• The Journal of Korean Physical Therapy
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• v.11 no.2
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• pp.81-92
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• 1999

중추신경계의 미주신경동쪽핵(DMV)내 유사핵분열후 신경세포로 외래 유전자를 전달하는 매개체로서 pseudorabies 바이러스(PRV)의 유전자 조작기술은 흰쥐의 결장내로 PRV를 주입시킨 후 복잡한 신경로 추적에 관한 연구에서 하나의 바이러스에 의해 얻어지는 것보다 더욱 유용한 결과산출이 가능하게 하였다. 본 연구에서는 흰쥐의 생체내 실험모델로 하나의 바이러스 또는 이중 바이러스 주입에 PRV의 유전자 조작된 2종 바이러스를 사용하였다. 이 2종의 바이러스는 PRV의 Bartha 종에서 유래되었지만 면역조직화학적으로 검출할 수 있는 동일한 유전산물을 산출할 수 있도록 구성되었다. PRV-BaBlu는 PRV 게놈의 Us 구역 중 gC 자리에 lacZ 유전자를 삽입하여 산출되었는데 $\beta-galactosidase$ 발현은 이 바이러스에 감염된 신경원의 독특한 표시자로 나타났다. PRV-D는 2가지 단계에 의해 조성되었는데 첫째, PRV-Bartha의 Us 구역의 일부 유전자를 제거하고, 야생형인 PRV-Be DNA로 복구시켰는데 이로써 PRV-D는 PRV-Bartha 또는 PRV-Bablu에 존재하고 있지 않는 외피 당단백질인 gE와 gI를 지니게 되었다. 본 연구의 결과는 다음과 같았다. 첫째, PRV-D의 개별적 접종에 의해 얻어진 감염은 PRV-BaBlu에 의한 동일 신경회로의 감염보다 유의하게 빨랐다. 둘째, 유전자 조작된 PRV의 변이종은 변이종 상호간 및 부모 바이러스와 상이하였다. PRV-D는 PRV-Bartha 또는 PRV-BaBlu보다 감염독성이 더 강했고, PRV-BaBlu는 PRV-Bartha보다 감염독성이 약했다. 셋째, 결장을 지배하는 미주신경동쪽핵내 신경원은 변이종 바이러스들을 동시에 접종하였을 경우 이중감염을 나타내었다.

• The Journal of Korean Physical Therapy
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• v.9 no.1
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• pp.59-70
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• 1997

The ability of neurotropic alpha herpesviruses to replicate within synaptically linked neurons has made these pathogens valuable tools for transneuronal analysis. Recent studies suggest that unique gene products expressed by genetically engineered strains of virus may permit the use of multiple strains in complex tracing paradigms. In the present study we have examined the invasiveness of two genetically engineered strains of the swine pathogen known as pseudorabies virus(PRV). The two strains were isogenic with the attenuated Bartha strain of PRV; in one strain a lacZ reporter gene was inserted into the gC locus (PRV-BaBlu; $4.75\times10^8pfu/ml$) contrained a PRV envelope glycoprotein gene that was absent in PRV-BaBlu. Simultaneous or temporally separated sequential injection of $4\mu\ell$ of each strain into the ventral wall of the stomach produced a predictale course of retrograde synaptic infection. The results were as follows: 1. PRV-BaBlu and PRV-D infected the dorsal motor nucleus of vagus nerve(DMV) and paraventricular nucleus(PVN). 2. Invasion and replication of PRV-D occured at a faster rate than the parental strain or PRV-BaBlu. 3. PRV-D was much more virulent than PRV-BaBlu or the parental strain. 4. Co-injection of PRV-D and PRV-BaBlu produced an infection that was more virulent than that produced by the parental strain (PRV-Bartha), 5. Neurons in DMV were permissive to co-infection with PRV-D and PRV-BaBlu when they were injected simultaneously into the same site. 6. Replication of PRV-BaBlu was compromised by prior infection of the same circuit with PRV-D. 7. Prior infection of neurons with PRV-D maked them resistant to infection with PRV-BaBlu.

