Effects of Acupuncture on Monoamine Neurotransmitters in Raphe Nuclei in Obese Rats

Effects of acupuncture on the levels of neurotransmitters in the raphe nuclei were investigated in obeserats.It was found that the levels of tryptophan (Trp) and 5-hydroxyindoleacetic acid (5-HIAA) wereincreased,and 5-hydroxytryptamine (5-HT) level and 5-HT/5-HIAA ratio decreased in the raphe nucleiof the obese group as compared with the normal group;and that acupuncture could produce weightreduction,increase the 5-HT level and 5-HT/5-HIAA ratio,and decrease the contents of Trp and5-HIAA,but did not change the levels of dopamine (DA) and noradrenaline (NA).It is indicated thatbenign regulative action of acupuncture on 5-HT and its metabolism in the raphe nuclei is possibly oneof the factors for reducing weight by acupuncture.

Ca^2＋ in the dorsal raphe nucleus promotes wakefulness via endogenous sleep-wake regulating pathway in the rats

Aim Serotonergic neurons in the dorsal raphe nucleus （DRN） are involved in the control of sleep- wake states. Our previous study indicated that calcium （Ca^2＋ ） modulation in the DRN plays an important role in rapid eye movement sleep （REMS） and non-REMS （NREMS） regulation during pentobarbital hypnosis. Presentstudy was designed to investigate the effects of Ca^2＋ in the DRN on sleep-wake regulation and related neuronal mechanism in freely moving rats. Methods CaC12 （25 or 50 nmol） was microinjected into the DRN and sleep pa- rameters were detective by electroencephalogram （EEG） Monoamine neurotransmitters were detected by HPLC- ECD. c-Fos expression in sleep-wake regulating nucleus was detected by immunohistochemistry. Results Our re- sults showed that microinjection of CaC12 （25 or 50 nmol） into the DRN promoted wakefulness and suppressed NREMS including slow wave sleep （SWS） and REMS in freely moving rats. Meanwhile intro-DRN application of CaC12 （25 or 50 nmol） significantly increased serotonin in DRN and hypothalamus, as well increased noradrenaline in locus coeruleus （LC） and hypothalamus. Immunohistochemistry study indicated that the application CaC12 （25 or 50 nmol） into the DRN significantly increased c-Fos expression ratio in wake-promoting neurons including seroton- ergic neurons in the DRN, noradrenergic neurons in the LC and orxinergic neurons in the perifornical nucleus （Pef） , but decreased c-Fos expression ratio of GABAergic sleep-promoting neurons in the ventrolateral preoptic nu- cleus （VLPO）. Conclusion These results suggest that Ca^2＋ in the DRN exert arousal effects via up-regulation of serotonergic function in endogenous sleep-wake regulating pathways.

Chinia gen.nov.—the second diatom genus simonsenioid raphe from mangroves in Fujian,China

A new monotypic diatom genus Chinia gen.nov.from estuarine mangrove habitats in Fujian,China is described.Light,scanning and transmission electron microscopy observations of its morphology show that this diatom differs from any established raphid diatom genus.The new genus is distinguished by a strongly undulate valve face,eccentric raphe canal,and perforated copulae.Its distinctly elevated and perforated raphe canal,classifi ed as simonsenioid type,is supported by sparse infundibulum-like portulae,and the portulae are separated by large fenestrae.The valves of Chinia also have hymenate areolar occlusions and the raphe lacks a central nodule.However,unlike most Bacillariaceae with two plastids,including Simonsenia,Chinia has a single plastid similar to those in Entomoneis and Surirella.Despite the similarities to Simonsenia,Chinia features porous raphe canal and loose striation,which is different from those of Simonsenia that has a solid raphe canal and distinct rows of areolae.These distinctive cytological and morphological characters suggest an affinity of the new genus to the Bacillariaceae.

