High-Resolution Optical Molecular Imaging of Changes in Choline Metabolism in Oral Neoplasia

Luo, Zhen-Ge; Loja, Melissa N.; Farwell, D. Greg; Luu, Quang; Donald, Paul J.; Amott, Deborah; Gandour‐Edwards, Regina; Nitin, Nitin

doi:10.1593/tlo.12289

Cited by 7 publications

(4 citation statements)

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“…Lipidomic profiling demonstrated strong incorporation of the analog, which substitutes for choline at a rate of up to 44% when a concentration of 500 μM was used, while showing no significant impact on the ratios of non-choline lipids [ 12 ]. This method has been applied to visualize changes in choline uptake in assaying the toxicity of cisplatin to cancer cells and in comparing choline uptake between cancerous and non-cancerous biopsy samples [ 15 , 16 ]. Propargylcholine has also been used to study the catabolism of choline phospholipids in the pathogen Pseudomonas aeruginosa [ 17 ].…”

Section: Introductionmentioning

confidence: 99%

Bioorthogonal click chemistry for fluorescence imaging of choline phospholipids in plants

2018

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BackgroundPhospholipids are important structural and signaling molecules in plant membranes. Some fluorescent dyes can stain general lipids of membranes, but labeling and visualization of specific lipid classes have yet to be developed for most components of the membrane. New techniques for visualizing membrane lipids are needed to further delineate their dynamic structural and signaling roles in plant cells. In this study we examined whether propargylcholine, a bioortholog of choline, can be used to label the major membrane lipid, phosphatidylcholine, and other choline phospholipids in plants. We established that propargylcholine is readily taken up by roots, and that its incorporation is not detrimental to plant growth. After plant tissue is harvested and fixed, a click-chemistry reaction covalently links the alkyne group of propargylcholine to a fluorescently-tagged azide, resulting in specific labeling of choline phospholipids.ResultsUptake of propargylcholine, followed by click chemistry with fluorescein or Alexa Fluor 594 azide was used to visualize choline phospholipids in cells of root, leaf, stem, silique and seed tissues from Arabidopsis thaliana. Co-localization with various subcellular markers indicated coinciding fluorescent signals in cell membranes, such as the tonoplast and the ER. Among different cell types in the leaf epidermis, guard cells displayed strong labeling. Mass spectrometry-based lipidomic analysis of the various plant tissues revealed that incorporation of propargylcholine was strongest in roots with approximately 50% of total choline phospholipids being labeled, but it was also incorporated in the other tissues including seeds. Phospholipid profiling confirmed that, in each tissue analyzed, incorporation of the bioortholog had little impact on the pool of choline plus choline-like phospholipids or other lipid species.ConclusionWe developed and validated a click-chemistry based method for fluorescence imaging of choline phospholipids using a bioortholog of choline, propargylcholine, in various cell-types and tissues from Arabidopsis. This click-chemistry method provides a direct way to metabolically tag and visualize specific lipid molecules in plant cells. This work paves the way for future studies addressing in situ localization of specific lipids in plants.Electronic supplementary materialThe online version of this article (10.1186/s13007-018-0299-2) contains supplementary material, which is available to authorized users.

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Section: Introductionmentioning

confidence: 99%

Bioorthogonal click chemistry for fluorescence imaging of choline phospholipids in plants

2018

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“…For example, this technique has been used to develop an optical molecular imaging approach to visualize changes associated with oral neoplasia. Here it was found that cell mean fluorescence intensities resulting from PCho fluorescence labeling of neoplastic tissues were 4-5-fold greater than normal samples (Luo et al, 2013). This labeling protocol was also used to determine the effects of different anti-cancer drugs on choline metabolism, and indicated that the inhibitors cisplatin (Luo et al, 2012), gefitinib, and carboplatin (Luo et al, 2016) were detectable based on decreased PCho uptake.…”

Section: Applications Of Lipid Metabolic Labeling To Cancer Imaging and Studymentioning

confidence: 87%

Metabolic labeling of glycerophospholipids via clickable analogs derivatized at the lipid headgroup

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2020

Chemistry and Physics of Lipids

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Metabolic labeling, in which substrate analogs containing diminutive tags can infiltrate biosynthetic pathways and generate labeled products in cells, has led to dramatic advancements in the means by which complex biomolecules can be detected and biological processes can be elucidated. Within this realm, metabolic labeling of lipid products, particularly in a manner that is headgroup-specific, brings about a number of technical challenges including the complexity of lipid metabolic pathways as well as the simplicity of biosynthetic precursors to headgroup functionality. As such, only a handful of strategies for metabolic labeling of lipids have thus far been reported. However, these approaches provide enticing examples of how strategic modifications to substrate structures, particularly by introducing clickable moieties, can enable the hijacking of lipid biosynthesis. Furthermore, early work in this field has led to an explosion in diverse applications by which these techniques have been exploited to answer key biological questions or detect and track various lipid-containing biological entities. In this article, we review these efforts and emphasize recent advancements in the development and application of lipid metabolic labeling strategies.

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“…Amino acids are used for the synthesis of tissue proteins and hormones and serve as energy-producing substances in organisms [ 27 ]. Phosphatidylcholine is an significant component of cell membranes and plays an important role in cell signaling pathways [ 28 , 29 ]. Acetyl-CoA plays an essential role in glycolysis and fatty acid oxidation [ 30 ].…”

Section: Discussionmentioning

confidence: 99%

Transcriptome Analysis Reveals Candidate Genes Involved in Light-Induced Primordium Differentiation in Pleurotus eryngii

et al. 2021

IJMS

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Pleurotus eryngii, a highly valued edible fungus, is one of the major commercially cultivated mushrooms in China. The development of P. eryngii, especially during the stage of primordium differentiation, is easily affected by light. However, the molecular mechanism underlying the response of primordium differentiation to light remains unknown. In the present study, primordium expression profiles under blue-light stimulation, red-light stimulation, and exposure to darkness were compared using high-throughput sequencing. A total of 16,321 differentially expressed genes (DEGs) were identified from three comparisons. GO enrichment analysis showed that a large number of DEGs were related to light stimulation and amino acid biosynthesis. KEGG analyses demonstrated that the MAPK signaling pathway, oxidative phosphorylation pathway, and RNA transport were most active during primordium differentiation. Furthermore, it was predicted that the blue-light photoreceptor WC-1 and Deoxyribodipyrimidine photolyase PHR play important roles in the primordium differentiation of P. eryngii. Taken together, the results of this study provide a speculative mechanism that light induces primordium differentiation and a foundation for further research on fruiting body development in P. eryngii.

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High-Resolution Optical Molecular Imaging of Changes in Choline Metabolism in Oral Neoplasia

Cited by 7 publications

References 45 publications

Bioorthogonal click chemistry for fluorescence imaging of choline phospholipids in plants

Bioorthogonal click chemistry for fluorescence imaging of choline phospholipids in plants

Metabolic labeling of glycerophospholipids via clickable analogs derivatized at the lipid headgroup

Transcriptome Analysis Reveals Candidate Genes Involved in Light-Induced Primordium Differentiation in Pleurotus eryngii

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