Temporomandibular inflammation mobilizes parvalbumin and FosB/deltaFosB neurons of amygdala and dorsal raphe

Pathophysiological mechanisms involved in orofacial pain and their relationship with emotional disorders have emerged as an important research area for multidisciplinary studies. In particular, temporomandibular disorders (TMD) have been evaluated clinically from both physiological and psychological perspectives. We hypothesized that an altered neuronal activity occurs in the amygdala and the dorsal raphe nucleus (DR), encephalic regions involved in the modulation of painful and emotional information. Adult male Wistar rats were used in an experimental complete Freund's adjuvant (CFA)-induced temporomandibular joint (TMJ) inflammation model. CFA was applied for 1 or 10 days, and the animals were euthanized for brain samples dissection for FosB/ΔFosB and parvalbumin (PV) immunostaining. Our results were consistent in showing that the amygdala and DR were activated in the persistent inflammatory phase (10 days) and that the expression of PV+ interneurons in the amygdala was decreased. In contrast, in the DR, the expression of PV+ interneurons was increased in persistent states of CFA-induced TMJ inflammation. Moreover, at 10 days of inflammation, there was an increased co-localization of PV+ and FosB/ΔFosB+ neurons in the basolateral and central nucleus of the amygdala. Different nuclei of the amygdala, as well as portions of the DR, were activated in the persistent phase (10 days) of TMJ inflammation. In conclusion, altered activity of the amygdala and DR was detected during persistent inflammatory nociception in the temporomandibular joint. These regions may be essential for both sensory and affective dimensions of orofacial pain.

Effects of Temporary Inactivation and Electrical Stimulation of the Dorsal Raphe Nucleus on Morphine-induced Conditioned Place Preference

Background: The dorsal raphe nucleus (DRN) influences a wide range of behavioral and reward function. In this study, we evaluated electrical stimulation and inactivation of DRN on morphine conditioned place preference (CPP). Methods: The rats were anesthetised (n = 7 for each group) and the electrode and cannula were implanted into the DRN by stereotaxic instrument. Electrical stimulation (100μA) and reversible inactivation by lidocaine were induced into DRN and then morphine-induced CPP was investigated. Results: The stimulation of DRN in combination with effective dose of morphine showed a significant decrease only on expression phases 20s (SD 33.7) when compared with morphine group 119.85s (SD 23.7) (One way ANOVA, Tukey’s; P = 0.036). Also, this stimulation in combination with ineffective dose of morphine showed a significant increase only on acquisition phases 67.5s (SD 41.2) of CPP compared with morphine group -46s (SD 18.51) (P = 0.034). Also, there were not significant differences in inactivation of DRN by lidocaine on different phase of CPP (P = 0.091). Conclusion: It is possible that electrical stimulation of the DRN with changes in concentration of serotonin or involving other transmitters such as glutamate and gamma amino butyric acid (GABA) would be involved to these changes of CPP.

Fos expression in the prefrontal cortex and mesencephalic dorsal raphe nucleus in lactating rats after social instigation

Females are often less aggressive than males, but they exhibit high levels of agonistic behavior against an intruder in the area of ​​the nest during lactation. This behavior is referred to as maternal aggression. In rats, maternal aggressive behavior occurs more often from postpartum day 3 (PPD 3) to PPD 12. Social instigation is an experimental protocol used to increase the levels of aggression that are typical of the species. In the present study we used social instigation to analyze the expression of a marker of neuronal activity, c-fos. Lactating rats on PPD 5, in the presence of their pups, were divided into four groups: (1) no social instigation and no aggressive behavior, (2) social instigation and no aggressive behavior, (3) no social instigation and aggressive behavior, and (4) social instigation and aggressive behavior. Sixty minutes after the aggression test we used immunohistochemistry to detect Fos in two brain regions, the ventral-orbital region of the prefrontal cortex (VO PFC) and dorsal raphe nucleus (DRN). Our results showed that rats with aggressive behavior that were provoked exhibited an increase in Fos expression in the VO PFC compared with the control group (i.e., no social instigation and no aggressive behavior). No change in Fos expression was found in the DRN. These results complement previous findings with microinjection of serotonin 5-hydroxytryptamine-1B receptor agonists into the same region, demonstrating that the VO PFC is an important region in the modulation of maternal aggressive behavior.

