The conventional perception of astrocytes as mere supportive cells within the brain has recently been called into question by empirical evidence, which has revealed their active involvement in regulating brain functio...The conventional perception of astrocytes as mere supportive cells within the brain has recently been called into question by empirical evidence, which has revealed their active involvement in regulating brain function and encoding behaviors associated with emotions.Specifically, astrocytes in the basolateral amygdala have been found to play a role in the modulation of anxiety-like behaviors triggered by chronic stress. Nevertheless, the precise molecular mechanisms by which basolateral amygdala astrocytes regulate chronic stress–induced anxiety-like behaviors remain to be fully elucidated. In this study, we found that in a mouse model of anxiety triggered by unpredictable chronic mild stress, the expression of excitatory amino acid transporter 2 was upregulated in the basolateral amygdala. Interestingly, our findings indicate that the targeted knockdown of excitatory amino acid transporter 2 specifically within the basolateral amygdala astrocytes was able to rescue the anxiety-like behavior in mice subjected to stress. Furthermore, we found that the overexpression of excitatory amino acid transporter 2 in the basolateral amygdala, whether achieved through intracranial administration of excitatory amino acid transporter 2agonists or through injection of excitatory amino acid transporter 2-overexpressing viruses with GfaABC1D promoters, evoked anxiety-like behavior in mice. Our single-nucleus RNA sequencing analysis further confirmed that chronic stress induced an upregulation of excitatory amino acid transporter 2 specifically in astrocytes in the basolateral amygdala. Moreover, through in vivo calcium signal recordings, we found that the frequency of calcium activity in the basolateral amygdala of mice subjected to chronic stress was higher compared with normal mice.After knocking down the expression of excitatory amino acid transporter 2 in the basolateral amygdala, the frequency of calcium activity was not significantly increased, and anxiety-like behavior was obviously mitigated. Additionally, administration of an excitatory amino acid transporter 2 inhibitor in the basolateral amygdala yielded a notable reduction in anxiety level among mice subjected to stress. These results suggest that basolateral amygdala astrocytic excitatory amino acid transporter 2 plays a role in in the regulation of unpredictable chronic mild stress-induced anxiety-like behavior by impacting the activity of local glutamatergic neurons, and targeting excitatory amino acid transporter 2 in the basolateral amygdala holds therapeutic promise for addressing anxiety disorders.展开更多
Aim Prenatal stress （PS） can lead to abnormal behavior of offspring and increase the incidence of mental illness. Previous researches have shown that levels of glutamate and its receptor expression are closely relat...Aim Prenatal stress （PS） can lead to abnormal behavior of offspring and increase the incidence of mental illness. Previous researches have shown that levels of glutamate and its receptor expression are closely relat- ed to the occurrence of this phenomenon. Furthermore, recent study has demonstrated that the expression levels of excitatory amino acid transporters 2 （EAAT2） in different brain regions of 1 month PS offspring rats have changed. Methods The SD pregnant rats were used restraint stress to imitate PS from gestation 14 -~ 19 days. Offspring rats were weaned 21 days after birth. The expression of EAAT2 of hippocampus was observed by Western blot. Results The expression of EAAT2 of 1 month PS offspring rats was significantly decreased in comparison to control group. However, the expression of EAAT2 of 2 month PS offspring rats was significantly increased in comparison to 1 month PS offspring rats. Conclusion These phenomena have illustrated that the expression of EAAT2 of PS off- spring rats could show time dependence or reversibility. The expression of EAAT2 may play an important role in the development of mental illness of offspring rats influenced by PS.展开更多
