Plastics such as polyvinyl chlorides （PVC） are widely used in many indoor constructed environments; however, their unbound chemicals, such as di-（2-ethylhexyl） phthalates （DEHP）, can leach into the surrounding e...Plastics such as polyvinyl chlorides （PVC） are widely used in many indoor constructed environments; however, their unbound chemicals, such as di-（2-ethylhexyl） phthalates （DEHP）, can leach into the surrounding environment. This study focused on DEHP＇s effect on the central nervous system by determining the precise DEHP content in mice brain tissue after exposure to the chemical, to evaluate the specific exposure range. Primary neuronal-astrocyte co-culture systems were used as in vitro models for chemical hazard identification of DEHP. Oxidative stress was hypothesized as a probable mechanism involved, and therefore the total reactive oxygen species （ROS） concentration was determined as a biomarker of oxidative stress. In addition, NeuriteTracer, a neurite tracing plugin with ImageJ, was used to develop an assay for neurotoxicity to provide quantitative measurements of neurological parameters, such as neuronal number, neuron count and neurite length, all of which could indicate neurotoxic effects. The results showed that with 1 nmol/L DEHP exposure, there was a significant increase in ROS concentrations, indicating that the neuronal-astrocyte cultures were injured due to exposure to DEHP. In response, astrocyte proliferation （gliosis） was initiated, serving as a mechanism to maintain a homeostatic environment for neurons and protect neurons from toxic chemicals. There is a need to assess the cumulative effects of DEHP in animals to evaluate the possible uotake and effects on the human neuronal system from exoosure to DEHP in the indoor environment.展开更多
Epilepsy is a chronic and severe neurological disorder that has negative effects on the autonomous activities of patients. Functionally, Trem2(triggering receptor expressed on myeloid cells-2) is an immunoglobulin rec...Epilepsy is a chronic and severe neurological disorder that has negative effects on the autonomous activities of patients. Functionally, Trem2(triggering receptor expressed on myeloid cells-2) is an immunoglobulin receptor that affects neurological and psychiatric genetic diseases. Based on this rationale, we aimed to assess the potential role of Trem2 integration with the PI3 K/Akt pathway in epilepsy. We used microarray-based gene expression profiling to identify epilepsy-related differentially-expressed genes. In a mouse hippocampal neuron model of epilepsy, neurons were treated with lowMg^2+ extracellular fluid, and the protein and mRNA expression of Trem2 were determined. Using a gain-offunction approach with Trem2, neuronal apoptosis and its related factors were assessed by flow cytometry, RT-qPCR,and Western blot analysis. In a pilocarpine-induced epileptic mouse model, the malondialdehyde(MDA) and8-hydroxy-20-deoxyguanosine(8-OHdG) content and superoxide dismutase(SOD) and glutathione-peroxidase(GSH-Px) activity in the hippocampus were determined,and the protein expression of Trem2 was measured. In addition, the regulatory effect of Trem2 on the PI3 K/Akt pathway was analyzed by inhibiting this pathway in both the cell and mouse models of epilepsy. Trem2 was found to occupy a core position and was correlated with epilepsy.Trem2 was decreased in the hippocampus of epileptic miceand epileptic hippocampal neurons. Of crucial importance,overexpression of Trem2 activated the PI3 K/Akt pathway to inhibit neuronal apoptosis. Moreover, activation of the PI3 K/Akt pathway through over-expression of Trem2 alleviated oxidative stress, as shown by the increased expression of SOD and GSH-Px and the decreased expression of MDA and 8-OHdG. The current study defines the potential role of Trem2 in inhibiting the development of epilepsy, indicating that Trem2 up-regulation alleviates hippocampal neuronal injury and oxidative stress, and inhibits neuronal apoptosis in epilepsy by activating the PI3 K/Akt pathway.展开更多
Efficient viral vectors for mapping and manipulating long-projection neuronal circuits are crucial in structural and functional studies of the brain. The SAD strain rabies virus with the glycoprotein gene deleted pseu...Efficient viral vectors for mapping and manipulating long-projection neuronal circuits are crucial in structural and functional studies of the brain. The SAD strain rabies virus with the glycoprotein gene deleted pseudotyped with the N2 C glycoprotein(SAD-RV(DG)-N2 C(G)) shows strong neuro-tropism in cell culture, but its in vivo efficiency for retrograde gene transduction and neuro-tropism have not been systematically characterized.We compared these features in different mouse brain regions for SAD-RV-N2 C(G) and two other widely-used retrograde tracers, SAD-RV(DG)-B19(G) and r AAV2-retro. We found that SAD-RV(DG)-N2 C(G) enhanced the infection efficiency of long-projecting neurons by^10 times but with very similar neuro-tropism, compared with SAD-RV(DG)-B19(G). On the other hand, SAD-RV(DG)-N2 C(G) had an infection efficiency comparable with r AAV2-retro, but a more restricted diffusion range, and broader tropism to different types and regions of longprojecting neuronal populations. These results demonstrate that SAD-RV(DG)-N2 C(G) can serve as an effective retrograde vector for studying neuronal circuits.展开更多
BACKGROUND： Electrophysiology can prove the integration of afferent information from the stomach meridian of Foot-Yangming in the nucleus tractus solitarius （NTS） and objectively describe the specific association b...BACKGROUND： Electrophysiology can prove the integration of afferent information from the stomach meridian of Foot-Yangming in the nucleus tractus solitarius （NTS） and objectively describe the specific association between meridian vessels and Zang Fu organs. OBJECTIVE： To investigate the effects of afferent information from acupuncture at Sibai （ST 2） acupoint on neuronal discharge in rat NTS. DESIGN, TIME AND SETFING： A randomized, controlled, animal experiment was performed at the Key Laboratory of Meridian-Vessels and Zang Fu Organs, Traditional Chinese Medicine University of Hunan, State Administration of Traditional Chinese Medicine, and Key Laboratory of Acupuncture, Moxibustion, and the Biological Information of Hunan Higher Education Institutes, between December 2005 and October 2008. MATERIALS： A total of 52 Sprague Dawley rats, of either gender, aged 4 months, were included in this study. Acupuncture needles of 0.32 mm （diameter） x 40 mm （length） were used. METHODS： An extracellular recording protocol was applied. The Sibai （ST 2） acupoint in the stomach meridian of Foot-Yangming was used as an acupuncture point （acupoint）. Simultaneously, Dicang （ST 4） and Neiting （ST 44） acupoints in the stomach meridian of Foot-Yangming, Quanliao （S118） acupoint in the small intestine meridian of Hand-Taiyang, and a non-acupoint lateral to Sibai （ST 2） acupoint, were selected as controls. The Sibai （ST 2） acupoint was stimulated for 30 seconds by hand acupuncture through twirling and rotating, to determine the neurons responding to body surface stimulation in the NTS. MAIN OUTCOME MEASURES： Frequency of responding NTS neurons after acupuncture at four acupoints including Sibai （ST 2）, Dicang （ST 4）, Neiting （ST 44） and Quanliao （SI 18） and one non-acupoint. RESULTS： The frequency of responding NTS neurons was significantly higher after acupuncture at Sibaithan at control sites including the Dicang （ST 4）, Neiting （ST 44） and Quanliao （S118） acupoints and at the non-acupoint （P 〈 0.01）. The frequency of responding NTS neurons at Dicang （ST 4） and Quanliao （SI 18） was significantly higher than at Quanliao （SI 18） and the non-acupoint （P 〈 0.05）. The rate of frequency change of responding NTS neurons for the Sibai （ST 2）, Dicang （ST 4）, Neiting （ST 44）, and Quanliao （S118） acupoints as well as the non-acupoint was （35.08±4.80） %, （28.25± 5.46） %, （27.57± 4.87） %, （20.02 ±4.23） %, and （18.55 ±2.49） % respectively. Simultaneously, significant differences existed between Sibai （ST 2） and the other acupoints （P 〈 0.05 or P〈 0.01）. CONCLUSION： Compared with the Dicang （ST 4） and Neiting （ST 44） acupoints in the stomach meridian of Foot-Yangming, Quanliao （SI 18） acupoint in the small intestine meridian of Hand-Taiyang, and the non-acupoint lateral to Sibai （ST 2） acupoint, the Sibai （ST 2） acupoint in the stomach meridian of Foot- Yangming is more closely related to the NTS. In the stomach meridian of Foot- Yangming, afferent information is different in distant and near Shu acupoints, indicating that each Shu acupoint has its own specificity.展开更多