5-HT1A autoreceptor modulation of locomotor activity induced by nitric oxide in the rat dorsal raphe nucleus

The dorsal raphe nucleus (DRN) is the origin of ascending serotonergic projections and is considered to be an important component of the brain circuit that mediates anxiety- and depression-related behaviors. A large fraction of DRN serotonin-positive neurons contain nitric oxide (NO). Disruption of NO-mediated neurotransmission in the DRN by NO synthase inhibitors produces anxiolytic- and antidepressant-like effects in rats and also induces nonspecific interference with locomotor activity. We investigated the involvement of the 5-HT1A autoreceptor in the locomotor effects induced by NO in the DRN of male Wistar rats (280-310 g, N = 9-10 per group). The NO donor 3-morpholinosylnomine hydrochloride (SIN-1, 150, and 300 nmol) and the NO scavenger S-3-carboxy-4-hydroxyphenylglycine (carboxy-PTIO, 0.1-3.0 nmol) were injected into the DRN of rats immediately before they were exposed to the open field for 10 min. To evaluate the involvement of the 5-HT1A receptor and the N-methyl-D-aspartate (NMDA) glutamate receptor in the locomotor effects of NO, animals were pretreated with the 5-HT1A receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT, 8 nmol), the 5-HT1A receptor antagonist N-(2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl)-N-2-pyridinyl-cyclohexanecarboxamide maleate (WAY-100635, 0.37 nmol), and the NMDA receptor antagonist DL-2-amino-7-phosphonoheptanoic acid (AP7, 1 nmol), followed by microinjection of SIN-1 into the DRN. SIN-1 increased the distance traveled (mean ± SEM) in the open-field test (4431 ± 306.1 cm; F7,63 = 2.44, P = 0.028) and this effect was blocked by previous 8-OH-DPAT (2885 ± 490.4 cm) or AP7 (3335 ± 283.5 cm) administration (P < 0.05, Duncan test). These results indicate that 5-HT1A receptor activation and/or facilitation of glutamate neurotransmission can modulate the locomotor effects induced by NO in the DRN.

Changes in firing rate and firing pattern of midbrain dopaminergic neurons after lesioning of the dorsal raphe nucleus by 5,7-drhydroxytryptamine in adult rats / 西安交通大学学报·英文版

Objective: To study the effect of serotonergic efferent projection of the dorsal raphe nucleus (DRN) on the activity of substantia nigra pars compacta (SNc) and ventral tegmenta area (VTA) dopaminergic neurons after lesioning of the DRN by the neurotoxin 5,7-drhydroxytryptamine (5,7-DHT) in rat. Methods: The changes in the firing rate and firing pattern of SNc and VTA dopaminergic neurons were observed with extracellular recording in control and the lesioned rats. Results: The results showed that the mean firing rates of the fast-firing dopaminergic neurons of the SNc in control and the lesioned rats were (5.34±0.13) Hz (n=23) and (7.13±0.49) Hz (n=37), respectively. The mean firing rate of the fast-firing dopaminergic neurons of the SNc in the lesioned rats was significantly increased when compared to that of control rats (P<0.01), while the mean firing rate of the slow-firing dopaminergic neurons of the SNc did not change. The mean firing rates of dopaminergic neurons of the VTA in control and the lesioned rats were (5.27±0.38) Hz (n=35) and (3.6±0.2) Hz (n=52), respectively. Lesioning of the DRN induced a significant decrease in the mean firing rate of dopaminergic neurons of the VTA. The firing pattern of SNc and VTA dopaminergic neurons changed towards a more bursting or irrgular firing after the lesioning. Conclusion: These data suggest that the serotonergic efferent projections of the DRN significantly affect the activity of SNc and VTA dopaminergic neurons.

Changes in firing rate and firing pattern of midbrain dopaminergic neurons after lesioning of the dorsal raphe nucleus by 5,7-drhydroxytryptamine in adult rats / 药物分析学报

Objective To study the effect of serotonergic efferent projection of the dorsal rophe nucleus (DRN) on the activity of substantia nigro pars compacta (SNc) and ventral tegmenta area (VTA) dopaminergic neurons after lesioning of the DRN by the neurotoxin 5,7-drhydroxytryptamine (5,7-DHT) in rot. Methods The changes in the firing rote and firing pattern of SNc and VTA dopaminergic neurons were observed with extrocellular recording in control and the lesioned rats. Results The results showed that the mean firing rotes of the fast-firing dopaminergic neurons of the SNc in control and the lesioned rots were (5.34±0. 13 ) Hz (n = 23 ) and ( 7.13±0. 49 ) Hz (n=37), respectively. The mean firing rote of the fast-firing dopaminergic neurons of the SNc in the lesioned rats was significantly increased when compared to that of control rots (P<0.01), while the mean firing rote of the slow-firing dopaminergic neurons of the SNc did not change. The mean firing rotes of dopaminergic neurons of the VTA in control and the lesioned rots were (5.27±0. 38)Hz (n=35) and (3.6±0.2)Hz (n=52), respectively. Lesioning of the DRN induced a significant decrease in the mean firing rote of dopaminergic neurons of the VTA. The firing pattern of SNc and VTA dopaminergic neurons changed towards a more bursting or irrgular firing after the lesioning. Conlusion These data suggest that the serotonergic efferent projections of the DRN significantly affect the activity of SNe and VTA dopaminergic neurons.

