This work was supported by funding from the Ellison Medical Foundation (SDP, <a href="http://www.ellisonfoundation.org/index.jsp" target="_blank">http://www.ellisonfoundation.org/index.jsp</a>), NIH K01AG031917 (NJL, <a href="http://www.nih.gov" target="_blank">http://www.nih.gov/</a>), NIH 5T32GM007315-35 (JR) and NIH R01AG030593 (SDP). This work utilized the resources of the Drosophila Aging Core (DAC) of the Nathan Shock Center of Excellence in the Biology of Aging funded by the National Institute of Aging P30-AG-013283 (<a href="http://www.nih.gov" target="_blank">http://www.nih.gov/</a>). The authors would like to thank the Pletcher Laboratory for helpful discussions and particularly Brian Chung for critical reading of the manuscript. We would like to acknowledge Nick Asher and Kathryn Borowicz for assistance with data collection.

We recommend storing experimental foods, yeast paste, and grape agar plates that appear in the protocol at 4 &deg;C and using them within 1-2 months as long as mold and dryness have not set in. Standard environmental conditions for both the larval and adult stage involve maintenance of flies in an incubator at 25 &deg;C with a 12:12 hr light dark cycle and 60% relative humidity.
1. Preparation of Experimental Food
<ol>
	<li>For larval growth, we use a modified Caltech Medium5, whi...

