• Animal cells and systems
• /
• v.6 no.2
• /
• pp.159-165
• /
• 2002

Projections from the prefrontal cortex to subdivisions of the dorsal raphe nucleus were investigated in the rat using retrograde and anterograde tracing methods. A retrograde tracer, gold-conjugated horseradish peroxidase (WGA-apo-HRP-gold), was injected into each subdivision of the dorsal raphe including lateral wing, dorsomedial, and ventromedial areas. The majority of retrogradely labeled cells were located in the prelimbic, infralim-bic, and dorsal peduncular areas of the medial prefrontal cortex. A few cells were also identified in the cingulate, various regions of the orbital, and agranular insular cortices. Secondly, an anterograde tracer, Phaseolus vulgaris leucoagglutinin (PHA-L), was injected into the medial prefrontal cortex involving the prelimbic or infralimbic areas. Axonal fibers with varicosities were identified in all subdivisions of the DR including the lateral wing, dorsomedial, and ventromedial areas. Projections were bilateral, with ipsilateral predominance. Axonal fibers were observed at the lateral border of medial longitudinal fasciculus or in the interfascicular region at the midline. The present findings demonstrate that both the midline and lateral wing regions of the dorsal raphe nucleus receive excitatory input from cognitive and emotional centers of the cerebral cortex.

• Journal of Acupuncture Research
• /
• v.17 no.2
• /
• pp.261-276
• /
• 2000

본 실험은 pseudorabies 바이러스 (PRV) 의 Bartha strain 을 안면신경의 측두지, 하지를 지배하는 신경 (좌골신경) 및 상지를 지배하는 신경 (요골, 척골, 정중신경) 에 주입한 후 4 일간의 생존시간이 경과한 후 척수와 뇌를 적출하여 동결절편을 제작한 후 면역조직화학적 염색기법과 X-gal 조직화학 염색법을 시행하여 염색된 신경세포체를 척수와 뇌에 투사된 공통영역을 관찰하고 두침의 영역중 하나인 운동구와 사지와의 관계에 대한 실험적 증거를 제시하고자 시행하였다. 위의 실험에서 얻어진 결과는 아래와 같다. 1. 안면신경의 측두지, 하지를 지배하는 신경 (좌골신경) 및 상지를 지배하는 신경 (요골, 측골, 정중신경) 에서 투사된 공통된 영역은 척수에서 경수의 층판 1-IV, 흉수의 intermediolateral nucleus(IML), dorsal nucleus(D) 및 층판 X, 요수의 층판 IV, V, 천수의 층판 IV, V, IX, X 등의 영역에서 관찰되었고, 뇌줄기에서는 caudoventrolateral reticular nucleus(CVL), nucleus solitary tract(Sol), rostroventrolateral nucleus(RVL), area postrema(AP), raphe nuclei(raphe pallidus, raphe obscurus, raphe magnus), inferior olivary nucleus 의 등쪽부분 (gigantocellular reticular nucleus, Gi), Kolliker-Fuse nucleus(KF), central gray(CG), dorsal raphe nucleus (DR) and A5 영역에 표지된다. 또한 paraventricular hypothalamic nucleus(PRV) 와 lateral hypothalamic reticular nucleus(LH)에서도 관찰되고 locus coeruleus(LC) 와 subcoeruleus nuc!eus(SubCA) 에서도 관찰된다.

• Korean Journal of Biological Psychiatry
• /
• v.3 no.2
• /
• pp.191-202
• /
• 1996

This study was aimed to charily the change of NADPH-diaphorase(NADPH-d) and neuropeptide Y(NPY) associated with aging of the rots. To verily the effect of aging of NPY and NADPH-d-positive neurons in the brain stem, the neurons were stained by the immunohistochemical and histochemical method. In the aged group, the number of NADPH-d-positive neurons was significantly decreased in substantia nigra lateralis, supragenual nucleus, raphe magnus nucleus and raphe obscurus nucleus as compared with control group. The number of NADPH-d-positive neurons was not significantly decreased in superior colliculus, interpeduncular nucleus, central gray. dorsal raphe nucleus, retrorubral nucleus, pedunculopontine tegmental nucleus, laterodorsal tegmental nucleus, pontine reticular nucleus, prepositus hypoglossal nucleus and nucleus solitarius of the aged rats. The NADPH-d and NPY-positive neurons were found in the interpeduncular nucleus, central gray, substantia nigra lateralis, laterodorsal tegmental nucleus, nucleus solitarius, raphe magnus nucleus, raphe obscurus nucleus of the control and aged groups. The coexistence of NADPH-d and NPY in the some cell was not found in the brain stem of both groups.

