{"title":"Drosophila melanogaster imaginal disc assays to study the polyamine transport system.","authors":"Shannon L Nowotarski, Justin R DiAngelo","doi":"10.1016/bs.mie.2025.01.038","DOIUrl":null,"url":null,"abstract":"<p><p>Polyamine metabolism in higher eukaryotes is well studied; however, the mechanism of how the polyamines putrescine, spermidine and spermine enter the cell remains unclear. An effective approach to investigate potential players that function in the uptake of polyamines involves using the Drosophila melanogaster imaginal disc assay. Leg imaginal discs dissected from Drosophila melanogaster wandering third star larvae can be assessed for leg development after 18 h of treatment with hormones to induce this process. The protocol described here details how to use genetically manipulated Drosophila melanogaster to test candidate genes involved in the polyamine transport system, how to dissect leg imaginal discs and how to assess the entry of polyamines into the cells of the imaginal disc.</p>","PeriodicalId":18662,"journal":{"name":"Methods in enzymology","volume":"715 ","pages":"351-361"},"PeriodicalIF":0.0000,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Methods in enzymology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1016/bs.mie.2025.01.038","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/2/6 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 0
Abstract
Polyamine metabolism in higher eukaryotes is well studied; however, the mechanism of how the polyamines putrescine, spermidine and spermine enter the cell remains unclear. An effective approach to investigate potential players that function in the uptake of polyamines involves using the Drosophila melanogaster imaginal disc assay. Leg imaginal discs dissected from Drosophila melanogaster wandering third star larvae can be assessed for leg development after 18 h of treatment with hormones to induce this process. The protocol described here details how to use genetically manipulated Drosophila melanogaster to test candidate genes involved in the polyamine transport system, how to dissect leg imaginal discs and how to assess the entry of polyamines into the cells of the imaginal disc.
期刊介绍：
The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 500 volumes the series contains much material still relevant today and is truly an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research and genetics-just to name a few. Five of the 2013 Nobel Laureates have edited or contributed to volumes of MIE.
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