Name: measurement of survival time in brachionus rotifers: synchronization of maternal conditions
Uploaded: 2016-07-22T13:00:00
Description: Watch this Scientific Journal Video about Measurement of Survival Time in Brachionus Rotifers: Synchronization of Maternal Conditions at JoVE.com

Vi er taknemmelige for George Jarvis, Martha Bock, og Bette Hecox-Lea, Marinbiologisk Laboratorium, for deres hjælp i optagelserne.

1. Udarbejdelse af Media Bemærk: Brug halvt fortyndet Brujewicz kunstigt havvand af saltholdigheden 16,5 ppt (PSU). Andre kunstige havvande også ofte brugt til kultur Brachionus hjuldyr 25,26. <ol><li> Tilføj 454 mM NaCl, 26 mM MgCl2, 27 mM MgSO4, 10 mM KCI og 10 mM CaCl2 til 4,5 I destilleret vand (slutvolumen vil være 5 I). Alternativt kan du bruge demineraliseret vand til fortynding i stedet for destilleret vand. Tilføj CaCl...

<ol>
	<li>Wallace, R. L., Snell, T. W., Ricci, C., Nogrady, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Rotifera Vol.1: Biology, ecology and systematics. , (2006).</li><li>Segers, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Annotated checklist of the rotifers (Phylum Rotifera), with notes on nomenclature, taxonomy and distribution. , (2007).</li><li>Mark Welch, D. B., Meselson, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+for+the+evolution+of+bdelloid+rotifers+without+sexual+reproduction+or+genetic+exchange.">Evidence for the evolution of bdelloid rotifers without sexual reproduction or genetic exchange.</a> Science. 288 (5469), 1211-1215 (2000).</li><li>Suga, K., Mark Welch, D., Tanaka, Y., Sakakura, Y., Hagiwara, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+expressed+sequence+tags+of+the+cyclically+parthenogenetic+rotifer+Brachionus+plicatilis.">Analysis of expressed sequence tags of the cyclically parthenogenetic rotifer Brachionus plicatilis.</a> PLoS ONE. 2, e671 (2007).</li><li>Denekamp, N. Y., 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=Discovering+genes+associated+with+dormancy+in+the+monogonont+rotifer+Brachionus+plicatilis.">Discovering genes associated with dormancy in the monogonont rotifer Brachionus plicatilis.</a> BMC Genomics. 10, 108 (2009).</li><li>Lee, J. -. S., 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=Sequence+analysis+of+genomic+DNA+(680+Mb)+by+GS-FLX-Titanium+sequencer+in+the+monogonont+rotifer,+Brachionus+ibericus.">Sequence analysis of genomic DNA (680 Mb) by GS-FLX-Titanium sequencer in the monogonont rotifer, Brachionus ibericus.</a> Hydrobiologia. 662 (1), 65-75 (2010).</li><li>Flot, J. -. 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=Genomic+evidence+for+ameiotic+evolution+in+the+bdelloid+rotifer+Adineta+vaga.">Genomic evidence for ameiotic evolution in the bdelloid rotifer Adineta vaga.</a> Nature. 500 (7463), 453-457 (2013).</li><li>Dunn, C. 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=Broad+phylogenomic+sampling+improves+resolution+of+the+animal+tree+of+life.">Broad phylogenomic sampling improves resolution of the animal tree of life.</a> Nature. 452 (7188), 745-749 (2008).</li><li>Yoshinaga, T., Kaneko, G., Kinoshita, S., Tsukamoto, K., Watabe, 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+molecular+mechanisms+of+life+history+alterations+in+a+rotifer:+a+novel+approach+in+population+dynamics.">The molecular mechanisms of life history alterations in a rotifer: a novel approach in population dynamics.</a> Comp. Biochem. Physiol. B Biochem. Mol. Biol. 136 (4), 715-722 (2003).</li><li>Dahms, H. -. U., Hagiwara, A., Lee, J. -. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ecotoxicology,+ecophysiology,+and+mechanistic+studies+with+rotifers.">Ecotoxicology, ecophysiology, and mechanistic studies with rotifers.</a> Aquat. Toxicol. 101 (1), 1-12 (2011).</li><li>Snell, T. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rotifers+as+models+for+the+biology+of+aging.">Rotifers as models for the biology of aging.</a> Int. Rev. Hydrobiol. 99 (1-2), 84-95 (2014).</li><li>Snell, T. W., Johnston, R. K., Gribble, K. E., Mark Welch, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rotifers+as+experimental+tools+for+investigating+aging.">Rotifers as experimental tools for investigating aging.</a> Invertebr. Reprod. Dev. 59, 5-10 (2015).</li><li>Kaneko, G., 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=Molecular+characterization+of+Mn-superoxide+dismutase+and+gene+expression+studies+in+dietary+restricted+Brachionus+plicatilis+rotifers.">Molecular characterization of Mn-superoxide dismutase and gene expression studies in dietary restricted Brachionus plicatilis rotifers.</a> Hydrobiologia. 546, 117-123 (2005).