Role of the serotoninergic system in the sodium appetite control

The present article reviews the role of the serotoninergic system in the regulation of the sodium appetite. Data from the peripheral and icv administration of serotoninergic (5-HTergic) agents showed the participation of 5-HT2/3 receptors in the modulation of sodium appetite. These observations were extended with the studies carried out after brain serotonin depletion, lesions of DRN and during blockade of 5-HT2A/2C receptors in lateral parabrachial nucleus (LPBN). Brain serotonin depletion and lesions of DRN increased the sodium appetite response, in basal conditions, after sodium depletion and hypovolemia or after beta-adrenergic stimulation as well. These observations raised the hypothesis that the suppression of ascending pathways from the DRN, possibly, 5-HTergic fibers, modifies the angiotensinergic or sodium sensing mechanisms of the subfornical organ involved in the control of the sodium appetite. 5-HTergic blockade in LPBN induced to similar results, particularly those regarded to the natriorexigenic response evoked by volume depletion or increase of the hypertonic saline ingestion induced by brain angiotensinergic stimulation. In conclusion, many evidences lead to acceptation of an integrated participation resulting of an interaction, between DRN and LPBN, for the sodium appetite control.

Este artigo revisa o papel do sistema serotoninérgico no controle do apetite ao sódio. Dados derivados da administração periférica e icv de agentes serotoninérgicos demonstraram a participação de receptores 5-HT2/3 na modulação do apetite ao sódio. Estas observações foram estendidas com os estudos realizados após a depleção cerebral de serotonina, lesões do NDR e durante o bloqueio 5-HT2A/2C no núcleo parabraquial lateral (NPBL). A depleção cerebral de serotonina e as lesões do NDR aumentaram o apetite ao sódio, em condições basais, após depleção de sódio, durante a hipovolemia ou após a estimulação beta-adrenérgica. Estas evidências suscitaram a hipótese de que a supressão de vias ascendentes do NDR, possivelmente 5-HT, alteram os mecanismos angiotensinérgicos e a atividade dos sensores de sódio do órgão subfornicial envolvidos no controle do apetite ao sódio. O bloqueio serotoninérgico no NPBL induziu a resultados similares, particularmente aqueles relacionados com a resposta natriorexigênica provocada pela depleção de volume ou o aumento da ingestão de salina hipertônica induzida pela estimulação angiotensinérgica cerebral. Em resumo, as evidências convergem para a admissão de uma participação integrada resultante da interação recíproca entre NDR e NPBL objetivando controlar o apetite ao sódio.

HYPOXIA INDUCED EXPRESSION OF FOS IN THE DISTAL CEREBROSPINAL FLUID CONTACTING NEURONS IN THE DORSAL RAPHE NUCLEUS OF THE RAT / 神经解剖学杂志

The effects of hypoxia on the expression of Fos was studied in the distal cerebrospinal fluid contacting neurons (CSF-CNs) in the dorsal raphe nucleus (DR) of the rat. The hypoxic models mimic 8000 m high level were established in a high-altitude decompression chamber. Injecting the tracer of CB ( chorela toxin B subunit) into the rat's lateral ventricles, we investigated the distribution of the distal CSF-CNs in the DR. The single and double immunohistochemistry staining with antibodies to Fos and CB were used to observe the expression of Fos in the distal CSF-CNs in the DR. The results showed that a large number of CB-like immunoreactive (-LI) neurons (31.16 ±3.36/per section) were located in the DR. In hypoxic experimental rats, the number of Fos-LI neurons was increased sharply (40.28 ±2.17/per section, P ＜0.05 compared to control rats), and a few CB/Fos-LI neurons (2.00 ±0.39/per section) could be observed in the DR. In control rats, several Fos-LI neurons (5.55 ±0.81/per section) and no CB/Fos-LI neuron could be observed in the DR. These results suggest that some of the distal CSF-CNs in the DR may play roles in transmitting information between brain and cerebrospinal fluid and modulating the function of brain following hypoxic stimulation.

