אנו מודים אוסמן Kaftanoglu לעצות מועילות וסיוע בזמן צילומים. ברצוננו להודות למבקרים האנונימיים על הערות תובנה. עבודה זו נתמכה על ידי המועצה למחקר של נורבגיה (מעניק 180,504, 191,699, ו213,976), מארי Curie/FP7 (נ&quot;צ פרויקט. 238,665), המכון הלאומי ל( AG22500 P01 NIA מענק) הזדקנות, ונאמנויות הצדקה Pew.

<ol>
	<li>Munch, D., Amdam, G. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+curious+case+of+aging+plasticity+in+honey+bees.">The curious case of aging plasticity in honey bees.</a> FEBS Lett. 584, 2496-2503 (2010).</li><li>Buffenstein, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Negligible+senescence+in+the+longest+living+rodent,+the+naked+mole-rat:+insights+from+a+successfully+aging+species.">Negligible senescence in the longest living rodent, the naked mole-rat: insights from a successfully aging species.</a> J Comp Physiol B. 178, 439-445 (2008).</li><li>Parker, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=What+are+social+insects+telling+us+about+aging?.">What are social insects telling us about aging?.</a> Myrmecological News. 13, 103-110 (2010).</li><li>Seeley, T. 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> The Wisdom of the Hive. , (1995).</li><li>Amdam, G. V., Omholt, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+regulatory+anatomy+of+honeybee+lifespan.">The regulatory anatomy of honeybee lifespan.</a> J Theor Biol. 216, 209-228 (2002).</li><li>Amdam, G. V., Norberg, K., Hagen, A., Omholt, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Social+exploitation+of+vitellogenin.">Social exploitation of vitellogenin.</a> Proc Natl Acad Sci U S A. 100, 1799-1802 (2003).</li><li>Guidugli, K. R., 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=Vitellogenin+regulates+hormonal+dynamics+in+the+worker+caste+of+a+eusocial+insect.">Vitellogenin regulates hormonal dynamics in the worker caste of a eusocial insect.</a> FEBS Lett. 579, 4961-4965 (2005).</li><li>Amdam, G. V., 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=Social+reversal+of+immunosenescence+in+honey+bee+workers.">Social reversal of immunosenescence in honey bee workers.</a> Exp Gerontol. 40, 939-947 (2005).</li><li>Seehuus, S. C., Norberg, K., Gimsa, U., Krekling, T., Amdam, G. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reproductive+protein+protects+functionally+sterile+honey+bee+workers+from+oxidative+stress.">Reproductive protein protects functionally sterile honey bee workers from oxidative stress.</a> Proc Natl Acad Sci U S A. 103, 962-967 (2006).</li><li>Scheiner, R., Amdam, G. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+tactile+learning+is+related+to+social+role+in+honeybees.">Impaired tactile learning is related to social role in honeybees.</a> J Exp Biol. 212, 994-1002 (2009).</li><li>Behrends, A., Scheiner, R., Baker, N., Amdam, G. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cognitive+aging+is+linked+to+social+role+in+honey+bees+(Apis+mellifera.">Cognitive aging is linked to social role in honey bees (Apis mellifera.</a> Exp Gerontol. 42, 1146-1153 (2007).</li><li>M&#252;nch, D., Baker, N., Kreibich, C. D., Braten, A. T., Amdam, G. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+the+laboratory+and+during+free-flight:+old+honey+bees+reveal+learning+and+extinction+deficits+that+mirror+mammalian+functional+decline.">In the laboratory and during free-flight: old honey bees reveal learning and extinction deficits that mirror mammalian functional decline.</a> PLoS One. 5, e13504 (2010).</li><li>Vance, J. T., Williams, J. B., Elekonich, M. M., Roberts, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+age+and+behavioral+development+on+honey+bee+(Apis+mellifera)+flight+performance.">The effects of age and behavioral development on honey bee (Apis mellifera) flight performance.</a> J Exp Biol. 212, 2604-2611 (2009).</li><li>Dukas, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mortality+rates+of+honey+bees+in+the+wild.">Mortality rates of honey bees in the wild.</a> Insect Soc. 55, (2008).</li><li>Behrends, A., Scheiner, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Learning+at+old+age:+a+study+on+winter+bees.">Learning at old age: a study on winter bees.