</li><li>Yoshinaga, T., 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=Insulin-like+growth+factor+signaling+pathway+involved+in+regulating+longevity+of+rotifers.">Insulin-like growth factor signaling pathway involved in regulating longevity of rotifers.</a> Hydrobiologia. 546, 347-352 (2005).</li><li>Ozaki, Y., Kaneko, G., Yanagawa, Y., Watabe, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calorie+restriction+in+the+rotifer+Brachionus+plicatilis+enhances+hypoxia+tolerance+in+association+with+the+increased+mRNA+levels+of+glycolytic+enzymes.">Calorie restriction in the rotifer Brachionus plicatilis enhances hypoxia tolerance in association with the increased mRNA levels of glycolytic enzymes.</a> Hydrobiologia. 649 (1), 267-277 (2010).</li><li>Kailasam, 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=Effects+of+calorie+restriction+on+the+expression+of+manganese+superoxide+dismutase+and+catalase+under+oxidative+stress+conditions+in+the+rotifer+Brachionus+plicatilis.">Effects of calorie restriction on the expression of manganese superoxide dismutase and catalase under oxidative stress conditions in the rotifer Brachionus plicatilis.</a> Fish. Sci. 77 (3), 403-409 (2011).</li><li>Garc&#236;a-Garc&#236;a, G., Sarma, S., N&#250;&#241;ez-Orti, A. R., Nandini, S. <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+the+mixture+of+two+endocrine+disruptors+(ethinylestradiol+and+levonorgestrel)+on+selected+ecological+endpoints+of+Anuraeopsis+fissa+and+Brachionus+calyciflorus+(Rotifera).">Effects of the mixture of two endocrine disruptors (ethinylestradiol and levonorgestrel) on selected ecological endpoints of Anuraeopsis fissa and Brachionus calyciflorus (Rotifera).</a> Int. Rev. Hydrobiol. 99 (1-2), 166-172 (2014).</li><li>Yang, J., Mu, Y., Dong, S., Jiang, Q., Yang, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+the+expression+of+four+heat+shock+proteins+during+the+aging+process+in+Brachionus+calyciflorus+(rotifera).">Changes in the expression of four heat shock proteins during the aging process in Brachionus calyciflorus (rotifera).</a> Cell Stress Chaperones. 19 (1), 33-52 (2014).</li><li>Han, J., 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=Sublethal+gamma+irradiation+affects+reproductive+impairment+and+elevates+antioxidant+enzyme+and+DNA+repair+activities+in+the+monogonont+rotifer+Brachionus+koreanus.">Sublethal gamma irradiation affects reproductive impairment and elevates antioxidant enzyme and DNA repair activities in the monogonont rotifer Brachionus koreanus.</a> Aquat. Toxicol. 155, 101-109 (2014).</li><li>Gribble, K. E., Jarvis, G., Bock, M., Mark Welch, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maternal+caloric+restriction+partially+rescues+the+deleterious+effects+of+advanced+maternal+age+on+offspring.">Maternal caloric restriction partially rescues the deleterious effects of advanced maternal age on offspring.</a> Aging Cell. 13 (4), 623-630 (2014).</li><li>Yoshinaga, T., Hagiwara, A., Tsukamoto, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+periodical+starvation+on+the+survival+of+offspring+in+the+rotifer+Brachionus+plicatilis.">Effect of periodical starvation on the survival of offspring in the rotifer Brachionus plicatilis.</a> Fish. Sci. 67 (2), 373-374 (2001).</li><li>Kaneko, G., 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=Calorie+restriction-induced+maternal+longevity+is+transmitted+to+their+daughters+in+a+rotifer.">Calorie restriction-induced maternal longevity is transmitted to their daughters in a rotifer.</a> Funct. Ecol. 25 (1), 209-216 (2011).</li><li>Lansing, A. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+transmissible,+cumulative,+and+reversible+factor+in+aging.">A transmissible, cumulative, and reversible factor in aging.</a> J. Gerontol. 2 (3), 228-239 (1947).</li><li>Yoshinaga, T., Hagiwara, A., Tsukamoto, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+periodical+starvation+on+the+life+history+of+Brachionus+plicatilis+O.+F.+M&#252;ller+(Rotifera):+a+possible+strategy+for+population+stability.">Effect of periodical starvation on the life history of Brachionus plicatilis O. F. M&#252;ller (Rotifera): a possible strategy for population stability.</a> J. Exp. Mar. Biol. Ecol. 253 (2), 253-260 (2000).</li><li>Gribble, K. E., Kaido, O., Jarvis, G., Mark Welch, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patterns+of+intraspecific+variability+in+the+response+to+caloric+restriction.">Patterns of intraspecific variability in the response to caloric restriction.</a> Exp. Gerontol. 51, 28-37 (2014).