흰쥐에서 WGA-HRP와 pseudorabies virus를 이용한 정관의 신경로에 대한 연구

이창현,정옥봉,고병문,이봉희,김수명,김인식,양홍현,Lee, Chang-Hyun,Chung, Ok-Bong,Ko, Byung-Moon,Lee, Bong-Hee,Kim, Soo-Myung,Kim, In-Shik,Yang, Hong-Hyun 대한수의학회 2003 大韓獸醫學會誌 Vol.43 No.1

This experimental studies was to investigate the location of PNS and CNS labeled neurons following injection of 2% WGA-HRP and pseudorabies virus (PRY), Bartha strain, into the ductus deferens of rats. After survival times 4-5 days following injection of 2% WGA-HRP and PRV, the rats were perfused, and their brain, spinal cord, sympathetic ganglia and spinal ganglia were frozen sectioned ($30{\mu}m$). These sections were stained by HRP histochemical and PRY inummohistochemical staining methods, and observed with light microscope. The results were as follows ; 1. The location of sympathetic ganglia projecting to the ductus deferens were observed in pelvic ganglion, inferior mesenteric ganglion and L1-6 lwnbar sympathetic ganglia. 2. The location of spinal ganglia projecting to the ductus deferens were observed in T13-L6 spinal ganglia. 3. The PRY labeled neurons projecting to the ductus deferens were observed in lateral spinal nucleus, lamina I, II and X of cervical segments. In thoracic segments, PRY labeled neurons were observed in dorsomedial part of lamina I, II and III, and dorsolateral part of lamina IV and V. Densely labeled neurons were observed in intermediolateral nucleus. In first lumbar segment, labeled neurons were observed in intermediolateral nucleus and dorsal commisural nucleus. In sixth lumbar segment and sacral segments, dense labeled neurons were observed in sacral parasympathetic nuc., lamina IX and X. 4. In the medulla oblongata, PRV labeled neurons projecting to the ductus deferens were observed in the trigeminal spinal nuc., A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nuc., rostroventrolateral reticular nuc., area postrema, nuc. tractus solitarius, raphe obscurus nuc., raphe pallidus nuc., raphe magnus nuc., parapyramidal nuc., lateral reticular nuc., gigantocellular reticular nuc.. 5. In the pons, PRV labeled neurons projecting to the ductus deferens were ohserved in parabrachial nuc., Kolliker-Fuse nuc., locus cooruleus, subcooruleus nuc. and AS noradrenalin cells. 6. In midbrain, PRV labeled neurons projecting to the ductus deferens were observed in periaqueductal gray substance, substantia nigra and dorsal raphe nuc.. 7. In the diencephalon, PRV labeled neurons projecting to the ductus deferens were observed in paraventricular hypahalamic nuc., lateral hypothalamic nuc., retrochiasmatic nuc. and ventromedial hypothalamic nuc.. 8. In cerebrum, PRV labeled neurons projecting to the ductus deferens were observed in area 1 of parietal cortex. These results suggest that WGA-HRP labeled neurons of the spinal cord projecting to the rat ductus deferens might be the first-order neurons related to the viscero-somatic sensory and sympathetic postganglionic neurons, and PRV labeled neurons of the brain and spinal cord may be the second and third-order neurons response to the movement of smooth muscles in ductus deferens. 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 monitaing the internal environment. These observations provide evidence for previously unknown projections from ductus deferens to spinal cord and brain which may be play an important neuroanatornical basic evidence in the regulation of ductus deferens function.

Central Localization of the Neurons Projecting to the Urinary Bladder Meridian

김정연,전홍재,이상룡,이창현,정옥봉,Kim Jeong-Yun,Jun Hong-Jae,Lee Sang-Ryoung,Lee Chang-Hyun,Chung Ok-Bong Society for Meridian and Acupoint 2000 대한침구의학회지 Vol.17 No.1