The Role of Dorsal Raphe Nucleus and 5-HT in the Electro-acupunctural Inhibition of the cAMP Induced ECoG Epileptiform Activities in Rabbits

Like electroacupuncture (EA),electrical stimulation of the dorsal raphe nucle-us (DRN)also inhibits the cAMP induced cortical epileptic waves and the EA inducedinhibition can be abolished or markedly reduced by iv cyproheptadine (CYP) or destruc-tion of DRN by electrolysis.The results suggest that 5-HT neurons in the DRN mayplay an important role in the EA inhibition of the generation of cAMP induced elec-trocorticogram (ECoG) epileptic waves.

Effects of various frequency electrical stimulation of the dorsal raphe nucleus on spontaneous firing activities in the rat subthalamic nucleus

BACKGROUND： Some investigations have demonstrated that exogenous 5-hydroxytryptamine increases the spontaneous firing rate of subthalamic nucleus （STN） neurons in the rat brain. OBJECTIVE： To validate the effect of electrical stimulation to the dorsal raphe nucleus （DRN） on the neuronal activities of the STN in rats, as well as analyze the differences in the effects of electrical stimulation at various frequencies. DESIGN, TIME AND SETTING： Experiments were performed from March 2007 to June 2007 in the Electrophysiology Laboratory of Liaoning Medical University with a randomized controlled animal study design. MATERIALS： Twenty-four healthy male Sprague-Dawley （SD） rats, weighing 250-350 g, were selected for this study. An A320R constant electrical stimulator was purchased from World Precision Instruments Company （USA）; a Spike 2 biological signal acquisition system was purchased from British CED Company. METHODS： Twenty-four SD rats were randomly assigned into a model group and a normal group, with 12 rats in each group. To mimic Parkinson＇s disease, rats in the model group were injected with 4μL of 6-hydroxydopamine into the right striatum, then received deep brain stimulation. Rats in the normal group received deep brain stimulation in same brain region without modeling. Electrical stimulation （width, 0.06 ms; intensity, 0.2-0.6 mA; frequency, 20-130 Hz; train duration, 5 seconds） was delivered to the DRN. MAIN OUTCOME MEASURES： The firing rates of STN neurons were observed by extracellular recording using a biological signal acquisition system. RESULTS： DRN-high-frequency stimulation （DRN-HFS） induced excitation in 59% of the STN neurons in the normal group and 50% of the STN neurons in the model group; mean firing rates increased significantly from （7.14±0.75） and （7.94 ± 0.61） Hz to （11.17 ±1.49） and （12.11 ± 1.05） Hz, respectively （P 〈 0.01）. Spontaneous firing rate increased significantly in 53% of neurons in normal rats in a frequency-dependent manner when stimulation frequency was in the range 80-130 Hz. CONCLUSION： DRN-HFS induced an excitatory effect on the spontaneous activity of STN neurons in both normal and PD rats. There was a frequency-dependent effect of electrical stimulation of the DRN on spontaneous firing activities in STN neurons.

Distributive characteristics of projection from vestibular nuclei to nucleus raphe magnus in rats

