{"id":18423,"date":"2020-02-10T17:43:40","date_gmt":"2020-02-10T16:43:40","guid":{"rendered":"https:\/\/news.embl.de\/?p=18423"},"modified":"2024-03-22T11:16:58","modified_gmt":"2024-03-22T10:16:58","slug":"new-technique-for-studying-living-fly-larvae","status":"publish","type":"post","link":"https:\/\/www.embl.org\/news\/science\/new-technique-for-studying-living-fly-larvae\/","title":{"rendered":"New technique for studying living fly larvae"},"content":{"rendered":"\n<p>Scientists led by the European Molecular Biology Laboratory in Heidelberg, the Institute for Genetics of the University of Cologne, the Max Planck Institute for Biology of Ageing, and CECAD and the CMMC in Cologne have developed a method to study processes taking place in the cells of a living fruit fly larva. This technique, published in&nbsp;<em>Nature Protocols<\/em>&nbsp;on 10 February, provides a simple but effective way to study the functions of organs in living animals.<\/p>\n\n\n\n<p>For decades, scientists have been using the fruit fly <em>Drosophila <\/em>to understand biological processes. Key biological processes and 60% of fruit fly genes are also seen in humans, having been evolutionarily conserved. The fruit fly is therefore a powerful model organism for studying genetics. Scientists have created many genetic tools and techniques to study molecular processes of development and human disease in the fruit fly.<\/p>\n\n\n\n<h2 class=\"wp-block-heading\">The challenge<\/h2>\n\n\n\n<p>Most previous studies have been carried out in the <em>Drosophila<\/em> embryo or in adult flies, but the fruit fly larva also offers enormous research potential. Its transparent body, with highly developed functional organs including brain, gut, and muscle, makes the larva an excellent system for observing the dynamics of cellular and molecular processes in living animals.<\/p>\n\n\n\n<p><em>\u201cThe cells in a fruit fly larva are much bigger than in the embryo. You can see all organelles and even the subcellular processes. That&#8217;s the beauty of the system,\u201d<\/em>&nbsp;says Parisa Kakanj, the scientist who led the study in Germany.<\/p>\n\n\n\n<p>However, the larva\u2019s continuous crawling makes live monitoring a challenge, and some cellular processes only reveal their secrets through long-term imaging. Kakanj, a researcher in the groups of Maria Leptin and Linda Partridge, tackled this challenge and developed a method for simple immobilisation of the larvae that allows long-term live imaging and the investigation of cellular events at high resolution.<\/p>\n\n\n<div\n  class=\"vf-embed vf-embed--custom-ratio\"\n\n  style=\"--vf-embed-max-width: 100%;\n    --vf-embed-custom-ratio-x: 640;\n    --vf-embed-custom-ratio-y: 360;\"><iframe loading=\"lazy\" width=\"640\" height=\"360\" src=\"https:\/\/www.youtube.com\/embed\/dFMoTW8kSnI\" frameborder=\"0\" allow=\"accelerometer; autoplay; encrypted-media; gyroscope; picture-in-picture\" allowfullscreen><\/iframe><\/div>\n\n\n\n<p class=\"vf-u-margin__bottom--sm\"><\/p>\n\n\n\n<p class=\"vf-figure__caption\"><strong>Live imaging of insulin signalling and formation of a contractile cable during wound healing in Drosophila larvae.