</a> Front Behav Neurosci. 4, 15 (2010).</li><li>Huang, Z. -. Y., Robinson, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Honeybee+colony+integration:+Worker-worker+interactions+mediate+hormonally+regulated+plasticity+in+division+of+labor.">Honeybee colony integration: Worker-worker interactions mediate hormonally regulated plasticity in division of labor.</a> Proc Natl Acad Sci USA. 89, 11726-11729 (1992).</li><li>Huang, Z. Y., Robinson, G. E. <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+honey+bee+division+of+labor+by+colony+age+demography.">Regulation of honey bee division of labor by colony age demography.</a> Behavioral Ecology and Sociobiology. 39, 147-158 (1996).</li><li>Double, K. L., 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=The+comparative+biology+of+neuromelanin+and+lipofuscin+in+the+human+brain.">The comparative biology of neuromelanin and lipofuscin in the human brain.</a> Cell Mol Life Sci. 65, 1669-1682 (2008).</li><li>Fonseca, D. B., Brancato, C. L., Prior, A. E., Shelton, P. M., Sheehy, 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=Death+rates+reflect+accumulating+brain+damage+in+arthropods.">Death rates reflect accumulating brain damage in arthropods.</a> Proc Biol Sci. 272, 1941-1947 (2005).</li><li>Baker, N., Wolschin, F., Amdam, G. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Age-related+learning+deficits+can+be+reversible+in+honeybees+Apis+mellifera.">Age-related learning deficits can be reversible in honeybees Apis mellifera.</a> Exp Gerontol. 47, 764-772 (2012).</li><li>Capaldi, E. A., 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=Ontogeny+of+orientation+flight+in+the+honeybee+revealed+by+harmonic+radar.">Ontogeny of orientation flight in the honeybee revealed by harmonic radar.</a> Nature. 403, 537-540 (2000).</li><li>Marco Antonio, D. S., Guidugli-Lazzarini, K. R., do Nascimento, A. M., Simoes, Z. L., Hartfelder, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=RNAi-mediated+silencing+of+vitellogenin+gene+function+turns+honeybee+(Apis+mellifera)+workers+into+extremely+precocious+foragers.">RNAi-mediated silencing of vitellogenin gene function turns honeybee (Apis mellifera) workers into extremely precocious foragers.</a> Naturwissenschaften. 95, 953-961 (2008).</li><li>Whitfield, C. W., Cziko, A. -. M., Robinson, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+expression+profiles+in+the+brain+predict+behavior+in+individual+honey+bees.">Gene expression profiles in the brain predict behavior in individual honey bees.</a> Science. 302, 296-299 (2003).</li><li>Schmidt, J. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Attraction+of+reproductive+honey+bee+swarms+to+artificial+nests+by+Nasonov+pheromone.">Attraction of reproductive honey bee swarms to artificial nests by Nasonov pheromone.</a> Journal of Chemical Ecology. 20, 1053-1056 (1994).</li><li>De Moraes, R., Bowen, I. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modes+of+cell+death+in+the+hypopharyngeal+gland+of+the+honey+bee+(Apis+mellifera+L).">Modes of cell death in the hypopharyngeal gland of the honey bee (Apis mellifera L).</a> Cell Biol Internat. 24, 737-743 (2000).</li><li>Sheehy, 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=A+flow-cytometric+method+for+quantification+of+neurolipofuscin+and+comparison+with+existing+histological+and+biochemical+approaches.">A flow-cytometric method for quantification of neurolipofuscin and comparison with existing histological and biochemical approaches.</a> Arch Gerontol Geriatr. 34, 233-248 (2002).</li><li>Hsieh, Y. S., Hsu, C. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Honeybee+trophocytes+and+fat+cells+as+target+cells+for+cellular+senescence+studies.">Honeybee trophocytes and fat cells as target cells for cellular senescence studies.</a> Exp Gerontol. 46, 233-240 (2011).</li><li>Wolschin, F., Munch, D., Amdam, G. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structural+and+proteomic+analyses+reveal+regional+brain+differences+during+honeybee+aging.">Structural and proteomic analyses reveal regional brain differences during honeybee aging.</a> J Exp Biol. 212, 4027-4032 (2009).</li></ol>

יש לנו מה למסור.