Transsynaptic tracer이며 신경친화성 virus인 pseudorabies virus(PRV)를 방광(膀胱), 방광유(膀胱兪), 위중(委中) 및 중추(中極)에 주입(注入)한 후 4일간의 생존기간이 경과한 후 희생시켜 면역조직화학침액법(免疫組織化學染色法)에 의하여 뇌척수에 표지된 공통된 영역들을 비교하여 관찰한 결과는 다음과 같다. 1. 방광벽(膀胱壁), 방광유(膀胱兪), 위중(委中) 및 중추(中極)에서 척수에 투사된 영역은 흉수(胸髓), 요수(腰髓) 및 천수(薦髓)에 모두 표지되었으며 공통적으로 표지된 부위는 척수(脊髓)의 층판 IV, V, VII, IX, X영역에 표지되었으나 주로 강하게 표지된 공통된 영역은 층판 VII의 중간외측핵, 가슴기둥 및 층판 X영역이었다. 2. 방광벽(膀胱壁), 방광유(膀胱兪), 위중(委中) 및 중추(中極)에서 뇌(腦)에 투사된 공통된 영역은 연수(延髓)에서는 A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nucleus에서 양성반응을 나타내었다. 솔기핵의 경우 아핵인 불명솔기핵, 창백솔기핵 및 큰솔기핵에서 양성반응을 보였다. 다리뇌에서는 청색반점, Barrington's nucleus, A5세포군 및 삼차신경운동핵에서 양성반응을 보였고, 중뇌에서는 눈돌림신경핵, 눈돌림신경섬유 및 다리핵에서 양성반응을 보였다. 간뇌에서는 시상하부(視床下部)의 뇌실(腦室)곁핵과 시상의 뇌실곁핵에서 양성반응을 보였고 대뇌(大腦)에서는 septal nucleus, 피질(皮質)의 뒷다리영역, 마루엽, 이마엽에서 양성반응을 보였다. 이상의 결과를 종합하면 방광(膀胱)에서 투사되는 뇌척수의 영역과 방광유(膀胱兪)나 위중(委中)에서 투사되는 공통된 표지영역들은 방광(膀胱)과 족태양방광경(足太陽膀胱經) 그리고 그 경락(經絡)의 경혈(經穴)들이 어떤 상관성(相關性)을 가지고 연결(連結)되어 있다는 사실을 실험적으로 알 수 있었다. 특히 방광(膀胱)과 방광유(膀胱兪), 위중(委中)에서 투사된 공통된 표지영역, 즉 배뇨중추인 Barrington's nucleus에 표지되는 것은 내장(內臟)-경락(經絡)이 central autonomic pathway에 의하여 서로 연결되었음을 입증하는 중요한 결과(結果)라고 사려(思慮)된다.

아연 전처리가 시간 경과에 따른 카드뮴 유도 고환 독성에미치는 보호 효과

제갈승주 ( Seung Joo Jekal ),이경선 ( Kyung Sun Lee ),정옥봉 ( Ok Bong Chung ),임효빈 ( Hyo Bin Im ) 대한임상검사과학회 2009 대한임상검사과학회지(KJCLS) Vol.41 No.3

Cadmium (Cd) is known to exert gonadotoxic and spermiotoxic effects. The present study was performedto investigate the morphological effects and metallothionein (MT) expression by zinc pretreatment in thecourse of time of cadmium-induced testicular injury in rat. Fifty male Spraque-Dawley rats weighing 160~180 g were divided into two groups : saline-pretreated cadmium group and zinc-pretreated cadmium group. Rats of two groups received subcutaneous injection of saline and 100 mg/kg ZnSO4 at 0, 2, 5 and 8 hrs intervals respectively. Cadmium chloride (4.5 mg/kg CdCl2) was administrated intraperitoneally at 2 hrs after zinc injection and rats were killed 0, 12, 24, 48 and 72 hrs later. Testicular tissue damages, interstitial (Leydig) cells status and MT expression were determined using hematoxylin-eosin stained sections and a computerized image analysis system on sections immunostained with a mouse anti-metallothionein respectively. Zinc pretreatment was significantly reduced testicular damages in five pathological categories after cadmium administation. The number of surviving interstitial cells was significantly higher in thezinc-pretreated group than in the saline-preatreated group at 48 and 72 hrs after cadmium administration. Non-damaged testis showed the positivity of MT staining in spermatogenic cells, Sertoli cells and endothelium of blood vessel, but not in the Leydig cells. The positivity of MT staining in saline-pretreated group was significantly reduced at 24 hrs after cadmium administration, whereas zinc-pretreated group showed strong MT positive staining similar to the 0 hr by 42 hrs after cadmium administration. In damaged testis, MT positive staining was also observed in the Leydig cells of both groups. These results suggest that a major preventive effect of zinc against cadmium-induced testicular toxicity may be due to its ability to reduce the cytotoxicity of cadmium in spermatogenic cells and Leydig cells by inhibiting the susceptibility of the testis to cadmium but not MT production by cadmium.