<\/strong> A sensor for an insulin signal is shown in green, and myosin, a motor protein, in magenta. When the skin is wounded, insulin signalling triggers the formation and contraction of an actomyosin cable around the wound, which constricts the wound and disappears when the wound is closed. VIDEO: Used with permission from Kakanj, P. et al., Nature Protocols (2020).<\/p>\n\n\n\n<h2 class=\"wp-block-heading\">Simple, fast, and efficient<\/h2>\n\n\n\n<p>In the past, scientists have used mechanical methods or anaesthetics to tranquilise fruit fly larvae. Both approaches had undesired side-effects, such as mechanical pressure, limited immobilisation, and high mortality. Further, simultaneous screening of many larvae was impossible.<\/p>\n\n\n\n<p><em>\u201cIn contrast to previous techniques, we developed a simple method for short-term treatment with <\/em><em>ether, a classical <\/em><em>anaesthetic. <\/em><em>This approach allowed undisrupted long-term immobilisation of many larvae in parallel,\u201d <\/em>explains Parisa Kakanj.<\/p>\n\n\n\n<figure class=\"vf-figure wp-block-image alignnone size-full wp-image-18425\"><img loading=\"lazy\" decoding=\"async\" width=\"620\" height=\"425\" class=\"vf-figure__image\" src=\"https:\/\/news.embl.de\/wp-content\/uploads\/2020\/02\/202002_secondary_image-Letin-ib.jpg\" alt=\"The head of Drosophila larva, seen with a scanning electron microscope. Credit: Parisa Kakanj\/EMBL\" class=\"wp-image-18425\" srcset=\"https:\/\/www.embl.org\/news\/wp-content\/uploads\/2020\/02\/202002_secondary_image-Letin-ib.jpg 620w, https:\/\/www.embl.org\/news\/wp-content\/uploads\/2020\/02\/202002_secondary_image-Letin-ib-300x206.jpg 300w\" sizes=\"auto, (max-width: 620px) 100vw, 620px\" \/><figcaption class=\"vf-figure__caption\">The head of Drosophila larva, seen with a scanning electron microscope. Credit: Parisa Kakanj\/EMBL<\/figcaption><\/figure>\n\n\n\n<h2 class=\"wp-block-heading\">A promising model for drug discovery<\/h2>\n\n\n\n<p>To prove the technique&#8217;s capabilities, the team investigated wound healing in larvae. They examined the influence of insulin and TOR \u2013 crucial signalling molecules for cell survival, growth, and proliferation \u2013 on this process. By studying the subcellular dynamics, the scientists created a temporal map of insulin and TOR pathway activity during wound closure. They found that lowering insulin signalling at the wound edge slows down the healing process.<\/p>\n\n\n\n<p>Long-term&nbsp;<em>in vivo<\/em>&nbsp;imaging, coupled with genetic manipulations, opens the path to many aspects of biology and physiology.&nbsp;<em>\u201cThis technique will help many scientists studying neuronal signalling, fat metabolism, or tumour formation, and will provide new opportunities for drug discovery,\u201d <\/em>says Parisa Kakanj.<\/p>\n\n\n\n<div\n\n  \n  \n  class=\"vf-box vf-box--inlay\">\n  <h3 class=\"vf-box__heading\">Source article<\/h3>\n  <p class=\"vf-box__text\"><a href=\"https:\/\/www.nature.com\/articles\/s41596-019-0282-z\">Kakanj, P. <em>et al<\/em>. Long-term in vivo imaging of Drosophila larvae. Published in <em>Nature Protocols<\/em> on 10 February 2020.