אנחנו כאן לאמץ בעבר תאר גישות 8,16,17,19,20, ולשלב אותם לתוך זרימת עבודה אחת שתאפשר לימוד הזדקנות גמישה בדבורי דבש. המטרה שלנו היא לספק למדענים כי הם טירון בתחום זה עם כלי סטנדרטי מוגדר להשיג חומר מדגם רלוונטי, וכדי לשפר את שחזור ניסיוני בקרב צוותי מחקר שונים. בעוד שהנ...

מבני המושבה המורכבים של בעלי חיים חברתיים מאוד נשמרים באמצעות האינטראקציה של כת רבייה, וכת עוזר של כלל עובדים הלא כפול עם התנהגויות משימה חברתית שונות. בפועלים השונים, התאמות פיסיולוגיות ספציפיות לאפשר התנהגויות טיפול SIB מובהקות, וגם קשורות להבדלי תוחלת חיים קיצוניים. דבורי דבש וחפרפרות מייצגים את המודלים של בעלי החיים הטובים ביותר שפותחו כדי ללמוד איך sociality קשור לדפוסי מואצים, זניח או הפוך הזדקנות 1-3. במושבות דבורי דבש, מלכת הטלה אחת נעזרת באלף עובדים שנוטים להרהר, ללקט מזון, ולעסוק בשמירה, ויסות חום או התנהגויות היגיינה 4. בין העובדים אלה מלקטי מזון קצרים מאוד, דבורי אחות עם ביניים, ודבורים בחורף (diutinus) עם תוחלת החיים הארוכים ביותר הם. אנשים, לעומת זאת, לא באופן קבוע מחויבים מסוים ווrker סוג, אבל להציג ontogeny התנהגותיים גמיש: הם משתנים ממשימת התנהגות חברתית אחד למשנו (&quot;קסטות זמניות&quot;). דבורי קאלו יכולים לשנות לדבורי אחות להרהר בטיפול, שסופו של דבר עשוי להשתנות לליקוט מזון בחוץ. עם זאת, דבורי קן חסר ניסיון יכולים גם להפוך לדבורים בחורף ארוך חיים, ומלקטי מזון קצר מועד יכול אפילו לחזור לאחיות בדרך כלל אורך חיים גדולים יותר. עובדים עם קיצוניות (דבורים בחורף) ואורך חיים בינוניים (דבורי אחות) פיתחו היטב איברי ייצור ואחסון מזון עם משאבים עצומות - בניגוד למלקטי מזון קצר מועד (הנסקרת ב 1,5). עם זאת, שהרגולציה של תוחלת חיים בודדות חורגת שינויים פשוטים באיזון המשאבים של פרט שהוצע על ידי מחקר על חלבון חלמון, שבה יש פונקציות מותאמות מגוונות בכת העובד שאינו מתרבה, כגון ייצור ריבת 6, בקרה הורמונלית 7, חיסוניים 8 ונוגד חמצון להגנה 9. דפוסים של fירידת unctional (הזדקנות) פערי תוחלת חיים מראה בקרב עובדים, כפי שנקבע על חוש הריח, וגם לתפקודי מוח או מנוע אחרים 10-13. באופן ספציפי, הירידה המשמעותית בתפקוד למידה רק לאחר שבועיים של ליקוט תואמת התקדמות תמותה דומה במלקטי מזון 14, בניגוד לחוסר ירידה לזיהוי (הזדקנות זניחה) בדבורים בחורף ארוכים, חי 15. כדי לזהות את טביעות האצבעות מולקולריות של הזדקנות גמישה שהותאמנו פרדיגמות ניסוי קבעו כי נאפשר לניטור וטיפול במעברי הזדקנות מסוג 8,16,17. להתנסות פרטים 1 איך להשיג דגימות שבו ניתן להפריד את ההשפעות של גיל כרונולוגי והתנהגויות חברתיות מסוימות עובד מסוג על הזדקנות. ניסוי 2 מתאר את ההיפוך של מלקטי מזון עם מואצים לתוך דבורי אחות עם דינמיקת הזדקנות מואטת. ניסוי 3 מספק גישה לחיטוט השפעות של הזדקנות תאית על ידי anatomicaכימות ליטר של סמן ביולוגי שהוקם להזדקנות תאים (lipofuscin) 18.

