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vps13d affects epileptic seizures by regulating mitochondrial fission and autophagy in epileptic rats

Genes & diseases 2024年第6期11卷 452-465页

作者： Jian Wang Fan Zhang Zhong Luo Haiqing Zhang Changyin Yu Zucai Xu department of Neurology Affiliated Hospital of Zunyi Medical UniversityZunyiGuizhou 563000China department of Neurology Affliated Aerospace Hospital of Zunyi Medical UniversityZunyiGuizhou 563000China department of Clinical Medicine Zunyi Medical and Pharmaceutical CollegeZunyiGuizhou 563000China

Abnormal mitochondrial dynamics can lead to seizures,and improved mitochon-drial dynamics can alleviate *** protein sorting 13d(vps13d)is closely associ-ated with regulating mitochondrial homeostasis and ***,further investigation is required to determine whether vps13d affects seizures by influencing mitochondrial dy-namics and *** aimed to investigate the influence of vps13d on behavior in a rat model of acute epileptic ***,we established an acute epileptic seizure rat model and employed the CRISPR/CAS9 technology to construct a lentivirus to silence the vps13d ***,we used the HT22 mouse hippocampal neuron cell line to establish a stable strain with suppressed expression of vps13d in ***,we performed quantitative prote-omic and bioinformatics analyses to confirm the mechanism by which vps13d influences mito-chondrial dynamics and autophagy,both in vitro and in vivo using the experimental acute epileptic seizure *** found that knockdown of vps13d resulted in reduced seizure la-tency and increased seizure frequency in the experimental *** staining and western blot analysis revealed a significant increase in mitochondrial dynamin-related pro-tein 1 expression following vps13d ***,we observed a significant reduction in LC3Il protein expression levels and the LC31/LC3l ratio(indicators for autophagy)accompa-nied by a significant increase in P62 expression(an autophagy adaptor protein).The proteomic analysis confirmed the up-regulation of P62 protein ***,we propose that vps13d plays a role in modulating seizures by influencing mitochondrial dynamics and autophagy.

关键词： Autophagy Mitochondrial dynamics Mitochondrial fission Seizures vps13d

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The lncRNA Snhg1-vps13d vesicle trafficking system modulates the dual effects of IL-7 signaling to control memory Cd8 T cell establishment

The lncRNA Snhg1-Vps13D vesicle trafficking system modulates...

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第十四届全国免疫学学术大会

作者： Yanyan Zhang Baohua Li Qiang Bai Lubin Jiang Ting Ni Lilin Ye Yuzhang Wu 中国科学院大学重庆医院陆军军医大学免疫所中国科学院上海巴斯德研究所复旦大学

The efficient induction and long-term persistence of pathogen-specific memory Cd8 T cells are pivotal to rapidly curb the reinfection. Recent studies indicated that long-noncoding RNAs expression is highly cell-and stage-specific during T cell development and differentiation, suggesting their potential roles in T cell programs. However, the key lncRNAs playing crucial roles in memory Cd8 T cell establishment remain to be clarified. Through Cd8 T cell subsets profiling of lncRNAs, this study found a key lncRNA-Snhg1 with the conserved na?vehi-effectorlo-memoryhi expression pattern in Cd8 T cells of both mice and human, that can promote memory formation while impeding effector Cd8 in acute viral infection. Further, Snhg1 was found physically interacting with the conserved vesicle trafficking protein vps13d to promote IL-7Rα membrane location specifically. With the deep mechanism probing, the Results show Snhg1-vps13d regulated IL-7 signaling with its dual-function and dose-effect in memory Cd8 generation, which not just because of the sustaining role of STAT5-BCL-2 axis for memory survival, but more through the STAT3-TCF1-Blimp1 axis for transcriptional launch program of memory differentiation. Moreover, we performed further study with finding the similar high-low-high expression pattern of human SNHG1/vps13d/IL7R/TCF7 in Cd8 T cell subsets from PBMC samples of the convalescent COVId-19 patients. The central role of Snhg1-vps13d-IL-7R-TCF1 axis in memory Cd8 establishment makes it a potencial target for improving the vaccination effects to control the ongoing pandemic.