Functional changes in synaptic transmission from the lateral entorhinal cortex to the dentate gyrus(LEC-DG)are considered responsible for the chronification of pain.However,the underlying alterations in fan cells,whic...Functional changes in synaptic transmission from the lateral entorhinal cortex to the dentate gyrus(LEC-DG)are considered responsible for the chronification of pain.However,the underlying alterations in fan cells,which are the predominant neurons in the LEC that project to the DG,remain elusive.Here,we investigated possible mechanisms using a rat model of complete Freund’s adjuvant(CFA)-induced inflammatory pain.We found a substantial increase in hyperpolarization-activated/cyclic nucleotide-gated currents(Ih),which led to the hyperexcitability of LEC fan cells of CFA slices.This phenomenon was attenuated in CFA slices by activating dopamine D2,but not D1,receptors.Chemogenetic activation of the ventral tegmental area-LEC projection had a D2 receptor-dependent analgesic effect.Intra-LEC microinjection of a D2 receptor agonist also suppressed CFA-induced behavioral hypersensitivity,and this effect was attenuated by pre-activation of the Ih.Our findings suggest that down-regulating the excitability of LEC fan cells through activation of the dopamine D2 receptor may be a strategy for treating chronic inflammatory pain.展开更多
BACKGROUND： Apoptosis plays an important role in central neural diseases and trauma. B-cell lymphoma/Leukemia-2 （Bcl-2） can inhibit apoptosis in a wide variety of cells including neurons. In this experiment, by stu...BACKGROUND： Apoptosis plays an important role in central neural diseases and trauma. B-cell lymphoma/Leukemia-2 （Bcl-2） can inhibit apoptosis in a wide variety of cells including neurons. In this experiment, by studying Bcl-2 over-expression transgenic （TG） mice subjected to spinal cord injury （SCI）, we investigated whether Bcl-2 could reduce posttraumatic neuronal apoptosis, reduce the range of damage, and improve the behavioral functional recovery after contusive SCI.METHODS： Nine Bcl-2 TG mice and nine control mice were subjected to SCI of moderate severity at T10, with the use of weight dropping （WD） method （impact force 2.5×3.0 g/cm）. At times up to 1 day, 7 days and 14 days after SCI, functional defi cits were evaluated with Basso, Beattie, and Bresnahan （BBB） scales, and apoptosis of neurons was investigated by using the TUNEL method. Another three control mice only underwent lamina opening, but were not subjected to SCI, to provide blank comparison.RESULTS： The mean functional scores for the control mice （5.05 ±0.35） were lower than those for the Bcl-2 TG mice （5.45 ±0.15）, although the unpaired T-test revealed no signifi cant difference （P=0.67）. On the other hand, the number of TUNEL positive neurons and integrated option density （IOD） scores for the Bcl-2 TG mice were both signifi cantly lower than those for the control mice （P〈0.05）.CONCLUSIONS： This experiment suggests that overexpression of Bcl-2 may suppress neuronal apoptosis after SCI. Bcl-2 may be an important factor within the central nervous system that can relieve the damage after trauma.展开更多
Astrocytes are increasingly recognized to play an active role in learning and memory,but whether neural inputs can trigger event-specific astrocytic Ca^(2+)dynamics in real time to participate in working memory remain...Astrocytes are increasingly recognized to play an active role in learning and memory,but whether neural inputs can trigger event-specific astrocytic Ca^(2+)dynamics in real time to participate in working memory remains unclear due to the difficulties in directly monitoring astrocytic Ca^(2+)dynamics in animals performing tasks.Here,using fiber photometry,we showed that population astrocytic Ca^(2+)dynamics in the hippocampus were gated by sensory inputs(centered at the turning point of the T-maze)and modified by the reward delivery during the encoding and retrieval phases.Notably,there was a strong inter-locked and antagonistic relationship between the astrocytic and neuronal Ca^(2+)dynamics with a 3-s phase difference.Furthermore,there was a robust synchronization of astrocytic Ca^(2+)at the population level among the hippocampus,medial prefrontal cortex,and striatum.The inter-locked,bidirectional communication between astrocytes and neurons at the population level may contribute to the modulation of information processing in working memory.展开更多
Epidemiological and animal studies indicate that pre-existing diabetes increases the risk of Parkinson's disease(PD).However,the mechanisms underlying this association remain unclear.In the present study,we found ...Epidemiological and animal studies indicate that pre-existing diabetes increases the risk of Parkinson's disease(PD).However,the mechanisms underlying this association remain unclear.In the present study,we found that high glucose(HG)levels in the cerebrospinal fluid(CSF)of diabetic rats might enhance the effect of a subthreshold dose of the neurotoxin 6-hydroxydopamine(6-OHDA)on the development of motor disorders,and the damage to the nigrostriatal dopaminergic neuronal pathway.In vitro,HG promoted the 6-OHDA-induced apoptosis in PC12 cells differentiated to neurons with nerve growth factor(NGF)(NGF-PC12).Metabolomics showed that HG promoted hyperglycolysis in neurons and impaired tricarboxylic acid cycle(TCA cycle)activity,which was closely related to abnormal mitochondrial fusion,thus resulting in mitochondrial loss.Interestingly,HG-induced upregulation of pyruvate kinase M2(PKM2)combined with 6-OHDA exposure not only mediated glycolysis but also promoted abnormal mitochondrial fusion by upregulating the expression of MFN2 in NGF-PC12 cells.In addition,we found that PKM2 knockdown rescued the abnormal mitochondrial fusion and cell apoptosis induced by HGþ6-OHDA.Furthermore,we found that shikonin(SK),an inhibitor of PKM2,restored the mitochondrial number,promoted TCA cycle activity,reversed hyperglycolysis,enhanced the tolerance of cultured neurons to 6-OHDA,and reduced the risk of PD in diabetic rats.Overall,our results indicate that diabetes promotes hyperglycolysis and abnormal mitochondrial fusion in neurons through the upregulation of PKM2,leading to an increase in the vulnerability of dopaminergic neurons to 6-OHDA.Thus,the inhibition of PKM2 and restoration of mitochondrial metabolic homeostasis/pathways may prevent the occurrence and development of diabetic PD.展开更多
Acknowledgments： I would like to express my appreciation to Professor Puro DG for leading me to this research topic during my stay as a research fellow in his laboratory at the University of Michigan in 2001, and als...Acknowledgments： I would like to express my appreciation to Professor Puro DG for leading me to this research topic during my stay as a research fellow in his laboratory at the University of Michigan in 2001, and also to Professor Ikeda T forgiving me the opportunity to study abroad and then to continue to investigate this topic in the Department of Ophthalmology at Osaka Medical College, lapan.展开更多