</li><li>Snell, T. W., Johnston, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glycerol+extends+lifespan+of+Brachionus+manjavacas+(Rotifera)+and+protects+against+stressors.">Glycerol extends lifespan of Brachionus manjavacas (Rotifera) and protects against stressors.</a> Exp. Gerontol. 57, 47-56 (2014).</li><li>Kim, H. -. J., Hagiwara, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+female+aging+on+the+morphology+and+hatchability+of+resting+eggs+in+the+rotifer+Brachionus+plicatilis+M&#252;ller.">Effect of female aging on the morphology and hatchability of resting eggs in the rotifer Brachionus plicatilis M&#252;ller.</a> Hydrobiologia. 662 (1), 107-111 (2011).</li><li>Kim, H. -. J., 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=Light-dependent+transcriptional+events+during+resting+egg+hatching+of+the+rotifer+Brachionus+manjavacas.">Light-dependent transcriptional events during resting egg hatching of the rotifer Brachionus manjavacas.</a> Mar. Genomics. 20, 25-31 (2015).</li><li>Gribble, K. E., Welch, D. B. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Life-span+extension+by+caloric+restriction+is+determined+by+type+and+level+of+food+reduction+and+by+reproductive+mode+in+Brachionus+manjavacas+(Rotifera).">Life-span extension by caloric restriction is determined by type and level of food reduction and by reproductive mode in Brachionus manjavacas (Rotifera).</a> J. Gerontol. A Biol. Sci. Med. Sci. 68 (4), 349-358 (2013).</li><li>Kaneko, G., Kinoshita, S., Yoshinaga, T., Tsukamoto, K., Watabe, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+expression+patterns+of+stress+protein+genes+during+population+growth+of+the+rotifer+Brachionus+plicatilis.">Changes in expression patterns of stress protein genes during population growth of the rotifer Brachionus plicatilis.</a> Fish. Sci. 68 (6), 1317-1323 (2002).</li><li>Kim, H. J., Sawada, C., Hagiwara, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavior+and+reproduction+of+the+rotifer+Brachionus+plicatilis+species+complex+under+different+light+wavelengths+and+intensities.">Behavior and reproduction of the rotifer Brachionus plicatilis species complex under different light wavelengths and intensities.</a> Int. Rev. Hydrobiol. 99 (1-2), 151-156 (2014).</li><li>Yoshinaga, T., Hagiwara, A., Tsukamoto, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+conditioned+media+on+the+asexual+reproduction+of+the+monogonont+rotifer+Brachionus+plicatilis+O.+F.+Müller.">Effect of conditioned media on the asexual reproduction of the monogonont rotifer Brachionus plicatilis O. F. Müller.</a> Hydrobiologia. 412, 103-110 (1999).</li><li>Ohmori, F., Kaneko, G., Saito, T., Watabe, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+growth-promoting+protein+in+the+conditioned+media+from+the+rotifer+Brachionus+plicatilis+at+an+early+exponential+growth+phase.">A novel growth-promoting protein in the conditioned media from the rotifer Brachionus plicatilis at an early exponential growth phase.</a> Hydrobiologia. 667 (1), 101-117 (2011).</li><li>Collet, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Modelling Survival Data in Medical Research. , 151-193 (1993).</li><li>Bouliotis, G., Billingham, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Crossing+survival+curves:+alternatives+to+the+log-rank+test.">Crossing survival curves: alternatives to the log-rank test.</a> Trials. 12, A137 (2011).</li><li>Yang, J., 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=Changes+in+expression+of+manganese+superoxide+dismutase,+copper+and+zinc+superoxide+dismutase+and+catalase+in+Brachionus+calyciflorus+during+the+aging+process.">Changes in expression of manganese superoxide dismutase, copper and zinc superoxide dismutase and catalase in Brachionus calyciflorus during the aging process.</a> PloS ONE. 8 (2), e57186 (2013).</li><li>Snell, T. W., Johnston, R. K., Rabeneck, B., Zipperer, C., Teat, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Joint+inhibition+of+TOR+and+JNK+pathways+interacts+to+extend+the+lifespan+of+Brachionus+manjavacas+(Rotifera).">Joint inhibition of TOR and JNK pathways interacts to extend the lifespan of Brachionus manjavacas (Rotifera).</a> Exp. Gerontol. 52, 55-69 (2014).</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+nematode+Caenorhabditis-elegans+-+major+biological+and+environmental-factors+influencing+life-span.">Aging in nematode Caenorhabditis-elegans - major biological and environmental-factors influencing life-span.</a> Mech. Ageing Dev. 6 (6), 413-429 (1977).</li><li>Priest, N. K., Mackowiak, B., Promislow, D. 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=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 (5), 927-935 (2002).</li></ol>