• Toxicological Research
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• v.22 no.4
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• pp.403-409
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• 2006

To localize the connection between intergeniculate nucleus and suprachisasmatic nucleus through geniculohypothalamic tract in postnatal mongolian gerbil, we injected lacZ inserted PRV-Bartha strain into suprachiasrnatic nucleus and tried to immunostain against it with Rb134 and mouse $anti-{\beta}-galactosidase$. The numbers of immunoreactive neurons in intergeniculate leaflet were $8{\pm}3.2$ in P1 Period, $10{\pm}4.1$ in P3 Period and $13{\pm}6.2$ in P7 Period, and was statistically significant (p<0.05) and had tendency to increase with time consuming. The results showed that intergeniculate leaflet had projected some axons into suprachiasrnatic nucleus through geniculohyptothalamic tract in postnatal mongolian gerbil. But we could not exclude the possibility of direct projections from dorsal and ventral geniculate nuclei into suprachisamatic nucleus completely.

• The Journal of Korean Physical Therapy
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• v.10 no.2
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• pp.113-125
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• 1998

The neurotropic psudorabies virus(PRV) to replicate within neurons is very useful pathogen for neuronal tracing. I carried out this study to investigate the parametric analysis on the viral infection in the rat circadian control center with two genetically engineered strains out of PRV. The two strains are isogenic with the attenuated Bartha strain of PRV ; in one strain a lacZ reporter gene was inserted into the gC locus (PRV-BaBlu ; $4.75\times10^8pfu/ml$) and the other strain contained a PRV envelope glycoprotein gene(PRV-D ; $2.5\times10^8pfu/ml$) theat is absent in PRV-BaBlu. simultaneous or temporally separated sequential injection of$2{\mu}l$ of each strain into the vetreous body of eye produced a course of transsynaptic infection of retinohypothalamic circuitry. The results were as follows; 1. PRV-BaBlu and PRV-D infected the suprachiasmatic nucleus in hypothalamus and intergeniculate leaflet in lateral geniculate nucleus of thalamus. 2. The rate of PRV infection was dependent upon PRV strain. 3. Pre-infected neurons by PRV-D were interfered with the replication of PRV-BaBlu. 4. Dual injection of PRV-D and PRV-BaBlu showed more virulent than the parental strain.

• The Journal of Korean Physical Therapy
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• v.23 no.5
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• pp.35-41
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• 2011

Purpose: This study was carried out to investigate the spatiotemporal localization of the amygdaloid nucleus innervating the rat stomach using PRV-BaBlu, which has been known to be an excellent type of neurotracer with the ability to transpass the neuronalsynaptic cleft. Methods: Ninety Sprague-Dawley rats (250~300 g) that were injected with PRV-BaBlu into the stomach were randomly divided into 3, 4 and 5 day groups (each group n=30). $2{\mu}l$ of PRV-BaBlu, a genetically modified strain of PRV-Bartha with the lac-Z gene,was injected into the rat stomach and immunostained with a mouse anti-${\beta}$-galactosidase at 3, 4 and 5 days after the virus injection. Results: The PRV-BaBlu infected the neurons in the amygdaloid nucleus, and the degree of viral infection in experimental animals showed a tendency to increase significantly with time (p<0.05). The neurons between the left and right amygdaloid nucleus significantly differ (p<0.05). Conclusion: This showed that PRV-BaBlu was an excellent neurotracer for localizing the amygdaloid nucleus, and the amygdaloid nucleus has a sensory input and motor output on stomach movement, influencing emotional behavior.

• Journal of Acupuncture Research
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• v.18 no.2
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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.