BACKGROUND Cognitive dysfunction in epileptic patients is a high-incidence complication.Its mechanism is related to nervous system damage during seizures,but there is no effective diagnostic biomarker.Neuronal pentrax...BACKGROUND Cognitive dysfunction in epileptic patients is a high-incidence complication.Its mechanism is related to nervous system damage during seizures,but there is no effective diagnostic biomarker.Neuronal pentraxin 2(NPTX2)is thought to play a vital role in neurotransmission and the maintenance of synaptic plasticity.This study explored how serum NPTX2 and electroencephalogram(EEG)slow wave/fast wave frequency ratio relate to cognitive dysfunction in patients with epilepsy.AIM To determine if serum NPTX2 could serve as a potential biomarker for diagnosing cognitive impairment in epilepsy patients.METHODS The participants of this study,conducted from January 2020 to December 2021,comprised 74 epilepsy patients with normal cognitive function(normal group),37 epilepsy patients with cognitive dysfunction[epilepsy patients with cognitive dysfunction(ECD)group]and 30 healthy people(control group).The minimental state examination(MMSE)scale was used to evaluate cognitive function.We determined serum NPTX2 levels using an enzyme-linked immunosorbent kit and calculated the signal value of EEG regions according to the EEG recording.Pearson correlation coefficient was used to analyze the correlation between serum NPTX2 and the MMSE score.RESULTS The serum NPTX2 level in the control group,normal group and ECD group were 240.00±35.06 pg/mL,235.80±38.01 pg/mL and 193.80±42.72 pg/mL,respectively.The MMSE score was lowest in the ECD group among the three,while no significant difference was observed between the control and normal groups.In epilepsy patients with cognitive dysfunction,NPTX2 level had a positive correlation with the MMSE score(r=0.367,P=0.0253)and a negative correlation with epilepsy duration(r=−0.443,P=0.0061)and the EEG slow wave/fast wave frequency ratio value in the temporal region(r=−0.339,P=0.039).CONCLUSION Serum NPTX2 was found to be related to cognitive dysfunction and the EEG slow wave/fast wave frequency ratio in patients with epilepsy.It is thus a potential biomarker for the diagnosis of cognitive impairment in patients with epilepsy.展开更多
c-Jun NH2-terminal kinase（JNK）-interacting protein 3 plays an important role in brain-derived neurotrophic factor/tropomyosin-related kinase B（Trk B） anterograde axonal transport. It remains unclear whether JNK-in...c-Jun NH2-terminal kinase（JNK）-interacting protein 3 plays an important role in brain-derived neurotrophic factor/tropomyosin-related kinase B（Trk B） anterograde axonal transport. It remains unclear whether JNK-interacting protein 1 mediates similar effects, or whether JNK-interacting protein 1 affects the regulation of Trk B anterograde axonal transport. In this study, we isolated rat embryonic hippocampus and cultured hippocampal neurons in vitro. Coimmunoprecipitation results demonstrated that JNK-interacting protein 1 formed Trk B complexes in vitro and in vivo. Immunocytochemistry results showed that when JNK-interacting protein 1 was highly expressed, the distribution of Trk B gradually increased in axon terminals. However, the distribution of Trk B reduced in axon terminals after knocking out JNK-interacting protein 1. In addition, there were differences in distribution of Trk B after JNK-interacting protein 1 was knocked out compared with not. However, knockout of JNK-interacting protein 1 did not affect the distribution of Trk B in dendrites. These findings confirm that JNK-interacting protein 1 can interact with Trk B in neuronal cells, and can regulate the transport of Trk B in axons, but not in dendrites.展开更多
Background:Parkinson’s disease(PD)is a common neurodegenerative disease,characterized by symptoms like tremors,muscle rigidity,and slowmovement.Themain cause of these symptoms is the loss of dopamineproducing neurons...Background:Parkinson’s disease(PD)is a common neurodegenerative disease,characterized by symptoms like tremors,muscle rigidity,and slowmovement.Themain cause of these symptoms is the loss of dopamineproducing neurons in a brain area called the substantia nigra.Various genetic and environmental factors contribute to this neuronal loss.Once symptoms of PD begin,they worsen with age,which also impacts several critical cellular processes.Leucine-rich repeat kinase 2(LRRK2)is a gene associated with PD.Certain mutations in LRRK2,such as G2019S,increase its activity,disrupting cellular mechanisms necessary for healthy neuron function,including autophagy and lysosomal activity.Exposure to rotenone(RTN)promotes LRRK2 activity in neurons and contributes to cellular senescence andα-syn accumulation.Methods:In this study,human dopaminergic progenitor cells were reprogrammed to study the effects of RTN with the co-treatment of LRRK2 inhibitor on cellular senescence.We measured the cellular senescence using quantifying proteins of senescence markers,such as p53,p21,Rb,phosphorylated Rb,andβ-galatocidase,and the enzymatic activity of senescence-associatedβ-galatocidase.And we estimated the levels of accumulatedα-synuclein(α-syn),which is increased via the impaired autophagy-lysosomal pathway by cellular senescence.Then,we evaluated the association of the G2019S LRRK2 mutation and senescence-associatedβ-galatocidase and the levels of accumulated or secretedα-syn,and the neuroinflammatory responses mediated by the secretedα-syn in rat primary microglia were determined using the release of pro-inflammatory cytokines.Results:RTN raised senescence markers and affected the phosphorylation of Rab10,a substrate of LRRK2.The inhibiting agent MLI2 reduced these senescence markers and Rab10 phosphorylations.Additionally,RTN increasedα-syn levels in the neurons,while MLI2 aided in degrading it.When focusing on cells from PD patients with the G2019S mutation,an increase in cellular senescence and release ofα-syn was observed,provoking neuroinflammation.Treatment with the LRRK2 inhibitor MLI2 decreased both cellular senescence andα-syn secretion,thereby mitigating inflammatory responses.Conclusion:Overall,inhibiting LRRK2 may provide a beneficial strategy formanaging PD.展开更多
Neuropathic pain,often featuring allodynia,imposes significant physical and psychological burdens on patients,with limited treatments due to unclear central mechanisms.Addressing this challenge remains a crucial unsol...Neuropathic pain,often featuring allodynia,imposes significant physical and psychological burdens on patients,with limited treatments due to unclear central mechanisms.Addressing this challenge remains a crucial unsolved issue in pain medicine.Our previous study,using protein kinase C gamma(PKCγ)-tdTomato mice,highlights the spinal feedforward inhibitory circuit involving PKCγ neurons in gating neuropathic allodynia.However,the regulatory mechanisms governing this circuit necessitate further elucidation.We used diverse transgenic mice and advanced techniques to uncover the regulatory role of the descending serotonin(5-HT)facilitation system on spinal PKCγ neurons.Our findings revealed that 5-HT neurons from the rostral ventromedial medulla hyperpolarize spinal inhibitory interneurons via 5-HT_(2C) receptors,disinhibiting the feedforward inhibitory circuit involving PKCγ neurons and exacerbating allodynia.Inhibiting spinal 5-HT_(2C) receptors restored the feedforward inhibitory circuit,effectively preventing neuropathic allodynia.These insights offer promising therapeutic targets for neuropathic allodynia management,emphasizing the potential of spinal 5-HT_(2C) receptors as a novel avenue for intervention.展开更多