흰쥐 숨뇌의 맨아래구역에서 Phenylalanine 처리에 따른 Monosodium Glutamate의 효과

이창현(Chang-Hyun Lee),고병문(Byoung-Moon Ko),김명순(Myoung-Soon Kim),정옥봉(Ok-Bong Chung) 대한체질인류학회 2000 해부·생물인류학 (Anat Biol Anthropol) Vol.13 No.2

간추림 Glutamate 는 수용체를 통하여 신경세포의 흥분을 전달하는 아미노산계 신경전달물질이다. 본 연구는 생리식염수, monosodium glutamate(MSG), phenylalanine, MSG-phenylalanine 을 투여하여 숨뇌의 맨아래구역에서 TH 신경세포를 조사하였다. 맨아래구역에서 catecholamine신경세포를 관찰하기 위하여 면역세포화학적 방법을 이용하였다. MSG(4mg/g bw)의 투여에 의해 흰쥐 맨아래구역의 TH 신경세포가 손상되었고, MSG에 이은 phenylalanine (15 mg/g bw)의 투여에 의해 TH 신경세포의 손상이 감소되었다. 이로서 phenylalanine 이 MSG 에 의한 신경세포 독성을 억제시키는 것으로 생각된다.

스트렙토조토신 유도 당뇨 흰쥐에서 전기자극이 상처치유와 피부 비만세포에 미치는 영향

제갈승주 ( Seung Joo Jekal ),이경선 ( Kyung Sun Lee ),정옥봉 ( Ok Bong Chung ),이재형 ( Jae Hyoung Lee ) 대한임상검사과학회 2008 대한임상검사과학회지(KJCLS) Vol.40 No.2

The aim of this study was to investigate the effect of electrical stimulation on healing of impaired wound and alteration of mast cells in experimental diabetic rats. Thirty male Sprague-Dawley rats were divided into three groups : incision (control), diabetes+incision (diabetes) and diabetes + incision + electrical stimulation (D/ES). Diabetes was induced in rats by streptozotocin (STZ) injection (60 mg/kg, one time) and 20 mm length incision wounds were created on the back after shaving hair. The electrical stimulation rats were treated with a current intensity of 30~50 V at 120 pps and 140 μs for 10 days from 3 days after STZ injection. The lesion and adjacent skin tissues were fixed with 10% buffered formalin, embedded with paraffin. For wound healing analysis, hematoxylin-eosin (HE) and picrosirius red staining were performed. Mast cells (MC) were stained with toluidine blue (pH 0.5) and quantified at ×200 using a light microscope. The density of keratinocyte proliferation and microvessels in skin tissues were analyzed using a computerized image analysis system on sections immunostained with proliferative cell nuclear antigen (PCNA) and α-smooth muscle actin (α-SMA), respectively. The results showed that the wound healing rate, collagen density and neoepidermis thickness, density of PCNA-positive cells and density of α-SMA-positive vessels were significantly higher in D/ES rats than in diabetic rats. The density of MCs and degranulated MCs in D/ES rats were also significantly higher than those in diabetic rats. These findings suggest that the electrical stimulation may promote the tissue repair process by accelerating collagen production, keratinocyte proliferation and angiogenesis in the diabetic rats, and MCs are required for wound healing of skin in rats.

아연 전처리가 시간 경과에 따른 카드뮴 유도 고환 독성에 미치는 보호 효과

제갈승주 ( Seung-joo Jekal ),이경선 ( Kyung-sun Lee ),정옥봉 ( Ok-bong Chung ),임효빈 ( Hyo-bin Im ) 대한임상검사과학회 2010 대한임상검사과학회지(KJCLS) Vol.42 No.3