<br \/>DOI: 10.1038\/s41596-019-0282-z<\/a><\/p>\n<\/div>\n\n\n\n<div\n\n  \n  \n  class=\"vf-box vf-box--inlay\">\n  <h3 class=\"vf-box__heading\">Related links<\/h3>\n  <p class=\"vf-box__text\"><a href=\"http:\/\/www.uni-koeln.de\/math-nat-fak\/genetik\/groups\/Leptin\/lep_lab.htm\">Leptin Group<br \/><\/a><a href=\"https:\/\/www.age.mpg.de\/science\/research-laboratories\/partridge\/\">Partridge Group<\/a><\/p>\n<\/div>\n\n\n<hr class=\"vf-divider\"\/>\n\n\n\n<h2 class=\"wp-block-heading\" id=\"de\">Eine innovative Technik: Visualisierung von Organen und Zellen in lebenden Fliegenlarven&nbsp;<\/h2>\n\n\n\n<p>Kleine Abweichungen in der Zusammensetzung einer Zelle k\u00f6nnen ihre Funktion radikal ver\u00e4ndern, und Krankheiten wie Diabetes, Krebs oder neuronale Funktionsst\u00f6rungen hervorrufen. Wissenschaftler des Europ\u00e4ischen Laboratoriums f\u00fcr Molekularbiologie, des Instituts f\u00fcr Genetik der Universit\u00e4t zu K\u00f6ln, des Max-Planck-Instituts f\u00fcr Biologie des Alterns, des CECAD und CMMC in K\u00f6ln haben jetzt eine Methode entwickelt, um die Zellen in der lebenden Fruchtfliegenlarve zu beobachten und zellul\u00e4re Prozessen zu verfolgen. Diese Technik, die am 10. Februar in&nbsp;<em>Nature Protocols<\/em>&nbsp;ver\u00f6ffentlicht wurde, bietet eine einfache, aber effektive M\u00f6glichkeit, die Funktionen von Organen in lebenden Tieren zu untersuchen.<\/p>\n\n\n\n<figure class=\"vf-figure wp-block-image size-large\"><img loading=\"lazy\" decoding=\"async\" width=\"620\" height=\"425\" class=\"vf-figure__image\" src=\"https:\/\/www.embl.org\/news\/wp-content\/uploads\/2020\/02\/202002_cover_image-Leptin-ib.jpg\" alt=\"\" class=\"wp-image-18427\" srcset=\"https:\/\/www.embl.org\/news\/wp-content\/uploads\/2020\/02\/202002_cover_image-Leptin-ib.jpg 620w, https:\/\/www.embl.org\/news\/wp-content\/uploads\/2020\/02\/202002_cover_image-Leptin-ib-300x206.jpg 300w\" sizes=\"auto, (max-width: 620px) 100vw, 620px\" \/><figcaption class=\"vf-figure__caption\">Zusammenstellung von Bildern aus einem Video, dass die Wundheilung in der Epidermis einer Drosophila Larve zeigt. Die Zellen exprimieren zwei fluoreszierende Marker: einen PIP3 Sensor (gr\u00fcn) und die leichte Myosin Kette (magenta). Credit: Parisa Kakanj\/EMBL<\/figcaption><\/figure>\n\n\n\n<p>Die Fruchtfliege&nbsp;<em>Drosophila<\/em>&nbsp;gilt als idealer Modelorganismus in der Genforschung. Schl\u00fcsselprozesse und 60% der Gene der Fruchtfliege sind identisch zum Menschen. Daher haben Wissenschaftler viele genetische Techniken kreiert, um molekulare Prozesse der Entwicklung und menschliche Krankheiten in der Fliege zu untersuchen.<\/p>\n\n\n\n<h3 class=\"wp-block-heading\">Die Herausforderung<\/h3>\n\n\n\n<p>Die meisten der bisherigen Studien wurden am Embryo oder an der ausgewachsenen Fliege durchgef\u00fchrt, aber auch die Fruchtfliegenlarve bietet ein enormes Forschungspotenzial. Ihr transparenter K\u00f6rper mit funktionsf\u00e4higen Organen \u2013 wie Gehirn, Darm und Muskeln \u2013 macht die Larve zu einem hervorragenden Forschungsobjekt, um die Dynamik von Zellen und molekularer Vorg\u00e4nge in lebenden Tieren zu beobachten.<\/p>\n\n\n\n<p><em>\u201eDie Zellen in einer Fruchtfliegenlarve sind viel gr\u00f6\u00dfer als im Embryo. Man kann daher alle Organellen und sogar die kleinsten Prozesse sehen. Das ist das Sch\u00f6ne an diesem System\u201c,<\/em>&nbsp;sagt Wissenschaftlerin Parisa Kakanj, die die Studie in Deutschland geleitet hat.