Abnormal synchronous neuronal activity has been widely detected by brain imaging of autistic patients,but its underlying neural mechanism remains unclear.Compared with wild-type mice,our in vivo two-photon imaging sho...Abnormal synchronous neuronal activity has been widely detected by brain imaging of autistic patients,but its underlying neural mechanism remains unclear.Compared with wild-type mice,our in vivo two-photon imaging showed that transgenic(Tgl)mice over-expressing human autism risk gene MeCP2 exhibited higher neuronal synchrony in the young but lower synchrony in the adult stage.Whole-cell recording of neuronal pairs in brain slices revealed that higher neuronal synchrony in young postnatal Tgl mice was atributed mainly to more prevalent giant slow inward currents(SICs).Both in vivo and slice imaging further demonstrated more dynamic activity and higher synchrony in astrocytes from young Tgl mice.Blocking astrocytic gap junctions markedly decreased the generation of SICs and overall cell synchrony in the Tgl brain.Furthermore,the expression level of Cx43 protein and the coupling efficiency of astrocyte gap junctions remained unchanged in Tgi mice.Thus,astrocytic gap junctions facilitate but do not act as a direct trigger for the abnormal neuronal synchrony in young Tgl mice,revealing the potential role of the astrocyte network in the pathogenesis of MeCP2 duplication syndrome.展开更多
Bromodomain containing 2 （Brd2） protein belongs to the Bromodomains and Extra Terminal domain （BET） family of chromatin adaptors characterized by the presence of two N-terminal tandem bromodomains and an exclusive...Bromodomain containing 2 （Brd2） protein belongs to the Bromodomains and Extra Terminal domain （BET） family of chromatin adaptors characterized by the presence of two N-terminal tandem bromodomains and an exclusive C-terminal extra terminal domain （ET） （Belkina and Denis, 2012; Shi and Vakoc, 2014）. Bromodomains are involved in recognizing acetylated histone tails and other acetylated proteins while the ET domain has been implicated in protein-protein interaction.展开更多
BACKGROUND: Ⅴ secretory phospholipase A2 (sPLA2-Ⅴ) is abundant in many mammal tissues. However, it remains unknown whether sPLA2-Ⅴ causes biological or pathological response in central nervous system. OBJECTIVE: To...BACKGROUND: Ⅴ secretory phospholipase A2 (sPLA2-Ⅴ) is abundant in many mammal tissues. However, it remains unknown whether sPLA2-Ⅴ causes biological or pathological response in central nervous system. OBJECTIVE: To observe the effect of phospholipase A2-Ⅴ (PLA2-Ⅴ) and its inhibitor (indoxam) on hippocampal neuron survival. DESIGN: A repetitive measurement. SETTING: The Animal Center of South Carolina University. MATERIALS: Sprague-Dawley pregnancy day-7, 14, 21 female rats were selected; Reagents: sPLA2- Ⅴ and indoxam were obtained from the Dennis Research Laboratories METHODS: The experiment was finished at the animal center in South Carolina University from January to December, 2004. 0, 12.5, 25, 50 and 100 μg/L sPLA2-Ⅴ were added to neuron with none-MgCl2 Eagle’s medium at 37 ℃, then changed to normal neuron culture medium after 3 hours. 1, 2.5, 5 and 10 μmol/L indoxam was added at 6 hours after 100 μg/L sPLA2-Ⅴwas put to Day-21 SD rat hippocampal embryonic neurons with none-MgCl2 Eagle’s medium at 37 ℃. After 3 hours in the inhibition experiment, it was changed to normal neuron culture medium. The embryonic hippocampal neurons were primarily cultured, and the neuron survival ratio was detected with morphological method. MAIN OUTCOME MEASURES: Survival ratio of hippocampal neurons. RESULTS: ① Effects of sPLA2-Ⅴon neuron survival: When sPLA2-Ⅴ was 0, 12.5, 25, 50 and 100 μg/L, the neuron survival ratios in embryonic neurons of day-7 SD rats were (95.3±1.1)%, (81.4±3.1)%, (74.2±2.2)%, (62.4±1.7)% and (48.9±1.6)%, those in embryonic neurons of day-14 rats were (93.2±1.4)%, (74.3±1.9)%, (68.1±1.7)%, (56.1±1.4)% and (42.5±1.1)%, and those in embryonic neurons of day-21 rats were (91.2±1.2)%, (69.4±2.1)%, (60.3±2.2)%, (49.1±1.2)% and (35.5±1.9)%. There were significant differences among different concentrations (P < 0.05). ② Effects of indoxam on neuron survival: In case of sPLA2-Ⅴ 100 μg/L, the neuron survival ratios were (58.65±1.4)%, (69.34±1.1)%, (82.11±1.2)% and (95.28±0.9)% when indoxam was 1, 2.5, 5 and 10 μmol/L, respectively. There were significant differences among different concentrations (P < 0.05). CONCLUSION: ① The of neuronal death ratio is in a concentration-dependent manner with sPLA2-Ⅴ, and increases as the embryonic aging. ② Indoxam inhibits the proapoptotic effect of sPLA2-Ⅴ.展开更多
This editorial summarizes the latest literature on the roles of neuronal PAS domain protein 2 and KN motif/ankyrin repeat domain 1 in type 2 diabetes(T2D).We highlight their involvement inβ-cell dysfunction,explore t...This editorial summarizes the latest literature on the roles of neuronal PAS domain protein 2 and KN motif/ankyrin repeat domain 1 in type 2 diabetes(T2D).We highlight their involvement inβ-cell dysfunction,explore their potential as therapeutic targets,and discuss the implications for new treatment strategies.We offer valuable insights into relevant gene regulation and cellular mechanisms relevant for the targeted management of T2D.展开更多
The NSC-34 cell line is a widely recognized motor neuron model and various neuronal differentiation protocols have been exploited. Under previously reported experimental conditions, only part of the cells resemble dif...The NSC-34 cell line is a widely recognized motor neuron model and various neuronal differentiation protocols have been exploited. Under previously reported experimental conditions, only part of the cells resemble differentiated neurons;however, they do not exhibit extensive and time-prolonged neuritogenesis, and maintain their duplication capacity in culture. The aim of the present work was to facilitate long-term and more homogeneous neuronal differentiation in motor neuron–like NSC-34 cells. We found that the antimitotic drug cytosine arabinoside promoted robust and persistent neuronal differentiation in the entire cell population. Long and interconnecting neuronal processes with abundant growth cones were homogeneously induced and were durable for up to at least 6 weeks in culture. Moreover, cytosine arabinoside was permissive, dispensable, and mostly irreversible in priming NSC-34 cells for neurite initiation and regeneration after mechanical dislodgement. Finally, the expression of the cell proliferation antigen Ki67 was inhibited by cytosine arabinoside, whereas the expression levels of neuronal growth associated protein 43, vimentin, and motor neuron–specific p75, Islet2, homeobox 9 markers were upregulated, as confirmed by western blot and/or confocal immunofluorescence analysis. Overall, these findings support the use of NSC-34 cells as a motor neuron model for properly investigating neurodegenerative mechanisms and prospectively identifying neuroprotective strategies.展开更多
基金supported by the Key Project of National Natural Science Foundation of China(No.51136002)China Key Technologies R&D Program(No.2012BAJ02B03)
文摘Plastics such as polyvinyl chlorides （PVC） are widely used in many indoor constructed environments; however, their unbound chemicals, such as di-（2-ethylhexyl） phthalates （DEHP）, can leach into the surrounding environment. This study focused on DEHP＇s effect on the central nervous system by determining the precise DEHP content in mice brain tissue after exposure to the chemical, to evaluate the specific exposure range. Primary neuronal-astrocyte co-culture systems were used as in vitro models for chemical hazard identification of DEHP. Oxidative stress was hypothesized as a probable mechanism involved, and therefore the total reactive oxygen species （ROS） concentration was determined as a biomarker of oxidative stress. In addition, NeuriteTracer, a neurite tracing plugin with ImageJ, was used to develop an assay for neurotoxicity to provide quantitative measurements of neurological parameters, such as neuronal number, neuron count and neurite length, all of which could indicate neurotoxic effects. The results showed that with 1 nmol/L DEHP exposure, there was a significant increase in ROS concentrations, indicating that the neuronal-astrocyte cultures were injured due to exposure to DEHP. In response, astrocyte proliferation （gliosis） was initiated, serving as a mechanism to maintain a homeostatic environment for neurons and protect neurons from toxic chemicals. There is a need to assess the cumulative effects of DEHP in animals to evaluate the possible uotake and effects on the human neuronal system from exoosure to DEHP in the indoor environment.