BACKGROUND： Morphological studies have confirmed that vestibular nuclei accepts serotoninergic projections from nucleus raphe magnus, nucleus raphes pallidus, etc. But it is still unclear whether there is bi-directional association between vestibular nuclei and nucleus raphe magnus. OBJECTIVE： To observe the characteristics of projective fibers from vestibular nuclei to nucleus raphe magnus using tetramethyl rhodamine （TMR） in rats, so as to provide more sufficient morphological evidence of neural association from vestibular nuclei. DESIGN： An observational experiment. SETTING： Department of Anatomy （K.K. Leung Brain Research Center）, the Fourth Military Medical University of Chinese PLA. MATERIALS： Eighteen male SD rats of clean degree, weighing 250-280 g, were provided by the Experimental Animal Center of the Fourth Military Medical University of Chinese PLA. METHODS： The experiments were carried out in the laboratory of Department of Anatomy （K.K. Leung Brain Research Center）, the Fourth Military Medical University of Chinese PLA from September 2006 to January 2007. All the rats were anesthetized with intraperitoneal injection of pentobarbital sodium, then according to the coordinates on the rat brain atlas, 0.1 μL TMR （100 g/L） was injected into nucleus raphes magnus via the tip of glass microtubule by means of microinjection. Seven days later, the rats were anesthetized, then perfused and fixed to remove brain, and then frozen coronal brain sections were prepared. The retrogradely labeled neurons in the injected and projected sites were observed under fluorescence microscope. Light filters with evoked wave length of 540-553 nm and emission wave length ≥ 1 580 nm were selected to observe the orange TMR-labeled neurons. All the sections were observed and counted under the fluorescence microscope. MAIN OUTCOME MEASURES： Characteristics and number of retrogradely labeled neurons at different sites of nuclei. RESULTS： Totally 18 SD rats were enrolled, 9 of them were excluded due to the deviation of injected site, and the other 9 were involved in the final analysis of results. The concentrated region of TMR injection was mainly restricted to nucleus raphes magnus, and diffused to the surrounding area to different extents. There were obvious differences in the distributions of the labeled neurons among the subdivisions in vestibular nuclei, as well as the distributions of the labeled neurons at different sites in the same subdivision. The majority of the labeled neurons distributed in the rostral levels of medial vestibular nucleus and the lateral vestibular nucleus, while fewer labeled neurons were observed in superior vestibular nucleus. CONCLUSION： ① There might be bi-directional association between vestibular nucli and nucleus raphe magnus, suggesting that nucleus raphe magnus played a role in the transmission and processing of vestibular information. ② The projection from nucleus raphe magnus to vestibular nucleus has certain distributive characteristics in the region.

Colocalization of SOM and NOS in the neurons of raphe nucleiinnervating the pharyngeal muscles in the rats: PRV,SOMandNADPH-d triplelabelingstudy

Objective: To investigate the colocalization of Somatostatin (SOM ) and NADPH--diaphorase (NADPHd ) of the neurons in raphe nuclei innervating pharyngeal muscles in the rats. Methods: After PRV was injected into the pharyngeal muscles. PRV and SOM immunofluorescence double labeling procedure was completed at first,then proceeded NADPH -d histochemistry. Results: PRV and SOM double--labled cells were present mainly in the nucleus raphe magnus. but some PRV and SOM double labled neurons were found in the other raphe nuclei as well, such as nucleus raphe pallidus. nucleus raphe obsurus, median raphe nucleus and dorsal raphe nucleus.NADPH -d positive neurons were also observed in the raphe nuclei. PRV. SOM and NADPH-d triple labeling neurons were found in the nucleus raphe magnus. Conclusion: It is suggested that the colocalization of SOM and NADPH--d of the neurons in the raphe nuclei innervating pharyngeal muscles may play an important role in the coordination of the pharyngeal motility.

THE NEURONS DOUBLE-LABELLED BY SEROTONIN AND GLUTAMATE-IMMUNOREACTIVITY IN MEDULLARY RAPHE NUCLEI PROJECTING TO THE CEREBELLAR CORTEX OF THE KITTEN

The neurons with coexistence of serotonin-and giutamate-immunoreactivity(5-HT/Glu cells)in medullary raphe nuclei were investigated in the present study by using immunofluorescence technique.On the basis or the investigation projections of the 5-HT/Glu cells to the cerebellar cortex were studied by means or a combined microsphere retrograde transport.The results showed that most of the 5-HT cells in medullary raphe nuclei exhibited Glu-immunoreactivity.The cell count indicated that 81.77% of the 5-HT cells contained Gin-immunoreactivity;40.35%of the Gin-cells contained 5-HT.The retrograde labeled cells by microspkere(MS cells)were found in all medullary rapke nuclei. 5-HT/MS cells were rare.The Gin/MS cells had a rate in total MS cells(40.31 %).Three trible-labeled cells (5-HT/Glu/MS cells)were found. The finding or the 5-HT/Glu cells in medullary raphe nuclei projecting to the cerebellar cortex may give an iportant significance to understanding the functional connection between medullary raphe nuclei and cerebellum.