<ol>
	<li>Toivonen, J. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=No+influence+of+Indy+on+lifespan+in+Drosophila+after+correction+for+genetic+and+cytoplasmic+background+effects.">No influence of Indy on lifespan in Drosophila after correction for genetic and cytoplasmic background effects.</a> PLoS Genet. 3, e95 (2007).</li><li>Spencer, C. C., Howell, C. E., Wright, A. R., Promislow, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Testing+an+'aging+gene'+in+long-lived+drosophila+strains:+increased+longevity+depends+on+sex+and+genetic+background.">Testing an 'aging gene' in long-lived drosophila strains: increased longevity depends on sex and genetic background.</a> Aging Cell. 2, 123-130 (2003).</li><li>Burnett, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Absence+of+effects+of+Sir2+overexpression+on+lifespan+in+C.+elegans+and+Drosophila.">Absence of effects of Sir2 overexpression on lifespan in C. elegans and Drosophila.</a> Nature. 477, 482-485 (2011).</li><li>Bokov, A. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Does+reduced+IGF-1R+signaling+in+Igf1r+/-+mice+alter+aging?.">Does reduced IGF-1R signaling in Igf1r+/- mice alter aging?.</a> PLoS One. 6, e26891 (2011).</li><li>Lewis, E. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+standard+food+medium.">A new standard food medium.</a> Drosophila Information Service. 34, 117-118 (1960).</li><li>Skorupa, D. A., Dervisefendic, A., Zwiener, J., Pletcher, S. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dietary+composition+specifies+consumption,+obesity,+and+lifespan+in+Drosophila+melanogaster.">Dietary composition specifies consumption, obesity, and lifespan in Drosophila melanogaster.</a> Aging Cell. 7, 478-490 (2008).</li><li>Rera, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+longevity+and+tissue+homeostasis+by+the+Drosophila+PGC-1+homolog.">Modulation of longevity and tissue homeostasis by the Drosophila PGC-1 homolog.</a> Cell Metab. 14, 623-634 (2011).</li><li>Kaplan, E. L., Meier, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonparametric+Estimation+from+Incomplete+Observations.">Nonparametric Estimation from Incomplete Observations.</a> Journal of the American Statistical Association. 53, 457-481 (1958).</li><li>Pletcher, S. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitigating+the+Tithonus+Error:+Genetic+Analysis+of+Mortality+Phenotypes.">Mitigating the Tithonus Error: Genetic Analysis of Mortality Phenotypes.</a> Sci. Aging Knowl. Environ. 2002, pe14 (2002).</li><li>Pletcher, S. D., Khazaeli, A. A., Curtsinger, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Why+do+life+spans+differ?+Partitioning+mean+longevity+differences+in+terms+of+age-specific+mortality+parameters.">Why do life spans differ? Partitioning mean longevity differences in terms of age-specific mortality parameters.</a> J. Gerontol. A Biol. Sci. Med. Sci. 55, 381-389 (2000).</li><li>Promislow, , Tatar, , Pletcher, , Carey, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Below-threshold+mortality:+implications+for+studies+in+evolution,+ecology+and+demography.">Below-threshold mortality: implications for studies in evolution, ecology and demography.</a> Journal of Evolutionary Biology. 12, 314-328 (1999).</li><li>Pletcher, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Model+fitting+and+hypothesis+testing+for+age-specific+mortality+data.">Model fitting and hypothesis testing for age-specific mortality data.</a> Journal of Evolutionary Biology. 12, 430-439 (1999).</li><li>Partridge, L., Gems, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Benchmarks+for+ageing+studies.">Benchmarks for ageing studies.</a> Nature. 450, 165-167 (2007).</li><li>Roman, G., Endo, K., Zong, L., Davis, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=P[Switch],+a+system+for+spatial+and+temporal+control+of+gene+expression+in+Drosophila+melanogaster.">P[Switch], a system for spatial and temporal control of gene expression in Drosophila melanogaster.</a> Proceedings of the National Academy of Sciences of the United States of America. 98, 12602-12607 (2001).</li><li>Ford, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alteration+of+Drosophila+life+span+using+conditional,+tissue-specific+expression+of+transgenes+triggered+by+doxycyline+or+RU486/Mifepristone.">Alteration of Drosophila life span using conditional, tissue-specific expression of transgenes triggered by doxycyline or RU486/Mifepristone.</a> Exp. Gerontol. 42, 483-497 (2007).</li><li>Priest, N. K., Mackowiak, B., Promislow, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+parental+age+effects+on+the+evolution+of+aging.">The role of parental age effects on the evolution of aging.</a> Evolution. 56, 927-935 (2002).</li><li>Smith, E. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Feeding+Drosophila+a+biotin-deficient+diet+for+multiple+generations+increases+stress+resistance+and+lifespan+and+alters+gene+expression+and+histone+biotinylation+patterns.">Feeding Drosophila a biotin-deficient diet for multiple generations increases stress resistance and lifespan and alters gene expression and histone biotinylation patterns.</a> J. Nutr. 137, 2006-2012 (2007).</li><li>Sorensen, J. G., Loeschcke, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Larval+crowding+in+Drosophila+melanogaster+induces+Hsp70+expression,+and+leads+to+increased+adult+longevity+and+adult+thermal+stress+resistance.">Larval crowding in Drosophila melanogaster induces Hsp70 expression, and leads to increased adult longevity and adult thermal stress resistance.</a> J. Insect Physiol. 47, 1301-1307 (2001).</li><li>Bass, T. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimization+of+dietary+restriction+protocols+in+Drosophila.">Optimization of dietary restriction protocols in Drosophila.</a> J. Gerontol. A Biol. Sci. Med. Sci. 62, 1071-1081 (2007).</li><li>Miquel, J., Lundgren, P. R., Bensch, K. G., Atlan, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+temperature+on+the+life+span,+vitality+and+fine+structure+of+Drosophila+melanogaster.">Effects of temperature on the life span, vitality and fine structure of Drosophila melanogaster.</a> Mechanisms of Ageing and Development. 5, 347-370 (1976).</li><li>Pittendrigh, C. S., Minis, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Circadian+systems:+longevity+as+a+function+of+circadian+resonance+in+Drosophila+melanogaster.">Circadian systems: longevity as a function of circadian resonance in Drosophila melanogaster.</a> Proceedings of the National Academy of Sciences of the United States of America. 69, 1537-1539 (1972).</li><li>Joshi, A., Mueller, L. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adult+crowding+effects+on+longevity+in+Drosophila+melanogaster:+Increase+in+age-dependent+mortality.">Adult crowding effects on longevity in Drosophila melanogaster: Increase in age-dependent mortality.</a> Current Science. 72, 255-260 (1997).</li><li>Ja, W. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prandiology+of+Drosophila+and+the+CAFE+assay.">Prandiology of Drosophila and the CAFE assay.</a> Proceedings of the National Academy of Sciences of the United States of America. 104, 8253-8256 (2007).</li><li>Lee, K. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lifespan+and+reproduction+in+Drosophila:+New+insights+from+nutritional+geometry.">Lifespan and reproduction in Drosophila: New insights from nutritional geometry.</a> Proceedings of the National Academy of Sciences of the United States of America. 105, 2498-2503 (2008).</li><li>Gargano, J. W., Martin, I., Bhandari, P., Grotewiel, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+iterative+negative+geotaxis+(RING):+a+new+method+for+assessing+age-related+locomotor+decline+in+Drosophila.">Rapid iterative negative geotaxis (RING): a new method for assessing age-related locomotor decline in Drosophila.</a> Experimental gerontology. 40, 386-395 (2005).</li></ol>

No conflicts of interest declared.

The protocol presented here describes a method for producing reproducible measurements of adult longevity in Drosophila that is adaptable for assessment of genetic, pharmacological, and environmental interventions. Crucial aspects of the protocol include carefully controlling the larval development environment, minimizing adult stress, and minimizing bias across experimental groups and controls. We also present the use of the dLife lifespan experiment management software. By simply attaching a bar code or RFID t...