基金supported by Beijing Key Laboratory of Neuromodulation(BZ0098)the Precision Medicine Project of the Ministry of Science and Technology of China(2016YFC0904400)
文摘Epilepsy is a chronic and severe neurological disorder that has negative effects on the autonomous activities of patients. Functionally, Trem2(triggering receptor expressed on myeloid cells-2) is an immunoglobulin receptor that affects neurological and psychiatric genetic diseases. Based on this rationale, we aimed to assess the potential role of Trem2 integration with the PI3 K/Akt pathway in epilepsy. We used microarray-based gene expression profiling to identify epilepsy-related differentially-expressed genes. In a mouse hippocampal neuron model of epilepsy, neurons were treated with lowMg^2+ extracellular fluid, and the protein and mRNA expression of Trem2 were determined. Using a gain-offunction approach with Trem2, neuronal apoptosis and its related factors were assessed by flow cytometry, RT-qPCR,and Western blot analysis. In a pilocarpine-induced epileptic mouse model, the malondialdehyde(MDA) and8-hydroxy-20-deoxyguanosine(8-OHdG) content and superoxide dismutase(SOD) and glutathione-peroxidase(GSH-Px) activity in the hippocampus were determined,and the protein expression of Trem2 was measured. In addition, the regulatory effect of Trem2 on the PI3 K/Akt pathway was analyzed by inhibiting this pathway in both the cell and mouse models of epilepsy. Trem2 was found to occupy a core position and was correlated with epilepsy.Trem2 was decreased in the hippocampus of epileptic miceand epileptic hippocampal neurons. Of crucial importance,overexpression of Trem2 activated the PI3 K/Akt pathway to inhibit neuronal apoptosis. Moreover, activation of the PI3 K/Akt pathway through over-expression of Trem2 alleviated oxidative stress, as shown by the increased expression of SOD and GSH-Px and the decreased expression of MDA and 8-OHdG. The current study defines the potential role of Trem2 in inhibiting the development of epilepsy, indicating that Trem2 up-regulation alleviates hippocampal neuronal injury and oxidative stress, and inhibits neuronal apoptosis in epilepsy by activating the PI3 K/Akt pathway.
基金the National Basic Research Program(973 Program)of China(2015CB755601)the Strategic Priority Research Program of the Chinese Academy of Sciences(XDB32030200)+1 种基金the National Natural Science Foundation of China(31771156,81661148053,91632303,31800885,31500868,31671120 and 91732304)the China Postdoctoral Science Foundation(2019M653118 and 2018M632946).
文摘Efficient viral vectors for mapping and manipulating long-projection neuronal circuits are crucial in structural and functional studies of the brain. The SAD strain rabies virus with the glycoprotein gene deleted pseudotyped with the N2 C glycoprotein(SAD-RV(DG)-N2 C(G)) shows strong neuro-tropism in cell culture, but its in vivo efficiency for retrograde gene transduction and neuro-tropism have not been systematically characterized.We compared these features in different mouse brain regions for SAD-RV-N2 C(G) and two other widely-used retrograde tracers, SAD-RV(DG)-B19(G) and r AAV2-retro. We found that SAD-RV(DG)-N2 C(G) enhanced the infection efficiency of long-projecting neurons by^10 times but with very similar neuro-tropism, compared with SAD-RV(DG)-B19(G). On the other hand, SAD-RV(DG)-N2 C(G) had an infection efficiency comparable with r AAV2-retro, but a more restricted diffusion range, and broader tropism to different types and regions of longprojecting neuronal populations. These results demonstrate that SAD-RV(DG)-N2 C(G) can serve as an effective retrograde vector for studying neuronal circuits.
基金Supported by:Major State Basic Research Development Program of China(973 Program),No. 2009CB522904
文摘BACKGROUND： Electrophysiology can prove the integration of afferent information from the stomach meridian of Foot-Yangming in the nucleus tractus solitarius （NTS） and objectively describe the specific association between meridian vessels and Zang Fu organs. OBJECTIVE： To investigate the effects of afferent information from acupuncture at Sibai （ST 2） acupoint on neuronal discharge in rat NTS. DESIGN, TIME AND SETFING： A randomized, controlled, animal experiment was performed at the Key Laboratory of Meridian-Vessels and Zang Fu Organs, Traditional Chinese Medicine University of Hunan, State Administration of Traditional Chinese Medicine, and Key Laboratory of Acupuncture, Moxibustion, and the Biological Information of Hunan Higher Education Institutes, between December 2005 and October 2008. MATERIALS： A total of 52 Sprague Dawley rats, of either gender, aged 4 months, were included in this study. Acupuncture needles of 0.32 mm （diameter） x 40 mm （length） were used. METHODS： An extracellular recording protocol was applied. The Sibai （ST 2） acupoint in the stomach meridian of Foot-Yangming was used as an acupuncture point （acupoint）. Simultaneously, Dicang （ST 4） and Neiting （ST 44） acupoints in the stomach meridian of Foot-Yangming, Quanliao （S118） acupoint in the small intestine meridian of Hand-Taiyang, and a non-acupoint lateral to Sibai （ST 2） acupoint, were selected as controls. The Sibai （ST 2） acupoint was stimulated for 30 seconds by hand acupuncture through twirling and rotating, to determine the neurons responding to body surface stimulation in the NTS. MAIN OUTCOME MEASURES： Frequency of responding NTS neurons after acupuncture at four acupoints including Sibai （ST 2）, Dicang （ST 4）, Neiting （ST 44） and Quanliao （SI 18） and one non-acupoint. RESULTS： The frequency of responding NTS neurons was significantly higher after acupuncture at Sibaithan at control sites including the Dicang （ST 4）, Neiting （ST 44） and Quanliao （S118） acupoints and at the non-acupoint （P 〈 0.01）. The frequency of responding NTS neurons at Dicang （ST 4） and Quanliao （SI 18） was significantly higher than at Quanliao （SI 18） and the non-acupoint （P 〈 0.05）. The rate of frequency change of responding NTS neurons for the Sibai （ST 2）, Dicang （ST 4）, Neiting （ST 44）, and Quanliao （S118） acupoints as well as the non-acupoint was （35.08±4.80） %, （28.25± 5.46） %, （27.57± 4.87） %, （20.02 ±4.23） %, and （18.55 ±2.49） % respectively. Simultaneously, significant differences existed between Sibai （ST 2） and the other acupoints （P 〈 0.05 or P〈 0.01）. CONCLUSION： Compared with the Dicang （ST 4） and Neiting （ST 44） acupoints in the stomach meridian of Foot-Yangming, Quanliao （SI 18） acupoint in the small intestine meridian of Hand-Taiyang, and the non-acupoint lateral to Sibai （ST 2） acupoint, the Sibai （ST 2） acupoint in the stomach meridian of Foot- Yangming is more closely related to the NTS. In the stomach meridian of Foot- Yangming, afferent information is different in distant and near Shu acupoints, indicating that each Shu acupoint has its own specificity.
