In vito fber photometry is a powerful technique to analyze the dy namics of population neurons during fiunctional study of neuroscience.Here,we introduced a detailed protocol for fiber photometry-based calciun reordin...In vito fber photometry is a powerful technique to analyze the dy namics of population neurons during fiunctional study of neuroscience.Here,we introduced a detailed protocol for fiber photometry-based calciun reording in freely moving mice,covering from virus injection,fiber stub insertion,optogenetical stimulation to data procurement and analysis.Furthemnore,we applied this protocol to explore neuronal activity of mice latenal-posterior(LP)thalaric nucleus in response to optogenetical stimulation of primary visual cortex(V1)neurons,and explore axon clusters activity of optogenetically evoked V1 neurons.Final confirmation of virus-based protein expression in V1 and precise fber insertion indicated that the surgery procedure of this protocol is reliable for functional calcium recording.The scripts for data analysis and some tips in our protocol are provided in details.Together,this protocol is simple,low-cost,and effective for neuronal activity detection by fiber photometry,which will hep neuroscience researchers to carry out fiunctional and behavioral study in vivo.展开更多
Generating diverse motor behaviors critical for survival is a challenge that confronts the central nervous system(CNS)of all animals.During movement execution,the CNS performs complex calculations to control a large n...Generating diverse motor behaviors critical for survival is a challenge that confronts the central nervous system(CNS)of all animals.During movement execution,the CNS performs complex calculations to control a large number of neuromusculoskeletal elements.The theory of modular motor control proposes that spinal interneurons are organized in discrete modules that can be linearly combined to generate a variety of behavioral patterns.These modules have been previously represented as stimulus-evoked force fields(FFs)comprising isometric limb-endpoint forces across workspace locations.Here,we ask whether FFs elicited by different stimulations indeed represent the most elementary units of motor control or are themselves the combination of a limited number of even more fundamental motor modules.To probe for potentially more elementary modules,we optogenetically stimulated the lumbosacral spinal cord of intact and spinalized Thy1-ChR2 transgenic mice(n=21),eliciting FFs from as many single stimulation loci as possible(20-70 loci per mouse)at minimally necessary power.We found that the resulting varieties of FFs defied simple categorization with just a few clusters.We used gradient descent to further decompose the FFs into their underlying basic force fields(BFFs),whose linear combination explained FF variability.Across mice,we identified 4-5 BFFs with partially localizable but overlapping representations along the spinal cord.The BFFs were structured and topographically distributed in such a way that a rostral-to-caudal traveling wave of activity across the lumbosacral spinal cord may generate a swing-to-stance gait cycle.These BFFs may represent more rudimentary submodules that can be flexibly merged to produce a library of motor modules for building different motor behaviors.展开更多
Optogenetic has been widely applied in various pathogenesis investigations of neuropathic diseases since its accurate and targeted regulation of neuronal activity.However,due to the mismatch between the soft tissues a...Optogenetic has been widely applied in various pathogenesis investigations of neuropathic diseases since its accurate and targeted regulation of neuronal activity.However,due to the mismatch between the soft tissues and the optical waveguide,the long-term neural regulation within soft tissue(such as brain and spinal cord)by implantable optical fibers is a large challenge.Herein,we designed a modulus selfadaptive hydrogel optical fiber(MSHOF)with tunable mechanical properties(Young’modulus was tunable in the range of 0.32-10.56MPa)and low light attenuation(0.12-0.21 dB/cm,472nm laser light),which adapts to light transmission under soft tissues.These advantages of MSHOF can ensure the effectiveness of optogenetic stimulation meanwhile safeguarding the safety of the brain/materials interaction interface.In addition,this work provides more design possibilities of MSHOF for photogenetic stimuli and has significant application prospects in photomedical therapy.展开更多
Optogenetics has revolutionized the field of neuroscience by enabling precise control of neural activity through light-sensitive proteins known as opsins.This review article discusses the fundamental principles of opt...Optogenetics has revolutionized the field of neuroscience by enabling precise control of neural activity through light-sensitive proteins known as opsins.This review article discusses the fundamental principles of optogenetics,including the activation of both excitatory and inhibitory opsins,as well as the development of optogenetic models that utilize recombinant viral vectors.A considerable portion of the article addresses the limitations of optogenetic tools and explores strategies to overcome these challenges.These strategies include the use of adeno-associated viruses,cell-specific promoters,modified opsins,and methodologies such as bioluminescent optogenetics.The application of viral recombinant vectors,particularly adeno-associated viruses,is emerging as a promising avenue for clinical use in delivering opsins to target cells.This trend indicates the potential for creating tools that offer greater flexibility and accuracy in opsin delivery.The adaptations of these viral vectors provide advantages in optogenetic studies by allowing for the restricted expression of opsins through cell-specific promoters and various viral serotypes.The article also examines different cellular targets for optogenetics,including neurons,astrocytes,microglia,and Schwann cells.Utilizing specific promoters for opsin expression in these cells is essential for achieving precise and efficient stimulation.Research has demonstrated that optogenetic stimulation of both neurons and glial cells-particularly the distinct phenotypes of microglia,astrocytes,and Schwann cells-can have therapeutic effects in neurological diseases.Glial cells are increasingly recognized as important targets for the treatment of these disorders.Furthermore,the article emphasizes the emerging field of bioluminescent optogenetics,which combines optogenetic principles with bioluminescent proteins to visualize and manipulate neural activity in real time.By integrating molecular genetics techniques with bioluminescence,researchers have developed methods to monitor neuronal activity efficiently and less invasively,enhancing our understanding of central nervous system function and the mechanisms of plasticity in neurological disorders beyond traditional neurobiological methods.Evidence has shown that optogenetic modulation can enhance motor axon regeneration,achieve complete sensory reinnervation,and accelerate the recovery of neuromuscular function.This approach also induces complex patterns of coordinated motor neuron activity and promotes neural reorganization.Optogenetic approaches hold immense potential for therapeutic interventions in the central nervous system.They enable precise control of neural circuits and may offer new treatments for neurological disorders,particularly spinal cord injuries,peripheral nerve injuries,and other neurodegenerative diseases.展开更多
