黑腹果蝇影像盘试验研究多胺转运系统。

4区生物学 Q3 Biochemistry, Genetics and Molecular Biology

Methods in enzymology Pub Date : 2025-01-01 Epub Date: 2025-02-06 DOI:10.1016/bs.mie.2025.01.038

Shannon L Nowotarski, Justin R DiAngelo

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引用次数: 0

摘要

高等真核生物的多胺代谢得到了很好的研究；然而，多胺腐胺、亚精胺和精胺如何进入细胞的机制尚不清楚。一种有效的方法来调查在多胺摄取中起作用的潜在参与者，包括使用黑腹果蝇想象盘试验。经激素诱导18小时后，从黑腹果蝇流浪第三星幼虫身上解剖的腿部影像椎间盘可以评估腿部发育。本文详细描述了如何使用基因操纵的黑腹果蝇来测试参与多胺运输系统的候选基因，如何解剖腿部影像椎间盘以及如何评估多胺进入影像椎间盘细胞的情况。

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Drosophila melanogaster imaginal disc assays to study the polyamine transport system.

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.

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来源期刊

Methods in enzymology 生物-生化研究方法

CiteScore

2.90

自引率

0.00%

发文量

308

审稿时长

3-6 weeks

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.