GLUTAMATE－IMMUNOREACTIVE NEURONS IN MEDULLARY RAPHE－CEREBELLUM PATHWAY IN THE KITTEN

Using a combined microsphere retrograde transport with immunofluorescence technique,the glutamate-immunoreactive neurons in the medullary raphe nuclei that project to the cerebellar cortex were investigated in the present study.The cells double-labeled by glutamate-immunoresctivity and microspheres were found in all medullary raphe nuclei.Among all the cerebellar projection neurons in the medullary raphe nuclei,(49. 49±2.78)% (±)in pallidus (NRP),(62.02±3. 62) % in obscures (NRD),(3. 03±3. 03) % in magnus (NRM) were immunoreactive to glutamate.The results raise the possibility that the cerehellar vermal cortex (lobules Ⅴ-Ⅶ) may receive some excitatory input from medullary raphe nuclei to affect the function of the cerebellum.

THE ROLE OF NUCLEUS RAPHE MAGNUS IN THE ANTINOCICEPTIVE EFFECT OF MUSCLE SPINDLE AFFERENTS IN THE RAT

Objective To investigate the role of NRM in the antinociceptive effect of muscle spindle afferents,the influence of NRM lesion on the inhibitory effect of muscle spindle afferents on the nociceptive responses of wide dynamic range (WDR) neurons and the effects of the muscle spindle afferents on the NRM neuronal activities were observed. Methods The single units of WDR neurons in the spinal dorsal horn were recorded extracellularly, and the inhibitory effects of activating muscle spindle afferents by intravenous administration of succinylcholine (SCH) on the C fibers evoked responses (C responses) of WDR neurons were tested before and after lesion of NRM.The effects of the muscle spindle afferents activated by administrating SCH on the single NRM neurons were also examined.Results ①It was found that the C responses of WDR neurons were significantly inhibited by intravenously administration of SCH, and the inhibitory effect was reduced after lesion of NRM;②The activities of most of the NRM neurons could be changed significantly by administrating SCH. According to their responses, NRM neurons could be classified into three types:excitatory, inhibitory and non responsive neurons, and the responses were dose dependent. Conclusion These results suggest that the muscle spindle afferents evoked by SCH may activate the NRM neurons, which plays an important role in the antinociception of muscle spindle afferents.

Oxytocin in rat nucleus raphe magnus influences pain modulation

The main function of Nucleus raphe magnus (NRM) is mostly pain mediation. Our previous study has demonstrated that oxytocin (OXT) regulates antinociception through the central nervous system rather than the peripheral organs, and pain stimulation increases OXT concentration in the NRM. The experiment was designed to investigate OXT in the rat NRM effect on pain modulation. The results showed that 1) pain stimulation increased OXT concentration in NRM perfusion liquid;2) Intra-NRM microinjection of OXT increased the pain threshold in a dose-dependent manner, whereas intra-NRM microinjection of OXT receptor antagonist, desGly-NH2, d(CH2)5[D-Tyr2, Thr-sup-4]OVT decreased the pain threshold;3) NRM pre-treatment with OXT receptor antagonist completely attenuated the pain threshold increase induced by intra-NRM administration of OXT. The data suggested that OXT in NRM was involved in antinociception via OXT receptors.

Congenital median raphe cysts: Coexistence of cystic lesions and canal-like lesions

Median raphe cyst of the perineum is a rare congenital anomaly and was first reported in 1895. We report a 9-year-old male with median raphe cysts in the perineum which had become swollen and then spontaneously ruptured several times since he was 3 years old. There were two separate lesions;one was a cystic lesion and the other was a canal-like lesion. They were completely excised and the inner layer of the cysts was composed of transitional epithelium.