In this study,we aimed to develop an in vivo electrophysiological bone-nerve preparation to record the activity of peripheral sensory neurons that innervate the murine tibia.A small nerve that innervates the tibial ma...In this study,we aimed to develop an in vivo electrophysiological bone-nerve preparation to record the activity of peripheral sensory neurons that innervate the murine tibia.A small nerve that innervates the tibial marrow cavity was identified in isoflurane-anesthetized C57BL/6 mice,and placed over a platinum hook electrode for extracellular recording.Whole-nerve activity was amplified,filtered and sampled at 20 kHz using PowerLab(ADInstruments).A cannula was placed into the marrow cavity to deliver mechanical stimuli(by pressurizing with injection of saline)and/or capsaicin.Optical stimulation was achieved by application of 473 nm blue light(1 Hz,0.25-0.5 ms,0-12.5 mW/mm)to the tibial marrow cavity in Wnt1-Cre;loxP-ChR2 mice.Murine bone afferent neurons responded to high threshold noxious mechanical stimulation,coded for the intensity of mechanical stimulation,could be sensitized by capsaicin,and did not suffer stimulus-evoked fatigue at 10-minute interstimulus intervals.Electrical and optical stimulation within the marrow cavity evoked action potentials with conduction velocities in the Aδand/or C fiber range.These new approaches to recording the activity of bone afferent neurons will allow us to take advantage of transgenic and optogenetic tools to further our understanding of mechanisms that generate and maintain bone pain in the future.展开更多
Depression is a multifaceted disorder with a largely unresolved etiology influenced by a complex interplay of pathogenic factors.Despite decades of research,it remains a major condition that significantly diminishes p...Depression is a multifaceted disorder with a largely unresolved etiology influenced by a complex interplay of pathogenic factors.Despite decades of research,it remains a major condition that significantly diminishes patients’quality of life.Advances in optogenetics have introduced a powerful tool for exploring the neural mechanisms underlying depression.By selectively expressing optogenes in specific cell types in mice,researchers can study the roles of these cells through targeted light stimulation,offering new insights into central nervous system disorders.The use of viral vectors to express opsins in distinct neuronal subtypes enables precise activation or inhibition of these neurons via light.When combined with behavioral,morphological,and electrophysiological analyses,optogenetics provides an invaluable approach to investigating the neural mechanisms of psychiatric conditions.This review synthesizes current research on the application of optogenetics to understand the mechanisms of depression.This study aims to enhance our knowledge of optogenetic strategies for regulating depression and advancing antidepressant research.展开更多
Epilepsy,a common neurological disorder,is characterized by recurrent seizures that can lead to cognitive,psychological,and neurobiological consequences.The pathogenesis of epilepsy involves neuronal dysfunction at th...Epilepsy,a common neurological disorder,is characterized by recurrent seizures that can lead to cognitive,psychological,and neurobiological consequences.The pathogenesis of epilepsy involves neuronal dysfunction at the molecular,cellular,and neural circuit levels.Abnormal molecular signaling pathways or dysfunction of specific cell types can lead to epilepsy by disrupting the normal functioning of neural circuits.The continuous emergence of new technologies and the rapid advancement of existing ones have facilitated the discovery and comprehensive understanding of the neural circuit mechanisms underlying epilepsy.Therefore,this review aims to investigate the current understanding of the neural circuit mechanisms in epilepsy based on various technologies,including electroencephalography,magnetic resonance imaging,optogenetics,chemogenetics,deep brain stimulation,and brain-computer interfaces.Additionally,this review discusses these mechanisms from three perspectives:structural,synaptic,and transmitter circuits.The findings reveal that the neural circuit mechanisms of epilepsy encompass information transmission among different structures,interactions within the same structure,and the maintenance of homeostasis at the cellular,synaptic,and neurotransmitter levels.These findings offer new insights for investigating the pathophysiological mechanisms of epilepsy and enhancing its clinical diagnosis and treatment.展开更多
Objective:This study aims to establish an economically viable and easily accessible adult animal model for optogenetic activation of auditory neurons using adeno-associated viruses(AAVs)carrying Ch R2(H134R)to explore...Objective:This study aims to establish an economically viable and easily accessible adult animal model for optogenetic activation of auditory neurons using adeno-associated viruses(AAVs)carrying Ch R2(H134R)to explore the potential of cochlear optogenetics as a hearing restoration technology.Methods:Healthy adult guinea pigs were used in the experiments.The viral vector AAV2/8-Ch R2(H134R)-h Syn-e YFP was administered to the right cochlea via the round window membrane.The confocal microscopy and reverse transcription polymerase chain reaction(RT-PCR)were utilized to analyze the Ch R2(H134R)expression localized to spiral ganglion neurons(SGNs).The auditory pathway activation was assessed by recording the optical compound action potential(oCAP)and acoustic compound action potential(a CAP)at various laser intensities.Results:The Ch R2(H134R)-e YFP expression was confirmed in 90%of the tested animals,localized to the SGNs of the injected ear.Higher m RNA levels of Ch R2(H134R)and e YFP were observed in the injected ear compared to the non-injected ear,while actin(Actb)m RNA levels were not significantly different.The o CAP was successfully elicited by a 470 nm blue light laser stimulus,with similar amplitudes and latency periods to those of a CAPs when the o CAP was evoked by 5.80 m W blue light and the a CAP was evoked by a 40 d B SPL click.The amplitudes of o CAPs increased with increasing laser intensity.Conclusion:This study demonstrates the viability of optogenetic activation of the auditory system in adult guinea pigs through the transduction of AAV-Ch R2(H134R)in SGNs.Cochlear optogenetics demonstrates potential as a hearing restoration technology,providing a basis for further clinical research and opening new avenues for investigation.展开更多
Spinal cord injury is a severe neurological disorder;however,current treatment methods often fail to restore nerve function effectively.Spinal cord stimulation via electrical signals is a promising therapeutic modalit...Spinal cord injury is a severe neurological disorder;however,current treatment methods often fail to restore nerve function effectively.Spinal cord stimulation via electrical signals is a promising therapeutic modality for spinal cord injury.Based on similar principles,this review aims to explore the potential of optical and acoustic neuromodulation techniques,emphasizing their benefits in the context of spinal cord injury.Photoacoustic imaging,renowned for its noninvasive nature,high-resolution capabilities,and cost-effectiveness,is well recognized for its role in early diagnosis,dynamic monitoring,and surgical guidance in stem cell therapies for spinal cord injury.Moreover,photoacoustodynamic therapy offers multiple pathways for tissue regeneration.Optogenetics and sonogenetics use genetic engineering to achieve precise neuronal activation,while photoacoustoelectric therapy leverages photovoltaic materials for electrical modulation of the nervous system,introducing an innovative paradigm for nerve system disorder management.Collectively,these advancements represent a transformative shift in the diagnosis and treatment of spinal cord injury,with the potential to significantly enhance nerve function remodeling and improve patient outcomes.展开更多
The hippocampus is the brain structure that is responsible for the formation of learning memories.Sleep disorders leading to cognitive impairment are strongly associated with the hippocampus.Phototherapy offers a new ...The hippocampus is the brain structure that is responsible for the formation of learning memories.Sleep disorders leading to cognitive impairment are strongly associated with the hippocampus.Phototherapy offers a new physical therapy for the treatment of sleep disorders,with the advantages of being noninvasive and having few side effects.However,the mechanism by which phototherapy improves cognitive impairment caused by sleep disorders remains unclear.In this study,we used phototherapy combined with optogenetic technology to investigate the effect of noninvasive phototherapy on cognitive functions in sleep-deprived mice.Our results suggest that phototherapy might improve cognitive functions in sleep-deprived mice by modulating the hippocampus.Our study expands the research progress on noninvasive phototherapy for the treatment of sleep disorders.展开更多