• Animal cells and systems
• /
• v.8 no.3
• /
• pp.221-229
• /
• 2004

The projections from the dorsal raphe (DR) to the locus coeruleus (LC) or vice versa were analyzed in the rat using an anterograde tracer, Phaseolus vulgaris leucoagglutinin (PHA-L) combined with serotonin (5-hydroxytryptamine, 5-HT) or dopamine-beta-hydroxylase (DBH) immunostaining. Following the injection of PHA-L into the middle DR, DR-originating fibers with varicosities have contacted DBH-immunolabeled cells in the rostral, middle, and caudal LC. Axon terminals were also observed in the subcoeruleus nucleus. When the PHA-L injection was confined within the caudal DR, axonal fibers with varicosities were observed mainly at the rostral pole of the LC. Following the injection of PHA-L into the caudal, principal LC, labeled fibers with varicosities have contacted 5-HT-immunolabeled neurons at dorsomedial, ventromedial, lateral wing, and caudal sub-divisions of the DR. The present anterograde study suggests that the DR or the LC nuclei communicate with each other in order to perform a variety of functions including vigilance, analgesia, and stress responses.

• The Korean Journal of Physiology
• /
• v.24 no.2
• /
• pp.403-411
• /
• 1990

The present investigation was performed to see a possible influence of the dorsal raphe nucleus (DRN) on pancreatic exocrine secretion in anesthetized rats since the DRN had been known to exert a regulatory mechanism on sympathetic activity which was known to be very important for pancreatic exocrine secretion, particularly in rats. Twenty-nine Sprague-Dawley rats fasted for 24 hours were anesthetized by i.p. injection of 1 g/kg of urethane. The pancreatic duct was cannulated to collect pancreatic juice while bile juice was diverted into the jejunum. The duodenopyloric junction was tightly ligated. After surgery for collection of pancreatic exocrine secretion and recording of carotid blood pressure, a coaxial electrode was stereotaxically inserted in the DRN with a guide of a brain atlas. And then, electrical stimulus of biphasic square wave with 2 v, 2 msec, 40 Hz was applied on the electrode for 10 minutes. Pancreatic volume flow and protein output secreted in 10 min were measured. Either bilateral cervical vagotomy or spinal cord transection at the level of $C4{\sim}C5$ was performed 20 min prior to stimulation of the DRN. 1) Electrical stimulation of the DRN resulted in significant (p<0.05) increase in pancreatic volume flow and protein output. These stimulatory effects were not affected by cervical vagotomy but completely abolished by cervical cord transection. 2) Electrical stimulation of the DRN also resulted in significant (p<0.05) rise of blood pressure of the carotid artery. The hypertensive effect was not affected by cervical vagotomy but completely abolished by cervical cord transection. The results strongly suggest that the DRN, a part of the central serotonergic system, could exert a stimulatory influence on pancreatic exocrine secretion by increasing the sympathetic activity in anesthetized rats.

• The Korean Journal of Physiology and Pharmacology
• /
• v.25 no.5
• /
• pp.449-457
• /
• 2021

The sleep-wake cycle is regulated by the alternating activity of sleep- and wake-promoting neurons. The dorsal raphe nucleus (DRN) secretes 5-hydroxytryptamine (5-HT, serotonin), promoting wakefulness. Melatonin secreted from the pineal gland also promotes wakefulness in rats. Our laboratory recently demonstrated that daily changes in nitric oxide (NO) production regulates a signaling pathway involving with-no-lysine kinase (WNK), Ste20-related proline alanine rich kinase (SPAK)/oxidative stress response kinase 1 (OSR1), and cation-chloride co-transporters (CCC) in rat DRN serotonergic neurons. This study was designed to investigate the effect of melatonin on NO-regulated WNK-SPAK/OSR1-CCC signaling in wake-inducing DRN neurons to elucidate the mechanism underlying melatonin's wake-promoting actions in rats. Ex vivo treatment of DRN slices with melatonin suppressed neuronal nitric oxide synthase (nNOS) expression and increased WNK4 expression without altering WNK1, 2, or 3. Melatonin increased phosphorylation of OSR1 and the expression of sodium-potassium-chloride co-transporter 1 (NKCC1), while potassium-chloride co-transporter 2 (KCC2) remained unchanged. Melatonin increased the expression of tryptophan hydroxylase 2 (TPH2, serotonin-synthesizing enzyme). The present study suggests that melatonin may promote its wakefulness by modulating NO-regulated WNK-SPAK/OSR1-KNCC1 signaling in rat DRN serotonergic neurons.