Den nuværende protokol beskriver en fremgangsmåde til måling af overlevelsestid i Brachionus hjuldyr. Det kritiske skridt er synkroniseringen af ​​hjuldyr forhold over flere generationer. Når eksperimentelle hjuldyr er godt synkroniseret, er en typisk type I overlevelse kurve observeret med meget lidt tidligt dødelighed som rapporteret i flere tidligere undersøgelser 18,24,37,38. Standardafvigelser af deres overlevelsestid således blevet mindre i forhold til dårligt synkroniserede hjuldyr...

Hjuldyr er mikroskopiske kosmopolitiske dyreplankton (&lt;1 mm), der udgør den phylum Rotifera en. De har en simpel krop plan sammensat af cirka 1.000 somatiske celler såvel som en karakteristisk hjul-lignende ciliære apparat kaldet koronaen, som anvendes til brug og fodring. De fleste hjuldyr tilhører klasserne Monogononta eller Bdelloidea, som indeholder omkring 1.600 og 500 arter, henholdsvis 2. Monogonont hjuldyr har generelt både seksuelle og aseksuelle reproduktive faser (cyklisk partenogenese), mens bdelloid hjuldyr reproducere ved obligatorisk partenogenese 3. Det er således muligt at opnå genetisk identiske hjuldyr individer, hvilket sikrer høj reproducerbarhed i forsøg. Derudover har de flere andre fordele som modelorganismer, såsom en kort levetid, let kultur, tilgængelighed af genomiske og transkriptomisk sekvensdata 4-7, og en unik fylogenetiske position fjernt fra enrthropods og nematoder 8. Hjuldyr er derfor lovende hvirvelløse modeller i økologiske, toksikologiske og aldrende undersøgelser 9-12. Overlevelsestiden under udsættelse for stress eller kemikalier er en ofte målt parameter i disse forskningsområder 13-19. Imidlertid er forsigtighed nødvendig, når måling overlevelsestiden af ​​hjuldyr, fordi det er modtagelig for miljøforhold deres mødre. Nemlig, i monogonont hjuldyr Brachionus manjavacas, kvindelige afkom fra alderen mødre har en kortere levetid end dem fra unge mødre; dog maternal kalorieindhold begrænsning (CR) delvis udligner de skadelige virkninger af fremskreden moderens alder 20. I B. plicatilis, maternal CR giver afkom levetid, lang overlevelsestid under sult, og høj oxidativt stress modstand forbundet med øget udtryk af antioxidant enzymer 21,22. Den moderens alder effekter også blevet observeret i bdelloid hjuldyr 23. Derfor bør betingelserne for eksperimentelle hjuldyr nøje synkroniseret over flere generationer, før målinger af overlevelsestid. Her giver vi en protokol til måling af overlevelsestid i Brachionus hjuldyr efter synkronisering af dyrkningsbetingelser over flere generationer. Intermitterende fastende (IF), en variation af CR hvor hjuldyr fodres regelmæssigt, blev påført afsløre virkningen af synkronisering som følge af de velkendte virkninger af IF på levetiden 22,24.

