Name: solid plate-based dietary restriction in caenorhabditis elegans
Uploaded: 2011-05-28T14:00:00
Description: Watch this Scientific Journal Video about Solid Plate-based Dietary Restriction in Caenorhabditis elegans at JoVE.com

Este trabalho foi financiado por um prêmio de piloto do Centro de Choque Nathan (P30 AG13283) e uma subvenção R01 (R01 AG028516) do Instituto Nacional do Envelhecimento (NIA) para A.-LH

<html> 1. Estabelecer uma curva de calibração para a estimativa das concentrações de bactérias Esta seção descreve um método para a estimativa da concentração de bactérias por qualquer estirpe de bactérias dada (por exemplo, OP50, HT115) que será usado como fonte de alimento para os experimentos A restrição alimentar em C. elegans. Curvas de calibração em separado devem ser estabelecidos para cada bactéria cepa utilizada. <ol><li> Inocular uma única colônia de <...

<ol>
	<li>Hsu, A. L., Murphy, C. T., Kenyon, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+aging+and+age-related+disease+by+DAF-16+and+heat-shock+factor.">Regulation of aging and age-related disease by DAF-16 and heat-shock factor.</a> Science. 300, 1142-1145 (2003).</li><li>Kenyon, C., Chang, J., Gensch, E., Rudner, A., Tabtiang, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+C.+elegans+mutant+that+lives+twice+as+long+as+wild+type.">A C. elegans mutant that lives twice as long as wild type.</a> Nature. 366, 461-464 (1993).</li><li>Apfeld, J., Kenyon, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+lifespan+by+sensory+perception+in+Caenorhabditis+elegans.">Regulation of lifespan by sensory perception in Caenorhabditis elegans.</a> Nature. 402, 804-809 (1999).</li><li>Ching, T. T., Paal, A. B., Mehta, A., Zhong, L., Hsu, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=drr-2+encodes+an+eIF4H+that+acts+downstream+of+TOR+in+diet-restriction-induced+longevity+of+C.+elegans.">drr-2 encodes an eIF4H that acts downstream of TOR in diet-restriction-induced longevity of C. elegans.</a> Aging Cell. 9, 545-557 (2010).</li><li>Weindruch, R., Walford, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Retardation of Aging and Disease by Dietary Restriction. , (1988).</li><li>Masoro, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subfield+history:+caloric+restriction,+slowing+aging,+and+extending+life.">Subfield history: caloric restriction, slowing aging, and extending life.</a> Sci Aging Knowledge Environ. , RE2-RE2 (2003).</li><li>Masoro, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Overview+of+caloric+restriction+and+ageing.">Overview of caloric restriction and ageing.</a> Mech Ageing Dev. 126, 913-922 (2005).</li><li>Houthoofd, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Axenic+growth+up-regulates+mass-specific+metabolic+rate,+stress+resistance,+and+extends+life+span+in+Caenorhabditis+elegans.">Axenic growth up-regulates mass-specific metabolic rate, stress resistance, and extends life span in Caenorhabditis elegans.</a> Exp Gerontol. 37, 1371-1378 (2002).</li><li>Houthoofd, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=No+reduction+of+metabolic+rate+in+food+restricted+Caenorhabditis+elegans.">No reduction of metabolic rate in food restricted Caenorhabditis elegans.</a> Exp Gerontol. 37, 1359-1369 (2002).</li><li>Klass, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aging+in+the+nematode+Caenorhabditis+elegans:+major+biological+and+environmental+factors+influencing+life+span.">Aging in the nematode Caenorhabditis elegans: major biological and environmental factors influencing life span.</a> Mech Ageing Dev. 6, 413-429 (1977).</li><li>Bishop, N. A., Guarente, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two+neurons+mediate+diet-restriction-induced+longevity+in+C.+elegans.">Two neurons mediate diet-restriction-induced longevity in C. elegans.</a> Nature. 447, 545-549 (2007).</li><li>Panowski, S. H., Wolff, S., Aguilaniu, H., Durieux, J., Dillin, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PHA-4/Foxa+mediates+diet-restriction-induced+longevity+of+C.+elegans.">PHA-4/Foxa mediates diet-restriction-induced longevity of C. elegans.</a> Nature. 447, 550-555 (2007).</li><li>Lakowski, B., Hekimi, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+genetics+of+caloric+restriction+in+Caenorhabditis+elegans.">The genetics of caloric restriction in Caenorhabditis elegans.</a> Proc Natl Acad Sci U S A. 95, 13091-13096 (1998).</li><li>Kaeberlein, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lifespan+extension+in+Caenorhabditis+elegans+by+complete+removal+of+food.">Lifespan extension in Caenorhabditis elegans by complete removal of food.</a> Aging Cell. 5, 487-494 (2006).</li><li>Greer, E. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+AMPK-FOXO+pathway+mediates+longevity+induced+by+a+novel+method+of+dietary+restriction+in+C.+elegans.">An AMPK-FOXO pathway mediates longevity induced by a novel method of dietary restriction in C. elegans.</a> Curr Biol. 17, 1646-1656 (2007).</li></ol>