<table border="1">
 <tbody>
 <tr>
 <td>Name of the reagent</td>
 <td>Company</td>
 <td>Catalogue number</td>
 <td>Comments (optional)</td>
 </tr>
 <tr>
 <td>Active Dry Yeast</td>
 <td>Fleishmann's Yeast</td>
 <td>2192</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Grape Agar Powder Premix</td>
 <td>Genesee Scientific</td>
 <td>47-102</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Large Embryo Collection Cages</td>
 <td>Genesee Scientific</td>
 <td>59-101</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Large Replacement End Caps</td>
 <td>Genesee Scientific</td>
 <td>59-103</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>6 oz Square Bottom Bottles, polypropylene</td>
 <td>Genesee Scientific</td>
 <td>32-130</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Flugs Closures for Stock Bottles</td>
 <td>Genesee Scientific</td>
 <td>49-100</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Drosophila Vials, Wide, Polystrene</td>
 <td>Genesee Scientific</td>
 <td>32-117</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Flugs Closures for Wide Vials</td>
 <td>Genesee Scientific</td>
 <td>49-101</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Wide Orifice Aardvark Pipet Tips, 200 ul</td>
 <td>Denville Scientific</td>
 <td>P1105-CP</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Flystuff Flypad, Standard Size</td>
 <td>Genesee Scientific</td>
 <td>59-114</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>BD Falcon 15 ml Conical Centrifuge Tubes</td>
 <td>Fisher Scientific</td>
 <td>14-959-70C</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Fisherbrand Petri Dishes with Clear Lids, Raised Ridge; 100 O.D. x 15 mm H;</td>
 <td>Fisher Scientific</td>
 <td>08-757-12</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Kimax* Colorware Flasks 1,000 ml yellow</td>
 <td>Fisher Scientific</td>
 <td>10-200-47</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>PBS pH 7.4 10x</td>
 <td>Invitrogen</td>
 <td>70011044</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Gelidium Agar</td>
 <td>Mooragar</td>
 <td>n/a</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Brewer's Yeast</td>
 <td>MP Biomedicals</td>
 <td>0290331280</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Granulated Sugar</td>
 <td>Kroger</td>
 <td>n/a</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Tegosept</td>
 <td>Genesee Scientific</td>
 <td>20-266</td>
 <td>Fly Food Preservative</td>
 </tr>
 <tr>
 <td>Propionic Acid, 99%</td>
 <td>Acros Organics</td>
 <td>149300025</td>
 <td>Fly Food Preservative</td>
 </tr>
 <tr>
 <td>Kanamycin Sulfate</td>
 <td>ISC BioExpress</td>
 <td>0408-10G</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Tetracycline HCl</td>
 <td>VWR</td>
 <td>80058-724</td>
 <td>&nbsp;</td>
 </tr>
 </tbody>
</table>

measurement of lifespan in drosophila melanogaster

Aging is a phenomenon that results in steady physiological deterioration in nearly all organisms in which it has been examined, leading to reduced physical performance and increased risk of disease. Individual aging is manifest at the population level as an increase in age-dependent mortality, which is often measured in the laboratory by observing lifespan in large cohorts of age-matched individuals. Experiments that seek to quantify the extent to which genetic or environmental manipulations impact lifespan in simple model organisms have been remarkably successful for understanding the aspects of aging that are conserved across taxa and for inspiring new strategies for extending lifespan and preventing age-associated disease in mammals.
The vinegar fly, Drosophila melanogaster, is an attractive model organism for studying the mechanisms of aging due to its relatively short lifespan, convenient husbandry, and facile genetics. However, demographic measures of aging, including age-specific survival and mortality, are extraordinarily susceptible to even minor variations in experimental design and environment, and the maintenance of strict laboratory practices for the duration of aging experiments is required. These considerations, together with the need to practice careful control of genetic background, are essential for generating robust measurements. Indeed, there are many notable controversies surrounding inference from longevity experiments in yeast, worms, flies and mice that have been traced to environmental or genetic artifacts1-4. In this protocol, we describe a set of procedures that have been optimized over many years of measuring longevity in Drosophila using laboratory vials. We also describe the use of the dLife software, which was developed by our laboratory and is available for download (<a href="http://sitemaker.umich.edu/pletcherlab/software">http://sitemaker.umich.edu/pletcherlab/software</a>). dLife accelerates throughput and promotes good practices by incorporating optimal experimental design, simplifying fly handling and data collection, and standardizing data analysis. We will also discuss the many potential pitfalls in the design, collection, and interpretation of lifespan data, and we provide steps to avoid these dangers.

A simplified scheme of the protocol is presented in Figure 1, where key steps are outlined. The synchronization part of the protocol can be used for various assays that require age-matched adult flies.
Typical survivorship curves of wild-type flies are shown in Figure 2a, using the dLife experiment management software (Figure 2b,c). Adult males usually live shorter, with both populations achieving a mean and median longevity of &gt;50 days on ...

Watch this Scientific Journal Video about Measurement of Lifespan in Drosophila melanogaster at JoVE.com

Measurement of Lifespan in Drosophila melanogaster

Drosophila melanogaster is a powerful model organism for exploring the molecular basis of longevity regulation. This protocol will discuss the steps involved in generating a reproducible, population-based measurement of longevity as well as potential pitfalls and how to avoid them.

Measurement of Lifespan in Drosophila melanogaster

Aging is a phenomenon that results in steady physiological deterioration in nearly all organisms in which it has been examined, leading to reduced ...

measurement-of-lifespan-in-drosophila-melanogaster

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33.7K Görüntüleme.  University of Michigan. Drosophila melanogaster uzun ömürlü düzenleme moleküler temelini keşfetmek için güçlü bir model organizmadır. Bu protokol bir tekrarlanabilir, uzun ömürlü yanı sıra potansiyel tuzaklar nüfus tabanlı ölçme ve bunlardan kaçınmanın yolları üreten katılan adımları görüşecek.