This study is based on wireless optogenetic technology,utilizing the CRY2/CIB1 photosensitive system to achieve spatiotemporal control of PD-L1 expression.In vitro experiments showed that the surface PD-L1 positivity ...This study is based on wireless optogenetic technology,utilizing the CRY2/CIB1 photosensitive system to achieve spatiotemporal control of PD-L1 expression.In vitro experiments showed that the surface PD-L1 positivity rate of cells increased from 28.6±3.1%to 67.3±5.4%(P<0.001).In animal experiments,the terminal tumor volume in the light exposure group was 450±90 mm3,with a tumor inhibition rate of approximately 49.4%(P<0.001),and the median survival was extended to 32 days(compared to 24 days in the control group,P=0.004).Immunological tests revealed a significant increase in CD8+T cell infiltration(112±18 vs 52±10 cells/HPF,P<0.01),a 30%decrease in the proportion of Tregs(P<0.05),and an increase in the M1/M2 macrophage ratio to 1.8.The results suggest that the wireless optogenetic system can not only precisely regulate PD-L1 but also remodel the tumor immune microenvironment,providing a new approach for precise immunotherapy of GBM.展开更多
Dear Editor,It is now well established that optogenetic stimulation can achieve precise intervention and modulate the activity of local neurons or neural circuits in the brain.Although this technique holds promise for...Dear Editor,It is now well established that optogenetic stimulation can achieve precise intervention and modulate the activity of local neurons or neural circuits in the brain.Although this technique holds promise for clinical therapy for neurological and psychiatric disorders,it requires the expression of lightsensitive proteins(such as channel rhodopsin)or photoactivatable chemicals(such as caged neurotransmitters)in the targeted brain regions[1].展开更多
Background:The development of ketamine-like rapid antidepressants holds promise for enhancing the therapeutic efficacy of depression,but the underlying cellular and molecular mechanisms remain unclear.Implicated in de...Background:The development of ketamine-like rapid antidepressants holds promise for enhancing the therapeutic efficacy of depression,but the underlying cellular and molecular mechanisms remain unclear.Implicated in depression regulation,the neuropeptide pituitary adenylate cyclase-activating polypeptide(PACAP)is investigated here to examine its role in mediating the rapid antidepressant response.Methods:The onset of antidepressant response was assessed through depression-related behavioral paradigms.The signaling mechanism of PACAP in the hippocampal dentate gyrus(DG)was evaluated by utilizing site-directed gene knockdown,pharmacological interventions,or optogenetic manipulations.Overall,446 mice were used for behavioral and molecular signaling testing.Mice were divided into control or experimental groups randomly in each experiment,and the experimental manipulations included:chronic paroxetine treatments(4 d,9 d,14 d)or a single treatment of ketamine;social defeat or lipopolysaccharides-injection induced depression models;different doses of PACAP(0.4 ng/site,2 ng/site,4 ng/site;microinjected into the hippocampal DG);pharmacological intra-DG interventions(CALM and PACAP6-38);intra-DG viral-mediated PACAP RNAi;and opotogenetics using channelrhodopsins 2(ChR2)or endoplasmic natronomonas halorhodopsine 3.0(eNpHR3.0).Behavioral paradigms included novelty suppressed feeding test,tail suspension test,forced swimming test,and sucrose preference test.Western blotting,ELISA,or quantitative real-time PCR(RT-PCR)analysis were used to detect the expressions of proteins/peptides or genes in the hippocampus.Results:Chronic administration of the slow-onset antidepressant paroxetine resulted in an increase in hippocampal PACAP expression,and intra-DG blockade of PACAP attenuated the onset of the antidepressant response.The levels of hippocampal PACAP expression were reduced in both two distinct depression animal models and intra-DG knockdown of PACAP induced depression-like behaviors.Conversely,a single infusion of PACAP into the DG region produced a rapid and sustained antidepressant response in both normal and chronically stressed mice.Optogenetic intra-DG excitation of PACAP-expressing neurons instantly elicited antidepressant responses,while optogenetic inhibition induced depression-like behaviors.The longer optogenetic excitation/inhibition elicited the more sustained antidepressant/depression-like responses.Intra-DG PACAP infusion immediately facilitated the signaling for rapid antidepressant response by inhibiting calcium/calmodulin-dependent protein kinaseⅡ(CaM KⅡ)-eukaryotic elongation factor 2(eEF2)and activating the mammalian target of rapamycin(mTOR).Pre-activation of CaMKⅡsignaling within the DG blunted PACAP-induced rapid antidepressant response as well as eEF2-mTOR-brain-derived neurotrophic factor(BDNF)signaling.Finally,acute ketamine treatment upregulated hippocampal PACAP expression,whereas intraDG blockade of PACAP signaling attenuated ketamine’s rapid antidepressant response.Conclusions:Activation of hippocampal PACAP signaling induces a rapid antidepressant response through the regulation of CaMKⅡinhibition-governed eEF2-mTOR-BDNF signaling.展开更多
Optogenetics,a technique that employs light for neuromodulation,has revolutionized the study of neural mechanisms and the treatment of neurological disorders due to its high spatiotemporal resolution and cell-type spe...Optogenetics,a technique that employs light for neuromodulation,has revolutionized the study of neural mechanisms and the treatment of neurological disorders due to its high spatiotemporal resolution and cell-type specificity.However,visible light,particularly blue and green light,commonly used in conventional optogenetics,has limited penetration in biological tissue.This limitation necessitates the implantation of optical fibers for light delivery,especially in deep brain regions,leading to tissue damage and experimental constraints.To overcome these challenges,the use of orange-red and infrared light with greater tissue penetration has emerged as a promising approach for tetherless optical neuromodulation.In this review,we provide an overview of the development and applications of tetherless optical neuromodulation methods with long wavelengths.We first discuss the exploration of orange-red wavelength-responsive rhodopsins and their performance in tetherless optical neuromodulation.Then,we summarize two novel tetherless neuromodulation methods using near-infrared light:upconversion nanoparticle-mediated optogenetics and photothermal neuromodulation.In addition,we discuss recent advances in mid-infrared optical neuromodulation.展开更多
Behavioral studies play a crucial role in unraveling the mechanisms underlying brain function.Recent advances in optogenetics,neuronal typing and labeling,and circuit tracing have facilitated the dissection of the neu...Behavioral studies play a crucial role in unraveling the mechanisms underlying brain function.Recent advances in optogenetics,neuronal typing and labeling,and circuit tracing have facilitated the dissection of the neural circuitry involved in various important behaviors.The identification of monosynaptic connections,both upstream and downstream of specific neurons,serves as the foundation for understanding complex neural circuits and studying behavioral mechanisms.However,the practical implementation and mechanistic understanding of monosynaptic connection tracing techniques and functional identification remain challenging,particularly for inexperienced researchers.Improper application of these methods and misinterpretation of results can impede experimental progress and lead to erroneous conclusions.In this paper,we present a comprehensive description of the principles,specific operational details,and key steps involved in tracing anterograde and retrograde monosynaptic connections.We outline the process of functionally identifying monosynaptic connections through the integration of optogenetics and electrophysiological techniques,providing practical guidance for researchers.展开更多