Regenerative Properties of Fetal n. Raphe Cells in Modeling Injuries of Serotonergic Neurite-Glial Fibers in Vitro

It is considered that renewal of growth in serotoninergic caudal fibers their sprouting to injury site and formation ofoutgrowth net is factor for locomotion recovery. The aim of our work was to study the influence of rat fetal neural cells enriched withserotonin-producing cells, on morphogenesis, gene expression of serotogenesis （Pet1, Nk2.2, Lmxlb, Tphl, Tph2, Sert） andserotonin level in organotypic n. raphe cultures in injury of neurite-glial fibers in vitro. Injury was modeled by the way of crossingneurite-glial fibers after their formation under long-term culture. The cell accumulation was observed in the injury zone when fetal n.raphe cells, previously enriched with serotonin-producing cells were added into organotypic culture of n. raphe with crossing fibersunder the culture conditions. As a result, crossing site was filled with these cells after 14 days of culture. The activation of expressionin cascade of regulatory genes-regulators of serotogenesis Pet1, Lmxlb, Tphl, Tph2, Sert and increased serotonin content in theculture material were also observed. The activation of gene expression of serotogenesis Pet1 and Sert, and increased serotonincontent by 1.7 times were revealed when conditioned medium from culture of fetal rat cells was used. An eviction of single cells wasobserved in control samples of organotypic n. raphe culture after injury. Crossing site was remained without signs of cell filling inthese samples at day 14 after culture. The activation of regulatory gene expression of serotogenesis Pet1, Nkx2.2, Lmxlb, Tphl,Tph2, Sert and restoration of serotonin content were also absent in the culture material.

AB010.A retinoraphe projection regulates looming-induced defensive behavior

Animals promote their survival by avoiding rapidly approaching objects that indicate threats.It is believed that looming cues are detected by retinal ganglion cells(RGCs)that project to the superior colliculus(SC).However,the exact type of RGC that transmits looming-related signals remains unclear.Here we identify a specific transient type of RGCs that controls mouse looming-evoked defensive response by sending axonal collaterals to the dorsal raphe nucleus(DRN)and SC.Looming signals transmitted by DRN-projecting RGCs activate DRN GABA neurons and in turn inhibit serotonin neurons.Moreover,optogenetically stimulating serotonin neurons reduces looming-evoked defensive behaviors.Thus,a dedicated population of RGCs detects rapidly approaching visual threats and their input to the DRN controls a serotonergic self-gating mechanism that regulates innate defensive responses.Our study provides new insights into how DRN and SC work in concert to extract and translate visual threats into defensive behavioral responses.

The raphe nuclei are the early lesion site of gastric α-synuclein propagation to the substantia nigra

Parkinson's disease(PD)is a neurodegeneration disease withα-synuclein accumulated in the substantia nigra pars compacta(SNpc)and most of the dopaminergic neurons are lost in SNpc while patients are diagnosed with PD.Exploring the pathology at an early stage contributes to the development of the disease-modifying strategy.Although the“gut–brain”hypothesis is proposed to explain the underlying mechanism,where the earlier lesioned site in the brain of gastricα-synuclein and howα-synuclein further spreads are not fully understood.Here we report that caudal raphe nuclei(CRN)are the early lesion site of gastricα-synuclein propagating through the spinal cord,while locus coeruleus(LC)and substantia nigra pars compacta(SNpc)were further affected over a time frame of 7 months.Pathologicalα-synuclein propagation via CRN leads to neuron loss and disordered neuron activity,accompanied by abnormal motor and non-motor behavior.Potential neuron circuits are observed among CRN,LC,and SNpc,which contribute to the venerability of dopaminergic neurons in SNpc.These results show that CRN is the key region for the gastricα-synuclein spread to the midbrain.Our study provides valuable details for the“gut–brain”hypothesis and proposes a valuable PD model for future research on early PD intervention.

Effects of 5-hydroxytryptamine ascending pathways of dorsal raphe nuclei and habenular nucleus on the respiration and blood pressure of rats

Obstructive sleep apnea syndrome （OSAS） was a disease of breath obstacle happened in the process of sleep. The central mechanism of OSAS has not yet been fully elucidated. Most of studies focused on raphe nuclei and 5-hydroxytryptamine （5-HT）, and showed that brain serotonergic activity might be decreased in OSAS. It is well known that the dorsal and medial raphe nuclei provide almost all the serotonergic innervation to the forebrain. A previous study evaluated the stimulation of the dorsal raphe nuclei （DRN） in the rat inducing mainly pressor and sympathoexcitatory responses.

