Characterization of genetic and molecular tools for studying the endogenous expression of Lactate dehydrogenase in Drosophila melanogaster.
Characterization of genetic and molecular tools for studying the endogenous expression of Lactate dehydrogenase in Drosophila melanogaster.
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研究果蝇乳酸脱氢酶内源表达的遗传和分子工具的特征。
DOI：
10.1101/2023.06.15.545165
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Tennessen,JasonM
中科院分区：
文献类型：
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作者：
Rai,Madhulika;Carter,SarahM;Shefali,ShefaliA;Chawla,Geetanjali;Tennessen,JasonM
果蝇幼虫的发育依赖于一种专门的代谢状态，利用碳水化合物和其他膳食营养素来促进快速生长。幼虫代谢程序的一个独特特征是，与果蝇生命周期的其他阶段相比，该生长阶段的乳酸脱氢酶 (Ldh) 活性高度升高，表明 Ldh 在促进幼虫发育中发挥着关键作用。先前对幼虫Ldh活性的研究主要集中在该酶在整个动物水平上的功能，然而，Ldh表达在幼虫组织之间存在显着差异，提出了该酶如何促进组织特异性生长程序的问题。在这里，我们描述了两种转基因报告基因和一种可用于研究体内 Ldh 表达的抗体。我们发现所有三个工具都产生相似的 Ldh 表达模式。此外，这些试剂表明幼虫 Ldh 表达模式很复杂，表明这种酶的用途因细胞类型而异。总的来说，我们的研究验证了一系列可用于研究果蝇糖酵解代谢的遗传和分子试剂。
Drosophila melanogasterlarval development relies on a specialized metabolic state that utilizes carbohydrates and other dietary nutrients to promote rapid growth. One unique feature of the larval metabolic program is that Lactate Dehydrogenase (Ldh) activity is highly elevated during this growth phase when compared to other stages of the fly life cycle, indicating that Ldh serves a key role in promoting juvenile development. Previous studies of larval Ldh activity have largely focused on the function of this enzyme at the whole animal level, however,Ldhexpression varies significantly among larval tissues, raising the question of how this enzyme promotes tissue-specific growth programs. Here we characterize two transgene reporters and an antibody that can be used to studyLdhexpressionin vivo. We find that all three tools produce similarLdhexpression patterns. Moreover, these reagents demonstrate that the larvalLdhexpression pattern is complex, suggesting the purpose of this enzyme varies across cell types. Overall, our studies validate a series of genetic and molecular reagents that can be used to study glycolytic metabolism in the fly.