• Applied Microscopy
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• 제30권4호
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• pp.377-387
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• 2000

뇌실을 감싸는 뇌실막층은 대부분이 일반적인 뇌실막 세포로 이루어졌으나, 이들 세포 사이에 간혹 띠뇌실막 세포(tanycyte)가 분포하고 있다. 띠뇌실막세포는 일반적인 뇌실막세포와는 달리 뇌의 실질로 뻗은 매우 긴 기저돌기를 갖고 있다. 특히 제3뇌실의 뇌실막층에서 주로 연구된 띠뇌실막세포는 뇌실과 뇌실질의 혈관 혹은 신경세포와의 사이에 물질교환을 담당하는 것으로 추측되고 있으며, 띠뇌실막세포는 일반적인 뇌실막세포와는 달리 glial fibrillary acidic protein(GFAP)항체에 대해 양성반응을 보이는 것으로 알려져 있다. 본 연구는 면역조직화학 및 면역금표지법을 이용하여 박쥐 맨아래구역을 감싸는 뇌실막층에서 GFAP 항체에 대해 양성반응을 보이는 세포의 분포여부 및 이의 미세 구조를 확인하고자 시행하여 다음과 같은 결론을 얻었다. 세로 절단한 절편을 대상으로 GFAP 항체를 이용하여 면역염색한 후 광학현미경으로 확인한 결과 맨아래구역을 감싸는 뇌실막층에 GFAP 양성반응을 보이는 세포가 관찰되었으며, 특히 맨아래구역의 목부분에서 양성반응 세포가 많이 모여 있었다. GFAP양성반응을 보이는 세포들은 매우 긴 기저돌기를 갖고 있었으며, 기저돌기에서도 매우 강한 GFAP 양성반응을 보였다. 세포체에서는 주변부에서 양성반응을 보였다. 전자현미경하에서 맨아래구역 띠뇌실막세포는 주로 ependymal tanycytes였으며, 자유면에 소수의 미세융모 및 세포질돌기가 관찰되었으나, 섬모는 관찰되지 않았다. 기저부에 특징적으로 긴 기저돌기를 갖고 있었으며 이 돌기에는 중간세사 및 세로로 길게 달리는 사립체가 발달되어 있었고, 세포체와 돌기에 지방방울이 산재되어 있었다. 금입자를 표지한 GFAP항체를 사용하여 면역 염색한 후 전자현미경으로 관찰한 결과 세포체에서 핵을 둘러싸는 중간세사에서 금입자를 관찰할 수 있었으며, 나머지 핵을 비롯한 다른 세포소기관이나 세포질에서는 관찰되지 많았다. 또한 기저돌기에서도 중간세사에서만 금입자를 관찰할 수 있었다. 이와 같은 본 실험의 결과는 박쥐 맨아래구역 뇌실막층에도 띠뇌실막세포가 존재하고 있으며, 이 세포의 발달된 긴 기저돌기는 띠뇌실막세포가 일반적인 뇌실막세포와는 다른 기능을 동면동물인 박쥐의 맨아래구역에서도 수행하고 있음을 암시하고 있으나 이를 확인하기 위해서 더욱 자세한 연구가 필요한 것으로 사료된다.

• Journal of Acupuncture Research
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• 제20권6호
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• pp.89-102
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• 2003