<table border="0" cellpadding="0" cellspacing="0" width="688">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:268px;">Sodium chloride</td>
			<td style="width:135px;">Wako</td>
			<td style="width:119px;">190-13921</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:268px;">Magnesium chloride</td>
			<td style="width:135px;">Wako</td>
			<td style="width:119px;">136-03995</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:268px;">Magnesium sulfate</td>
			<td style="width:135px;">Wako</td>
			<td>131-00427</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:268px;">Potassium chloride</td>
			<td style="width:135px;">Wako</td>
			<td style="width:119px;">168-22111</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:268px;">Calcium chloride</td>
			<td style="width:135px;">Wako</td>
			<td style="width:119px;">035-00455</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:268px;">Sodium bicarbonate</td>
			<td style="width:135px;">Wako</td>
			<td style="width:119px;">199-05985</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:268px;">Sodium bromide</td>
			<td style="width:135px;">Wako</td>
			<td style="width:119px;">190-01515</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:268px;">Membrane filter (0.45 &#181;m pore size)</td>
			<td style="width:135px;">Millipore</td>
			<td style="width:119px;">HAWP04700</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:268px;">Culture plate, 6-well, non-treated</td>
			<td style="width:135px;">Thomas Scientific</td>
			<td style="width:119px;">6902D01</td>
			<td>Flat bottom</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:268px;">Culture plate, 48-well, non-treated</td>
			<td style="width:135px;">Thomas Scientific</td>
			<td style="width:119px;">6902D07</td>
			<td>Flat bottom</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:268px;">Tetraselmis, Living</td>
			<td style="width:135px;">Carolina Biological Supply Company</td>
			<td style="width:119px;">152610</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:268px;">PRISM 6</td>
			<td style="width:135px;">GraphPad Software</td>
			<td style="width:119px;">Version 6.0d</td>
			<td></td>
		</tr>
	</tbody>
</table>

measurement of survival time in brachionus rotifers: synchronization of maternal conditions

Rotifers are microscopic cosmopolitan zooplankton used as models in ecotoxicological and aging studies due to their several advantages such as short lifespan, ease of culture, and parthenogenesis that enables clonal culture. However, caution is required when measuring their survival time as it is affected by maternal age and maternal feeding conditions. Here we provide a protocol for powerful and reproducible measurement of the survival time in Brachionus rotifers following a careful synchronization of culture conditions over several generations. Empirically, poor synchronization results in early mortality and a gradual decrease in survival rate, thus resulting in weak statistical power. Indeed, under such conditions, calorie restriction (CR) failed to significantly extend the lifespan of B. plicatilis although CR-induced longevity has been demonstrated with well-synchronized rotifer samples in past and present studies. This protocol is probably useful for other invertebrate models, including the fruitfly Drosophila melanogaster and the nematode Caenorhabditis elegans, because maternal age effects have also been reported in these species.

