Lung branching morphogenesis is accompanied by temporal metabolic changes towards a glycolytic preference.
Lung branching morphogenesis is accompanied by temporal metabolic changes towards a glycolytic preference.
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DOI：
10.1186/s13578-021-00654-w
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发表时间：
2021-07-17
影响因子：
7.5
通讯作者：
Moura RS
中科院分区：
文献类型：
--
作者：
Fernandes-Silva H;Alves MG;Araújo-Silva H;Silva AM;Correia-Pinto J;Oliveira PF;Moura RS
肺分支形态发生的特征在于上皮-间充质相互作用，其最终定义了气道传导系统。在整个过程中，能量和结构大分子是维持高增殖率所必需的。对肺发育的分子机制的广泛了解与缺乏关于胚胎肺代谢要求的数据形成对比。在这里，我们研究了与鸡肺分支的早期阶段相关的代谢谱。在这项研究中，我们使用了一个离体肺外植体培养系统，并分析了消耗/生产的细胞外代谢中间体与葡萄糖催化剂（丙氨酸，乳酸盐和乙酸盐）的1H-NMR光谱在培养基中。然后，我们通过qPCR表征了代谢物膜转运蛋白（mct 1、mct 3、mct 8、mct 1、mct 3、mct 4和mct 8）和糖酵解酶（hk 1、hk 2、pfk 1、ldha、ldhb、pdha和pdhb）的转录水平。用原位杂交法测定ldha和ldhb mRNA的空间定位。通过使用基于EdU的测定直接评估DNA合成来分析增殖。此外，我们进行蛋白质印迹分析LDHA和LDHT蛋白水平。最后，我们使用Clark型电极来评估肺移植物的呼吸能力。葡萄糖消耗减少，而丙氨酸，乳酸和乙酸的生产逐步增加的分支形态发生的进行。mRNA分析揭示了糖酵解途径中关键酶和转运蛋白表达水平的变化。LDH A和LDH B显示类似于近端-远端标记的区室特异性表达模式。此外，在活性分支位点检测到高增殖水平。LDH蛋白表达水平表明，LDHB可能是乳酸进行性升高的原因。同时，有一个稳定的耗氧率在整个分支形态建成。本报告描述了时间代谢的变化，伴随着鸡肺分支形态发生的早期阶段。总体而言，胚胎鸡肺似乎转移到糖酵解乳酸为基础的代谢肺分支发生。此外，这种代谢重新布线可能在肺发育过程中起着至关重要的作用。在线版本包含补充材料，可通过10.1186/s13578-021-00654-w获得。
Lung branching morphogenesis is characterized by epithelial-mesenchymal interactions that ultimately define the airway conducting system. Throughout this process, energy and structural macromolecules are necessary to sustain the high proliferative rates. The extensive knowledge of the molecular mechanisms underlying pulmonary development contrasts with the lack of data regarding the embryonic lung metabolic requirements. Here, we studied the metabolic profile associated with the early stages of chicken pulmonary branching. In this study, we used an ex vivo lung explant culture system and analyzed the consumption/production of extracellular metabolic intermediates associated with glucose catabolism (alanine, lactate, and acetate) by 1H-NMR spectroscopy in the culture medium. Then, we characterized the transcript levels of metabolite membrane transporters (glut1, glut3, glut8, mct1, mct3, mct4, and mct8) and glycolytic enzymes (hk1, hk2, pfk1, ldha, ldhb, pdha, and pdhb) by qPCR. ldha and ldhb mRNA spatial localization was determined by in situ hybridization. Proliferation was analyzed by directly assessing DNA synthesis using an EdU-based assay. Additionally, we performed western blot to analyze LDHA and LDHT protein levels. Finally, we used a Clark-Type Electrode to assess the lung explant's respiratory capacity. Glucose consumption decreases, whereas alanine, lactate, and acetate production progressively increase as branching morphogenesis proceeds. mRNA analysis revealed variations in the expression levels of key enzymes and transporters from the glycolytic pathway. ldha and ldhb displayed a compartment-specific expression pattern that resembles proximal–distal markers. In addition, high proliferation levels were detected at active branching sites. LDH protein expression levels suggest that LDHB may account for the progressive rise in lactate. Concurrently, there is a stable oxygen consumption rate throughout branching morphogenesis. This report describes the temporal metabolic changes that accompany the early stages of chicken lung branching morphogenesis. Overall, the embryonic chicken lung seems to shift to a glycolytic lactate-based metabolism as pulmonary branching occurs. Moreover, this metabolic rewiring might play a crucial role during lung development. The online version contains supplementary material available at 10.1186/s13578-021-00654-w.
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影响因子：
4
作者：
Delbaere, Joke;Vancamp, Pieter;Darras, Veerle M.
通讯作者：
Darras, Veerle M.
影响因子：
2.7
作者：
Fernandes-Silva H;Correia-Pinto J;Moura RS
通讯作者：
Moura RS
DOI：
10.3390/microarrays6020007
发表时间：
2017-04-05
期刊：
Microarrays (Basel, Switzerland)
影响因子：
--
作者：
Byers MS;Howard C;Wang X
通讯作者：
Wang X
影响因子：
64.5
作者：
Homem, Catarina C. F.;Steinmann, Victoria;Knoblich, Juergen A.
通讯作者：
Knoblich, Juergen A.
DOI：
10.1016/0006-291x(87)91335-0
发表时间：
1987-05-29
影响因子：
3.1
作者：
ENGLE, MJ;DOOLEY, M;BROWN, DJ
通讯作者：
BROWN, DJ