Restrição dietética (DR) é conhecido por aumentar a longevidade ea retardar o aparecimento de várias doenças relacionadas à idade em uma ampla gama de espécies, incluindo mamíferos 5-7. Manipulações ambientais e genéticos têm sido usados ​​para modelar DR em C. elegans, incluindo worms cultura em um meio semi-definido líquido 8,9, diluição da fonte de alimento de bactérias em meio líquido 10-12, e uso de mutantes comportamento que se alimentam em um ritmo m...

<table border="1"><tbody><tr><td> Nome do reagente </td><td> Companhia </td><td> Número de catálogo </td><td> Comentários </td></tr><tr><td> Carbenicilina </td><td> Duchefa Biochemie </td><td> C0109.25 </td><td> Dissolvido em água. </td></tr><tr><td> Canamicina </td><td> Roche </td><td> 10106801001 </td><td> Dissolvido em água. </td></tr><tr><td> Bacto-peptona </td><td> Becton, Dickinson e companhia </td><td> DF0118072 </td><td></td></tr><tr><td> Bacto-triptona </td><td> Becton, Dickinson e companhia </td><td> DF0123075 </td><td></td></tr><tr><td> Bacto levedura-extrato de </td><td> Becton, Dickinson e companhia </td><td> DF0127071 </td><td></td></tr><tr><td> Agar </td><td> Becton, Dickinson e companhia </td><td> DF00145170 </td><td></td></tr><tr><td> Tubo de cultura estéril </td><td> EUA Scientific </td><td> 1485-0810 </td><td></td></tr><tr><td> Espectrofotômetro </td><td> Amersham Biosciences </td><td> Gene Quant pro </td><td> Amersham agora faz parte da GE Healthcare. </td></tr><tr><td> Microscópio de dissecação </td><td> Zeiss </td><td> Stemi 2000 </td><td></td></tr><tr><td> Centrifugador </td><td> Beckman Coulter </td><td> Accuspin FR </td><td></td></tr></tbody></table>

solid plate-based dietary restriction in caenorhabditis elegans

Redução da ingestão alimentar sem desnutrição ou fome é conhecido por aumentar a longevidade ea retardar o aparecimento de várias doenças relacionadas à idade em uma ampla gama de espécies, incluindo mamíferos. Ela também provoca uma diminuição no peso corporal e fertilidade, bem como níveis mais baixos de glicose plasmática, insulina e IGF-1 nesses animais. Este tratamento é muitas vezes referida como restrição dietética (DR) ou a restrição calórica (CR). O nemátodo Caenorhabditis elegans tem emergido como um organismo modelo importante para o estudo da biologia do envelhecimento. Manipulações ambientais e genéticos têm sido usados ​​para modelar DR e têm mostrado que prolonga a vida em C. elegans. No entanto, muitos dos estudos relatados DR em C. elegans foram feitas por animais propagação em meios líquidos, enquanto a maioria dos estudos genéticos na área do envelhecimento foram feitos no ágar padrão sólido em placas de petri. Aqui apresentamos um protocolo DR usando o padrão sólido NGM placa de ágar-base com as bactérias mortas.