Cadmium (Cd) is known to exert gonadotoxic and spermiotoxic effects. The present study was performed to investigate the morphological effects and metallothionein (MT) expression by zinc pretreatment in thecourse of time of cadmium-induced testicular injury in rat. Fifty male Spraque-Dawley rats weighing 160-180 g were divided into two groups: saline-pretreated cadmium group and zinc-pretreated cadmium group. Rats of two groups received subcutaneous injection of saline and 100 mg/kg ZnSO4 at 0, 2, 5 and 8 hrs intervals respectively. Cadmium chloride (4.5 mg/kg CdCl2) was administrated intraperitoneally at 2 hr after zinc injection and rats were killed 0, 12, 24, 48 and 72 hrs later. Testicular tissue damages, Interstitial (Leydig) cells status and MT expression were determined using hematoxylin-eosin stained sections and acomputerized image analysis system on sections immunostained with a mouse anti-metallothionein respectively. Zinc pretreatment was significantly reduced testicular damages in five pathological categories after cadmium administation. The number of surviving interstitial cells was significantly higher in the zinc-pretreated group than in the saline-preatreated group at 48 and 72 hrs after cadmium administration. Non-damaged testis showed the positivity of MT staining in spermatogenic cells, Sertoli cells and endothelium of blood vessel, but not in the Leydig cells. The potitivity of MT staining in saline-pretreated group was significantly reduced at 24 hrs after cadmium administration, whereas zinc-pretreated group showed strong MT positive staining similar to the 0 hr by 42 hrs after cadmium administration. In damaged testis, MT positive staining was also observed in the Leydig cells of both groups. These results suggest a major preventive effect of zinc against cadmium-induced testiculat toxicity may be due to its ability to reduce the cytotoxicity of cadmium in spermatogenic cells and Leydig cells by inhibiting the susceptibility of the testis to cadmium but not MT production by cadmium

족태양방광경(足太陽膀胱經)에서 투사(投射)되는 신경원(神經元)의 표식부위(標識部位)에 대(對)한 연구(硏究)

김정연 ( Jeong Yun Kim ),전홍재 ( Hong Jae Jun ),이상룡 ( Sang Ryoung Lee ),이창현 ( Chang Hyun Lee ),정옥봉 ( Ok Bong Chung ) 대한경락경혈학회 2000 Korean Journal of Acupuncture Vol.17 No.1

Transsynaptic tracer이며 신경친화성 virus인 pseudorabies virus(PRV)를 膀胱, 膀胱兪, 委中 및 中極에 注入한 후 4일간의 생존기간이 경과한 후 희생시켜 免疫組織化學染色法에 의하여 뇌척수에 표지된 공통된 영역들을 비교하여 관찰한 결과는 다음과 같다. 1. 膀胱壁, 膀胱兪, 委中 및 中極에서 척수에 투사된 영역은 胸髓, 腰髓 및 薦髓에 모두 표지되었으며 공통적으로 표지된 부위는 脊髓의 층판 IV, V, VII, IX, X영역에 표지되었으나 주로 강하게 표지된 공통된 영역은 층판 VII의 중간외측핵, 가슴기둥 및 층판 X영역이었다. 2. 膀胱壁, 膀胱兪, 委中 및 中極에서 腦에 투사된 공통된 영역은 延髓에서는 A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nucleus에서 양성반응을 나타내었다. 솔기핵의 경우 아핵인 불명솔기핵, 창백솔기핵 및 큰솔기핵에서 양성반응을 보였다. 다리뇌에서는 청색반점, Barrington`s nucleus, A5세포군 및 삼차신경운동핵에서 양성반응을 보였고, 중뇌에서는 눈돌림신경핵, 눈돌림신경섬유 및 다리핵에서 양성반응을 보였다. 간뇌에서는 視床下部의 腦室곁핵과 시상의 뇌실곁핵에서 양성반응을 보였고 大腦에서는 septal nucleus, 皮質의 뒷다리영역, 마루엽, 이마엽에서 양성반응을 보였다. 이상의 결과를 종합하면 膀胱에서 투사되는 뇌척수의 영역과 膀胱兪나 委中에서 투사되는 공통된 표지영역들은 膀胱과 足太陽膀胱經 그리고 그 經絡의 經穴들이 어떤 相關性을 가지고 連結되어 있다는 사실을 실험적으로 알 수 있었다. 특히 膀胱과 膀胱兪, 委中에서 투사된 공통된 표지영역, 즉 배뇨중추인 Barrington`s nucleus에 표지되는 것은 內臟-經絡이 central autonomic pathway에 의하여 서로 연결되었음을 입증하는 중요한 結果라고 思慮된다.