关键词： memory Cd8 T cell lncRNA Snhg1 vps13d IL-7R TCF-1 viral infection STAT3 Blimp1

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vps13d调控HT22细胞线粒体动力学及其自噬的机制研究

VPS13D调控HT22细胞线粒体动力学及其自噬的机制研究

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作者：张帆遵义医科大学

学位级别：硕士

目的:通过小鼠海马神经元(HT22细胞)研究vps13d是否通过影响线粒体自噬,从而调节线粒体动力学相关蛋白的表达而影响线粒体动力学功能。方法:培养HT22细胞,构建vps13d基因敲减慢病毒质粒,将质粒转染至HT22细胞,嘌呤霉素筛选培养,使用RT-q... 详细信息

目的:通过小鼠海马神经元(HT22细胞)研究vps13d是否通过影响线粒体自噬,从而调节线粒体动力学相关蛋白的表达而影响线粒体动力学功能。方法:培养HT22细胞,构建vps13d基因敲减慢病毒质粒,将质粒转染至HT22细胞,嘌呤霉素筛选培养,使用RT-q PCR和免疫荧光半定量验证转染是否成功;转染成功后将细胞分为5个组:正常对照组、空病毒组(LV-vector)、vps13d敲减组(LV-sh2)、自噬抑制剂3-MA组(LV-vector+3-MA)、vps13d敲减合并3-MA组(LV-sh2+3-MA);免疫荧光半定量检测自噬相关蛋白LC3的表达水平;Western blot检测自噬相关蛋白P62和线粒体动力学相关蛋白MFN1、FIS1、dRP1的表达水平;使用透射电镜观察干预后各组细胞线粒体形态;选择正常对照组和vps13d敲减组细胞进行液相色谱-质谱联用技术(LC-MS),并进行蛋白质组学分析,通过统计学分析筛选差异蛋白并进行生物信息学分析。结果:1.荧光显微镜下观察发现,HT22细胞经过慢病毒转染和筛选后EGFP阳性细胞达90%以上。RT-q PCR结果显示LV-sh2组的vps13d的m RNA表达与LVvector组相比显著降低(Pvps13d表达与LV-vector组相比显著降低(PdRP1表达与LV-vector组和WT组相比显著升高(Pvps13d主要与HT22细胞的dNA合成、基因表达和蛋白质翻译的调控作用相关,其中自噬相关蛋白有Sqstm1(P62)、Nsfl1c、Rab7a、Park7、Hspa9、Plaa和Map2k1共7个,并都在vps13d敲减组细胞中表达升高。结论:vps13d可能通过dNA合成、基因表达和蛋白质翻译等方面调控HT22的细胞功能,其中vps13d功能减弱,可增强线粒体分裂并抑制自噬。

关键词： vps13d HT22 自噬线粒体动力学蛋白质组学

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Studies on the function of cocoon quality traits related genes of Bombyx mori

Studies on the function of cocoon quality traits related gen...

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中国蚕学会2021年学术年会

作者： Luochao Zhao Xia Sun Shen Qin Kaixiang Zhang Muwang Li Jiangsu Key Laboratory of Sericultural Biology and Biotechnology School of BiotechnologyJiangsu University of Science and Technology The Key Laboratory of Silkworm and Mulberry Genetic Improvement Ministry of AgricultureSericultural Research InstituteChinese Academy of Agricultural Science

The mechanism of regulating silk protein synthesis provides a basis for silkworm seed selection and silk gland bioreactor ***,we aim to explore the genetic basis for different silkworm varieties’ different silkworm *** recent years,the rapid development of high-throughput sequencing technology has promoted the research of genomics and functional *** this study,silk glands and silk production of different strains of Bombyx mori were *** on the whole genome sequencing data of silk glands from these different strains,combined with the QTL mapping information of silkworm cocoon silk traits,gene knockout technology was used to excavate the functional genes affecting the yield traits of silkworm cocoon silk,in order to further study the determining mechanism of silkworm cocoon silk *** results we can not only promote the cocoon silk in the silk gland to generate understanding of regulation,including the role of ribosomal proteins and genes involved in silk protein synthesis of aggregative distribution,from the Angle of the whole genome identified the main metabolic pathways may be associated with silk production,and gain a lot of silk gland differentially expressed genes of data,we can use these data to the silkworm cocoon,silk production to the gene enrichment related pathways have a whole *** study will lay a foundation for the molecular mechanism of determining cocoon silk yield,and will be of great significance for the breeding of silkworm with multi-silk yield,and will also provide guidance for the research.

关键词： Bombyx mori Silk Character Silk Gland vps13d Genome Sequencing

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