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

Aqui apresentamos um protocolo para a realização de restrição placa baseada em sólidos na dieta<em&gt; C. elegans</em&gt; Com bactérias mortas.

Placa sólida baseada na restrição dietética Caenorhabditis elegans</em

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 ...

Research

JoVE Journal

Biology

Solid Plate-based Dietary Restriction in Caenorhabditis elegans

Measuring Caenorhabditis elegans Life Span on Solid Media

Measuring Caenorhabditis elegans Life Span on Solid Media

Medição Caenorhabditis elegans Expectativa de Vida em meios sólidos

Aging is a degenerative process characterized by a progressive deterioration of cellular components and organelles resulting in mortality. The nematode ...

Biologia

Isolation and In vitro Activation of Caenorhabditis elegans Sperm

Isolation and In vitro Activation of Caenorhabditis elegans Sperm

Isolamento e In vitro A ativação do Caenorhabditis elegans Sperm

Males and hermaphrodites are the two naturally found sexual forms in the nematode C. elegans. The amoeboid sperm are produced by both males and ...

Time-lapse Microscopy of Early Embryogenesis in Caenorhabditis elegans

Time-lapse Microscopy of Early Embryogenesis in Caenorhabditis elegans

Lapso de tempo Microscopia de embriogênese inicial em Caenorhabditis elegans

Caenorhabditis elegans has often been used as a model system in studies of early developmental processes.  The transparency of the 
embryos, the genetic ...

Basic Caenorhabditis elegans Methods: Synchronization and Observation

Basic Caenorhabditis elegans Methods: Synchronization and Observation

Básico<em&gt; Caenorhabditis elegans</em&gt; Métodos: Sincronização e observação

Research into the molecular and developmental biology of the nematode Caenorhabditis elegans was begun in the early seventies by Sydney Brenner and it has ...

Prostaglandin Extraction and Analysis in Caenorhabditis elegans

Prostaglandin Extraction and Analysis in Caenorhabditis elegans

Extração de prostaglandina e Análise em<em&gt; Caenorhabditis elegans</em

Caenorhabditis elegans is emerging as a powerful animal model to study the biology of lipids1-9. Prostaglandins are an important class of eicosanoids, ...

Dietary Supplementation of Polyunsaturated Fatty Acids in Caenorhabditis elegans

Dietary Supplementation of Polyunsaturated Fatty Acids in Caenorhabditis elegans

A suplementação dietética de ácidos graxos poliinsaturados em<em&gt; Caenorhabditis elegans</em

Fatty acids are essential for numerous cellular functions. They serve as efficient energy storage molecules, make up the hydrophobic core of membranes, ...

Cultivation of Caenorhabditis elegans in Three Dimensions in the Laboratory

Cultivation of Caenorhabditis elegans in Three Dimensions in the Laboratory

cultivo de<em&gt; Caenorhabditis elegans</em&gt; Em três dimensões em Laboratório

The use of genetic model organisms such as Caenorhabditis elegans has led to seminal discoveries in biology over the last five decades. Most of what we ...

Measurement of Oxygen Consumption Rates in Intact Caenorhabditis elegans

Measurement of Oxygen Consumption Rates in Intact Caenorhabditis elegans

Medição das taxas de consumo de oxigênio em intacta Caenorhabditis elegans

Optimal mitochondrial function is critical for healthy cellular activity, particularly in cells that have high energy demands like those in the nervous ...

Quantifying Tissue-Specific Proteostatic Decline in Caenorhabditis elegans

Quantifying Tissue-Specific Proteostatic Decline in Caenorhabditis elegans

Quantificando o declínio proteostático específico do tecido em elegans caenorhabditis

The ability to maintain proper function and folding of the proteome (protein homeostasis) declines during normal aging, facilitating the onset of a ...

Modeling Age-Associated Neurodegenerative Diseases in Caenorhabditis elegans

Modeling Age-Associated Neurodegenerative Diseases in Caenorhabditis elegans

Modelagem de Doenças Neurodegenerativas Associadas à Idade em Caenorhabditis elegans

Battling human neurodegenerative pathologies and managing their pervasive socioeconomic impact is becoming a global priority. Notwithstanding their ...