基金supported by the National Natural Science Foundation of China(81901119 and 81901142)Special Project on Innovation and Generation of Medical Support Capacity,and the Natural Science Foundation of Tibet(XZ2019ZRG-119),China.
文摘Functional changes in synaptic transmission from the lateral entorhinal cortex to the dentate gyrus(LEC-DG)are considered responsible for the chronification of pain.However,the underlying alterations in fan cells,which are the predominant neurons in the LEC that project to the DG,remain elusive.Here,we investigated possible mechanisms using a rat model of complete Freund’s adjuvant(CFA)-induced inflammatory pain.We found a substantial increase in hyperpolarization-activated/cyclic nucleotide-gated currents(Ih),which led to the hyperexcitability of LEC fan cells of CFA slices.This phenomenon was attenuated in CFA slices by activating dopamine D2,but not D1,receptors.Chemogenetic activation of the ventral tegmental area-LEC projection had a D2 receptor-dependent analgesic effect.Intra-LEC microinjection of a D2 receptor agonist also suppressed CFA-induced behavioral hypersensitivity,and this effect was attenuated by pre-activation of the Ih.Our findings suggest that down-regulating the excitability of LEC fan cells through activation of the dopamine D2 receptor may be a strategy for treating chronic inflammatory pain.
文摘BACKGROUND： Apoptosis plays an important role in central neural diseases and trauma. B-cell lymphoma/Leukemia-2 （Bcl-2） can inhibit apoptosis in a wide variety of cells including neurons. In this experiment, by studying Bcl-2 over-expression transgenic （TG） mice subjected to spinal cord injury （SCI）, we investigated whether Bcl-2 could reduce posttraumatic neuronal apoptosis, reduce the range of damage, and improve the behavioral functional recovery after contusive SCI.METHODS： Nine Bcl-2 TG mice and nine control mice were subjected to SCI of moderate severity at T10, with the use of weight dropping （WD） method （impact force 2.5×3.0 g/cm）. At times up to 1 day, 7 days and 14 days after SCI, functional defi cits were evaluated with Basso, Beattie, and Bresnahan （BBB） scales, and apoptosis of neurons was investigated by using the TUNEL method. Another three control mice only underwent lamina opening, but were not subjected to SCI, to provide blank comparison.RESULTS： The mean functional scores for the control mice （5.05 ±0.35） were lower than those for the Bcl-2 TG mice （5.45 ±0.15）, although the unpaired T-test revealed no signifi cant difference （P=0.67）. On the other hand, the number of TUNEL positive neurons and integrated option density （IOD） scores for the Bcl-2 TG mice were both signifi cantly lower than those for the control mice （P〈0.05）.CONCLUSIONS： This experiment suggests that overexpression of Bcl-2 may suppress neuronal apoptosis after SCI. Bcl-2 may be an important factor within the central nervous system that can relieve the damage after trauma.
基金This work was supported by Start-up Funds from Wenzhou Medical University(89211010 and 89212012)the National Natural Science Foundation of China(81630040,31771178,and 81600991)the Natural Science Foundation of Zhejiang Province of China(LY21H090014 and LQ18C090002).
文摘Astrocytes are increasingly recognized to play an active role in learning and memory,but whether neural inputs can trigger event-specific astrocytic Ca^(2+)dynamics in real time to participate in working memory remains unclear due to the difficulties in directly monitoring astrocytic Ca^(2+)dynamics in animals performing tasks.Here,using fiber photometry,we showed that population astrocytic Ca^(2+)dynamics in the hippocampus were gated by sensory inputs(centered at the turning point of the T-maze)and modified by the reward delivery during the encoding and retrieval phases.Notably,there was a strong inter-locked and antagonistic relationship between the astrocytic and neuronal Ca^(2+)dynamics with a 3-s phase difference.Furthermore,there was a robust synchronization of astrocytic Ca^(2+)at the population level among the hippocampus,medial prefrontal cortex,and striatum.The inter-locked,bidirectional communication between astrocytes and neurons at the population level may contribute to the modulation of information processing in working memory.
基金the National Natural Science Foundation of China(Grant Nos.:82074039 and 82204584).
文摘Epidemiological and animal studies indicate that pre-existing diabetes increases the risk of Parkinson's disease(PD).However,the mechanisms underlying this association remain unclear.In the present study,we found that high glucose(HG)levels in the cerebrospinal fluid(CSF)of diabetic rats might enhance the effect of a subthreshold dose of the neurotoxin 6-hydroxydopamine(6-OHDA)on the development of motor disorders,and the damage to the nigrostriatal dopaminergic neuronal pathway.In vitro,HG promoted the 6-OHDA-induced apoptosis in PC12 cells differentiated to neurons with nerve growth factor(NGF)(NGF-PC12).Metabolomics showed that HG promoted hyperglycolysis in neurons and impaired tricarboxylic acid cycle(TCA cycle)activity,which was closely related to abnormal mitochondrial fusion,thus resulting in mitochondrial loss.Interestingly,HG-induced upregulation of pyruvate kinase M2(PKM2)combined with 6-OHDA exposure not only mediated glycolysis but also promoted abnormal mitochondrial fusion by upregulating the expression of MFN2 in NGF-PC12 cells.In addition,we found that PKM2 knockdown rescued the abnormal mitochondrial fusion and cell apoptosis induced by HGþ6-OHDA.Furthermore,we found that shikonin(SK),an inhibitor of PKM2,restored the mitochondrial number,promoted TCA cycle activity,reversed hyperglycolysis,enhanced the tolerance of cultured neurons to 6-OHDA,and reduced the risk of PD in diabetic rats.Overall,our results indicate that diabetes promotes hyperglycolysis and abnormal mitochondrial fusion in neurons through the upregulation of PKM2,leading to an increase in the vulnerability of dopaminergic neurons to 6-OHDA.Thus,the inhibition of PKM2 and restoration of mitochondrial metabolic homeostasis/pathways may prevent the occurrence and development of diabetic PD.