<table border="1">
 <tbody>
 <tr>
 <td>Name of reagent</td>
 <td>Company</td>
 <td>Catalogue number</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>Apifonda</td>
 <td>S&uuml;dzucker AG, Mannheim/Ochsenfurt, Germany</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>paraformaldehyde</td>
 <td>Sigma-Aldrich</td>
 <td>158127</td>
 <td></td>
 </tr>
 <tr>
 <td>phosphate-buffered saline</td>
 <td>Sigma-Aldrich</td>
 <td>P4417</td>
 <td></td>
 </tr>
 <tr>
 <td>Glycerol</td>
 <td>Merck</td>
 <td>1.04094.1000</td>
 <td></td>
 </tr>
 </tbody>
</table>

obtaining specimens with slowed, accelerated and reversed aging in the honey bee model

אגודות בעלי החיים חברתיים מאוד וכוללות הבדלי תוחלת חיים עצומים בין יחידים הקשורים באופן הדוק. בקרב חרקים חברתיים, דבורת הדבש היא המודל הטוב ביותר שהוקם במטרה ללמוד כיצד פלסטיות בתוחלת חיים והזדקנות מוסברת על ידי גורמים חברתיים. כת העובד של דבורי דבש דבורים כוללת אחות, אשר נוטים להרהר, ודבורים בוזזים, אשר אוספים צוף ואבקת פרחים. העבודות קודמות הראו כי תפקודי מוח וביצועי טיסה להזדקן במהירות רבה יותר במלקטי מזון מאשר באחיות. עם זאת, תפקודי מוח יכולים להתאושש, כאשר מלקטי מזון לחזור למשימות סיעוד. דפוסים כאלה של הזדקנות מואצת ופונקציונלית ההפוכה צמודים לרמות משאבי חילוף חומרים שהשתנו, לשינויים בשפע חלבון ואת תפקוד מערכת החיסון. Vitellogenin, חלבון חלמון עם פונקציות מותאמות בבקרה הורמונלית והגנה תאית, עשוי לשמש כאלמנט רגולטורים מרכזי ברשת ששולטת בדינמיקת ההזדקנות שונה בעובדים. כאן אנו מתארים כיצד הופעתם של אחיות ומלקטי מזון יכול להיות במעקב, ומניפולציות, ובם ההיפוך ממלקטי מזון קצר בדרך כלל לאחיות אורך חיים גדולות יותר. תוצאות הנציג שלנו להראות כיצד אנשים עם גיל כרונולוגי דומה להתמיין למלקטי מזון ודבורים אחות תחת תנאי ניסוי. אנו מדגימים כיצד ניתן תוקף היפוך התנהגותיות ממלקטי מזון בחזרה לאחיות. לאחרונה, אנו מראים כיצד שונים הזדקנות תאית יכולה להיות מוערכת על ידי מדידת ההצטברות של lipofuscin, סמן ביולוגי אוניברסלי של הזדקנות. ללימוד מנגנונים העשויים לקשר את ההשפעות חברתיות והזדקנות פלסטיות, פרוטוקול זה מספק כלי סטנדרטי שנקבע לרכישת חומר מדגם רלוונטי, וכדי לשפר את יכולת השוואת נתונים בין מחקרים עתידיים.

סעיפי 1 ו -2 לפרוטוקול פירוט כיצד ניתן להשיג קבוצות מבחן ללמוד תכונות של מואץ, האטו והתהפכו הזדקנות במושבות עם קבוצת גיל אחת. כדי לפקח על בידול עובד מהסוג שמלווה את ontogeny הנורמלי שאנחנו הערכנו ספירות בוזזות (&quot;ספירת כניסה&quot;) במשך 6 מושבות (איור 1, השווה סעיף 1). ?...

