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Knockout of butyrophilin subfamily 1 member A1(BTN1A1) alters lipid droplet formation and phospholipid composition in bovine mammary epithelial cells

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Journal of Animal Science and Biotechnology 2020年第4期11卷 975-985页

作者： Liqiang Han Menglu Zhang Zhiyang Xing Danielle N.Coleman Yusheng Liang Juan J.Loor Guoyu Yang College of Animal Science and Veterinary Medicine Henan Agricultural UniversityZhengzhou 450002PR China Department of Animal Sciences and Division of Nutritional Sciences University of IllinoisUrbanaIllinois 61801USA.

Background: Milk lipids originate from cytoplasmic lipid droplets(LD) that are synthesized and secreted from mammary epithelial cells by a unique membrane-envelopment process. Butyrophilin 1 A1(BTN1 A1) is one of the membrane proteins that surrounds LD, but its role in bovine mammary lipid droplet synthesis and secretion is not well ***: The objective was to knockout BTN1 A1 in bovine mammary epithelial cells(BMEC) via the CRISPR/Cas9 system and evaluate LD formation, abundance of lipogenic enzymes, and content of cell membrane phospholipid(PL) species. Average LD diameter was determined via Oil Red O staining, and profiling of cell membrane phospholipid species via liquid chromatography-tandem mass spectrometry(LC-MS/MS).Results: Lentivirus-mediated infection of the Cas9/sg RNA expression vector into BMEC resulted in production of a homozygous clone BTN1 A1^((-/-)). The LD size and content decreased following BTN1 A1 gene knockout. The m RNA abundance of fatty acid synthase(FASN) and peroxisome proliferator-activated receptor-gamma(PPARG) was downregulated in the BTN1 A1^((-/-))clone. Subcellular analyses indicated that BTN1 A1 and LD were co-localized in the cytoplasm. BTN1 A1 gene knockout increased the percentage of phosphatidylethanolamine(PE) and decreased phosphatidylcholine(PC), which resulted in a lower PC/PE ***: Results suggest that BTN1 A1 plays an important role in regulating LD synthesis via a mechanism involving membrane phospholipid composition.

关键词： lipid droplet Mammary epithelial cell Milk fat globule Phospholipid

OPA3 overexpression modulates lipid droplet production and sensitizes colorectal cancer cells to bevacizumab treatment

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BIOCELL 2024年第6期48卷 971-980页

作者： HONGBIAO WU DONGFANG LIU Department of Colorectal and Anal Surgery Ningbo First HospitalNingbo315010China

Background:Colorectal cancer(CRC)represents a substantial risk to public ***,theﬁrst US FDA-approved antiangiogenic drug(AAD)for human CRC treatment,faces resistance in *** role of lipid metabolism,particularly through OPA3-regulated lipid droplet production,in overcoming this resistance is under ***:The protein expression pattern of OPA3 in CRC primary/normal tissues was evaluated by bioinformatics ***3-overexpressed SW-480 and HCT-116 cell lines were established,and bevacizumab resistance and OPA3 effects on cell malignancy were ***3 protein/mRNA expression and lipid droplet-related genes were measured with Western blot and ***3 subcellular localization was detected using immunoﬂ*** and apoptosis were assessed via colony formation andﬂow *** formation assays were conducted to assess the angiogenic potential of human umbilical vein endothelial cells(HUVECs).lipid analysis was used to measure the phosphatidylcholine(PC)and lysophosphatidylcholine(LPC)levels in CRC ***:Bioinformatics analysis revealed that OPA3 was downregulated in *** of OPA3 inhibited CRC cell proliferation,stimulated apoptosis,and suppressed the angiogenic ability of ***3 effectively reversed the resistance of CRC cells to bevacizumab and decreased lipid droplet production in CRC ***,OPA3 reversed the bevacizumab-induced lipid droplet production in CRC cells,thereby increasing CRC cell sensitivity to bevacizumab ***:This study suggests that OPA3 modulates lipid metabolism in CRC cells and reduces resistance to bevacizumab in CRC ***,OPA3 may be a potential therapeutic target against the AAD resistance in CRC.

关键词： Colorectal cancer OPA3 Bevacizumab lipid droplet Antiangiogenic drug

Visualizing nitric oxide-dependent HIF-1 activity under hypoxia with a lipid droplet-targeting fluorescent probe

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Chinese Chemical Letters 2021年第12期32卷 3895-3898页

作者： Ying-Hao Pan Xiao-Xiao Chen Lei Dong Na Shao Li-Ya Niu Qing-Zheng Yang Key Laboratory of Radiopharmaceuticals Ministry of EducationCollege of ChemistryBeijing Normal UniversityBeijing 100875China

Evaluating the correlation between hypoxia inducible factor 1(HIF-1) and nitric oxide(NO) generated under hypoxia is of great significance. In this work, we developed a fluorescent probe for the monitor of HIF-1 activity influenced by NO under hypoxia in hepatoma cells with dual-targeting for hepatocyte and lipid droplet(LD). The probe shows excellent selectivity to NO and high sensitivity with 6000-fold fluorescence enhancement. Live cell imaging experiments revealed the probe’s capability of imaging exogenous and endogenous NO with specific in LDs of Hep G2 cells. For cells under hypoxia, HIF-1 induced LD level is observed to correlate with NO level. This work provides the in-situ visualization of NO-dependent HIF-1 upregulation through LD accumulation.

