OBJECTIVE The present study was aimed to investigate the role of Wnt/β-catenin sig.naling in spinal VGLUT2 regulation and neuropathic pain.METHODS To elucidate the association be.tween VGLUT2 and neuropathic pain,we ...OBJECTIVE The present study was aimed to investigate the role of Wnt/β-catenin sig.naling in spinal VGLUT2 regulation and neuropathic pain.METHODS To elucidate the association be.tween VGLUT2 and neuropathic pain,we determined the expression and distribution characteristics of VGLUT2 in mice subjected to spared nerve injury(SNI),and then observed the effects of two VGLUT2 targeting shRNAs on mechanical allodynia and glutamate release.The effects of Wnt/β-catenin signal.ing on VGLUT2 expression and pain behavior were investigated by using Wnt agonist,Wnt1,and Wnt/β-catenin pathway inhibitor XAV939 in SNI mice.RESULTS SNI surgery induced significant up-regula.tion of VGLUT2 on postoperative days 7,14,and 21.Double immunofluorescence labeling of VGLUT2 with NeuN,MAP2,Iba-1,or GFAP showed that VGLUT2 was mainly expressed in neurons in the dor.sal horn of the spinal cord after SNI(NeuN,MAP2).Intrathecal administration of VGLUT2 shRNAs be.fore or after SNI surgery significantly decreased mechanical allodynia and glutamate release.Mean.while,Wnt1/β-catenin signaling increased significantly after SNI surgery.Over-expression of β-catenin in PC12 cells increased VGLUT2 protein level,intrathecal administration of Wnt agonist or Wnt1 signifi.cantly increased VGLUT2 protein expression in spinal cord,while Wnt/β-catenin pathway inhibitor XAV939 decreased VGLUT2 expression in PC12 cells and spinal cord.Additionally,intrathecal admin.istration of XAV939 7 days after SNI significantly attenuated mechanical allodynia in mice,which was in accordance with down-regulation of VGLUT2 protein levels.VGLUT2 shRNAs significantly attenuat.ed Wnt agonist or Wnt1 induced mechanical allodynia.CONCLUSION Wnt1/β-catenin signaling path.way up-regu-lates the spinal VGLUT2 expression,and this regulation is involved in neuropathic pain behavior.展开更多
Adult male mice emit highly complex ultrasonic vocalizations(USVs)in response to female conspecifics.Such US Vs,thought to facilitate courtship behaviors,are routinely measured as a behavioral index in mouse models of...Adult male mice emit highly complex ultrasonic vocalizations(USVs)in response to female conspecifics.Such US Vs,thought to facilitate courtship behaviors,are routinely measured as a behavioral index in mouse models of neurodevelopmental and psychiatric disorders such as autism.While the regulation of US Vs by genetic factors has been extensively characterized,the neural mechanisms that control USV production remain largely unknown.Here,we report that optogenetic activation of the medial preoptic area(mPOA)elicited the production of USVs that were acoustically similar to courtship US Vs in adult mice.Moreover,mPOA vesicular GABA transporter-positive(Vgat +)neurons were more effective at driving USV production than vesicular glutamate transporter 2-positive neurons.Furthermore,ablation of mPOA Vgat+ neurons resulted in altered spectral features and syllable usage of USVs in targeted males.Together,these results demonstrate that the mPOA plays a crucial role in modulating courtship USVs and this may serve as an entry point for future dissection of the neural circuitry underlying USV production.展开更多
The lateral habenula(LHb),which is a critical neuroanatomical hub and a regulator of midbrain monoaminergic centers,is activated by events resulting in negative valence and contributes to the expression of both appeti...The lateral habenula(LHb),which is a critical neuroanatomical hub and a regulator of midbrain monoaminergic centers,is activated by events resulting in negative valence and contributes to the expression of both appetitive and aversive behaviors.However,whole-brain cell-type-specific monosynaptic inputs to the LHb in both sexes remain incompletely elucidated.In this study,we used viral tracing combined with in situ hybridization targeting vesicular glutamate transporter 2(vGlut2)and glutamic acid decarboxylase 2(Gad2)to generate a comprehensive whole-brain atlas of inputs to glutamatergic andγ-aminobutyric acid(GABA)ergic neurons in the LHb.We found>30 ipsilateral and contralateral brain regions that projected to the LHb.Of these,there were significantly more monosynaptic LHb-projecting neurons from the lateral septum,anterior hypothalamus,dorsomedial hypothalamus,and ventromedial hypothalamus in females than in males.More interestingly,we found a stronger GABAergic projection from the medial septum to the LHb in males than in females.Our results reveal a comprehensive connectivity atlas of glutamatergic and GABAergic inputs to the LHb in both sexes,which may facilitate a better understanding of sexual dimorphism in physiological and pathological brain functions.展开更多
基金supported by National Natural Science Foundation of China(81200850) Beijing Natural Science Foundation(7123224) National Science and Technology Major Project of China(2012ZX09301003-001)
