AIM: To investigate the regulation of Eaf2 protein in mouse lens cells apoptosis induced by ultraviolet (UV) radiation. METHODS: An eye of Eaf2 gene knockout mice or normal control mice was exposed to UV radiation, an...AIM: To investigate the regulation of Eaf2 protein in mouse lens cells apoptosis induced by ultraviolet (UV) radiation. METHODS: An eye of Eaf2 gene knockout mice or normal control mice was exposed to UV radiation, and the other one was non-exposed. All of lenses were analyzed by TUNEL and caspase 3 activity assays to determine the difference of the apoptosis induced by UV radiation. In addition, exposed and non-exposed lenses were analyzed by quantified p53 expression and real-time reverse transcription-polymerase chain reaction (RT-PCR) of Bax, Bid, Apaf-1, Puma and Noxa, to compare Eaf2 gene knockout mice and normal control mice. RESULTS: UV radiation caused apoptosis of lens cells in normal control mice and Eaf2 knockout mice. Activity of caspase 3 was significantly higher in normal control mice than Eaf2 knockout mice. Expression of p53 protein was significantly higher in lenses exposed to UV radiation than nonexposed lenses, but was similar between Eaf2 gene knockout mice and normal control mice in the same UV condition. After exposing to UV radiation, the analysis of real-time RT-PCR demonstrated that mRNA levels of Puma and Noxa were significantly higher in lenses of normal control mice than Eaf2 gene knockout mice, and that mRNA levels of Bax, Bid and Apaf-1 were not significantly different between gene knockout mice and normal control mice. CONCLUSION: Eaf2 increases lens cells apoptosis induced by ultraviolet radiation. And Eaf2 up-regulates expression of the Puma and the Noxa to act on lens cells apoptosis after UV radiation.展开更多
EAF1 and EAF2,the eleven-nineteen lysine-rich leukemia(ELL)-associated factors which can assemble to the super elongation complex(AFF1/4,AF9/ENL,ELL,and P-TEFb),are reported to participate in RNA polymeraseⅡto active...EAF1 and EAF2,the eleven-nineteen lysine-rich leukemia(ELL)-associated factors which can assemble to the super elongation complex(AFF1/4,AF9/ENL,ELL,and P-TEFb),are reported to participate in RNA polymeraseⅡto actively regulate a variety of biological processes,including leukemia and embryogenesis,but whether and how EAF1/2 function in hematopoietic system related hypoxia tolerance during embryogenesis remains unclear.Here,we unveiled that deletion of EAF1/2(eaf1^(-/-)and eaf2^(-/-))caused reduction in hypoxia tolerance in zebrafish,leading to reduced erythropoiesis during hematopoietic processes.Meanwhile,eaf1^(-/-)and eaf2^(-/-)mutants showed significant reduc-tion in the expression of key transcriptional regulators scl,lmo2,and gata1a in erythropoiesis at both 24 h post fertilization(hpf)and 72 hpf,with gata1a downregulated while scl and lmo2 upregulated at 14 hpf.Mechanistically,eaf1^(-/-)and eaf2^(-/-)mutants exhibited significant changes in the expression of epigenetic modified histones,with a significant increase in the binding enrichment of modified histone H3K27me3 in gata1a promoter rather than scl and lmo2 promoters.Additionally,eaf1^(-/-)and eaf2^(-/-)mutants exhibited a dynamic expression of canonical WNT/β-catenin signaling during erythropoiesis,with significant reduction in p-β-Catenin level and in the binding enrichment of both scl and lmo2 promoters with the WNT transcriptional factor TCF4 at 24 hpf.These findings demonstrate an important role of Eaf1/2 in erythropoiesis in zebrafish and may have shed some light on regeneration medicine for anemia and related diseases and on molecular basis for fish economic or productive traits,such as growth,disease resistance,hypoxia tolerance,and so on.展开更多