The nNOS expression in rat distal CSF contacting neurons and its relationship with morphine withdrawal / 中国药理学通报

Aim To observe the neuron nitric oxide synthase(nNOS) expression in the distal cerebrospinal fluid contacting neurons(CSF-CNs) of rat brain parenchyma, and investigate the role of CSF-CNs in the development of morphine dependence and withdrawal.Methods Male adult Sprague-Dawley rats, weighed 260?20 g,were experimented with A 3 ?l volume of 30% cholera toxin subunit B with horseradish peroxidase(CB-HRP) was injected into one of the rats′lateral ventricles to trace and locate the distal CSF-CNs of rat brain parenchyma 48 hours before the animals were killed. All animals were perfused and the relative tissue of rats′brain was removed.Frozen serial coronal sections (40 ?m) were cut. Then TMB-ST reaction procedure was used to stain the CB-HRP positive neurons,followed by immunohistochemistry double-labeling of the nNOS with CB-HRP positive neurons. The withdrawal symptoms were observed and scored. The numbers of the CB-HRP, and CB-HRP/nNOS positive neurons on the same segmental brain sections were counted.Results The withdrawal symptoms of the withdrawal group were significant, scores of all signs were significantly higher than those of the dependence groups and control group(P

Cytotoxic Effect of 6-OHDA on the Dopaminergic Neurons and Astrocytes in the Dorsal Raphe Nucleus of the Rat / 대한해부학회지

This study was designed to clarify the cytotoxic effects of 6-hydroxydopamine (6-OHDA) on the dopaminergic neurons and astrocytes in the dorsal raphe nucleus (DRN), and to investigate neurodegenerative changes by immuno-histochemistry. Adult male rats (Sprague-Dawley strain) weighing from 250 to 350 g were used as experimental animals. 6-OHDA (100 micrometer dissolved in 0.1% ascorbic acid) was injected into the lateral ventricle of the rat brain with the Hamilton syringe. The control rats were treated with the similar volume of 0.1 % ascorbic acid. The rats were sacrificed at the 3rd, 5th, 10th and 20th day, respectively, after the injection of 6-OHDA. The cytotoxicity of 6-OHDA resulted in severe neurodegeneration of the dopaminergic neurons in the DRN. In the 3rd day, the dopaminergic fibers were dilated. In the 5th and 10th days, the dopaminergic fibers were depleted, and dopaminergic cell bodies were shrunken. In the 20th day, the dopaminergic cell bodies were almost completely disappeared. Astroglial reactions induced by 6-OHDA were also observed in the DRN. In the 5th day, astrocytes were significantly increased as compared with that of the control value. The value were reached at its maximum by the 20th day. Based on the present results, it suggests that 6-OHDA may act as a specific neurotoxin to dopaminergic neurons in the DRN, and induce severe neurodegenerative changes. Also, it suggests that the astroglial reaction in the DRN is gradually activated during the neurodegerative changes.

Cytotoxic Effect of 5, 7-DHT on the Endogenous Glial Cells in the Dorsal Raphe Nucleus of the Rat / 대한해부학회지

This study was designed to clarify the cytotoxic effects of 5, 7-dihydroxytryptamine (5, 7-DHT) on the serotonergic neurons in the dorsal raphe nucleus, and to investigate the glial reaction during the neurodegenerative changes by light and electron microscopy. Adult male rats (Sprague-Dawley strain) weighing from 250 g to 350 g each were used as experimental animal. 5, 7-DHT (200 mg dissolved in 0.9% NaCl) was injected into the lateral ventricle of the rat brain with the Hamiton syringe fixed on stereotaxic apparatus. The control rats were given with the similar volume of 0.9% NaCl. The rats were sacrified on the 1st, 3rd, 5th, 10th and 20th day after the injection of 5, 7-DHT. The results were as follows : Glial reactions induced by 5, 7-DHT were also observed in DRN. In early experimental stage, microglial reactions prevailed, whereas astroglial reactions were prevailing in later stage. In addition, microglial cells phagocytosed and removed the degenerated cells. However, astrocytes in DRN did not show phagocytotic activities such as microglial cells. Based on the results, author thought that 5, 7-DHT act as a specific neurotoxin to serotonergic neurons in DRN, and induces severe neurodegenerative changes. The glial reactions in DRN are activated during the neurodegerative changes, and show characteristic patterns of glial reactions.