关键词： Bodipy Fluorescent probe HIF-1 lipid droplet Nitric oxide

Porcine enteric alphacoronavirus infection increases lipid droplet accumulation to facilitate the virus replication

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Journal of Integrative Agriculture 2024年第3期23卷 988-1005页

作者： Qi Gao Yongzhi Feng Ting Gong Dongdong Wu Xiaoyu Zheng Yizhuo Luo Yunlong Yang Zebu Song Lang Gong Guihong Zhang Guangdong Provincial Key Laboratory of Zoonosis Prevention and Control College of Veterinary MedicineSouth China Agricultural UniversityGuangzhou 510642China Maoming Branch Guangdong Laboratory for Lingnan Modern AgricultureMaoming 525000China Key Laboratory of Animal Vaccine Development Ministry of Agriculture and Rural AffairsGuangzhou 510000China Guangdong Laboratory for Lingnan Modern Agriculture Guangzhou 510642China

Coronaviruses are widely transmissible between humans and animals, causing diseases of varying severity. Porcine enteric alphacoronavirus(PEAV) is a newly-discovered pathogenic porcine enteric coronavirus in recent years, which causes watery diarrhea in newborn piglets. The host inflammatory responses to PEAV and its metabolic regulation mechanisms remain unclear, and no antiviral studies have been reported. Therefore, we investigated the pathogenic mechanism and antiviral drugs of PEAV. The transcriptomic analysis of PEAV-infected host cells revealed that PEAV could upregulate lipid metabolism pathways. In lipid metabolism, steady-state energy processes, which can be mediated by lipid droplets(LDs), are the main functions of organelles. LDs are also important in viral infection and inflammation. In infected cells, PEAV increased LD accumulation, upregulated NF-κB signaling, promoted the production of the inflammatory cytokines IL-1β and IL-8, and induced cell death. Inhibiting LD accumulation with a DGAT-1 inhibitor significantly inhibited PEAV replication, downregulated the NF-κB signaling pathway, reduced the production of IL-1β and IL-8, and inhibited cell death. The NF-κB signaling pathway inhibitor BAY11-7082 significantly inhibited LD accumulation and PEAV replication. Metformin hydrochloride also exerted anti-PEAV effects and significantly inhibited LD accumulation, downregulated the NF-κB signaling pathway, reduced the production of IL-1β and IL-8, and inhibited cell death. LD accumulation in the lipid metabolism pathway therefore plays an important role in the replication and pathogenesis of PEAV, and metformin hydrochloride inhibits LD accumulation and the inflammatory response to exert anti-PEAV activity and reducing pathological injury. These findings contribute new targets for developing treatments for PEAV infections.

关键词： porcine enteric alphacoronavirus NF-κB inflammatory pathway lipid droplet metformin hydrochloride

A lipid droplet-Associated GFP Reporter-Based Screen Identifies New Fat Storage Regulators in ***

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Journal of Genetics and Genomics 2014年第5期41卷 305-313页

作者： Zhenglong Liu Xia Li Qinlan Ge Mei Ding Xun Huang State Key Laboratory of Molecular Developmental Biology Institute of Genetics and Developmental Biology Chinese Academy of Sciences University of the Chinese Academy of Sciences

Fat storage disorders including obesity are pandemic human health problems. As a genetically amenable model organism, Caeno- rhabditis elegans has often been used to explore the molecular mechanisms of fat storage regulation. Dye staining of fixed animals and stimulated Raman scattering （SRS） microscopy methods have been used successfully to study fat storage, but a genetic screening system that takes full advantage of C. elegans transparency to perform live imaging of fluorescent protein reporters has not yet been reported. Here, we investigated the tissue-specific expression of the GFP fusion of Perilipin 1 （PLIN1）, a Drosophila lipid droplet-associated protein, in C. elegans. Our results indicate that PLINI：：GFP labels lipid droplets and can be used as a fat storage indicator in live worms. Through an RNAi screen, we further identified several previously uncharacterized new fat storage regulators.