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Obtaining Specimens with Slowed, Accelerated and Reversed Aging in the Honey Bee Model

בפועלי דבורת דבש, הזדקנות תלויה בהתנהגויות חברתיות ולא על גיל כרונולוגי. כאן אנו מראים כיצד ניתן להשיג עובד סוגים עם דפוסי הזדקנות שונים מאוד ונותחו להזדקנות תאית.

קבלת דגימות עם מואט, מואצת והפוך הזדקנות של דבורת דבש הדגם

Societies of highly social animals feature vast lifespan differences between closely related individuals. Among social insects, the honey bee is the best ...

obtaining-specimens-with-slowed-accelerated-reversed-aging-honey-bee

Research

JoVE Journal

Biology

11.2K Views.  Norwegian University of Life Sciences.  בפועלי דבורת דבש, הזדקנות תלויה בהתנהגויות חברתיות ולא על גיל כרונולוגי. כאן אנו מראים כיצד ניתן להשיג עובד סוגים עם דפוסי הזדקנות שונים מאוד ונותחו להזדקנות תאית. 

Video: Obtaining Specimens with Slowed, Accelerated and Reversed Aging in the Honey Bee Model

Brain Banking: Making the Most of your Research Specimens

Unbiased stereology is a method for accurately and efficiently estimating the total neuron number (or other cell type) in a given area of interest1. To achieve this goal 6-10 systematic sections should be probed covering the entire structure. Typically this involves processing 1/5 sections which leaves a significant amount of material unprocessed. In order to maximize the material, we propose an inexpensive method for preserving fixed tissue as part of a long-term storage research plan. As tissue is sliced and processed for the desired stain or antibody, alternate sections should be systematically placed in antigen preserve at -20&deg;C for future processing. Using 24-well plates, sections can be placed in order for future retrieval. Using this method, tissue can be stored and processed for immunohistochemistry over the course of years.

Brain banking and systematic sampling of biological material provides the basis for unbiased stereology and maximizes the potential data obtained from each specimen.

בנקאות המוח: הפיכת רוב דוגמאות המחקר שלך

Unbiased stereology is a method for accurately and efficiently estimating the total neuron number (or other cell type) in a given area of interest1.  To ...

In vivo Ca2+- Imaging of Mushroom Body Neurons During Olfactory Learning in the Honey Bee

The in vivo and semi-in vivo preparation for Calcium imaging has been developed in our lab by Joerges, K&uuml;ttner and Galizia over ten years ago, to measure odor evoked activity in the antennal lobe1. From then on, it has been continuously refined and applied to different neuropiles in the bee brain. Here, we describe the preparation currently used in the lab to measure activity in mushroom body neurons using a dextran coupled calcium-sensitive dye (Fura-2). We retrogradely stain mushroom body neurons by injecting dye into their axons or soma region. We focus on reducing the invasiveness, to achieve a preparation in which it is still possible to train the bee using PER conditioning. We are able to monitor and quantify the behavioral response by recording electro-myograms from the muscle which controls the PER (M17)2.
After the physiological experiment the imaged structures are investigated in greater detail using confocal scanning microscopy to address the identity of the neurons.

In vivo Ca2+- Imaging of Mushroom Body Neurons During Olfactory Learning in the Honey Bee

Bees can be conditioned in an appetitive olfactory learning paradigm (PER-conditioning). Using odors as stimuli, we established a method in which behavior is recorded while simultaneously Calcium Imaging is used to measure odor evoked activity in mushroom body neurons in vivo.

בשנת vivo Ca 2 + - הדמיה של נוירונים גוף אינפקטד במהלך הלמידה חוש הריח של דבורת הדבש

The in vivo and semi-in vivo preparation for Calcium imaging has been developed in our lab by Joerges, K&uuml;ttner and Galizia over ten years ago, to ...

Visualizing the Beating Heart in Drosophila

The Drosophila heart has recently emerged as a good model system for examining the genetic, cellular, and molecular mechanisms underlying function in myogenic hearts. A key step in examining heart function in the fly is finding a way to access the heart in a manner that preserves its myogenic function while still allowing the beating heart organ to be observed and recorded. Two different methods for observing and recording the beating heart in both larva and adult Drosophila are described here. Our semi-intact preparation using adult flies allows clear visualization of the abdominal heart without interference from the pigmented cuticle and overlying fat bodies. To record larval heart beats it is necessary to immobilize the larva, which minimizes body wall movements thereby reducing heart movements that are not associated with myocardial contractions. Our methodologies produce stable adult and larval heart preparations that can beat for hours at rates of 1-3 Hz.