文摘Acknowledgments： I would like to express my appreciation to Professor Puro DG for leading me to this research topic during my stay as a research fellow in his laboratory at the University of Michigan in 2001, and also to Professor Ikeda T forgiving me the opportunity to study abroad and then to continue to investigate this topic in the Department of Ophthalmology at Osaka Medical College, lapan.
基金Supported by 2022 Educational Research Program for Young and Middle-aged Teachers in Fujian Province(Science and Technology),No.JAT220107.
文摘BACKGROUND Cognitive dysfunction in epileptic patients is a high-incidence complication.Its mechanism is related to nervous system damage during seizures,but there is no effective diagnostic biomarker.Neuronal pentraxin 2(NPTX2)is thought to play a vital role in neurotransmission and the maintenance of synaptic plasticity.This study explored how serum NPTX2 and electroencephalogram(EEG)slow wave/fast wave frequency ratio relate to cognitive dysfunction in patients with epilepsy.AIM To determine if serum NPTX2 could serve as a potential biomarker for diagnosing cognitive impairment in epilepsy patients.METHODS The participants of this study,conducted from January 2020 to December 2021,comprised 74 epilepsy patients with normal cognitive function(normal group),37 epilepsy patients with cognitive dysfunction[epilepsy patients with cognitive dysfunction(ECD)group]and 30 healthy people(control group).The minimental state examination(MMSE)scale was used to evaluate cognitive function.We determined serum NPTX2 levels using an enzyme-linked immunosorbent kit and calculated the signal value of EEG regions according to the EEG recording.Pearson correlation coefficient was used to analyze the correlation between serum NPTX2 and the MMSE score.RESULTS The serum NPTX2 level in the control group,normal group and ECD group were 240.00±35.06 pg/mL,235.80±38.01 pg/mL and 193.80±42.72 pg/mL,respectively.The MMSE score was lowest in the ECD group among the three,while no significant difference was observed between the control and normal groups.In epilepsy patients with cognitive dysfunction,NPTX2 level had a positive correlation with the MMSE score(r=0.367,P=0.0253)and a negative correlation with epilepsy duration(r=−0.443,P=0.0061)and the EEG slow wave/fast wave frequency ratio value in the temporal region(r=−0.339,P=0.039).CONCLUSION Serum NPTX2 was found to be related to cognitive dysfunction and the EEG slow wave/fast wave frequency ratio in patients with epilepsy.It is thus a potential biomarker for the diagnosis of cognitive impairment in patients with epilepsy.
基金supported by the Henan Province Education Department Key Project of Science and Technology Research in China,No.12A350006
文摘c-Jun NH2-terminal kinase（JNK）-interacting protein 3 plays an important role in brain-derived neurotrophic factor/tropomyosin-related kinase B（Trk B） anterograde axonal transport. It remains unclear whether JNK-interacting protein 1 mediates similar effects, or whether JNK-interacting protein 1 affects the regulation of Trk B anterograde axonal transport. In this study, we isolated rat embryonic hippocampus and cultured hippocampal neurons in vitro. Coimmunoprecipitation results demonstrated that JNK-interacting protein 1 formed Trk B complexes in vitro and in vivo. Immunocytochemistry results showed that when JNK-interacting protein 1 was highly expressed, the distribution of Trk B gradually increased in axon terminals. However, the distribution of Trk B reduced in axon terminals after knocking out JNK-interacting protein 1. In addition, there were differences in distribution of Trk B after JNK-interacting protein 1 was knocked out compared with not. However, knockout of JNK-interacting protein 1 did not affect the distribution of Trk B in dendrites. These findings confirm that JNK-interacting protein 1 can interact with Trk B in neuronal cells, and can regulate the transport of Trk B in axons, but not in dendrites.
基金supportedby a grant from the Korean Fundfor Regenerative Medicine(KFRM),which is funded by the Korean government’s Ministry of Science and ICT and the Ministry of Health&Welfare(23A0102L1 to Janghwan Kim)by KRIBB Research Initiative Program(KGM5362521 to Janghwan Kim)+1 种基金supported by a grant fromthe National Research Foundation of Korea(NRF)which is funded by theMinistry of Science and ICT(MSIT)of the Korean government(RS-2023-NR077070 to SungWoo Park).
文摘Background:Parkinson’s disease(PD)is a common neurodegenerative disease,characterized by symptoms like tremors,muscle rigidity,and slowmovement.Themain cause of these symptoms is the loss of dopamineproducing neurons in a brain area called the substantia nigra.Various genetic and environmental factors contribute to this neuronal loss.Once symptoms of PD begin,they worsen with age,which also impacts several critical cellular processes.Leucine-rich repeat kinase 2(LRRK2)is a gene associated with PD.Certain mutations in LRRK2,such as G2019S,increase its activity,disrupting cellular mechanisms necessary for healthy neuron function,including autophagy and lysosomal activity.Exposure to rotenone(RTN)promotes LRRK2 activity in neurons and contributes to cellular senescence andα-syn accumulation.Methods:In this study,human dopaminergic progenitor cells were reprogrammed to study the effects of RTN with the co-treatment of LRRK2 inhibitor on cellular senescence.We measured the cellular senescence using quantifying proteins of senescence markers,such as p53,p21,Rb,phosphorylated Rb,andβ-galatocidase,and the enzymatic activity of senescence-associatedβ-galatocidase.And we estimated the levels of accumulatedα-synuclein(α-syn),which is increased via the impaired autophagy-lysosomal pathway by cellular senescence.Then,we evaluated the association of the G2019S LRRK2 mutation and senescence-associatedβ-galatocidase and the levels of accumulated or secretedα-syn,and the neuroinflammatory responses mediated by the secretedα-syn in rat primary microglia were determined using the release of pro-inflammatory cytokines.Results:RTN raised senescence markers and affected the phosphorylation of Rab10,a substrate of LRRK2.The inhibiting agent MLI2 reduced these senescence markers and Rab10 phosphorylations.Additionally,RTN increasedα-syn levels in the neurons,while MLI2 aided in degrading it.When focusing on cells from PD patients with the G2019S mutation,an increase in cellular senescence and release ofα-syn was observed,provoking neuroinflammation.Treatment with the LRRK2 inhibitor MLI2 decreased both cellular senescence andα-syn secretion,thereby mitigating inflammatory responses.Conclusion:Overall,inhibiting LRRK2 may provide a beneficial strategy formanaging PD.
基金supported by the National Natural Science Foundation of China(81971058,82371226,82101295,82301398)the National Funded Postdoctoral Researcher Program(GZC20233585)The Boost Plan of Xijing Hospital(XJZT24QN25,XJZT25CX22).