Figur 1 viser repræsentative overlevelseskurverne for dårligt synkroniserede populationer (ud af to gentagelser). I dette forsøg hjuldyr blev enten fodret hverdag [ad libitum (AL) gruppe] eller hver anden dag (IF gruppe). Median overlevelse var 13 og 18 dage i AL og IF grupper, hhv. Selv om det er velkendt, at IF forlænger levetiden for hjuldyr, dette eksperiment ikke påvise en statistisk signifikant forskel mellem livslængde i AL og IF grupper. Empi...

Watch this Scientific Journal Video about Measurement of Survival Time in Brachionus Rotifers: Synchronization of Maternal Conditions at JoVE.com

Measurement of Survival Time in Brachionus Rotifers: Synchronization of Maternal Conditions

Rotifers are microscopic zooplankton used as models in ecotoxicological and aging studies. Here we provide a protocol for powerful and reproducible measurement of survival time in Brachionus rotifers. Synchronization of culture conditions over several generations is of particular importance because maternal condition affects life history of offspring.

Måling af overlevelse Tid i<em&gt; Brachionus</em&gt; Hjuldyr: Synkronisering af Maternal betingelser

Rotifers are microscopic cosmopolitan zooplankton used as models in ecotoxicological and aging studies due to their several advantages such as short ...

Research

JoVE Journal

Biology

Measurement of Survival Time in Brachionus Rotifers: Synchronization of Maternal Conditions

Time-lapse Microscopy of Early Embryogenesis in Caenorhabditis elegans

Time-lapse Microscopy of Early Embryogenesis in 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

Grundlæggende<em&gt; Caenorhabditis elegans</em&gt; Metoder: Synkronisering og Observation

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

Formulations for Freeze-drying of Bacteria and Their Influence on Cell Survival

Formuleringer til frysetørring af bakterier og deres indflydelse på Cell Survival

Cellular water can be removed to reversibly inactivate microorganisms to facilitate storage. One such method of removal is freeze-drying, which is ...

Quantitative Measurement of Invadopodia-mediated Extracellular Matrix Proteolysis in Single and Multicellular Contexts

Kvantitativ måling af Invadopodia-medieret ekstracellulær matrix Proteolyse i Single og flercellede Contexts

Cellular invasion into local tissues is a process important in development and homeostasis.  Malregulated invasion and subsequent cell movement is ...

Measurement of Lifespan in Drosophila melanogaster

Measurement of Lifespan in Drosophila melanogaster

Måling af Levetid i<em&gt; Drosophila melanogaster</em

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

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs

Probe-baseret real-time PCR metoder til kvantitativ måling af microRNA

Probe-based quantitative PCR (qPCR) is a favoured method for measuring transcript abundance, since it is one of the most sensitive detection methods that ...

Synchronization of Caulobacter Crescentus for Investigation of the Bacterial Cell Cycle

Synchronization of Caulobacter Crescentus for Investigation of the Bacterial Cell Cycle

Synkronisering af<em&gt; Caulobacter Crescentus</em&gt; Til undersøgelse af bakteriecellen Cycle

The cell cycle is important for growth, genome replication, and development in all cells.  In bacteria, studies of the cell cycle have focused largely on ...

Temporal Tracking of Cell Cycle Progression Using Flow Cytometry without the Need for Synchronization

Temporal Sporing af cellecyklusprogression anvendelse af flowcytometri uden behov for synkronisering

This protocol describes a method to permit the tracking of cells through the cell cycle without requiring the cells to be synchronized. Achieving cell ...

High-resolution Time-lapse Imaging and Automated Analysis of Microtubule Dynamics in Living Human Umbilical Vein Endothelial Cells

Høj opløsning Time-lapse Imaging og Automatiseret analyse af mikrotubulusdynamik i levende menneske navlestrengsveneendotelceller

The physiological process by which new vasculature forms from existing vasculature requires specific signaling events that trigger morphological changes ...