<\/p>\n\n\n\n<p>Einige molekulare Prozesse und Interaktionen in einer Zelle k\u00f6nnen nur durch langzeitige Beobachtung entschl\u00fcsselt werden. Das kontinuierliche Krabbeln der Larve macht solche Beobachtung jedoch schwierig. Kakanj, eine Forscherin in den Gruppen von Maria Leptin und Linda Partridge, hat sich dieser Herausforderung gestellt und eine Methode zur einfachen Immobilisierung der Larven entwickelt. Diese erm\u00f6glicht Langzeit-Live-Beobachtungen von Zellen in hoher Aufl\u00f6sung.<\/p>\n\n\n<div\n  class=\"vf-embed vf-embed--custom-ratio\"\n\n  style=\"--vf-embed-max-width: 100%;\n    --vf-embed-custom-ratio-x: 640;\n    --vf-embed-custom-ratio-y: 360;\"><iframe loading=\"lazy\" width=\"640\" height=\"360\" src=\"https:\/\/www.youtube.com\/embed\/dFMoTW8kSnI\" frameborder=\"0\" allow=\"accelerometer; autoplay; encrypted-media; gyroscope; picture-in-picture\" allowfullscreen><\/iframe><\/div>\n\n\n\n<p class=\"vf-figure__caption\"><strong>Live-Aufnahme der Insulinsignale und Bildung eines kontraktilen Kabels w\u00e4hrend der Wundheilung bei Drosophila-Larven.<\/strong>&nbsp;Ein Sensor f\u00fcr Insulinsignale ist gr\u00fcn dargestellt, und Myosin, ein Motorprotein, in Magenta. Wenn die Haut verletzt wird, l\u00f6st das Insulinsignal die Bildung und Kontraktion eines Actomyosinkabels um die Wunde herum aus. Dieses zieht die R\u00e4nder der Wunde zusammen und verschwindet nachdem die Verletzung verschlossen ist. Video: Benutzung mit Genehmigung von Kakanj, P.&nbsp;et al.,&nbsp;Nature Protocols&nbsp;(2020).<\/p>\n\n\n\n<h3 class=\"wp-block-heading\">Einfach, schnell und effizient<\/h3>\n\n\n\n<p>In der Vergangenheit haben Wissenschaftler mechanische Methoden oder Bet\u00e4ubungsmittel eingesetzt, um Fruchtfliegenlarven zu immobilisieren. Beide Ans\u00e4tze hatten jedoch unerw\u00fcnschte Nebenwirkungen. Zudem war ein gleichzeitige mikroskopische Aufnahme vieler Larven nicht m\u00f6glich.<\/p>\n\n\n\n<p><em>\u201eIm Gegensatz zu den bisherigen Techniken haben wir eine einfache Methode zur Kurzzeitbehandlung mit \u00c4ther, einem klassischen An\u00e4sthetikum, entwickelt. Dieser Ansatz erm\u00f6glichte eine problemlose Langzeitimmobilisierung vieler Larven<\/em>\u201c, erkl\u00e4rt Parisa Kakanj.<\/p>\n\n\n\n<figure class=\"vf-figure wp-block-image size-large\"><img loading=\"lazy\" decoding=\"async\" width=\"620\" height=\"425\" class=\"vf-figure__image\" src=\"https:\/\/www.embl.org\/news\/wp-content\/uploads\/2020\/02\/202002_secondary_image-Letin-ib.jpg\" alt=\"The head of Drosophila larva, seen with a scanning electron microscope. Credit: Parisa Kakanj\/EMBL\" class=\"wp-image-18425\" srcset=\"https:\/\/www.embl.org\/news\/wp-content\/uploads\/2020\/02\/202002_secondary_image-Letin-ib.jpg 620w, https:\/\/www.embl.org\/news\/wp-content\/uploads\/2020\/02\/202002_secondary_image-Letin-ib-300x206.jpg 300w\" sizes=\"auto, (max-width: 620px) 100vw, 620px\" \/><figcaption class=\"vf-figure__caption\">Der Kopf einer Drosophila Larve unter einem Rasterelektronenmikroskop. Credit: Parisa Kakanj\/EMBL<\/figcaption><\/figure>\n\n\n\n<h3 class=\"wp-block-heading\">Ein vielversprechendes Modell f\u00fcr die Medikamentenentwicklung<\/h3>\n\n\n\n<p>Um die Leistungsf\u00e4higkeit der Technik zu beweisen, untersuchte das Team die Wundheilung bei Fruchtfliegenlarven. Sie analysierten den Einfluss von Insulin und TOR \u2013 einem entscheidenden Signalmolek\u00fcl f\u00fcr \u00dcberleben, Wachstum und Vermehrung \u2013 auf diesen Prozess. In ihrer Studie erstellten die Wissenschaftler eine Karte mit dem zeitlichen Ablauf der Insulin- und TOR-Signalweg-Aktivit\u00e4t w\u00e4hrend der Wundheilung. Sie stellten fest, dass eine Senkung der Insulinsignalaktivit\u00e4t am Wundrand die Heilungsprozesse verlangsamt.