Cytotoxic Effect of 5, 7-DHT on the Serotonergic Neurons in the Dorsal Raphe Nucleus of the Rat / 대한해부학회지

This study was designed to clarify the cytotoxic effects of 5, 7-dihydroxytryptamine (5, 7-DHT) on the serotonergic neurons in the dorsal raphe nucleus, and to investigate the glial reaction during the neurodegenerative changes by light and electron microscopy. Adult male rats (Sprague-Dawley strain) weighing from 250 g to 350 g each were used as experimental animal. 5, 7-DHT (200 microgram dissolved in 0.9% NaCl) was injected into the lateral ventricle of the rat brain with the Hamiton syringe fixed on stereotaxic apparatus. The control rats were given with the similar volume of 0.9% NaCl. The rats were sacrified on the 1st, 3rd, 5th, 10th and 20th day after the injection of 5, 7-DHT. The results were as follows : The cytotoxicity of 5, 7-DHT resulted in severe neurodegenerations of the serotonergic neurons. Most degenerated cells mainly showed necrotic findings, but a few of them exhibited apoptotic features. That is, in early stage of this experiment, the degenerated cells showed edematic changes of cytoplasm, but their nuclei were relatively seen intact. In late stage, the cells showed dark degenerative changes both in their cytoplasm and nuclei. Thereafter the cells were autolysed or phagocytosed by neighboring glial cells. Based on the results, author thought that 5, 7-DHT act as a specific neurotoxin to serotonergic neurons in DRN, and induces severe neurodegenerative changes. The glial reactions in DRN are activated during the neurodegerative changes, and show characteristic patterns of glial reactions.

SUBSTANCE P-LIKE PROJECTIONS FROM THE MIDBRAIN RAPHE NUCLEI AND PERIAQUEDUCTAL GRAY TO THE THALAMIC VENTRAL POSTERIOR NUCLEUS IN THE RATS—A COMBINED STUDY OF HRP TRACING AND IMMUNOCYTOCHEMICAL METHOD / 解剖学报

The afferant projections and their chemical nature from the midbrain raphe nuclei and periaqueductal gray to the thalamic ventral posterior nucleus were st- udied by HRP retrograde tracing combined with PAP immunocytochemical method Following injections of HRP into the thalamic ventral posterior nucleus, and reactions by HRP and PAP were performed subsequently. The double labelling cells containing HRP-positive granules and substance P-like immunoreactivity were found in various areas of brain, that is, the contralateral principal sensory nucleus of trigeminal nerve,nucleus gracilis,nucleus cuneatus, bilateral ventrola- teral divisions of the periaqueductal gray and midbrain raphe nuclei.A comparison of cell counts of double labeled cells with the total HRP-labeled cells indicated that the cells observed in the dorsal raphe nuclei were 21?7 in each rat, which constituted 48% of total HRP-labeling cells; The double labeling neurons in nuclei raphe centralis superior were found in two cases only, the cell bodies were very sparse and distributed mainly in the caudal area of these nuclei The double labeling cells were detected in bilateral ventrolateral divisions of periaqu- eductal gray, predominantly ipsilaterally. The double labeling cells observed in each rat were 26?9 ipsilaterally and 11?4 contralateraIly. The numbers of double labeling cells were 38% of the total counts of HRP-labeling cells in each side. These results above indicate that the thalamic ventral posterior nucleus receives afferent projections from midbrain raphe nuclei and ventrolateral division of periaqueductal gray and in the pathway, a part of substance P-positive neurons are exhibited. Therefore, this study provides evidence for existance ascending projection of substance P-like immunoreactivity.

DISTRIBUTION OF SUBSTANCE P-LIKE AND LEUCINE-E NKEPHALIN-LIKE IMMUNORE-ACTIVE NEURONS IN THE MIDBRAIN RAPHE NUCLEI OF THE RAT / 解剖学报

The different distributions of substance P(SP)-and leucine-enkephalin (L-ENK)-like immunoreactive neurons in the midbrain raphe nuclei of the rats have been studied, using PAP immunocytochemical technique. The results showed four main points: 1. SP-containing immunoreactive cell bodies were only seen in the most caudal part of the dorsal raphe nucleus, the average number of SP-positive cell bodies was about 167 in each animal. The L-ENK-like immunoreactive positive cell bodies were found almost in the whole length of the dorsal raphe nucleus, the L-ENK-positive cell bodies were about 132 in each animal. 2. L-ENK-Iike immunoreactive perikarya were mainly located in the midline area of the dorsal raphe nucleus. Besides the above areas, SP-containing immunoreactive cell bodies were also located bilaterally in the extended area from the raphe. 3. L-ENK-containing immunoreactive cell bodies constantly appeared in the median raphe nuclei, the average number was about 199 in each animal, but SP-containing immunoreactive bodies were only observed in this nucleus of two rats. 4. Some SP-like immunoreactive cell bodies or processes were observed in the vincinty of vascular walls, but the L-ENK positive neurons were not found similar results. The present experiment showed that the SP-like immunoreactive perikarya is located in the median raphe nuclei of the rat.