Objective: The expression of CRF(corticotropin releasing factor), CRF-R(receptor) and CRF-BP(binding protein) in CNS neurons projecting to the stomach and ST36 using the pseudorabies virus in the rat was investigated. Methods: After survival times of 5 days following injection of PRV-Ba-Gal, The thirty rats were perfused, and their brain were frozen sectioned($30{\mu}m$). These sections were stained by PRV-Ba-Gal histochemical staining method and(or) CRF, CRF-R and CRF-BP immunohistochemical method. The common expressed areas of the brain projecting to the stomach and zusanli(ST36) following injection of PRV-Ba-Gal were observed with light microscope. Results: 1) The dense accumulation of CRF-immunoreactive terminals is seen in the area postrema, n. tractus solitarius, external zone of the median eminence, with some immunoreactive CRF also present in the internal zone. 2) Aggregates of CRF-R immunoreactive perikarya are found in area postrema, n. tractus solitarius, lateral reticular n., gigantocellular reticular n., locus coeruleues, paraventricular n. of hypothalamus, median eminence, preoptic n., arcuate n. and hind limb area of cerebral cortex. 3) Aggregates of CRF-BP immunoreactive perikarya are found in area postrema, n. tractus solitarius, lateral reticular nucleus, gigantocellular reticular n., locus coeruleues, paraventricular n. of hypothalamus, median eminence and arcuate n.. Conclusions : These results suggest that PRV-Ba-Gal labeled areas projecting to stomach and ST36 may be related to the central autonomic pathways. A part of CNS neurons projecting to the stomach and ST36 were related to expression of CRF, CRF-R and CRF-BP related to the stress in central autonomic center.

• 대한한의학회지
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• 제18권1호
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• pp.58-71
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• 1997

In order to the location and local arrangement of nerve cell bodies and nerve fibers projecting to the meridian points related to facial nerve paralysis in the rat using the neural tracers, CTB and WGA-HRP, labeled neurons the were investigated by immunohistochemical and HRP histochemical methods following injection of 2.5% WGA-HRP and 1% CTB into Hyopko$(S_6)$. Chichang$(S_4)$, Sugu$(GV_{26})$, Sajukkong$(TE_{23})$ and Yangbaek$(G_{14})$. Following injection of Hyopko$(S_6)$, Chichang$(S_4)$, labeled motor neurons were founded in facial nucleus, trigeminal motor nucleus, reticular nucleus and hypoglossal nucleus. labeled sensory neurons were founded in trigeminal ganglia and $C_{1-2}$ spinal ganglia. sympathetic motor neurons were found in superior cervical ganglia. Sensory fibers labeled in brainstem were found in mesencephalic trigeminal tract, sensory root of trigeminal nerve, oral, interpolar and caudal part of trigeminal nucleus, area postrema, nucleus tractus solitarius, lateral reticular nucleus and $C_{1-2}$ spinal ganglia. Following injection of Sugu$(GV_{26})$, labeled motor neurons were founded in facial nucleus. Labeled sensory neurons were founded in trigeminal ganglia and $C_{1-2}$ spinal ganglia. Sympathetic motor neurons were found in superior cervical ganglia. Sensory fibers labeled in brainstem were found in spinal trigeminal tract, trigeminal motor nucleus, mesencephalic trigeminal tract, oral. interpolar and caudal parts of trigeminal nucleus, area postrema, nucleus tractus solitarius, lateral reticular nucleus, dorsal part of reticular part and $C_{1-2}$ spinal ganglia. Following injection of Sajukkong$(TE_{23})$ and Yangbaek$(G_{14})$, labeled motor neurons were founded in facial nucleus, trigeminal motor nucleus. Labeled sensory neurons were founded in trigeminal ganglia and $C_{1-2}$ spinal ganglia. sympathetic motor neurons were found in superior cervical ganglia. Sensory fibers labeled in brainstem were found in oral, interpolar and caudal parts of trigeminal nucleus, area postrema, nucleus tractus solitarius, inferior olovary nucleus, medullary reticular field and lamina I-IV of $C_{1-2}$ spinal cord. Location of nerve cell body and nerve fibers projecting to the meridian points related to the facial nerve paralysis in the rats were found in facial nucleus and trigeminal motor nucleus. Sensory neurone were found in trigeminal ganglia and $C_{1-2}$ spinal ganglia. Sympathetic motor neurons were found in superior cervical ganglia. Sensory fibers labeled in brainstem were found in mesencephalic trigeminal tract, oral, interpolar and caudal parts of trigeminal nucleus, area postrema, nucleus tractus solitarius. lateral reticular nucleus, medullary reticular field.