• Korean Journal of Veterinary Research
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• v.40 no.3
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• pp.463-470
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• 2000

Until now peudorabies virus(PRV) has been used a neurotracer, because of it's properties of retrograde & anterograde transport. But it's anterograde transfort is not perfect, so we tested the applicability of the Bartha strain of PRV(PRV-Ba) isolated from South Korea as a neurotracer in the visual system. We performed immunohistochemical study of the rabbit brain after intravitreal injection of the PRV-Ba. After given survival time(24, 48, 72, 96, 120, 144hrs), the brain was removed and processed immunohistochemical stain for PRV-Ba. The strong PRV immunoreactivity(PRV-ir) were almost oberserved contralaterally in oculomotor neurons, fro example Edinger-Westphal nucleus, trigerminal nucleus of pons and peritrigerminal zone but locus of innervating sensitive neurons. The latter were weak positive and selective. PRV-Ba immunoreactive neurons were stained strongly in nucleus compared to cytoplasm. This study suggests that PRV-Ba isolated from South Korea is also a useful neurotracer in the motor innervated system like other PRV-strain.

• The Journal of Korean Medicine
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• v.22 no.2
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• pp.31-40
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• 2001

This experimental studies was to investigate location of labeled neurons in CNS following injection of pseudorabies virus(PRV), Bartha strain, into the uterus and Sanyinjiao(Sp6) of rats. After survival times of 4-5 days following the injection of PRV, the rats were perfused, and their brain and spinal cord were frozen sectioned($30\mu\textrm{m}$). These sections were stained by PRV immunohistochemical staining methods, and observed with light microscope. The results were as follows: 1. In the spinal cord, overlap areas of PRV labeled neurons projecting to uterus and Sp6 were observed in lamina VII, IX and X areas of cervical segments. In thoracic segments, overlap areas were observed in lamina IV, VII, X and intermediolateral n.. In lumbar segments, overlap area of PRV labeled neurons were observed in lamina I, V-VII, IX, X and intermediolateral n.. In sacral segments, overlap areas of PRY labeled neurons were observed in lamina N, V, VII, X and sacral parasympathetic n.. 2. In the brain, overlap areas of PR V labeled neurons projecting to the uterus and Sp6 were observed in lateral paragigantocellular n., rostroventrolateral reticular n., raphe obscurus n., raphe pallidus n., raphe magnus n., locus coeruleus n., Barrington's n., A5 cell group, central gray n., paraventricular hypothalamic n. and arcuate n. This results suggest that overlap areas of PRV labeled neurons of the spinal cord projecting to the uterus and Sp6 might be the first-order neurons related to the viscera-somatic sensory and sympathetic preganglionic neurons. PRV labeled neurons of the brain may be the second and third-order neurons response to the movement of smooth muscle of uterus. These PRV labeled neurons may be central autonomic center related to the integration and modulation of reflex control linked to the sensory and motor system monitoring the internal environment. These overlap areas of spinal cord and brainmay be related to autonomic centers related to regulation of uterus.

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

• Toxicological Research
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• v.22 no.4
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• pp.411-416
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• 2006

This study was carried out to investigate the spaciotemporal plasticity of intergeniculate leaflet in postnatal mongolian gerbil using genetically modified pseudorabies virus recombinant, which was a kind of excellent neurotracer with the ability to transpass the neuronal synaptic cleft. In addition, we tried to evaluate the special role of intergeniculate leaflet as a signal controler of circardian rhythm by expression of various nourotransmitters in suprachiasrnatic nucleus. The PRV-BaBlu, a genetically modified strain of PRV-Bartha with lac-Z gene, was injected into vitreous body of postnatal mongolian gerbil, and immunostained. The PRV-Bablu infected the neurons in intergeniculate leaflet of postnatal mongolian gerbil, and the degree of viral infection in postnatal period of experimental animals had tendency to increase with time consuming. This results showed that the mutant PRV-Bar-tha strain with lac-Z gene, PRV-BaBlu, was a very excellent neurotracer to localize the retinogeniculate tract with infection of neurons in intergeniculate leaflet specially.