Optical-neural stimulation,which encompasses cutting-edge techniques such as optogenetics and infrared neurostimulation,employs distinct mechanisms to modulate brain function and behavior.These advanced neuromodulatio...Optical-neural stimulation,which encompasses cutting-edge techniques such as optogenetics and infrared neurostimulation,employs distinct mechanisms to modulate brain function and behavior.These advanced neuromodulation techniques offer accurate manipulation of targeted areas,even selectively modulating specific neurons,in the brain.This makes it possible to investigate the cause-and-effect connections between neural activity and behavior,allowing for a better comprehension of the intricate brain dynamics towards complex environments.Non-human primates serve as an essential animal model for investigating these complex functions in brain research,bridging the gap between the basic research and clinical applications.One of the earliest optical studies utilizing optogenetic neuromodulation in monkeys was conducted in 2009.Since then,the optical-neural stimulations have been effectively applied in non-human primates.This review summarises recent research that employed optogenetics or infrared neurostimulation techniques to regulate brain function and behavior in non-human primates.The current state of optical-neural stimulations discussed here demonstrates their efficacy in advancing the understanding of brain systems.Nevertheless,there are still challenges that need to be addressed before they can fully achieve their potential.展开更多
Retrograde adeno-associated viruses(AAVs)are capable of infecting the axons of projection neurons and serve as a powerful tool for the anatomical and functional characterization of neural networks.However,few retro-gr...Retrograde adeno-associated viruses(AAVs)are capable of infecting the axons of projection neurons and serve as a powerful tool for the anatomical and functional characterization of neural networks.However,few retro-grade AAV capsids have been shown to offer access to cor-tical projection neurons across different species and enable the manipulation of neural function in non-human primates(NHPs).Here,we report the development of a novel retro-grade AAV capsid,AAV-DJ8R,which efficiently labeled cortical projection neurons after local administration into the striatum of mice and macaques.In addition,intrastriatally injected AAV-DJ8R mediated opsin expression in the mouse motor cortex and induced robust behavioral alterations.Moreover,AAV-DJ8R markedly increased motor cortical neuron firing upon optogenetic light stimulation after viral delivery into the macaque putamen.These data demonstrate the usefulness of AAV-DJ8R as an efficient retrograde tracer for cortical projection neurons in rodents and NHPs and indicate its suitability for use in conducting functional interrogations.展开更多
Clinical experiments have proven that the pedunculopontine nucleus(PPN)plays a crucial role in the modulation of beta oscillations in Parkinson’s disease(PD).Here,we propose a new computational framework by introduci...Clinical experiments have proven that the pedunculopontine nucleus(PPN)plays a crucial role in the modulation of beta oscillations in Parkinson’s disease(PD).Here,we propose a new computational framework by introducing the PPN and related synaptic connections to the classic basal ganglia-thalamo-cortical model.Fascinatingly,the improved model can not only simulate the basic saturated and beta activities mentioned in previous studies but also produce the normal alpha rhythm that is much closer to physiological phenomena.Specifically,the results show that Parkinsonian oscillation activities can be controlled and modulated by the connection strength between the PPN and the globus pallidus internal nucleus(GPi)and the subthalamic nucleus(STN),supporting the fact that PPN is overinhibited in PD.Meanwhile,the internal mechanism underlying these state transitions is further explained from the perspective of dynamics.Additionally,both deep brain stimulation(DBS)and optogenetic technology are considered effective in terms of abnormal oscillations.Especially when a low-frequency DBS is added to the PPN,beta oscillations can be suppressed,but it is excited again as the DBS’s frequency gradually increases to a larger value.These results coincide with the experimental results that low-frequency stimulation of the PPN is effective,and verify the rationality of the model.Furthermore,we show that optogenetic stimulation of the globus pallidus external(GPe)expressing excitatory channelrhodopsin(ChR2)can effectively inhibit beta oscillations,whereas exciting the STN and PPN has a limited effect.These results are consistent with experimental reports suggesting that the symptoms of PD’s movement disorder can be alleviated under the GPe-ChR2,but not STN-ChR2,situation.Although the functional role of the PPN and the feasibility of optogenetic stimulation remain to be clinically explored,the results obtained help us understand the mechanisms of beta oscillations in PD.展开更多
The basal ganglia(BG) act as a cohesive functional unit that regulates motor function,habit formation,and reward/addictive behaviors. However,it is still not well understood how the BG maintains wakefulness and suppre...The basal ganglia(BG) act as a cohesive functional unit that regulates motor function,habit formation,and reward/addictive behaviors. However,it is still not well understood how the BG maintains wakefulness and suppresses sleep to achieve al these fundamental functions until genetical y engineered systems developed these years. Significant research efforts have recently been directed at developing genetic-molecular tools to achieve reversible and cell-type specific in vivo silencing or activation of neurons in behaving animals. Optogenetic tools can be used both to specifically activate or inhibit neurons of interest and identify functional synaptic connectivity between specific neuronal populations,both in vivo and in brain slices. Another recently developed system by Roth and colleagues permits the selective and ″remote″ manipulation(activation and silencing) of neuronal activity via all 3 major GPCR signaling pathways(G_i,G_s and G_q). These so-called ″ designer receptors exclusively activated by designer drugs″(DREADD) involve mutant GPCRs that do not respond to their endogenous ligands but are responsive to otherwise inert biological compounds. Recently,we demonstrated the essential roles and the neural pathways of the neurons expressing adenosine A_(2A) receptors or dopamine D_1 receptors in the BG for sleep-wake regulation using the genetically engineered systems including optogenetics and DREADD. We proposed a plausible model in which the caudate-putamen and the nucleus accumbens integrates behavioral processes with sleep/wakefulness through adenosine and dopamine receptors.展开更多
Many complex behaviors that do not require learning are displayed and are termed innate. Although traditionally the subject matter of ethology, innate behaviors offer a unique entry point for neuroscientists to dissec...Many complex behaviors that do not require learning are displayed and are termed innate. Although traditionally the subject matter of ethology, innate behaviors offer a unique entry point for neuroscientists to dissect the physiological mechanisms governing complex behaviors. Since the last century, converging evidence has implicated the hypothalamus as the central brain area that controls innate behaviors. Recent studies using cutting-edge tools have revealed that genetically-defined populations of neurons residing in distinct hypothalamic nuclei and their associated neural pathways regulate the initiation and maintenance of diverse behaviors including feeding, sleep, aggression, and parental care. Here, we review the newly-defined hypothalamic pathways that regulate each innate behavior. In addition, emerging general principles of the neural control of complex behaviors are discussed.展开更多
基金supported by the National Natural Science Foundation of China (Grant Nos.31371106 and 91632110)HZAU Independent Innovation Fund (2014BQ019).