<\/p>\n\n\n\n<p>Die langfristige mikroskopische Live-Aufnahme gekoppelt mit genetischen Manipulationen er\u00f6ffnet den Weg zu neuen Aspekten der Biologie und Physiologie.<\/p>\n\n\n\n<p>\u201e<em>Diese Technik wird vielen Wissenschaftlern bei der Untersuchung der neuronalen Signal\u00fcbertragung, des Fettstoffwechsels oder der Tumorbildung helfen und neue M\u00f6glichkeiten f\u00fcr die Medikamentenentwicklung er\u00f6ffnen<\/em>\u201c, sagt Parisa Kakanj.<\/p>\n\n\n\n<div\n\n  \n  \n  class=\"vf-box vf-box--inlay\">\n  <h3 class=\"vf-box__heading\">QUELLE<\/h3>\n  <p class=\"vf-box__text\"><a href=\"https:\/\/www.nature.com\/articles\/s41596-019-0282-z\">Kakanj, P. <em>et al<\/em>. Long-term in vivo imaging of Drosophila larvae. Published in <em>Nature Protocols<\/em> on 10 February 2020.<br \/>DOI: 10.1038\/s41596-019-0282-z<\/a><\/p>\n<\/div>\n\n\n\n<div\n\n  \n  \n  class=\"vf-box vf-box--inlay\">\n  <h3 class=\"vf-box__heading\">Links<\/h3>\n  <p class=\"vf-box__text\"><a href=\"http:\/\/www.uni-koeln.de\/math-nat-fak\/genetik\/groups\/Leptin\/lep_lab.htm\">Leptin Group<br \/><\/a><a href=\"https:\/\/www.age.mpg.de\/science\/research-laboratories\/partridge\/\">Partridge Group<\/a><\/p>\n<\/div>\n","protected":false},"excerpt":{"rendered":"<p>This technique provides a simple but effective way to study the functions of organs in living animals<\/p>\n","protected":false},"author":63,"featured_media":18427,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[2,17591],"tags":[1017,1018,478,833,43,1748],"embl_taxonomy":[],"class_list":["post-18423","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-science","category-science-technology","tag-cecad","tag-cmmc","tag-drosophila","tag-fruit-fly","tag-heidelberg","tag-press-release"],"acf":{"article_intro":"<p>Small changes in a cell\u2019s composition can radically transform its function and drive the development of diseases like diabetes, cancer, or neuronal dysfunction<\/p>\n","related_links":false,"article_sources":false,"vf_locked":false,"featured":false,"color":"#007B53","show_featured_image":false,"in_this_article":false,"youtube_url":"","mp4_url":"","video_caption":"","translations":[{"translation_language":"German","translation_anchor":"#de"}],"press_contact":"EMBL Generic"},"embl_taxonomy_terms":[],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v26.2 - https:\/\/yoast.com\/wordpress\/plugins\/seo\/ -->\n<title>New technique for studying living fly larvae | EMBL<\/title>\n<meta name=\"description\" content=\"Small changes in a cell\u2019s composition can radically transform its function and drive the development of diseases like diabetes, cancer, or neuronal dysfunction.\" \/>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/www.embl.org\/news\/science\/new-technique-for-studying-living-fly-larvae\/\" 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The cells express two fluorescently labelled markers: a PIP3 sensor (green) and the myosin light chain (magenta). 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