Background Meningeal lymphatic drainage is crucial for the clearance of amyloidβ(Aβ),supporting the maintenance of brain homeostasis.This makes it a promising therapeutic target for Alzheimer’s disease(AD).Long-ter...Background Meningeal lymphatic drainage is crucial for the clearance of amyloidβ(Aβ),supporting the maintenance of brain homeostasis.This makes it a promising therapeutic target for Alzheimer’s disease(AD).Long-term exercise can reduce the risk of AD;however,the underlying mechanism is not fully understood.In this study,we investigated whether exercise alleviates AD-related pathological changes by improving meningeal lymphatic drainage and its potential mechanisms.Methods The morphological and functional features of meningeal lymphatic vessels,as well as Aβand reactive gliosis in the brain,were compared between 6.5-month-old 5×FAD mice with or without 1 month of treadmill exercise.RNA sequencing,protein interactions analysis,gene knockdown mediated by adeno-associated virus,and lymphatic endothelial cell culture were conducted to investigate the mechanism underlying exercise-induced meningeal lymphatic vessel plasticity in 5×FAD mice.Results The structural integrity of meningeal lymphatic vessels was compromised in 5×FAD mice,compared with the wild-type mice.Treadmill exercise increased the diameter and the drainage capacity of the meningeal lymphatic vessels,reduced Aβdeposition,reactive gliosis and astrocyte senescence in the hippocampus and frontal cortex,and improved cognitive function of 5×FAD mice.Mechanistically,thrombospondin-1(TSP-1)exacerbated the inhibitory effect of Aβon lymphatic vessel formation and plasticity through interactions with CD36 and CD47,respectively.Exercise decreased the expression of TSP-1 in reactive astrocytes of AD mice by downregulating elevennineteen lysine-rich leukemia-associated factor 2(EAF2),a protein that facilitates the transcription of the TSP-1-encoding gene Thbs-1 by binding p53.Ultimately,we found that hippocampal astrocyte-specific knockdown of Thbs-1 or Eaf2 enhanced meningeal lymphatic drainage and alleviated AD-like pathology in the hippocampus of 5×FAD mice.Conclusions Long-term exercise protects against AD by enhancing the plasticity and drainage of meningeal lymphatic vessels through downregulation of the EAF2–p53–TSP-1 pathway associated with reactive astrocytes.展开更多
文摘AIM: To investigate the regulation of Eaf2 protein in mouse lens cells apoptosis induced by ultraviolet (UV) radiation. METHODS: An eye of Eaf2 gene knockout mice or normal control mice was exposed to UV radiation, and the other one was non-exposed. All of lenses were analyzed by TUNEL and caspase 3 activity assays to determine the difference of the apoptosis induced by UV radiation. In addition, exposed and non-exposed lenses were analyzed by quantified p53 expression and real-time reverse transcription-polymerase chain reaction (RT-PCR) of Bax, Bid, Apaf-1, Puma and Noxa, to compare Eaf2 gene knockout mice and normal control mice. RESULTS: UV radiation caused apoptosis of lens cells in normal control mice and Eaf2 knockout mice. Activity of caspase 3 was significantly higher in normal control mice than Eaf2 knockout mice. Expression of p53 protein was significantly higher in lenses exposed to UV radiation than nonexposed lenses, but was similar between Eaf2 gene knockout mice and normal control mice in the same UV condition. After exposing to UV radiation, the analysis of real-time RT-PCR demonstrated that mRNA levels of Puma and Noxa were significantly higher in lenses of normal control mice than Eaf2 gene knockout mice, and that mRNA levels of Bax, Bid and Apaf-1 were not significantly different between gene knockout mice and normal control mice. CONCLUSION: Eaf2 increases lens cells apoptosis induced by ultraviolet radiation. And Eaf2 up-regulates expression of the Puma and the Noxa to act on lens cells apoptosis after UV radiation.
基金supported by the Nation Natural Science Foundation of China(Program No.32070807)by the National Key R&D Program of China(2018YFD0900101)by the project 2020SKLBC-KF06 of State Key Laboratory of Biocontrol.