文摘In vito fber photometry is a powerful technique to analyze the dy namics of population neurons during fiunctional study of neuroscience.Here,we introduced a detailed protocol for fiber photometry-based calciun reording in freely moving mice,covering from virus injection,fiber stub insertion,optogenetical stimulation to data procurement and analysis.Furthemnore,we applied this protocol to explore neuronal activity of mice latenal-posterior(LP)thalaric nucleus in response to optogenetical stimulation of primary visual cortex(V1)neurons,and explore axon clusters activity of optogenetically evoked V1 neurons.Final confirmation of virus-based protein expression in V1 and precise fber insertion indicated that the surgery procedure of this protocol is reliable for functional calcium recording.The scripts for data analysis and some tips in our protocol are provided in details.Together,this protocol is simple,low-cost,and effective for neuronal activity detection by fiber photometry,which will hep neuroscience researchers to carry out fiunctional and behavioral study in vivo.
基金supported by the CUHK Faculty of Medicine Faculty Innovation Award FIA2016/A/04(to V.C.K.C.)Group Research Scheme NL/JW/rc/grs1819/0426/19hc(to V.C.K.C.)The Hong Kong Research Grants Council 24115318,CUHK-R4022-18,14114721,and 14119022(to V.C.K.C)。
文摘Generating diverse motor behaviors critical for survival is a challenge that confronts the central nervous system(CNS)of all animals.During movement execution,the CNS performs complex calculations to control a large number of neuromusculoskeletal elements.The theory of modular motor control proposes that spinal interneurons are organized in discrete modules that can be linearly combined to generate a variety of behavioral patterns.These modules have been previously represented as stimulus-evoked force fields(FFs)comprising isometric limb-endpoint forces across workspace locations.Here,we ask whether FFs elicited by different stimulations indeed represent the most elementary units of motor control or are themselves the combination of a limited number of even more fundamental motor modules.To probe for potentially more elementary modules,we optogenetically stimulated the lumbosacral spinal cord of intact and spinalized Thy1-ChR2 transgenic mice(n=21),eliciting FFs from as many single stimulation loci as possible(20-70 loci per mouse)at minimally necessary power.We found that the resulting varieties of FFs defied simple categorization with just a few clusters.We used gradient descent to further decompose the FFs into their underlying basic force fields(BFFs),whose linear combination explained FF variability.Across mice,we identified 4-5 BFFs with partially localizable but overlapping representations along the spinal cord.The BFFs were structured and topographically distributed in such a way that a rostral-to-caudal traveling wave of activity across the lumbosacral spinal cord may generate a swing-to-stance gait cycle.These BFFs may represent more rudimentary submodules that can be flexibly merged to produce a library of motor modules for building different motor behaviors.
基金supported by the National Key Research and Development Program of China(Nos.2021YFA1201302 and 2021YFA1201300)the National Natural Science Foundation of China(Nos.52303033,52173029)+1 种基金Shanghai Sailing Program(No.23YF1400400)the Natural Science Foundation of Shanghai(No.21ZR1400500).
文摘Optogenetic has been widely applied in various pathogenesis investigations of neuropathic diseases since its accurate and targeted regulation of neuronal activity.However,due to the mismatch between the soft tissues and the optical waveguide,the long-term neural regulation within soft tissue(such as brain and spinal cord)by implantable optical fibers is a large challenge.Herein,we designed a modulus selfadaptive hydrogel optical fiber(MSHOF)with tunable mechanical properties(Young’modulus was tunable in the range of 0.32-10.56MPa)and low light attenuation(0.12-0.21 dB/cm,472nm laser light),which adapts to light transmission under soft tissues.These advantages of MSHOF can ensure the effectiveness of optogenetic stimulation meanwhile safeguarding the safety of the brain/materials interaction interface.In addition,this work provides more design possibilities of MSHOF for photogenetic stimuli and has significant application prospects in photomedical therapy.
基金supported by a grant from the Russian Science Foundation,No.23-75-10041(to MY)。
文摘Optogenetics has revolutionized the field of neuroscience by enabling precise control of neural activity through light-sensitive proteins known as opsins.This review article discusses the fundamental principles of optogenetics,including the activation of both excitatory and inhibitory opsins,as well as the development of optogenetic models that utilize recombinant viral vectors.A considerable portion of the article addresses the limitations of optogenetic tools and explores strategies to overcome these challenges.These strategies include the use of adeno-associated viruses,cell-specific promoters,modified opsins,and methodologies such as bioluminescent optogenetics.The application of viral recombinant vectors,particularly adeno-associated viruses,is emerging as a promising avenue for clinical use in delivering opsins to target cells.This trend indicates the potential for creating tools that offer greater flexibility and accuracy in opsin delivery.The adaptations of these viral vectors provide advantages in optogenetic studies by allowing for the restricted expression of opsins through cell-specific promoters and various viral serotypes.The article also examines different cellular targets for optogenetics,including neurons,astrocytes,microglia,and Schwann cells.Utilizing specific promoters for opsin expression in these cells is essential for achieving precise and efficient stimulation.Research has demonstrated that optogenetic stimulation of both neurons and glial cells-particularly the distinct phenotypes of microglia,astrocytes,and Schwann cells-can have therapeutic effects in neurological diseases.Glial cells are increasingly recognized as important targets for the treatment of these disorders.Furthermore,the article emphasizes the emerging field of bioluminescent optogenetics,which combines optogenetic principles with bioluminescent proteins to visualize and manipulate neural activity in real time.By integrating molecular genetics techniques with bioluminescence,researchers have developed methods to monitor neuronal activity efficiently and less invasively,enhancing our understanding of central nervous system function and the mechanisms of plasticity in neurological disorders beyond traditional neurobiological methods.Evidence has shown that optogenetic modulation can enhance motor axon regeneration,achieve complete sensory reinnervation,and accelerate the recovery of neuromuscular function.This approach also induces complex patterns of coordinated motor neuron activity and promotes neural reorganization.Optogenetic approaches hold immense potential for therapeutic interventions in the central nervous system.They enable precise control of neural circuits and may offer new treatments for neurological disorders,particularly spinal cord injuries,peripheral nerve injuries,and other neurodegenerative diseases.
基金Australian National Health and Medical Research Council。
文摘In this study,we aimed to develop an in vivo electrophysiological bone-nerve preparation to record the activity of peripheral sensory neurons that innervate the murine tibia.A small nerve that innervates the tibial marrow cavity was identified in isoflurane-anesthetized C57BL/6 mice,and placed over a platinum hook electrode for extracellular recording.Whole-nerve activity was amplified,filtered and sampled at 20 kHz using PowerLab(ADInstruments).A cannula was placed into the marrow cavity to deliver mechanical stimuli(by pressurizing with injection of saline)and/or capsaicin.Optical stimulation was achieved by application of 473 nm blue light(1 Hz,0.25-0.5 ms,0-12.5 mW/mm)to the tibial marrow cavity in Wnt1-Cre;loxP-ChR2 mice.Murine bone afferent neurons responded to high threshold noxious mechanical stimulation,coded for the intensity of mechanical stimulation,could be sensitized by capsaicin,and did not suffer stimulus-evoked fatigue at 10-minute interstimulus intervals.Electrical and optical stimulation within the marrow cavity evoked action potentials with conduction velocities in the Aδand/or C fiber range.These new approaches to recording the activity of bone afferent neurons will allow us to take advantage of transgenic and optogenetic tools to further our understanding of mechanisms that generate and maintain bone pain in the future.