Excitatory amino acid transporters(EAATs) are responsible for excitatory amino acid transportation and are associated with auto-immune diseases in the central nervous system and peripheral tissues.However, the subcell...Excitatory amino acid transporters(EAATs) are responsible for excitatory amino acid transportation and are associated with auto-immune diseases in the central nervous system and peripheral tissues.However, the subcellular location and function of EAAT2 in macrophages are still obscure. In this study,we demonstrated that LPS stimulation increases expression of EAAT2(coded by Slc1a2) via NF-κB signaling. EAAT2 is necessary for inflammatory macrophage polarization through sustaining mTORC1 activation. Mechanistically, lysosomal EAAT2 mediates lysosomal glutamate and aspartate efflux to maintain V-ATPase activation, which sustains macropinocytosis and mTORC1. We also found that mice with myeloid depletion of Slc1a2 show alleviated inflammatory responses in LPS-induced systemic inflammation and high-fat diet induced obesity. Notably, patients with type Ⅱ diabetes(T2D) have a higher level of expression of lysosomal EAAT2 and activation of mTORC1 in blood macrophages. Taken together, our study links the subcellular location of amino acid transporters with the fate decision of immune cells,which provides potential therapeutic targets for the treatment of inflammatory diseases.展开更多
Glutamate is the predominant excitatory neurotransmitter in the human brain and it has been shown that prolonged activation of the glutamatergic system leads to nerve damage and cell death. Following release from the ...Glutamate is the predominant excitatory neurotransmitter in the human brain and it has been shown that prolonged activation of the glutamatergic system leads to nerve damage and cell death. Following release from the pre-synaptic neuron and synaptic transmission, glutamate is either taken up into the presynaptic neuron or neighbouring glia by transmembrane glutamate transporters. Excitatory amino acid transporter(EAAT) 1 and EAAT2 are Na+-dependant glutamate transporters expressed predominantly in glia cells of the central nervous system. As the most abundant glutamate transporters, their primary role is to modulate levels of glutamatergic excitability and prevent spill over of glutamate beyond the synapse. This role is facilitated through the binding and transportation of glutamate into astrocytes and microglia. The function of EAAT1 and EAAT2 is heavily regulated at the levels of gene expression, post-transcriptional splicing, glycosylation states and cell-surface trafficking of the protein. Both glutamatergic dysfunction and glial dysfunction have been proposed to be involved in psychiatric disorder. This review will present an overview of the roles that EAAT1 and EAAT2 play in modulating glutamatergic activity in the human brain, and mount an argument that these two transporters could be involved in the aetiologies of schizophrenia and affective disorders as well as represent potential drug targets for novel therapies for those disorders.展开更多
The present study established a rat model of global cerebral ischemia induced by chest compression for six minutes to dynamically observe expressional changes of three glutamate transporters in the cerebral cortex and...The present study established a rat model of global cerebral ischemia induced by chest compression for six minutes to dynamically observe expressional changes of three glutamate transporters in the cerebral cortex and hippocampus. After 24 hours of ischemia, expression of glutamate transporter-1 significantly decreased in the cerebral cortex and hippocampus, which was accompanied by neuronal necrosis. At 7 days post-ischemia, expression of excitatory amino acid carrier 1 decreased in the hippocampal CA1 region and cortex, and was accompanied by apoptosis Expression of glutamate-aspartate transporter remained unchanged at 6 hours 7 days after ischemia. These results suggested that glutamate transporter levels were altered at different periods of cerebral ischemia.展开更多
基金supported by the National Natural Science Foundation of China,Nos.32371070 (to JT),31761163005 (to JT),32100824 (to QX)the Shenzhen Science and Technology Program,Nos.RCBS20210609104606024 (to QX),JCY20210324101813035 (to DL)+4 种基金the Guangdong Provincial Key S&T Program,No.2018B030336001 (to JT)the Key Basic Research Program of Shenzhen Science and Technology Innovation Commission,Nos.JCYJ20200109115405930 (to JT),JCYJ20220818101615033 (to DL),JCYJ20210324115811031 (to QX),JCYJ20200109150717745 (to QX)Shenzhen Key Laboratory of Neuroimmunomodulation for Neurological Diseases,No.ZDSYS20220304163558001 (to JT)Guangdong Provincial Key Laboratory of Brain Connectome and Behavior,No.2023B1212060055 (to JT)the China Postdoctoral Science Foundation,No.2021M693298 (to QX)。
