实心板为基础的饮食限制在线虫</em
Summary
在这里，我们目前执行固体板为主的饮食限制在一个协议<em> C。线虫</em>与杀害细菌。
Abstract
没有营养不良或饥饿，减少食物的摄入，增加寿命和延迟发病的各种与年龄有关的疾病，在广泛的物种，包括哺乳动物。这也导致在体重和生育能力下降，以及血浆葡萄糖，胰岛素和IGF – 1在这些动物的较低水平。这种治疗方法通常被称为饮食限制（DR）或热量限制（CR）。 线虫已经成为一个重要的模式生物，研究衰老的生物学。环境和遗传操作已被用于模型的DR，并已显示出在 C的寿命延长线虫 。但是，许多报道DR 在 C的研究线虫是通过在液体介质中传播的动物，而在老龄问题领域的遗传研究的最标准在固体琼脂培养皿板。在这里，我们目前使用标准的固体NGM杀害细菌的琼脂板DR的协议。
Protocol
1。细菌浓度的估计建立校准曲线本节描述为任何给定的菌种（如OP50，HT115），将作为食物来源C.限制饮食实验使用细菌浓度的估算方法线虫 。每一株用于细菌，应建立单独的校准曲线。 接种单菌落的E.大肠杆菌 OP50细菌捡到3毫升的液体LB培养基中的细菌从一个新鲜的LB琼脂连胜。 广场OP50文化在37℃摇床，并允许细菌的生长过夜。 …
Discussion
饮食限制（DR）是已知的，以增加寿命和延迟在广泛的物种，包括哺乳动物5-7中各种与年龄有关的的疾病的发病。已用于环境和遗传操作模式在 C DR 线虫，包括在定义一个半液体介质中8,9，稀释10-12日在液体介质中的细菌的食物来源和使用行为的突变体，饲料以较慢的速率13培养蠕虫。然而，DR的遗传模拟可能产生的表型无关的DR，而在液体培养基中培?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
这项工作是支持到A – LH从弥敦道冲击中心（P30 AG13283）和授予R01（R01 AG028516）从国家老化研究所（NIA）的试点奖
Materials
| Name of the reagent | Company | Catalogue number | Comments |
| Carbenicillin | Duchefa Biochemie | C0109.25 | Dissolved in water. |
| Kanamycin | Roche | 10106801001 | Dissolved in water. |
| Bacto-peptone | Becton, Dickinson and company | DF0118072 | |
| Bacto-tryptone | Becton, Dickinson and company | DF0123075 | |
| Bacto-yeast extract | Becton, Dickinson and company | DF0127071 | |
| Agar | Becton, Dickinson and company | DF00145170 | |
| Sterile culture tube | USA Scientific | 1485-0810 | |
| Spectrophotometer | Amersham Biosciences | Gene Quant pro | Amersham is now part of GE Healthcare. |
| Dissecting Microscope | Zeiss | Stemi 2000 | |
| Centrifuge | Beckman Coulter | Accuspin FR |
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Tags
Solid Plate-based Dietary RestrictionCaenorhabditis ElegansFood Intake ReductionLifespan ExtensionAge-related DiseasesBody Weight DecreaseFertility DecreasePlasma Glucose DecreaseInsulin DecreaseIGF-1 DecreaseDietary Restriction (DR)Caloric Restriction (CR)Model OrganismEnvironmental ManipulationsGenetic ManipulationsLiquid Media PropagationSolid Agar In Petri PlatesDR ProtocolNGM Agar-based Plate
Cite This Article
Ching, T., Hsu, A. Solid Plate-based Dietary Restriction in Caenorhabditis elegans. J. Vis. Exp. (51), e2701, doi:10.3791/2701 (2011).