Visualizing the Beating Heart in Drosophila

Technique required for visualizing the beating heart in larval and adult Drosophila are presented. Each life stage requires a different methodology.

לדמיין את הלב הפועם תסיסנית</em

The Drosophila heart has recently emerged as a good model system for examining the genetic, cellular, and molecular mechanisms underlying function in ...

Methods for Comparing Nutrients in Beebread Made by Africanized and European Honey Bees and the Effects on Hemolymph Protein Titers

Honey bees obtain nutrients from pollen they collect and store in the hive as beebread. We developed methods to control the pollen source that bees collect and convert to beebread by placing colonies in a specially constructed enclosed flight area. Methods were developed to analyze the protein and amino acid composition of the pollen and beebread. We also describe how consumption of the beebread was measured and methods used to determine adult worker bee hemolymph protein titers after feeding on beebread for 4, 7 and 11 days after emergence. Methods were applied to determine if genotype affects the conversion of pollen to beebread and the rate that bees consume and acquire protein from it. Two subspecies (European and Africanized honey bees; EHB and AHB respectively) were provided with the same pollen source. Based on the developed methods, beebread made by both subspecies had lower protein concentrations and pH values than the pollen. In general, amino acid concentrations in beebread made by either EHB or AHB were similar and occurred at higher levels in beebread than in pollen. Both AHB and EHB consumed significantly more of the beebread made by AHB than by EHB. Though EHB and AHB consumed similar amounts of each type of beebread, hemolymph protein concentrations in AHB were higher than in EHB. Differences in protein acquisition between AHB and EHB might reflect environmental adaptations related to the geographic region where each subspecies evolved. These differences could contribute to the successful establishment of AHB populations in the New World because of the effects on brood rearing and colony growth.

Here are methods to quantify nutrients in pollen before and after its conversion to beebread by two subspecies of honeybees. We describe techniques to measure beebread consumption and resulting protein titers in both subspecies.

שיטות להשוואה בין חומרים מזינים בBeebread נוצרו על ידי דבורים דבש Africanized ואירופאי והשפעות על titers חלבון Hemolymph

Honey bees obtain nutrients from pollen they collect and store in the hive as beebread. We developed methods to control the pollen source that bees ...

Dissection and Observation of Honey Bee Dorsal Vessel for Studies of Cardiac Function

The European honey bee, Apis mellifera L., is a valuable agricultural and commercial resource noted for producing honey and providing crop pollination services, as well as an important model social insect used to study memory and learning, aging, and more. Here we describe a detailed protocol for the dissection of the dorsal abdominal wall of a bee in order to visualize its dorsal vessel, which serves the role of the heart in the insect. A successful dissection will expose a functional heart that, under the proper conditions, can maintain a steady heartbeat for an extended period of time. This allows the investigator to manipulate heart rate through the application of cardiomodulatory compounds to the dorsal vessel. By using either a digital microscope or a microscope equipped with a digital camera, the investigator can make video recordings of the dorsal vessel before and after treatment with test compounds. The videos can then be scored at a time convenient to the user in order to determine changes in heart rate, as well as changes in the pattern of heartbeats, following treatment. The advantages of this protocol are that it is relatively inexpensive to set up, easy to learn, requires little space or equipment, and takes very little time to conduct.

The abdominal dorsal vessel of the honey bee and other insects serves as the functional equivalent of the mammalian heart and plays an important role in nutrient transport, waste removal, immune function, and more. Here we describe a protocol for the visualization and pharmacological manipulation of bee heart rate.

דיסקציה תצפית של כלי דבש דבורים הגבי ללימודים של תפקוד הלב

The European honey bee, Apis mellifera L., is a valuable agricultural and commercial resource noted for producing honey and providing crop pollination ...