文摘EAF1 and EAF2,the eleven-nineteen lysine-rich leukemia(ELL)-associated factors which can assemble to the super elongation complex(AFF1/4,AF9/ENL,ELL,and P-TEFb),are reported to participate in RNA polymeraseⅡto actively regulate a variety of biological processes,including leukemia and embryogenesis,but whether and how EAF1/2 function in hematopoietic system related hypoxia tolerance during embryogenesis remains unclear.Here,we unveiled that deletion of EAF1/2(eaf1^(-/-)and eaf2^(-/-))caused reduction in hypoxia tolerance in zebrafish,leading to reduced erythropoiesis during hematopoietic processes.Meanwhile,eaf1^(-/-)and eaf2^(-/-)mutants showed significant reduc-tion in the expression of key transcriptional regulators scl,lmo2,and gata1a in erythropoiesis at both 24 h post fertilization(hpf)and 72 hpf,with gata1a downregulated while scl and lmo2 upregulated at 14 hpf.Mechanistically,eaf1^(-/-)and eaf2^(-/-)mutants exhibited significant changes in the expression of epigenetic modified histones,with a significant increase in the binding enrichment of modified histone H3K27me3 in gata1a promoter rather than scl and lmo2 promoters.Additionally,eaf1^(-/-)and eaf2^(-/-)mutants exhibited a dynamic expression of canonical WNT/β-catenin signaling during erythropoiesis,with significant reduction in p-β-Catenin level and in the binding enrichment of both scl and lmo2 promoters with the WNT transcriptional factor TCF4 at 24 hpf.These findings demonstrate an important role of Eaf1/2 in erythropoiesis in zebrafish and may have shed some light on regeneration medicine for anemia and related diseases and on molecular basis for fish economic or productive traits,such as growth,disease resistance,hypoxia tolerance,and so on.
基金supported by grants from the National Natural Science Foundation of China(82471608,82204365,82071199 and 81871117)the Natural Science Foundation of Jiangsu Province(BK20230057)+2 种基金Shandong Postdoctoral Science Foundation(SDCX-ZG-202400044)Shandong Postdoctoral Innovative Talents Program(SDBX2023056)the Natural Science Foundation of the Jiangsu Higher Education Institutions of China(23KJB310009).
文摘Background Meningeal lymphatic drainage is crucial for the clearance of amyloidβ(Aβ),supporting the maintenance of brain homeostasis.This makes it a promising therapeutic target for Alzheimer’s disease(AD).Long-term exercise can reduce the risk of AD;however,the underlying mechanism is not fully understood.In this study,we investigated whether exercise alleviates AD-related pathological changes by improving meningeal lymphatic drainage and its potential mechanisms.Methods The morphological and functional features of meningeal lymphatic vessels,as well as Aβand reactive gliosis in the brain,were compared between 6.5-month-old 5×FAD mice with or without 1 month of treadmill exercise.RNA sequencing,protein interactions analysis,gene knockdown mediated by adeno-associated virus,and lymphatic endothelial cell culture were conducted to investigate the mechanism underlying exercise-induced meningeal lymphatic vessel plasticity in 5×FAD mice.Results The structural integrity of meningeal lymphatic vessels was compromised in 5×FAD mice,compared with the wild-type mice.Treadmill exercise increased the diameter and the drainage capacity of the meningeal lymphatic vessels,reduced Aβdeposition,reactive gliosis and astrocyte senescence in the hippocampus and frontal cortex,and improved cognitive function of 5×FAD mice.Mechanistically,thrombospondin-1(TSP-1)exacerbated the inhibitory effect of Aβon lymphatic vessel formation and plasticity through interactions with CD36 and CD47,respectively.Exercise decreased the expression of TSP-1 in reactive astrocytes of AD mice by downregulating elevennineteen lysine-rich leukemia-associated factor 2(EAF2),a protein that facilitates the transcription of the TSP-1-encoding gene Thbs-1 by binding p53.Ultimately,we found that hippocampal astrocyte-specific knockdown of Thbs-1 or Eaf2 enhanced meningeal lymphatic drainage and alleviated AD-like pathology in the hippocampus of 5×FAD mice.Conclusions Long-term exercise protects against AD by enhancing the plasticity and drainage of meningeal lymphatic vessels through downregulation of the EAF2–p53–TSP-1 pathway associated with reactive astrocytes.
