Clusterin, a protein associated with multiple functions, is expressed in a wide variety of mammalian tissues. Although clusterin is known to be involved in neurodegenerative diseases, ageing, and tumorigenesis, a deta...Clusterin, a protein associated with multiple functions, is expressed in a wide variety of mammalian tissues. Although clusterin is known to be involved in neurodegenerative diseases, ageing, and tumorigenesis, a detailed analysis of the consequences of gain- or loss-of- function approaches has yet to be performed to understand the underlying mechanisms of clusterin functions. Since clusterin levels change in neurological diseases, it is likely that clusterin contributes to cell death and degeneration in general. Zebrafish was investigated as a model system to study human diseases. During development, zebrafish clusterin was expressed in the notochord and nervous system. Embryonic overexpression of clusterin by mRNA microinjection did not affect axis formation, whereas its knock-down by anti-sense morpholino treatment resulted in neuronal cell death. To analyze the function of clusterin in neurodegeneration, a transgenic zebrafish was investigated, in which nitroreductase expression is regulated under the control of a neuron-specifc huC promoter which is active between the stages of early neuronal precursors and mature neurons. Nitroreductase turns metronidazole into a cytotoxic agent that induces cell death within 12 h. After metronidazole treatment, transgenic zebrafish showed neuron-specific cell death. Interestingly, we also observed a dramatic induction of clusterin expression in the brain and spinal cord in these fish, suggesting a direct or indirect role of clusterin in neuronal cell death and thus, more generally, in neurodegeneration.展开更多
Reactive oxygen species(ROS)are believed to be inevitable and toxic by-products produced during cellular respiration.However,several lines of evidence show that diverse extracellular signals can activate ROS productio...Reactive oxygen species(ROS)are believed to be inevitable and toxic by-products produced during cellular respiration.However,several lines of evidence show that diverse extracellular signals can activate ROS production and provide important function for various biological processes.展开更多
Williams syndrome(WS)is a rare multisystemic disorder caused by recurrent microdeletions on 7q11.23,characterized by intellectual disability,distinctive craniofacial and dental features,and cardiovascular problems.Pre...Williams syndrome(WS)is a rare multisystemic disorder caused by recurrent microdeletions on 7q11.23,characterized by intellectual disability,distinctive craniofacial and dental features,and cardiovascular problems.Previous studies have explored the roles of individual genes within these microdeletions in contributing to WS phenotypes.Here,we report five patients with WS with 1.4 Mb-1.5 Mb microdeletions that include RFC2,as well as one patient with a 167-kb microdeletion involving RFC2 and six patients with intragenic variants within RFC2.To investigate the potential involvement of RFC2 in WS pathogenicity,we generate a rfc2 knockout(KO)zebrafish using CRISPR-Cas9 technology.Additionally,we generate a KO zebrafish of its paralog gene,rfc5,to better understand the functions of these RFC genes in development and disease.Both rfc2 and rfc5 KO zebrafish exhibit similar phenotypes reminiscent of WS,including small head and brain,jaw and dental defects,and vascular problems.RNA-seq analysis reveals that genes associated with neural cell survival and differentiation are specifically affected in rfc2 KO zebrafish.In addition,heterozygous rfc2 KO adult zebrafish demonstrate an anxiety-like behavior with increased social cohesion.These results suggest that RFC2 may contribute to the pathogenicity of WS,as evidenced by the zebrafish model.展开更多
基金supported by the research fund of Chungnam National University,Republic of Koreasupported by the Li Kashing Foundation at Shantou University Medical College,China
文摘Clusterin, a protein associated with multiple functions, is expressed in a wide variety of mammalian tissues. Although clusterin is known to be involved in neurodegenerative diseases, ageing, and tumorigenesis, a detailed analysis of the consequences of gain- or loss-of- function approaches has yet to be performed to understand the underlying mechanisms of clusterin functions. Since clusterin levels change in neurological diseases, it is likely that clusterin contributes to cell death and degeneration in general. Zebrafish was investigated as a model system to study human diseases. During development, zebrafish clusterin was expressed in the notochord and nervous system. Embryonic overexpression of clusterin by mRNA microinjection did not affect axis formation, whereas its knock-down by anti-sense morpholino treatment resulted in neuronal cell death. To analyze the function of clusterin in neurodegeneration, a transgenic zebrafish was investigated, in which nitroreductase expression is regulated under the control of a neuron-specifc huC promoter which is active between the stages of early neuronal precursors and mature neurons. Nitroreductase turns metronidazole into a cytotoxic agent that induces cell death within 12 h. After metronidazole treatment, transgenic zebrafish showed neuron-specific cell death. Interestingly, we also observed a dramatic induction of clusterin expression in the brain and spinal cord in these fish, suggesting a direct or indirect role of clusterin in neuronal cell death and thus, more generally, in neurodegeneration.
基金supported by grants from the Korean Ministry of Trade,Industry and Energy(10063396)the National Research Foundation(NRF)of Korea(2018M3A9B8021980)
文摘Reactive oxygen species(ROS)are believed to be inevitable and toxic by-products produced during cellular respiration.However,several lines of evidence show that diverse extracellular signals can activate ROS production and provide important function for various biological processes.
基金supported by the National Research Foundation of Korea grant funded by the Korean government(MIST)(2020R1A5A8017671,2022R1A2C100677813,and RS-2024-00349650)。
文摘Williams syndrome(WS)is a rare multisystemic disorder caused by recurrent microdeletions on 7q11.23,characterized by intellectual disability,distinctive craniofacial and dental features,and cardiovascular problems.Previous studies have explored the roles of individual genes within these microdeletions in contributing to WS phenotypes.Here,we report five patients with WS with 1.4 Mb-1.5 Mb microdeletions that include RFC2,as well as one patient with a 167-kb microdeletion involving RFC2 and six patients with intragenic variants within RFC2.To investigate the potential involvement of RFC2 in WS pathogenicity,we generate a rfc2 knockout(KO)zebrafish using CRISPR-Cas9 technology.Additionally,we generate a KO zebrafish of its paralog gene,rfc5,to better understand the functions of these RFC genes in development and disease.Both rfc2 and rfc5 KO zebrafish exhibit similar phenotypes reminiscent of WS,including small head and brain,jaw and dental defects,and vascular problems.RNA-seq analysis reveals that genes associated with neural cell survival and differentiation are specifically affected in rfc2 KO zebrafish.In addition,heterozygous rfc2 KO adult zebrafish demonstrate an anxiety-like behavior with increased social cohesion.These results suggest that RFC2 may contribute to the pathogenicity of WS,as evidenced by the zebrafish model.