Monitoring Colony-level Effects of Sublethal Pesticide Exposure on Honey Bees

The effects of sublethal pesticide exposure to honey bee colonies may be significant but difficult to detect in the field using standard visual assessment methods. Here we describe methods to measure the quantities of adult bees, brood, and food resources by weighing hives and hive parts, by photographing frames, and by installing hives on scales and with internal sensors. Data from these periodic evaluations are then combined with running average and daily detrended data on hive weight and internal hive temperature. The resulting datasets have been used to detect colony-level effects of imidacloprid applied in a sugar syrup as low as 5 parts per billion. The methods are objective, require little training, and provide permanent records in the form of sensor output and photographs.

Sublethal doses of pesticides may affect colonies in ways that are difficult to detect using only periodic or visual methods. Methods to measure total adult bee mass, brood, and food resources by weighing hives and hive parts, photographing frames, and installing sensors, are provided. Data analysis is also addressed.

ניטור המושבה ברמת ההשפעה של ההדברה לא קטלני על דבש דבורים

The effects of sublethal pesticide exposure to honey bee colonies may be significant but difficult to detect in the field using standard visual assessment ...

Methods to Study Changes in Inherent Protein Aggregation with Age in Caenorhabditis elegans

In the last decades, the prevalence of neurodegenerative disorders, such as Alzheimer's disease (AD) and Parkinson's disease (PD), has grown. These age-associated disorders are characterized by the appearance of protein aggregates with fibrillary structure in the brains of these patients. Exactly why normally soluble proteins undergo an aggregation process remains poorly understood. The discovery that protein aggregation is not limited to disease processes and instead part of the normal aging process enables the study of the molecular and cellular mechanisms that regulate protein aggregation, without using ectopically expressed human disease-associated proteins. Here we describe methodologies to examine inherent protein aggregation in Caenorhabditis elegans through complementary approaches. First, we examine how to grow large numbers of age-synchronized C. elegans to obtain aged animals and we present the biochemical procedures to isolate highly-insoluble-large aggregates. In combination with a targeted genetic knockdown, it is possible to dissect the role of a gene of interest in promoting or preventing age-dependent protein aggregation by using either a comprehensive analysis with quantitative mass spectrometry or a candidate-based analysis with antibodies. These findings are then confirmed by in vivo analysis with transgenic animals expressing fluorescent-tagged aggregation-prone proteins. These methods should help clarify why certain proteins are prone to aggregate with age and ultimately how to keep these proteins fully functional.

Methods to Study Changes in Inherent Protein Aggregation with Age in Caenorhabditis elegans

The goal of the method presented here is to explore protein aggregation during normal aging in the model organism C. elegans. The protocol represents a powerful tool to study the highly insoluble large aggregates that form with age and to determine how changes in proteostasis impact protein aggregation.

שיטות מחקר לשינויים הגלום צבירת חלבון עם הגיל ב Caenorhabditis elegans

In the last decades, the prevalence of neurodegenerative disorders, such as Alzheimer's disease (AD) and Parkinson's disease (PD), has grown. These ...

SA-&#946;-Galactosidase-Based Screening Assay for the Identification of Senotherapeutic Drugs

Cell senescence is one of the hallmarks of aging known to negatively influence a healthy lifespan. Drugs able to kill senescent cells specifically in cell culture, termed senolytics, can reduce the senescent cell burden in vivo and extend healthspan. Multiple classes of senolytics have been identified to date including HSP90 inhibitors, Bcl-2 family inhibitors, piperlongumine, a FOXO4 inhibitory peptide and the combination of Dasatinib/Quercetin. Detection of SA-&#946;-Gal at an increased lysosomal pH is one of the best characterized markers for the detection of senescent cells. Live cell measurements of senescence-associated &#946;-galactosidase (SA-&#946;-Gal) activity using the fluorescent substrate C12FDG in combination with the determination of the total cell number using a DNA intercalating Hoechst dye opens the possibility to screen for senotherapeutic drugs that either reduce overall SA-&#946;-Gal activity by killing of senescent cells (senolytics) or by suppressing SA-&#946;-Gal and other phenotypes of senescent cells (senomorphics). Use of a high content fluorescent image acquisition and analysis platform allows for the rapid, high throughput screening of drug libraries for effects on SA-&#946;-Gal, cell morphology and cell number.