基金funded by the Natural Science Foundation of China(No.82305049).
文摘Depression is a multifaceted disorder with a largely unresolved etiology influenced by a complex interplay of pathogenic factors.Despite decades of research,it remains a major condition that significantly diminishes patients’quality of life.Advances in optogenetics have introduced a powerful tool for exploring the neural mechanisms underlying depression.By selectively expressing optogenes in specific cell types in mice,researchers can study the roles of these cells through targeted light stimulation,offering new insights into central nervous system disorders.The use of viral vectors to express opsins in distinct neuronal subtypes enables precise activation or inhibition of these neurons via light.When combined with behavioral,morphological,and electrophysiological analyses,optogenetics provides an invaluable approach to investigating the neural mechanisms of psychiatric conditions.This review synthesizes current research on the application of optogenetics to understand the mechanisms of depression.This study aims to enhance our knowledge of optogenetic strategies for regulating depression and advancing antidepressant research.
基金supported by Basic Research Programs of Science and Technology Commission Foundation of Shanxi Province,No.20210302123486(to WJ).
文摘Epilepsy,a common neurological disorder,is characterized by recurrent seizures that can lead to cognitive,psychological,and neurobiological consequences.The pathogenesis of epilepsy involves neuronal dysfunction at the molecular,cellular,and neural circuit levels.Abnormal molecular signaling pathways or dysfunction of specific cell types can lead to epilepsy by disrupting the normal functioning of neural circuits.The continuous emergence of new technologies and the rapid advancement of existing ones have facilitated the discovery and comprehensive understanding of the neural circuit mechanisms underlying epilepsy.Therefore,this review aims to investigate the current understanding of the neural circuit mechanisms in epilepsy based on various technologies,including electroencephalography,magnetic resonance imaging,optogenetics,chemogenetics,deep brain stimulation,and brain-computer interfaces.Additionally,this review discusses these mechanisms from three perspectives:structural,synaptic,and transmitter circuits.The findings reveal that the neural circuit mechanisms of epilepsy encompass information transmission among different structures,interactions within the same structure,and the maintenance of homeostasis at the cellular,synaptic,and neurotransmitter levels.These findings offer new insights for investigating the pathophysiological mechanisms of epilepsy and enhancing its clinical diagnosis and treatment.
基金supported by the Beijing Natural Science Foundation of China under Grant 7222185。
文摘Objective:This study aims to establish an economically viable and easily accessible adult animal model for optogenetic activation of auditory neurons using adeno-associated viruses(AAVs)carrying Ch R2(H134R)to explore the potential of cochlear optogenetics as a hearing restoration technology.Methods:Healthy adult guinea pigs were used in the experiments.The viral vector AAV2/8-Ch R2(H134R)-h Syn-e YFP was administered to the right cochlea via the round window membrane.The confocal microscopy and reverse transcription polymerase chain reaction(RT-PCR)were utilized to analyze the Ch R2(H134R)expression localized to spiral ganglion neurons(SGNs).The auditory pathway activation was assessed by recording the optical compound action potential(oCAP)and acoustic compound action potential(a CAP)at various laser intensities.Results:The Ch R2(H134R)-e YFP expression was confirmed in 90%of the tested animals,localized to the SGNs of the injected ear.Higher m RNA levels of Ch R2(H134R)and e YFP were observed in the injected ear compared to the non-injected ear,while actin(Actb)m RNA levels were not significantly different.The o CAP was successfully elicited by a 470 nm blue light laser stimulus,with similar amplitudes and latency periods to those of a CAPs when the o CAP was evoked by 5.80 m W blue light and the a CAP was evoked by a 40 d B SPL click.The amplitudes of o CAPs increased with increasing laser intensity.Conclusion:This study demonstrates the viability of optogenetic activation of the auditory system in adult guinea pigs through the transduction of AAV-Ch R2(H134R)in SGNs.Cochlear optogenetics demonstrates potential as a hearing restoration technology,providing a basis for further clinical research and opening new avenues for investigation.
基金supported by the National Key R&D Program of China,No.2023YFC2509700the Beijing Natural Science Foundation-Haidian Original Innovation Joint Fund,No.L232141the Research and Application of Clinical Characteristic Diagnosis and Treatment Program,No.Z221100007422019(all to WD)。
文摘Spinal cord injury is a severe neurological disorder;however,current treatment methods often fail to restore nerve function effectively.Spinal cord stimulation via electrical signals is a promising therapeutic modality for spinal cord injury.Based on similar principles,this review aims to explore the potential of optical and acoustic neuromodulation techniques,emphasizing their benefits in the context of spinal cord injury.Photoacoustic imaging,renowned for its noninvasive nature,high-resolution capabilities,and cost-effectiveness,is well recognized for its role in early diagnosis,dynamic monitoring,and surgical guidance in stem cell therapies for spinal cord injury.Moreover,photoacoustodynamic therapy offers multiple pathways for tissue regeneration.Optogenetics and sonogenetics use genetic engineering to achieve precise neuronal activation,while photoacoustoelectric therapy leverages photovoltaic materials for electrical modulation of the nervous system,introducing an innovative paradigm for nerve system disorder management.Collectively,these advancements represent a transformative shift in the diagnosis and treatment of spinal cord injury,with the potential to significantly enhance nerve function remodeling and improve patient outcomes.
基金Supported by the Tianjin Municipal Education Commission Scientic Research Project No.2023YXZD10。
文摘The hippocampus is the brain structure that is responsible for the formation of learning memories.Sleep disorders leading to cognitive impairment are strongly associated with the hippocampus.Phototherapy offers a new physical therapy for the treatment of sleep disorders,with the advantages of being noninvasive and having few side effects.However,the mechanism by which phototherapy improves cognitive impairment caused by sleep disorders remains unclear.In this study,we used phototherapy combined with optogenetic technology to investigate the effect of noninvasive phototherapy on cognitive functions in sleep-deprived mice.Our results suggest that phototherapy might improve cognitive functions in sleep-deprived mice by modulating the hippocampus.Our study expands the research progress on noninvasive phototherapy for the treatment of sleep disorders.
文摘This study is based on wireless optogenetic technology,utilizing the CRY2/CIB1 photosensitive system to achieve spatiotemporal control of PD-L1 expression.In vitro experiments showed that the surface PD-L1 positivity rate of cells increased from 28.6±3.1%to 67.3±5.4%(P<0.001).In animal experiments,the terminal tumor volume in the light exposure group was 450±90 mm3,with a tumor inhibition rate of approximately 49.4%(P<0.001),and the median survival was extended to 32 days(compared to 24 days in the control group,P=0.004).Immunological tests revealed a significant increase in CD8+T cell infiltration(112±18 vs 52±10 cells/HPF,P<0.01),a 30%decrease in the proportion of Tregs(P<0.05),and an increase in the M1/M2 macrophage ratio to 1.8.The results suggest that the wireless optogenetic system can not only precisely regulate PD-L1 but also remodel the tumor immune microenvironment,providing a new approach for precise immunotherapy of GBM.