VGLUT3 neurons in median raphe control the efficacy of spatial memory retrieval via ETV4 regulation of VGLUT3 transcription

The raphe nucleus is critical for feeding, rewarding and memory. However, how the heterogenous raphe neurons are molecularly and structurally organized to engage their divergent functions remains unknown. Here, we genetically target a subset of neurons expressing VGLUT3. VGLUT3 neurons control the efficacy of spatial memory retrieval by synapsing directly with parvalbumin-expressing GABA interneurons(PGIs) in the dentate gyrus. In a mouse model of Alzheimer's disease(AD mice),VGLUT3→PGIs synaptic transmission is impaired by ETV4 inhibition of VGLUT3 transcription. ETV4 binds to a promoter region of VGLUT3 and activates VGLUT3 transcription in VGLUT3 neurons. Strengthening VGLUT3→PGIs synaptic transmission by ETV4 activation of VGLUT3 transcription upscales the efficacy of spatial memory retrieval in AD mice. This study reports a novel circuit and molecular mechanism underlying the efficacy of spatial memory retrieval via ETV4 inhibition of VGLUT3 transcription and hence provides a promising target for therapeutic intervention of the disease progression.

INFLUENCE OF ACUPUNCTURE,RAPHE STIMULATION OR MORPHINE ON INTERCOSTAL DISCHARGES ELICITED BY STIMULATION OF MOTOR CORTEX

Previous work showed that acupuncture activated bulbar medial reticular formation (including the nucleus raphe magnus) which in turn sent descending impulses via the dorsolateral funiculi (DLF) to inhibit the viscerosomatic reflex (VSR). The inhibitory effect of acupuncture on the splanchnically evoked potential in the or-

A novel complex-high-order graph convolutional network paradigm:ChyGCN

In recent years,there has been a growing interest in graph convolutional networks(GCN).However,existing GCN and variants are predominantly based on simple graph or hypergraph structures,which restricts their ability to handle complex data correlations in practical applications.These limitations stem from the difficulty in establishing multiple hierarchies and acquiring adaptive weights for each of them.To address this issue,this paper introduces the latest concept of complex hypergraphs and constructs a versatile high-order multi-level data correlation model.This model is realized by establishing a three-tier structure of complexes-hypergraphs-vertices.Specifically,we start by establishing hyperedge clusters on a foundational network,utilizing a second-order hypergraph structure to depict potential correlations.For this second-order structure,truncation methods are used to assess and generate a three-layer composite structure.During the construction of the composite structure,an adaptive learning strategy is implemented to merge correlations across different levels.We evaluate this model on several popular datasets and compare it with recent state-of-the-art methods.The comprehensive assessment results demonstrate that the proposed model surpasses the existing methods,particularly in modeling implicit data correlations(the classification accuracy of nodes on five public datasets Cora,Citeseer,Pubmed,Github Web ML,and Facebook are 86.1±0.33,79.2±0.35,83.1±0.46,83.8±0.23,and 80.1±0.37,respectively).This indicates that our approach possesses advantages in handling datasets with implicit multi-level structures.

THE ROLE OF LOCUS COERULEUS-NORADRENERGIC AND DORSAL RAPHE NUCLEUS-SEROTONERGIC SYSTEMS IN ANALGESIC EFFECT OF HYPOTHA-LAMIC ARCUATE NUCLEUS STIMULATION

Previous studies ia our laboratory have shown that hypothalamic arcuate nucleus (ARC),locus coeruleus (LG) and dorsal raphe nucleus (DR) play an important role in pain modulation and acupuncture analgesia, There is sufficient evidence that LG and DR are main sites concentrated with noradrenergic (NE) and serotonergic (5-HT) neuronal peri-