文摘Neuropathic pain,often featuring allodynia,imposes significant physical and psychological burdens on patients,with limited treatments due to unclear central mechanisms.Addressing this challenge remains a crucial unsolved issue in pain medicine.Our previous study,using protein kinase C gamma(PKCγ)-tdTomato mice,highlights the spinal feedforward inhibitory circuit involving PKCγ neurons in gating neuropathic allodynia.However,the regulatory mechanisms governing this circuit necessitate further elucidation.We used diverse transgenic mice and advanced techniques to uncover the regulatory role of the descending serotonin(5-HT)facilitation system on spinal PKCγ neurons.Our findings revealed that 5-HT neurons from the rostral ventromedial medulla hyperpolarize spinal inhibitory interneurons via 5-HT_(2C) receptors,disinhibiting the feedforward inhibitory circuit involving PKCγ neurons and exacerbating allodynia.Inhibiting spinal 5-HT_(2C) receptors restored the feedforward inhibitory circuit,effectively preventing neuropathic allodynia.These insights offer promising therapeutic targets for neuropathic allodynia management,emphasizing the potential of spinal 5-HT_(2C) receptors as a novel avenue for intervention.
基金supported by the Strategic Priority Research Program of the Chinese Academy of Sciences(XDB32010100)a National Natural Science Foundation of China project(31671113)+1 种基金a Shanghai Municipal Science and Technology Major Project(2018SHZDZX05)and the State Key Laboratory of Neuroscience.
文摘Abnormal synchronous neuronal activity has been widely detected by brain imaging of autistic patients,but its underlying neural mechanism remains unclear.Compared with wild-type mice,our in vivo two-photon imaging showed that transgenic(Tgl)mice over-expressing human autism risk gene MeCP2 exhibited higher neuronal synchrony in the young but lower synchrony in the adult stage.Whole-cell recording of neuronal pairs in brain slices revealed that higher neuronal synchrony in young postnatal Tgl mice was atributed mainly to more prevalent giant slow inward currents(SICs).Both in vivo and slice imaging further demonstrated more dynamic activity and higher synchrony in astrocytes from young Tgl mice.Blocking astrocytic gap junctions markedly decreased the generation of SICs and overall cell synchrony in the Tgl brain.Furthermore,the expression level of Cx43 protein and the coupling efficiency of astrocyte gap junctions remained unchanged in Tgi mice.Thus,astrocytic gap junctions facilitate but do not act as a direct trigger for the abnormal neuronal synchrony in young Tgl mice,revealing the potential role of the astrocyte network in the pathogenesis of MeCP2 duplication syndrome.
基金supported by Ministry of Economy and Competitiveness,Spain(MINECO),grant number BFU2012-37304by Junta de Andalucía,Spain,grant number P12-CTS-2064
文摘Bromodomain containing 2 （Brd2） protein belongs to the Bromodomains and Extra Terminal domain （BET） family of chromatin adaptors characterized by the presence of two N-terminal tandem bromodomains and an exclusive C-terminal extra terminal domain （ET） （Belkina and Denis, 2012; Shi and Vakoc, 2014）. Bromodomains are involved in recognizing acetylated histone tails and other acetylated proteins while the ET domain has been implicated in protein-protein interaction.
文摘BACKGROUND: Ⅴ secretory phospholipase A2 (sPLA2-Ⅴ) is abundant in many mammal tissues. However, it remains unknown whether sPLA2-Ⅴ causes biological or pathological response in central nervous system. OBJECTIVE: To observe the effect of phospholipase A2-Ⅴ (PLA2-Ⅴ) and its inhibitor (indoxam) on hippocampal neuron survival. DESIGN: A repetitive measurement. SETTING: The Animal Center of South Carolina University. MATERIALS: Sprague-Dawley pregnancy day-7, 14, 21 female rats were selected; Reagents: sPLA2- Ⅴ and indoxam were obtained from the Dennis Research Laboratories METHODS: The experiment was finished at the animal center in South Carolina University from January to December, 2004. 0, 12.5, 25, 50 and 100 μg/L sPLA2-Ⅴ were added to neuron with none-MgCl2 Eagle’s medium at 37 ℃, then changed to normal neuron culture medium after 3 hours. 1, 2.5, 5 and 10 μmol/L indoxam was added at 6 hours after 100 μg/L sPLA2-Ⅴwas put to Day-21 SD rat hippocampal embryonic neurons with none-MgCl2 Eagle’s medium at 37 ℃. After 3 hours in the inhibition experiment, it was changed to normal neuron culture medium. The embryonic hippocampal neurons were primarily cultured, and the neuron survival ratio was detected with morphological method. MAIN OUTCOME MEASURES: Survival ratio of hippocampal neurons. RESULTS: ① Effects of sPLA2-Ⅴon neuron survival: When sPLA2-Ⅴ was 0, 12.5, 25, 50 and 100 μg/L, the neuron survival ratios in embryonic neurons of day-7 SD rats were (95.3±1.1)%, (81.4±3.1)%, (74.2±2.2)%, (62.4±1.7)% and (48.9±1.6)%, those in embryonic neurons of day-14 rats were (93.2±1.4)%, (74.3±1.9)%, (68.1±1.7)%, (56.1±1.4)% and (42.5±1.1)%, and those in embryonic neurons of day-21 rats were (91.2±1.2)%, (69.4±2.1)%, (60.3±2.2)%, (49.1±1.2)% and (35.5±1.9)%. There were significant differences among different concentrations (P < 0.05). ② Effects of indoxam on neuron survival: In case of sPLA2-Ⅴ 100 μg/L, the neuron survival ratios were (58.65±1.4)%, (69.34±1.1)%, (82.11±1.2)% and (95.28±0.9)% when indoxam was 1, 2.5, 5 and 10 μmol/L, respectively. There were significant differences among different concentrations (P < 0.05). CONCLUSION: ① The of neuronal death ratio is in a concentration-dependent manner with sPLA2-Ⅴ, and increases as the embryonic aging. ② Indoxam inhibits the proapoptotic effect of sPLA2-Ⅴ.
文摘This editorial summarizes the latest literature on the roles of neuronal PAS domain protein 2 and KN motif/ankyrin repeat domain 1 in type 2 diabetes(T2D).We highlight their involvement inβ-cell dysfunction,explore their potential as therapeutic targets,and discuss the implications for new treatment strategies.We offer valuable insights into relevant gene regulation and cellular mechanisms relevant for the targeted management of T2D.
基金supported by FATALSDrug Project [Progetti di Ricerca@CNR SAC.AD002.173.058] from National Research Council,Italy (to CV)。
文摘The NSC-34 cell line is a widely recognized motor neuron model and various neuronal differentiation protocols have been exploited. Under previously reported experimental conditions, only part of the cells resemble differentiated neurons;however, they do not exhibit extensive and time-prolonged neuritogenesis, and maintain their duplication capacity in culture. The aim of the present work was to facilitate long-term and more homogeneous neuronal differentiation in motor neuron–like NSC-34 cells. We found that the antimitotic drug cytosine arabinoside promoted robust and persistent neuronal differentiation in the entire cell population. Long and interconnecting neuronal processes with abundant growth cones were homogeneously induced and were durable for up to at least 6 weeks in culture. Moreover, cytosine arabinoside was permissive, dispensable, and mostly irreversible in priming NSC-34 cells for neurite initiation and regeneration after mechanical dislodgement. Finally, the expression of the cell proliferation antigen Ki67 was inhibited by cytosine arabinoside, whereas the expression levels of neuronal growth associated protein 43, vimentin, and motor neuron–specific p75, Islet2, homeobox 9 markers were upregulated, as confirmed by western blot and/or confocal immunofluorescence analysis. Overall, these findings support the use of NSC-34 cells as a motor neuron model for properly investigating neurodegenerative mechanisms and prospectively identifying neuroprotective strategies.