基金supported by grants from the Key Strategic Science and Technology Cooperation Project of the Ministry of Science and Technology of China(2023YFE0206800)the National Natural Science Foundation of China(81625006,31820103005,32200620,32170976,81971874)+1 种基金Zhejiang Province Natural Science Foundation of China(LZ24C090003 and LY21C090003)the Peak Discipline Cultivation Program of Zhejiang University School of Basic Medicine.
文摘Dear Editor,It is now well established that optogenetic stimulation can achieve precise intervention and modulate the activity of local neurons or neural circuits in the brain.Although this technique holds promise for clinical therapy for neurological and psychiatric disorders,it requires the expression of lightsensitive proteins(such as channel rhodopsin)or photoactivatable chemicals(such as caged neurotransmitters)in the targeted brain regions[1].
基金supported by the National Key Research and Development Program of China(2022YFE0201000)the National Natural Science Foundation of China(82174002,82104416,82204652)the High-Level University Development Program of Guangdong Province,and the Guangzhou Key Science and Technology Research and Development Project(202206010109)。
文摘Background:The development of ketamine-like rapid antidepressants holds promise for enhancing the therapeutic efficacy of depression,but the underlying cellular and molecular mechanisms remain unclear.Implicated in depression regulation,the neuropeptide pituitary adenylate cyclase-activating polypeptide(PACAP)is investigated here to examine its role in mediating the rapid antidepressant response.Methods:The onset of antidepressant response was assessed through depression-related behavioral paradigms.The signaling mechanism of PACAP in the hippocampal dentate gyrus(DG)was evaluated by utilizing site-directed gene knockdown,pharmacological interventions,or optogenetic manipulations.Overall,446 mice were used for behavioral and molecular signaling testing.Mice were divided into control or experimental groups randomly in each experiment,and the experimental manipulations included:chronic paroxetine treatments(4 d,9 d,14 d)or a single treatment of ketamine;social defeat or lipopolysaccharides-injection induced depression models;different doses of PACAP(0.4 ng/site,2 ng/site,4 ng/site;microinjected into the hippocampal DG);pharmacological intra-DG interventions(CALM and PACAP6-38);intra-DG viral-mediated PACAP RNAi;and opotogenetics using channelrhodopsins 2(ChR2)or endoplasmic natronomonas halorhodopsine 3.0(eNpHR3.0).Behavioral paradigms included novelty suppressed feeding test,tail suspension test,forced swimming test,and sucrose preference test.Western blotting,ELISA,or quantitative real-time PCR(RT-PCR)analysis were used to detect the expressions of proteins/peptides or genes in the hippocampus.Results:Chronic administration of the slow-onset antidepressant paroxetine resulted in an increase in hippocampal PACAP expression,and intra-DG blockade of PACAP attenuated the onset of the antidepressant response.The levels of hippocampal PACAP expression were reduced in both two distinct depression animal models and intra-DG knockdown of PACAP induced depression-like behaviors.Conversely,a single infusion of PACAP into the DG region produced a rapid and sustained antidepressant response in both normal and chronically stressed mice.Optogenetic intra-DG excitation of PACAP-expressing neurons instantly elicited antidepressant responses,while optogenetic inhibition induced depression-like behaviors.The longer optogenetic excitation/inhibition elicited the more sustained antidepressant/depression-like responses.Intra-DG PACAP infusion immediately facilitated the signaling for rapid antidepressant response by inhibiting calcium/calmodulin-dependent protein kinaseⅡ(CaM KⅡ)-eukaryotic elongation factor 2(eEF2)and activating the mammalian target of rapamycin(mTOR).Pre-activation of CaMKⅡsignaling within the DG blunted PACAP-induced rapid antidepressant response as well as eEF2-mTOR-brain-derived neurotrophic factor(BDNF)signaling.Finally,acute ketamine treatment upregulated hippocampal PACAP expression,whereas intraDG blockade of PACAP signaling attenuated ketamine’s rapid antidepressant response.Conclusions:Activation of hippocampal PACAP signaling induces a rapid antidepressant response through the regulation of CaMKⅡinhibition-governed eEF2-mTOR-BDNF signaling.
基金supported by China Postdoctoral Science Foundation(2022M723356),"From 0 to 1"Original Innovation Project of the Basic Frontier Scientific Research Program of the Chinese Academy of Sciences(29J20-015-Ⅲ)Chinese Academy of Sciences 100 Talents Project:Research on Task oriented Functional Brain Development of Infants(29J20-052-Ⅲ)Natural Science Basic Research Plan in Shaanxi Province of China(2022JQ544).
文摘Optogenetics,a technique that employs light for neuromodulation,has revolutionized the study of neural mechanisms and the treatment of neurological disorders due to its high spatiotemporal resolution and cell-type specificity.However,visible light,particularly blue and green light,commonly used in conventional optogenetics,has limited penetration in biological tissue.This limitation necessitates the implantation of optical fibers for light delivery,especially in deep brain regions,leading to tissue damage and experimental constraints.To overcome these challenges,the use of orange-red and infrared light with greater tissue penetration has emerged as a promising approach for tetherless optical neuromodulation.In this review,we provide an overview of the development and applications of tetherless optical neuromodulation methods with long wavelengths.We first discuss the exploration of orange-red wavelength-responsive rhodopsins and their performance in tetherless optical neuromodulation.Then,we summarize two novel tetherless neuromodulation methods using near-infrared light:upconversion nanoparticle-mediated optogenetics and photothermal neuromodulation.In addition,we discuss recent advances in mid-infrared optical neuromodulation.
基金National Key Research and Development Program of China(2022YFC2402703)Scientific and Technological Innovation 2030 Program of China,major projects(2021ZD0200408)+3 种基金National Natural Science Foundation of China(32171007 and 31800880)Non-profit Central Research Institute Fund of the Chinese Academy of Medical Sciences(2018PT31041)Science and Technology Planning Project of Guangdong Province(2018B030331001)Fundamental Research Funds for the Central Universities(K20210089).
文摘Behavioral studies play a crucial role in unraveling the mechanisms underlying brain function.Recent advances in optogenetics,neuronal typing and labeling,and circuit tracing have facilitated the dissection of the neural circuitry involved in various important behaviors.The identification of monosynaptic connections,both upstream and downstream of specific neurons,serves as the foundation for understanding complex neural circuits and studying behavioral mechanisms.However,the practical implementation and mechanistic understanding of monosynaptic connection tracing techniques and functional identification remain challenging,particularly for inexperienced researchers.Improper application of these methods and misinterpretation of results can impede experimental progress and lead to erroneous conclusions.In this paper,we present a comprehensive description of the principles,specific operational details,and key steps involved in tracing anterograde and retrograde monosynaptic connections.We outline the process of functionally identifying monosynaptic connections through the integration of optogenetics and electrophysiological techniques,providing practical guidance for researchers.