文摘The conventional perception of astrocytes as mere supportive cells within the brain has recently been called into question by empirical evidence, which has revealed their active involvement in regulating brain function and encoding behaviors associated with emotions.Specifically, astrocytes in the basolateral amygdala have been found to play a role in the modulation of anxiety-like behaviors triggered by chronic stress. Nevertheless, the precise molecular mechanisms by which basolateral amygdala astrocytes regulate chronic stress–induced anxiety-like behaviors remain to be fully elucidated. In this study, we found that in a mouse model of anxiety triggered by unpredictable chronic mild stress, the expression of excitatory amino acid transporter 2 was upregulated in the basolateral amygdala. Interestingly, our findings indicate that the targeted knockdown of excitatory amino acid transporter 2 specifically within the basolateral amygdala astrocytes was able to rescue the anxiety-like behavior in mice subjected to stress. Furthermore, we found that the overexpression of excitatory amino acid transporter 2 in the basolateral amygdala, whether achieved through intracranial administration of excitatory amino acid transporter 2agonists or through injection of excitatory amino acid transporter 2-overexpressing viruses with GfaABC1D promoters, evoked anxiety-like behavior in mice. Our single-nucleus RNA sequencing analysis further confirmed that chronic stress induced an upregulation of excitatory amino acid transporter 2 specifically in astrocytes in the basolateral amygdala. Moreover, through in vivo calcium signal recordings, we found that the frequency of calcium activity in the basolateral amygdala of mice subjected to chronic stress was higher compared with normal mice.After knocking down the expression of excitatory amino acid transporter 2 in the basolateral amygdala, the frequency of calcium activity was not significantly increased, and anxiety-like behavior was obviously mitigated. Additionally, administration of an excitatory amino acid transporter 2 inhibitor in the basolateral amygdala yielded a notable reduction in anxiety level among mice subjected to stress. These results suggest that basolateral amygdala astrocytic excitatory amino acid transporter 2 plays a role in in the regulation of unpredictable chronic mild stress-induced anxiety-like behavior by impacting the activity of local glutamatergic neurons, and targeting excitatory amino acid transporter 2 in the basolateral amygdala holds therapeutic promise for addressing anxiety disorders.
文摘Aim Prenatal stress （PS） can lead to abnormal behavior of offspring and increase the incidence of mental illness. Previous researches have shown that levels of glutamate and its receptor expression are closely relat- ed to the occurrence of this phenomenon. Furthermore, recent study has demonstrated that the expression levels of excitatory amino acid transporters 2 （EAAT2） in different brain regions of 1 month PS offspring rats have changed. Methods The SD pregnant rats were used restraint stress to imitate PS from gestation 14 -~ 19 days. Offspring rats were weaned 21 days after birth. The expression of EAAT2 of hippocampus was observed by Western blot. Results The expression of EAAT2 of 1 month PS offspring rats was significantly decreased in comparison to control group. However, the expression of EAAT2 of 2 month PS offspring rats was significantly increased in comparison to 1 month PS offspring rats. Conclusion These phenomena have illustrated that the expression of EAAT2 of PS off- spring rats could show time dependence or reversibility. The expression of EAAT2 may play an important role in the development of mental illness of offspring rats influenced by PS.
基金supported by the National Natural Science Foundation of China (32225047 and U22A20510)Laboratory of Lingnan Modern Agriculture Project (NT2021005)+1 种基金National Center of Technology Innovation for Pigs (NCTIP-XD/B13)double first-class discipline promotion project (2023B10564001)。
文摘Excitatory amino acid transporters(EAATs) are responsible for excitatory amino acid transportation and are associated with auto-immune diseases in the central nervous system and peripheral tissues.However, the subcellular location and function of EAAT2 in macrophages are still obscure. In this study,we demonstrated that LPS stimulation increases expression of EAAT2(coded by Slc1a2) via NF-κB signaling. EAAT2 is necessary for inflammatory macrophage polarization through sustaining mTORC1 activation. Mechanistically, lysosomal EAAT2 mediates lysosomal glutamate and aspartate efflux to maintain V-ATPase activation, which sustains macropinocytosis and mTORC1. We also found that mice with myeloid depletion of Slc1a2 show alleviated inflammatory responses in LPS-induced systemic inflammation and high-fat diet induced obesity. Notably, patients with type Ⅱ diabetes(T2D) have a higher level of expression of lysosomal EAAT2 and activation of mTORC1 in blood macrophages. Taken together, our study links the subcellular location of amino acid transporters with the fate decision of immune cells,which provides potential therapeutic targets for the treatment of inflammatory diseases.