文摘OBJECTIVE The present study was aimed to investigate the role of Wnt/β-catenin sig.naling in spinal VGLUT2 regulation and neuropathic pain.METHODS To elucidate the association be.tween VGLUT2 and neuropathic pain,we determined the expression and distribution characteristics of VGLUT2 in mice subjected to spared nerve injury(SNI),and then observed the effects of two VGLUT2 targeting shRNAs on mechanical allodynia and glutamate release.The effects of Wnt/β-catenin signal.ing on VGLUT2 expression and pain behavior were investigated by using Wnt agonist,Wnt1,and Wnt/β-catenin pathway inhibitor XAV939 in SNI mice.RESULTS SNI surgery induced significant up-regula.tion of VGLUT2 on postoperative days 7,14,and 21.Double immunofluorescence labeling of VGLUT2 with NeuN,MAP2,Iba-1,or GFAP showed that VGLUT2 was mainly expressed in neurons in the dor.sal horn of the spinal cord after SNI(NeuN,MAP2).Intrathecal administration of VGLUT2 shRNAs be.fore or after SNI surgery significantly decreased mechanical allodynia and glutamate release.Mean.while,Wnt1/β-catenin signaling increased significantly after SNI surgery.Over-expression of β-catenin in PC12 cells increased VGLUT2 protein level,intrathecal administration of Wnt agonist or Wnt1 signifi.cantly increased VGLUT2 protein expression in spinal cord,while Wnt/β-catenin pathway inhibitor XAV939 decreased VGLUT2 expression in PC12 cells and spinal cord.Additionally,intrathecal admin.istration of XAV939 7 days after SNI significantly attenuated mechanical allodynia in mice,which was in accordance with down-regulation of VGLUT2 protein levels.VGLUT2 shRNAs significantly attenuat.ed Wnt agonist or Wnt1 induced mechanical allodynia.CONCLUSION Wnt1/β-catenin signaling path.way up-regu-lates the spinal VGLUT2 expression,and this regulation is involved in neuropathic pain behavior.
基金the National Natural Science Foundation of China (31871066, 31471065)the National Basic Research Development Program (973 Program) of China (2015CB559201)+2 种基金the Thousand Young Talents Program of Chinathe Strategic Priority Research Program of the Chinese Academy of Sciences (XDB32010200)as part of the Chinese Academy of Science interdisciplinary innovation team
文摘Adult male mice emit highly complex ultrasonic vocalizations(USVs)in response to female conspecifics.Such US Vs,thought to facilitate courtship behaviors,are routinely measured as a behavioral index in mouse models of neurodevelopmental and psychiatric disorders such as autism.While the regulation of US Vs by genetic factors has been extensively characterized,the neural mechanisms that control USV production remain largely unknown.Here,we report that optogenetic activation of the medial preoptic area(mPOA)elicited the production of USVs that were acoustically similar to courtship US Vs in adult mice.Moreover,mPOA vesicular GABA transporter-positive(Vgat +)neurons were more effective at driving USV production than vesicular glutamate transporter 2-positive neurons.Furthermore,ablation of mPOA Vgat+ neurons resulted in altered spectral features and syllable usage of USVs in targeted males.Together,these results demonstrate that the mPOA plays a crucial role in modulating courtship USVs and this may serve as an entry point for future dissection of the neural circuitry underlying USV production.
基金This work was supported by the NSFC-Guangdong Joint Fund(U20A6005)the National Natural Science Foundation of China(31630031 and 31930047)+2 种基金the Key-Area Research and Development Program of Guangdong Province(2018B030340001 and 2018B030331001)the Guangdong Provincial Key Laboratory of Brain Connectome and Behavior(2017B030301017)the Shenzhen Key Laboratory of Translational Research for Brain Diseases(ZDSYS20200828154800001).
文摘The lateral habenula(LHb),which is a critical neuroanatomical hub and a regulator of midbrain monoaminergic centers,is activated by events resulting in negative valence and contributes to the expression of both appetitive and aversive behaviors.However,whole-brain cell-type-specific monosynaptic inputs to the LHb in both sexes remain incompletely elucidated.In this study,we used viral tracing combined with in situ hybridization targeting vesicular glutamate transporter 2(vGlut2)and glutamic acid decarboxylase 2(Gad2)to generate a comprehensive whole-brain atlas of inputs to glutamatergic andγ-aminobutyric acid(GABA)ergic neurons in the LHb.We found>30 ipsilateral and contralateral brain regions that projected to the LHb.Of these,there were significantly more monosynaptic LHb-projecting neurons from the lateral septum,anterior hypothalamus,dorsomedial hypothalamus,and ventromedial hypothalamus in females than in males.More interestingly,we found a stronger GABAergic projection from the medial septum to the LHb in males than in females.Our results reveal a comprehensive connectivity atlas of glutamatergic and GABAergic inputs to the LHb in both sexes,which may facilitate a better understanding of sexual dimorphism in physiological and pathological brain functions.