文摘Optical-neural stimulation,which encompasses cutting-edge techniques such as optogenetics and infrared neurostimulation,employs distinct mechanisms to modulate brain function and behavior.These advanced neuromodulation techniques offer accurate manipulation of targeted areas,even selectively modulating specific neurons,in the brain.This makes it possible to investigate the cause-and-effect connections between neural activity and behavior,allowing for a better comprehension of the intricate brain dynamics towards complex environments.Non-human primates serve as an essential animal model for investigating these complex functions in brain research,bridging the gap between the basic research and clinical applications.One of the earliest optical studies utilizing optogenetic neuromodulation in monkeys was conducted in 2009.Since then,the optical-neural stimulations have been effectively applied in non-human primates.This review summarises recent research that employed optogenetics or infrared neurostimulation techniques to regulate brain function and behavior in non-human primates.The current state of optical-neural stimulations discussed here demonstrates their efficacy in advancing the understanding of brain systems.Nevertheless,there are still challenges that need to be addressed before they can fully achieve their potential.
基金supported by Ministry of Science and Technology of China(2019YFA0903803 and 2018YFA0801404)National Natural Science Foundation of China(31871090,32000730,81961128019,and 81901397)+7 种基金Shenzhen Science and Technology Innovation Commission(JCYJ20190809171003698,JCYJ202103243001018,JCYJ20180507182505475,and JCYJ20180504165804015)Shenzhen Technological Research Center for Primate Translational Medicine(F-2021-Z99-504979)Youth Innovation Promotion Association(CAS 2017120)Chinese Academy of Sciences International Partnership Program(172644KYSB20170004)China Postdoctoral Science Foundation(2019M653115.)CAS Key Laboratory of Brain Connectome and Manipulation(2019DP173024)Guangdong Provincial Key Laboratory of Brain Connectome and Behavior(2017B030301017)International Science and Technology Cooperation Base of Guangdong Province(2019A050505008).
文摘Retrograde adeno-associated viruses(AAVs)are capable of infecting the axons of projection neurons and serve as a powerful tool for the anatomical and functional characterization of neural networks.However,few retro-grade AAV capsids have been shown to offer access to cor-tical projection neurons across different species and enable the manipulation of neural function in non-human primates(NHPs).Here,we report the development of a novel retro-grade AAV capsid,AAV-DJ8R,which efficiently labeled cortical projection neurons after local administration into the striatum of mice and macaques.In addition,intrastriatally injected AAV-DJ8R mediated opsin expression in the mouse motor cortex and induced robust behavioral alterations.Moreover,AAV-DJ8R markedly increased motor cortical neuron firing upon optogenetic light stimulation after viral delivery into the macaque putamen.These data demonstrate the usefulness of AAV-DJ8R as an efficient retrograde tracer for cortical projection neurons in rodents and NHPs and indicate its suitability for use in conducting functional interrogations.
基金supported by the National Natural Science Foundation of China(Grant Nos.12072265 and 12372064).
文摘Clinical experiments have proven that the pedunculopontine nucleus(PPN)plays a crucial role in the modulation of beta oscillations in Parkinson’s disease(PD).Here,we propose a new computational framework by introducing the PPN and related synaptic connections to the classic basal ganglia-thalamo-cortical model.Fascinatingly,the improved model can not only simulate the basic saturated and beta activities mentioned in previous studies but also produce the normal alpha rhythm that is much closer to physiological phenomena.Specifically,the results show that Parkinsonian oscillation activities can be controlled and modulated by the connection strength between the PPN and the globus pallidus internal nucleus(GPi)and the subthalamic nucleus(STN),supporting the fact that PPN is overinhibited in PD.Meanwhile,the internal mechanism underlying these state transitions is further explained from the perspective of dynamics.Additionally,both deep brain stimulation(DBS)and optogenetic technology are considered effective in terms of abnormal oscillations.Especially when a low-frequency DBS is added to the PPN,beta oscillations can be suppressed,but it is excited again as the DBS’s frequency gradually increases to a larger value.These results coincide with the experimental results that low-frequency stimulation of the PPN is effective,and verify the rationality of the model.Furthermore,we show that optogenetic stimulation of the globus pallidus external(GPe)expressing excitatory channelrhodopsin(ChR2)can effectively inhibit beta oscillations,whereas exciting the STN and PPN has a limited effect.These results are consistent with experimental reports suggesting that the symptoms of PD’s movement disorder can be alleviated under the GPe-ChR2,but not STN-ChR2,situation.Although the functional role of the PPN and the feasibility of optogenetic stimulation remain to be clinically explored,the results obtained help us understand the mechanisms of beta oscillations in PD.
文摘The basal ganglia(BG) act as a cohesive functional unit that regulates motor function,habit formation,and reward/addictive behaviors. However,it is still not well understood how the BG maintains wakefulness and suppresses sleep to achieve al these fundamental functions until genetical y engineered systems developed these years. Significant research efforts have recently been directed at developing genetic-molecular tools to achieve reversible and cell-type specific in vivo silencing or activation of neurons in behaving animals. Optogenetic tools can be used both to specifically activate or inhibit neurons of interest and identify functional synaptic connectivity between specific neuronal populations,both in vivo and in brain slices. Another recently developed system by Roth and colleagues permits the selective and ″remote″ manipulation(activation and silencing) of neuronal activity via all 3 major GPCR signaling pathways(G_i,G_s and G_q). These so-called ″ designer receptors exclusively activated by designer drugs″(DREADD) involve mutant GPCRs that do not respond to their endogenous ligands but are responsive to otherwise inert biological compounds. Recently,we demonstrated the essential roles and the neural pathways of the neurons expressing adenosine A_(2A) receptors or dopamine D_1 receptors in the BG for sleep-wake regulation using the genetically engineered systems including optogenetics and DREADD. We proposed a plausible model in which the caudate-putamen and the nucleus accumbens integrates behavioral processes with sleep/wakefulness through adenosine and dopamine receptors.
基金supported by the Strategic Priority Research Program of the Chinese Academy of Sciences, China (XDB02030005)
文摘Many complex behaviors that do not require learning are displayed and are termed innate. Although traditionally the subject matter of ethology, innate behaviors offer a unique entry point for neuroscientists to dissect the physiological mechanisms governing complex behaviors. Since the last century, converging evidence has implicated the hypothalamus as the central brain area that controls innate behaviors. Recent studies using cutting-edge tools have revealed that genetically-defined populations of neurons residing in distinct hypothalamic nuclei and their associated neural pathways regulate the initiation and maintenance of diverse behaviors including feeding, sleep, aggression, and parental care. Here, we review the newly-defined hypothalamic pathways that regulate each innate behavior. In addition, emerging general principles of the neural control of complex behaviors are discussed.