Cellular senescence is the key factor in the development of chronic age-related pathologies. Identification of therapeutics that target senescent cells show promise for extending healthy aging. Here, we present a novel assay to screen for the identification of senotherapeutics based on measurement of senescence associated &#946;-Galactosidase activity in single cells.

א-β-גלטוסידאז-מבוסס על הקרנת ההקרנה לזיהוי תרופות הסאותרפיה

Cell senescence is one of the hallmarks of aging known to negatively influence a healthy lifespan. Drugs able to kill senescent cells specifically in cell ...

A 3D Printed Pollen Trap for Bumble Bee (Bombus) Hive Entrances

To verify the plant sources from which bumble bees forage for pollen, individuals must be collected to remove their corbicular pollen loads for analysis. This has traditionally been done by netting foragers at nest entrances or on flowers, chilling the bees on ice, and then removing the pollen loads from the corbiculae with forceps or a brush. This method is time and labor intensive, may alter normal foraging behavior, and can result in stinging incidents for the worker performing the task. Pollen traps, such as those used on honey bee hives, collect pollen by dislodging corbicular pollen loads from the legs of workers as they pass through screens at the nest entrance. Traps can remove a large quantity of pollen from returning forager bees with minimal labor, yet to date no such trap is available for use with bumble bee colonies. Workers within a bumble bee colony can vary in size making size selection of entrances difficult to adapt this mechanism to commercially reared bumble bee hives. Using 3D printing design programs, we created a pollen trap that successfully removes the corbicular pollen loads from the legs of returning bumble bee foragers. This method significantly reduces the amount of time required by researchers to collect pollen from bumble bee foragers returning to the colony. We present the design, results of pollen removal efficiency tests, and suggest areas of modifications for investigators to adapt traps to a variety of bumble bee species or nest box designs.

A 3D Printed Pollen Trap for Bumble Bee Bombus Hive Entrances

We present a non-lethal and automated mechanism to collect pollen from bumble bee (Bombus) workers returning to a hive. Instructions for producing, preparing, installing and using the devices are included. By using 3D-printed objects, modification to the design was timely, efficient and allowed for quick turnaround for testing.

מלכודת אבקה מודפסת תלת-ממדית לכניסות דבורת באמבל(בומבוס)

To verify the plant sources from which bumble bees forage for pollen, individuals must be collected to remove their corbicular pollen loads for analysis.  ...

Gonadectomy and Blood Sampling Procedures in the Small Size Teleost Model Japanese Medaka (Oryzias latipes)

Sex steroids, produced by the gonads, play an essential role in brain and pituitary tissue plasticity and in the neuroendocrine control of reproduction in all vertebrates by providing feedback to the brain and pituitary. Teleost fishes possess a higher degree of tissue plasticity and variation in reproductive&#160;strategies compared to mammals and appear to be useful models to investigate the role of sex steroids and the mechanisms by which they act. The removal of the main source of sex steroid production using gonadectomy together with blood sampling to measure steroid levels has been well-established and fairly feasible in bigger fish and is a powerful technique to investigate the role and effects of sex steroids. However, these techniques raise challenges when implemented in small size teleost models. Here, we describe the step-by-step procedures of gonadectomy in both males and female Japanese medaka followed by blood sampling. These protocols are shown to be highly feasible in medaka indicated by a high survival rate, safety for the life span and phenotype of the fish, and reproducibility in terms of sex steroid clearance. The use of these procedures combined with the other advantages of using this small teleost model will greatly improve the understanding of feedback mechanisms in the neuroendocrine control of reproduction and tissue plasticity provided by sex steroids in vertebrates.

Gonadectomy and Blood Sampling Procedures in the Small Size Teleost Model Japanese Medaka Oryzias latipes

The article describes a quick protocol to gonadectomize and sample blood from the small teleost fish, using Japanese medaka (Oryzias latipes) as a model, to investigate the role of sex steroids in animal physiology.

הליכי גונדקטומיה ודגימה בדם במודל טלוסט קטן מדאקה(Oryzias latipes)

Sex steroids, produced by the gonads, play an essential role in brain and pituitary tissue plasticity and in the neuroendocrine control of reproduction in ...

קבלת דגימות עם מואט, מואצת והפוך הזדקנות של דבורת דבש הדגם

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