• Journal of Acupuncture Research
• /
• v.15 no.2
• /
• pp.117-133
• /
• 1998

The purpose of this morphological study was to investigate the relationship to facial nerve and LI4 related to the large intestine meridian. The common locations of the spinal cord and brain projecting to the LI4 and facial nerve were observed fallowing injection of transsynaptic neurotropic virus, pseudorabis virus(PRV), into the LI4 and facial nerve of the rat. After survival times of 96 hours following injection of PRV, the rats were perfused, and their spinal cord and brain were frozen sectioned(30${\mu}m$). These sections were stained by PRV immunohistochemical staining method, and observed with light microscope The results were as follows: 1. The PRV labeled spinal cord segments projecting to the LI4 and facial nerve were founded in cervical, thoracic, lumbar and sacral segments. Dense labeled areas of each spinal cord segment were founded in lamina IV, V, X, lateral spinal nucleus, intermediolateral nucleus and dorsal nucleus. 2. The PRV labeled medulla oblongata projecting to the LI4 and facial nerve were founded in the A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nucleus, rostroventrolateral reticular nucleus, medullary reticular nucleus, nucleus tractus solitarius, raphe obscurus nucleus, raphe pallidus nucleus, raphe magnus nucleus, gigantocellular nucleus, lateral paragigantocellular nucleus, and spinal trigeminal nucleus.

• Korean Journal of Acupuncture
• /
• v.17 no.1
• /
• pp.101-121
• /
• 2000

The purpose of this morphological studies was to investigate the relation to the meridian, acupoint and nerve. The common locations of the spinal cord and brain projecting to the the gallbladder, GB34 and common peroneal nerve were observed following injection of transsynaptic neurotropic virus, pseudorabies virus(PRV), into the gallbladder, GB34 and common peroneal nerve of the rabbit. After survival times of 96 hours following injection of PRV, the thirty rabbits were perfused, and their spinal cord and brain were frozen sectioned($30{\mu}m$). These sections were stained by PRV immunohistochemical staining method, and observed with light microscope. The results were as follows: 1. In spinal cord, PRV labeled neurons projecting to the gallbladder, GB34 and common peroneal nerve were founded in thoracic, lumbar and sacral spinal segments. Densely labeled areas of each spinal cord segment were founded in lamina V, VII, X, intermediolateral nucleus and dorsal nucleus. 2. In medulla oblongata, The PRV labeled neurons projecting to the gallbladder, GB34 and common peroneal nerve were founded in the A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nucleus, rostroventrolateral reticular nucleus, medullary reticular nucleus, dorsal motor nucleus of vagus nerve, nucleus tractus solitarius, raphe obscurus nucleus, raphe pallidus nucleus, raphe magnus nucleus, gigantocellular nucleus, lateral paragigantocellular nucleus, principal sensory trigeminal nucleus and spinal trigeminal nucleus. 3. In Pons, PRV labeled neurons were parabrachial nucleus, Kolliker-Fuse nucleus and cochlear nucleus. 4. In midbrain, PRV labeled neurons were founded in central gray matter and substantia nigra. 5. In diencephalon, PRV labeled neurons were founded in lateral hypothalamic nucleus, suprachiasmatic nucleus and paraventricular hypothalamic nucleus. 6. In cerebral cortex, PRV labeled neuron were founded in hind limb area.This results suggest that PRV labeled common areas of the spinal cord projecting to the gallbladder, GB34 and common peroneal nerve may be first-order neurons related to the somatic sensory, viscero-somatic sensory and symapathetic preganglionic neurons, and PRV labeled common area of the brain may be first, second and third-order neurons response to the movement of smooth muscle in gallbladder and blood vessels.These PRV labeled neurons may be central autonomic center related to the integration and modulation of reflex control linked to the sensory system monitoring the internal environment, including both visceral sensation and various chemical and physical qualities of the bloodstream. The present morphological results provide that gallbladder meridian and acupoint may be related to the central autonomic pathways.

