This work was supported by a pilot award from the Nathan Shock Center (P30 AG13283) and a R01 grant (R01 AG028516) from the National Institute on Aging (NIA) to A.-L.H.

1. Establishing a calibration curve for the estimation of bacteria concentrations
This section describes a method for the estimation of bacteria concentration for any given bacteria strain (e.g. OP50, HT115) that will be used as a food source for Dietary restriction experiments in C. elegans. Separate calibration curves should be established for each bacteria strain used. 
<ol>
 <li>Inoculate a single colony of E. coli OP50 bacteria picked from a fresh streak of bacteria...

<ol>
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Dietary restriction (DR) is known to increase lifespan and delay the onset of various age-related diseases in a wide range of species, including mammals 5-7. Both environmental and genetic manipulations have been used to model DR in C. elegans, including culturing worms in a semi-defined liquid medium 8,9, dilution of the bacterial food source in liquid medium 10-12, and use of behavior mutants that feed at a slower rate 13. However, genetic mimics of DR may produce ...

<table border="1">
	<tbody>
		<tr>
			<td>Name of the reagent</td>
			<td>Company</td>
			<td>Catalogue number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Carbenicillin</td>
			<td>Duchefa Biochemie</td>
			<td>C0109.25</td>
			<td>Dissolved in water.</td>
		</tr>
		<tr>
			<td>Kanamycin</td>
			<td>Roche</td>
			<td>10106801001</td>
			<td>Dissolved in water.</td>
		</tr>
		<tr>
			<td>Bacto-peptone</td>
			<td>Becton, Dickinson and company</td>
			<td>DF0118072</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Bacto-tryptone</td>
			<td>Becton, Dickinson and company</td>
			<td>DF0123075</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Bacto-yeast extract</td>
			<td>Becton, Dickinson and company</td>
			<td>DF0127071</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Agar</td>
			<td>Becton, Dickinson and company</td>
			<td>DF00145170</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Sterile culture tube</td>
			<td>USA Scientific</td>
			<td>1485-0810</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Spectrophotometer</td>
			<td>Amersham Biosciences</td>
			<td>Gene Quant pro</td>
			<td>Amersham is now part of GE Healthcare.</td>
		</tr>
		<tr>
			<td>Dissecting Microscope</td>
			<td>Zeiss</td>
			<td>Stemi 2000</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>Beckman Coulter</td>
			<td>Accuspin FR</td>
			<td>&nbsp;</td>
		</tr>
	</tbody>
</table>

solid plate-based dietary restriction in caenorhabditis elegans

Reduction of food intake without malnutrition or starvation is known to increase lifespan and delay the onset of various age-related diseases in a wide range of species, including mammals. It also causes a decrease in body weight and fertility, as well as lower levels of plasma glucose, insulin, and IGF-1 in these animals. This treatment is often referred to as dietary restriction (DR) or caloric restriction (CR). The nematode Caenorhabditis elegans has emerged as an important model organism for studying the biology of aging. Both environmental and genetic manipulations have been used to model DR and have shown to extend lifespan in C. elegans. However, many of the reported DR studies in C. elegans were done by propagating animals in liquid media, while most of the genetic studies in the aging field were done on the standard solid agar in petri plates. Here we present a DR protocol using standard solid NGM agar-based plate with killed bacteria.

Watch this Scientific Journal Video about Solid Plate-based Dietary Restriction in Caenorhabditis elegans at JoVE.com

Solid Plate-based Dietary Restriction in Caenorhabditis elegans

Here we present a protocol for performing solid plate-based dietary restriction in C. elegans with killed bacteria.

Solid Plate-based Dietary Restriction in Caenorhabditis elegans

Reduction of food intake without malnutrition or starvation is known to increase lifespan and delay the onset of various age-related diseases in a wide ...

solid-plate-based-dietary-restriction-in-caenorhabditis-elegans

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JoVE Journal

Biology

16.5K visualizações.  University of Michigan. Aqui apresentamos um protocolo para a realização de restrição placa baseada em sólidos na dieta C. elegans Com bactérias mortas.