文摘Glutamate is the predominant excitatory neurotransmitter in the human brain and it has been shown that prolonged activation of the glutamatergic system leads to nerve damage and cell death. Following release from the pre-synaptic neuron and synaptic transmission, glutamate is either taken up into the presynaptic neuron or neighbouring glia by transmembrane glutamate transporters. Excitatory amino acid transporter(EAAT) 1 and EAAT2 are Na+-dependant glutamate transporters expressed predominantly in glia cells of the central nervous system. As the most abundant glutamate transporters, their primary role is to modulate levels of glutamatergic excitability and prevent spill over of glutamate beyond the synapse. This role is facilitated through the binding and transportation of glutamate into astrocytes and microglia. The function of EAAT1 and EAAT2 is heavily regulated at the levels of gene expression, post-transcriptional splicing, glycosylation states and cell-surface trafficking of the protein. Both glutamatergic dysfunction and glial dysfunction have been proposed to be involved in psychiatric disorder. This review will present an overview of the roles that EAAT1 and EAAT2 play in modulating glutamatergic activity in the human brain, and mount an argument that these two transporters could be involved in the aetiologies of schizophrenia and affective disorders as well as represent potential drug targets for novel therapies for those disorders.
基金supported by the National Natural Science Foundation of China, No. 81171168Shanghai Science and Technology Committee, No. 10140903200
文摘The present study established a rat model of global cerebral ischemia induced by chest compression for six minutes to dynamically observe expressional changes of three glutamate transporters in the cerebral cortex and hippocampus. After 24 hours of ischemia, expression of glutamate transporter-1 significantly decreased in the cerebral cortex and hippocampus, which was accompanied by neuronal necrosis. At 7 days post-ischemia, expression of excitatory amino acid carrier 1 decreased in the hippocampal CA1 region and cortex, and was accompanied by apoptosis Expression of glutamate-aspartate transporter remained unchanged at 6 hours 7 days after ischemia. These results suggested that glutamate transporter levels were altered at different periods of cerebral ischemia.
基金This work was supported by grants from National Natural Science Foundation of China ( No. 39825109) and National Key Project of Basic Science Research (No. G1999054007).
文摘目的:探讨阿利吉仑对氧糖剥夺(OGD)损伤的神经母细胞瘤SH-SY5Y细胞的保护作用及可能机制。方法:将SH-SY5Y细胞随机分为对照组、OGD组及阿利吉仑低、中和高剂量(5.0、10.0和20.0 μmol/L)组。CCK-8法检测细胞活力;ELISA检测兴奋性氨基酸转运蛋白2(EAAT2/GLT-1)、兴奋性氨基酸转运蛋白3(EAAT3/EAAC1)、兴奋性氨基酸转运蛋白4(EAAT4)、内皮素1(ET-1)和S100钙结合蛋白β亚基(S-100β)的表达;Hoechst 33258染色观察SH-SY5Y细胞形态变化;乳酸(LD)测试盒和超微量Na +-K +-ATP酶测试盒检测LD含量和Na +-K +-ATPase活性。结果: OGD损伤4 h时,细胞相对活力不足60%,因此4 h可作为后续实验OGD造模时间。与对照组相比,OGD组的GLT-1、EAAC1和EAAT4表达显著下调( P <0.05),ET-1和S-100β的表达显著上调( P <0.05);与OGD组相比,阿利吉仑组的GLT-1、EAAC1和EAAT4表达呈剂量依赖性上调, ET-1和S-100β表达呈剂量依赖性下调( P <0.05)。Hochest 33258染色结果表明,阿利吉仑可明显减少OGD引起的SH-SY5Y细胞凋亡。与对照组相比,OGD组的LD含量显著升高( P <0.05),Na +-K +-ATPase活性显著降低( P <0.05);与OGD组相比,阿利吉仑组的LD含量呈剂量依赖性降低,Na +-K +-ATPase活性呈剂量依赖性升高( P <0.05)。结论:阿利吉仑对OGD损伤的SH-SY5Y细胞有一定的保护作用,其机制可能与阿利吉仑上调GLT-1、EAAC1和EAAT4的水平,提高Na +-K +-ATPase的活性,下调ET-1和S-100β的表达及LD含量有关。