• 대한수의학회지
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• 제37권3호
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• pp.479-487
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• 1997

The present study was carried out to localize the central nuclei innervating the rat colon using pseudorabies virus-Bartha strain which has been known as a very useful neurotracer. The results were as follows. The central nuclei innervating the proximal colon were premotor area, subfornical organ, preoptic area in telencephalon, and paraventricular nucleus, bed nucleus of stria terminalis, retrochiasmatic area in the diencephalon, and periaqueductal gray, Edinger-Westphal nucleus, tegmental nucleus in the mesencephalon, and parabrachial nucleus, locus ceruleus, A5 area, $K{\ddot{o}}lliker$-Fuse nucleus, magnocellular reticular nucleus in the metencephalon, and nucleus tractus solitarius, A1 noradrenergic cell group, dorsal motor nucleus of vagus nerve, nucleus ambiguus, area postrema in the myelencephalon. In the spinal cord, the thoracic division had some nuclei innervating the proximal colon. The nuclei innervating the distal colon were paraventricular nucleus of the diencephalon, Edinger-Westphal nucleus of midbrain, and parabrachial nucleus, locus ceruleus, A5 area, $K{\ddot{o}}lliker$-Fuse nucleus, magnocellular reticular nucleus of the metencephalon, and nucleus tractus solitarius, dorsal motor nucleus of vagus nerve, nucleus ambiguus, area postrema in the myelencephalon. In the spinal cord, thoracic, lumbar and sacral division innervated the distal colon.

• 동의생리병리학회지
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• 제23권4호
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• pp.805-817
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• 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.

• 동의생리병리학회지
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• 제25권2호
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• pp.217-226
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• 2011

The aim of this study is to identify central neural pathway of neurons following the projection to the large intestine and Hapgok(LI4) which is Won acupoint of the large intestine meridian of hand-yangmyeong. In this experiment, Bartha's strain of pseudorabies virus was used to trace central localization of neurons related with large intestine and acupoint(LI4) which has been known to be able to regulate intestinal function. The animals were divided into 3 groups: group 1, injected into the large intestine; group 2, injected into the acupoint(LI4); group 3, injected into the acupoint(LI4) after severing the radial, ulnar, median nerve. After four days survival of rats, PRV labeled neurons were identified in the spinal cord and brain by immunohistochemical method. First-order PRV labeled neurons following the projection to large intestine, acupoint(LI4) and acupoint(LI4) after cutting nerve were found in the cervical, thoracic, lumbar and sacral spinal cord. Commonly labeled neurons were labeled in the lumbosacral spinal cord and thoracic spinal cord. They were found in lamina V- X, intermediomedial nucleus and dorsal column area. The area of sensory neurons projecting 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 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, periaqueductal gray matter. In the diencephalon, PRV labeled neurons were marked mostly in the arcuate nucleus and median eminence. These results suggest that overlapped CNS locations are related with autonomic nuclei which regulate the functions of large intestine-related organs and it was revealed by tracing PRV labeled neurons projecting large intestine and related acupoint(LI4).

• The Korean Journal of Pain
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• 제13권2호
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• pp.143-148
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• 2000

Background: Trigeminovascular system is implicated in the pathophysiology of the headache in migraine. This study was designed to evaluate the pattern of Fos protein expression in trigeminal nociceptive central pathway after meningeal stimulation of rats by capsaicin. Methods: The expression of Fos protein was examined by immunohistochemistry in thalamus, brainstem and upper cervical cord (at three levels corresponding to obex, 0.8 mm and 2 mm below obex) 2 hours after intracisternal injection of either diluted capsaicin solution (0.1 ml, $61{\mu}g/ml$) or normal saline (0.1 ml) through a catheter placed in the cisterna magna, or following epidural instillation of diluted capsaicin solution in urethane-anesthetized Sprague-Dawley rats. Results: Fos immunoreactivity was strongly expressed within lamina I, II of bilateral trigeminal nucleus caudalis (TNC) after cisternal capsaicin injection and magnitude of expression was greatest at level 2.0 mm below obex. Epidural capsaicin caused much less labelling than cisternal capsaicin. Fos positive cells were also observed in area postrema, nucleus of the solitary tract, medullary reticular nucleus and midline nuclear groups of the thalamus with similar intensity between capsaicin and control group. Conclusions: These results indicate that the injection of capsaicin into the cisterna magna is an effective stimulus for the induction of Fos protein within TNC through activation of trigeminovascular afferents and this animal model can be useful for the evaluation of the pathophysiology and drug development in migraine and related headache.