关键词： Fat storage lipid droplet PLIN1 C. elegans

Golgi-localized MORN1 promotes lipid droplet abundance and enhances tolerance to multiple stresses in Arabidopsis

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Journal of Integrative Plant Biology 2023年第8期65卷 1890-1903页

作者： Zhan Li Yue Gao Jiapei Yan Shuai Wang Shu Wang Yuanyuan Liu Shaokui Wang Jian Hua Guangdong Provincial Key Laboratory of Plant Molecular Breeding State Key Laboratory for Conservation and Utilization of Subtropical Agro-BioresourcesSouth China Agricultural UniversityGuangzhou 510640China School of Integrative Plant Science Plant Biology SectionCornell UniversityIthacaNY 14853USA

lipid droplet(LD)in vegetative tissues has recently been implicated in environmental responses in plants,but its regulation and its function in stress tolerance are not well ***,we identified a Membrane Occupation and Recognition Nexus 1(MORN1)gene as a contributor to natural variations of stress tolerance through genome-wide association study in Arabidopsis *** of its loss-of-function mutant and natural variants revealed that the MORN1 gene is a positive regulator of plant growth,disease resistance,cold tolerance,and heat *** MORN1 protein is associated with the Golgi and is also partly associated with *** truncations that disrupt these associations abolished the biological function of the MORN1 ***,the MORN1 gene is a positive regulator of LD abundance,and its role in LD number regulation and stress tolerance is highly ***,this study identifies MORN1 as a positive regulator of LD abundance and a contributor to natural variations of stress *** implicates a potential involvement of Golgi in LD biogenesis and strongly suggests a contribution of LD to diverse processes of plant growth and stress responses.

关键词： cold Golgi heat immunity lipid droplet MORN1 natural variation stress tolerance

The size matters： regulation of lipid storage by lipid droplet dynamics

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Science China(Life Sciences) 2017年第1期60卷 46-56页

作者： Jinhai Yu Peng Li Tsinghua-Peking Center for Life Sciences School of Life Sciences Tsinghua University Beijing 100084 China

Adequate energy storage is essential for sustaining healthy life. lipid droplet （LD） is the subcellular organelle that stores energy in the form of neutral lipids and releases fatty acids under energy deficient conditions. Energy storage capacity of LDs is primarily dependent on the sizes of LDs. Enlargement and growth of LDs is controlled by two molecular pathways： neutral lipid synthesis and atypical LD fusion. Shrinkage of LDs is mediated by the degradation of neutral lipids under energy demanding conditions and is controlled by neutral cytosolic lipases and lysosomal acidic lipases. In this review, we summarize recent progress regarding the regulatory pathways and molecular mechanisms that control the sizes and the energy storage capacity of LDs.

关键词： lipid storage lipid droplet TAG synthesis atypical LD fusion lipolysis

BODIPY 493 acts as a bright buffering fluorogenic probe for super-resolution imaging of lipid droplet dynamics

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Chinese Chemical Letters 2022年第12期33卷 5042-5046页

作者： Jie Chen Wenjuan Liu Xiangning Fang Qinglong Qiao Zhaochao Xu CAS Key Laboratory of Separation Science for Analytical Chemistry Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian 116023China University of Chinese Academy of Sciences Beijing 100049China

The need for temporal resolution and long-term stability in super-resolution fluorescence imaging has motivated research to improve the photostability of fluorescent *** to the inevitable photobleaching of fluorophores,it is difficult to obtain long-term super-resolution imaging regardless of the self-healing strategy of introducing peroxide scavengers or the strategy of fluorophore structure modification to suppress TICT *** buffered fluorogenic probe uses the intact probes in the buffer pool to continuously replace the photobleached ones in the target,which greatly improves the photostability and enables stable dynamic super-resolution imaging for a long *** the buffering capacity comes at the expense of reducing the number of fluorescent probes in targets,resulting in low staining fluorescence *** this paper,we selected BODIPY 493,a lipid droplet probe with high fluorescence brightness,to explore the dynamic process of lipid droplet staining of this probe in *** found that BODIPY 493 only needs very low laser power for lipid droplet imaging due to the high molecular accumulation in lipid droplets and the high brightness,and the spatiotemporal resolution is greatly *** importantly,we found that BODIPY 493 also has a certain buffering capacity,which enables BODIPY 493 to be used for super-resolution imaging of lipid droplet *** work reminds researchers to coordinate the buffering capacity and brightness of fluorogenic probes.

关键词： lipid droplet Super-resolution imaging Fluorescent probe Brightness Buffering

The lipid droplet： A conserved cellular organelle

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Protein & Cell 2017年第11期8卷 796-800页

作者： Congyan Zhang Pingsheng Liu National Laboratory of Biomacromolecules CAS Center for Excellence in Biomacromolecules Institute of Biophysics Chinese Academy of Sciences Beijing 100101 China University of Chinese Academy of Sciences Beijing-100049 China

The lipid droplet （LD） is a unique multi-functional orga- nelle that contains a neutral lipid core covered with a phospholipid monolayer membrane. The LDs have been found in almost all organisms from bacteria to humans with similar shape. Several conserved functions of LDs have been revealed by recent studies, including lipid metabolism and trafficking, as well as nucleic acid binding and protection. We summarized these findings and proposed a hypothesis that the LD is a conserved organelle.

关键词： lipid droplet conserved organelle lipidmetabolism nucleic acid handling