• Journal of Acupuncture Research
• /
• v.17 no.2
• /
• pp.95-117
• /
• 2000

The purpose of this morphological studies was to investigate the relation between the meridian, acupoints and viscera using neuroanatomical tracers. The common locations of the spinal ganglia, sympathetic chain ganglia, spinal cord and brain projecting to the large intestine meridian were observed following injection of transganglionic tracer, WGA-HRP and transsynaptic neurotropic virus, pseudorabies virus(PRV), Bartha strain(Ba) and PRV-Ba-Gal (Galactosidase)) into the the large intestine(cecum, colon and rectum), ST37 and LI4. After survival times of 96 hours following injection into the thirty rats with WGA-HRP, PRV-Ba and PRV-Ba-Gal. They were perfused, and their spinal ganglia, sympathetic chain ganglia, spinal cord and brain were frozen sectioned($30{\mu}m$). These sections were stained by HRP and X-gal histochemical and PRV immunohistochemical staining method, and observed with a light microscope. The results were as follows : 1. WGA-HRP labeled neurons innervating the large intestine were observed bilaterally within the T13-L4 sympathetic chain ganglia, and T9-11 spinal ganglia. WGA-HRP labeled neurons innervating ST37 were observed within the L3-5 sympathetic chain ganglia, and L2-4 spinal ganglia. WGA-HRP labeled neurons innervating LI4 were observed in the middle cervical ganglion and stellate ganglion, and C5-8 spinal ganglia. 2. In spinal cord, PRV-Ba labeled neurons projecting to the large intestine, ST37 and LI4 were found in thoracic, lumbar and sacral spinal segments. Densely labeled areas of each spinal cord segment were founded in lamina N, V, VII(intermediolateral nucleus), Ⅸ, X and dorsal nucleus. 3. In medulla oblongata, PRV-Ba and PRV-Ba-Gal labeled neurons projecting to the large intestine, ST37 and LI4 were commonly found in the A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nucleus, dorsal motor nucleus of vagus nerve, nucleus tractus solitarius, raphe obscurus nucleus, raphe pallidus nucleus, raphe magnus nucleus and gigantocellular nucleus. 4. In pons, PRV-Ba and PRV-Ba-Gal labeled neurons were commonly found in locus coeruleus, Kolliker-Fuse nucieus and A5 cell group. 5. In midbrain, PRV-Ba and PRV-Ba-Gal labeled neurons were commonly found in central gray matter. 6. In diencephalon, PRV-Ba and PRV-Ba-Gal labeled neurons were commonly found in paraventricular hypothalamic nucleus. These results suggest that PRV-Ba and PRV-Ba-Gal labeled common areas projecting to the large intestine may be correlated to that of the large intestine meridian, ST37 and LI4. Especially, These morphological results provide that interrelationship of meridian-acupoints -viscera may be related to the central autonomic pathways.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.23 no.4
• /
• pp.805-817
• /
• 2009

This study was to investigate central localization of neurons projecting to the urinary bladder and urinary bladder-related acupoints(Wijung, $BL_{40}$) and neurons of immunoreactive to hormones and hormone receptors regulating urinary bladder function by using peudorabies virus(PRV). In this experiment, Bartha's strain of pseudorabies virus was used in rats to trace central localization of urinary bladder-related neurons and urinary bladder-related acupoints($BL_{40}$) which can regulate urinary system. PRV was injected into the urinary bladder and acupoints($BL_{40}$) related urinary system. After six days survival of rats, mainly common labeled neurons projecting to the urinary bladder and urinary bladder-related acupoints were identified in spinal cord, medulla, pons and diencephalon by PRV immunohistochemical staining method. First-order PRV labeled neurons projecting to urinary bladder and urinary bladder-related acupoints were found in the cervical, thoracic, lumbar and sacral spinal cord. Commonly labeled preganglionic neurons were labeled in the lumbosacral spinal cord and thoracic spinal cord. They were found in the lateral horn area(sacral parasympathetic nucleus and intermediolateral nucleus), lamina V-X, intermediomedial nucleus and dorsal column area. The area of sensory neurons projecting to urinary bladder and Wijung($BL_{40}$) was L5-S2 spinal ganglia and T12-L1 spinal ganglia, respectively. In the brainstem, the neurons were labeled most evidently and consistently in the nucleus of tractus solitarius, area postrema, dorsal motor nucleus of vagus nerve, reticular nucleus, raphe nuclei(obscurus, magnus and pallidus), C3 adrenalin cells, parapyramidal area(lateral paragigantocellular nucleus), locus coeruleus, subcoeruleus nucleus, A5 cell group, Barrington's nucleus and periaqueductal gray matter. In the diencephalon, PRV labeled neurons were marked mostly in the paraventricular nucleus and a few ones were in the lateral hypothalamic nucleus, posterior hypothalamic nucleus, ventromedial hypothalamic nucleus, arcuate nucleus, median eminence, perifornical nucleus, periventricular nucleus and suprachiasmatic nucleus. In cerebral cortex, PRV labeled neurons were marked mostly in the frontal cortex, 1,2 area, hind limb area, agranular insular cortex. Immunoreactive neurons to Corticotropin releasiing factor(CRF), Corticotropin releasiing factor-receptor(CRF-R), c-fos and serotonin were a part of labeled areas among the virus-labeled neurons of urinary bladder and Wijung($BL_{40}$). The commonly labeled areas were nucleus tractus solitarius, area postrema, reticular nucleus, raphe nuclei(obscurus, magnus and pallidus), locus coeruleus, A5 cell group, Barrington,s nucleus, arcuate nucleus, paraventricular nucleus, frontal cortex 1, 2 area, hind limb, and perirhinal(agranular insular) cortex. These results suggest that overlapped CNS locations are related with autonomic nuclei which regulate the functions of urinary bladder-relate organs and it was revealed by tracing PRV labeled neurons projecting urinary bladder and urinary bladder-related acupoints. These commonly labeled areas often overlap with the neurons connected with hormones and hormone receptors related to urination.