• Journal of Acupuncture Research
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• 제17권2호
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• pp.261-276
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• 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.

• BMB Reports
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• 제57권3호
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• pp.149-154
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• 2024

The stomach has emerged as a crucial endocrine organ in the regulation of feeding since the discovery of ghrelin. Gut-derived hormones, such as ghrelin and cholecystokinin, can act through the vagus nerve. We previously reported the satiety effect of hypothalamic clusterin, but the impact of peripheral clusterin remains unknown. In this study, we administered clusterin intraperitoneally to mice and observed its ability to suppress fasting-driven food intake. Interestingly, we found its synergism with cholecystokinin and antagonism with ghrelin. These effects were accompanied by increased c-fos immunoreactivity in nucleus tractus solitarius, area postrema, and hypothalamic paraventricular nucleus. Notably, truncal vagotomy abolished this response. The stomach expressed clusterin at high levels among the organs, and gastric clusterin was detected in specific enteroendocrine cells and the submucosal plexus. Gastric clusterin expression decreased after fasting but recovered after 2 hours of refeeding. Furthermore, we confirmed that stomachspecific overexpression of clusterin reduced food intake after overnight fasting. These results suggest that gastric clusterin may function as a gut-derived peptide involved in the regulation of feeding through the gut-brain axis.

• Journal of Acupuncture Research
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• 제18권2호
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• pp.51-66
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• 2001

The purpose of this morphological studies was to investigate the central neural pathway projecting to the acupoints $B_{62}$ and $K_6$ using the neuroanatomical method following injection of transsynaptic neurotropic virus, pseudorabies virus(PRV-Ba and PRV-Ga) into the $B_{62}$ and $K_6$. After survival times of 96 hours following injection into the twenty rats with PRV-Ba(Bartha strain) and PRV-Ga(Bartha strain, ${\beta}$-galacidodase insertion). They were perfused, and their spinal cord and brain were frozen sectioned($30{\mu}m$). These sections were stained by X-gal histochemical and PRV immunohistochemical staining method, and observed with light microscope. The results were as follows : 1. In spinal cord, overlaped PRV-Ba and PRV-Ga labeled neurons projecting to the $B_{62}$ and $K_6$ were founded in thoracic, lumbar and sacral spinal segments. In thoracic spinal segments, Densely labeled areas were founded in lamina IV, V, VII(intermediolateral nucleus) and X areas. In lumbar segemnts, labeled areas were founded in lamina II, IV, V and X areas. In sacral spinal segments, labeled areas were founded in lamina IV, V and VI areas. 2. In brain, overlaped PRV-Ba and PRV-Ga labeled neurons projecting to the $B_{62}$ and $K_6$ were founded in the $A_1$ noradrenalin cells/$C_1$ adrenalin cells/caudoventrolateral reticular nucleus, rostroventrolateral reticular nuclens, nucleus tractus solitarius, area postrema, raphe obscurus nucleus, raphe paltidus nucleus, raphe magnus nucleus, lateral paragigantoceltular nucleus, lateral rcticular nucleus, gigantocellular nucleus, locus coeruleus, subcoeruleus nucleus, motor trigeminal nucleus, Kolliker-Fuse nucleus, $A_5$ cell group, central gray matter, oculomotor nerve, paraventricular hypothalamic nucleus, median eminence, amygdaloid nucleus, frontal cortex, forelimb area, hindlimb area, 1, 2 areas of parietal cortex and granular and agranular cortex. This results were suggest that overlaped PRV-Ba and PRV-Ga labeled areas projecting to the $B_{62}$ and $K_6$ may be related to the emotional relay pathway in the central autonomic center.