-3 (211705) | |
| BactoPeptone | Reagent | Fisher Scientific (BD Diagnostic Systems) | DF0118-17-0 (211677) | |
| CaCl2 | Reagent | Fisher Scientific (JT Baker) | NC9699248 (1332-01) | |
| Carbenicillin | Reagent | Gold BioTechnology (GBT) | C0109 | |
| Cholesterol | Reagent | Sigma-Aldrich | C75209 | |
| Fluorodeoxyuridine (FUDR) | Reagent | Sigma-Aldrich | F0503 | |
| Isopropyl β-d-Thiogalactopyranoside (IPTG) | Reagent | Gold BioTechnology (GBT) | I2481C5 | |
| KPi Dibasic | Reagent | Fisher Scientific (JT Baker) | 5087862 (3252-01) | |
| KPi Monobasic | Reagent | Fisher Scientific (JT Baker) | 5087861 (3246-01) | |
| MgSO4 | Reagent | Fisher Scientific (JT Baker) | NC9561800 (2500-01) | |
| NaCl | Reagent | Fisher Scientific | S251 | |
| Tris | Reagent | Sigma-Aldrich | T1503 | |
| Yeast Extract | Reagent | Fisher Scientific (BD Diagnostic Systems) | DF0886-17-0 (288620) |
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Cite This Article
Sutphin, G. L., Kaeberlein, M. Measuring Caenorhabditis elegans Life Span on Solid Media. J. Vis. Exp. (27), e1152, doi:10.3791/1152 (2009).