고슴도치 십이지장 점막의 장내분비세포의 미세구조

최월봉,원무호,서지은,Choi, Wol-Bong,Won, Moo-Ho,Seo, Ji-Eun 한국현미경학회 1987 Applied microscopy Vol.17 No.1

In order to discriminate the enteroendocrine cell types in the mucosal epithelium of the normal duodenum of the Korean hedgehog (Erinaceus koreanus). The tissues were fixed in the mixture of 1% paraformaldehyde and 1% glutaraldehyde in phosphate buffer (pH 7.2), and postfixed in 2% osmium tetroxide (phosphate buffer, pH 7.2). They were embedded in Araldite, and the ultrathin sections were made by LKB-V ultratome following the inspection of semithin sections stained with toluidine blue-borax solutions. Ultrathin sections contrasted with uranyl acetate and lead citrate were observed with JEM 100B electron microscope. At least six types of enteroendocrine cells distributed in the mucosal epithelium of the duodenum were identified according to their morphological characteristics mainly based on the size, shape, number and electron density of the secretory granules. Type I cells had moderately developed organelles. The secretory granules were pleomorphic ($370X510nm$), and the granule cores with high electron density were enveloped in limiting membrane and characterized by a narrow halo. Type II cells contained an indented nucleus and well-developed organelles. The secretory granules were round (350 nm) and classified in two kinds by electron density, moderate and high. Both granules were surrounded by limiting membrane and those with high electron density showed often a wide halo. Type III cells had an indented nucleus. The secretory granules with various electron density were round (220 nm) in shape. The granules with high electron density were enveloped in limiting membrane and characterized by a narrow halo, but those with low or moderate electron density had not been observed the limiting membrane. Type IV cells contained an indented nucleus and moderately developed organelles. The secretory granules were round (180 nm) in shape, and the granule cores with high electron density were enveloped in limiting membrane and showed often a wide halo. Type V cells had a large amount of rough endoplasmic reticulum. Secretory granules with low or moderate electron density were round (230 nm) in shape, and surrounded by limiting membrane and showed a narrow halo. Type VI cells contained an oval nucleus and well-developed organelles, especially Golgi complex. The secretory granules with high electron density were round (210 nm) in shape. The granules were enveloped in limiting membrane and showed often a wide halo.

WKY 랫드의 척수신경절 내에서 CGRP 면역반응세포의 분포에 대한 연구

신재원(Jae-Won Shin),원무호(Moo-Ho Won),이재철(Jae-Chul Lee),오양석(Yang-Seok Oh),성제경(Je-Kyung Seong),윤여성(Yeo-Sung Yoon) 한국실험동물학회 1998 Laboratory Animal Research Vol.14 No.2

척수 허혈 후 토끼 척수 전각에서 Glutamate와 GABA 면역반응구조의 변화

송경원(Kyung Won Song),박인헌(In Heon Park),원무호(Moo Ho Won),송영선(Young Sun Song),이승용(Seung Yong Lee),오원석(Won Seok Oh),박재용(Jae Yong Park),최대은(Dae Eun Choi) 대한정형외과학회 2004 대한정형외과학회지 Vol.39 No.6

목적: 척수 허혈(spinal ischemia)이 시간 경과에 따라 신경전달물질인 glutamate와 γ-aminobutyric acid (GABA) 면역 반응양상의 변화에 미치는 영향을 면역조직학적으로 규명하고자 본 연구를 수행하였다. 대상 및 방법: 토끼를 실험동물로 하여 정상군과 15분간 척수 허혈 후 재관류한 실험군(허혈-재관류 후 0.5, 1, 3, 6, 12, 24 및 48시간)의 척수 제7요수분절 전각에서 glutamate 및 GABA 면역조직화학반응의 변화를 관찰하였다. 결과: Glutamate 면역반응양상은 30분 군의 제 Ⅸ층의 신경세포들에서 면역반응을 보였고, 주변의 신경섬유들에서 염색성이 증가한 후 감소하였으며, 6시간 군 이후부터 혈관 주변에서 면역염색성이 나타나기 시작하여 12시간 군까지 증가하였다. 그러나 24시간 군 이후부터 다시 감소하여 48시간 군에서는 면역염색이 거의 관찰되지 않았다. GABA 면역반응양상은 6시간 군부터 12시간 군에 걸쳐 신경세포가 소실된 부위를 중심으로 증가하였고, 이러한 증가 양상은 제 Ⅶ 및 제 Ⅷ층에서도 관찰되었다. 그러나 24시간 군 이후부터 GABA 면역염색성이 다시 감소하여 48시간 군에서는 면역염색성이 관찰되지 않았다. 결론: 척수 허혈-재관류 후 척수 전각에서 glutamate 면역반응양상의 변화는 허혈-재 관류 후 짧은 시간대부터 일어나며, GABA 면역반응양상은 glutamate에 비해 늦은 시간대에서 변화가 일어나는 것으로 사료된다. Purpose: This study was carried out to investigate chronological changes of glutamate and γ-aminobutyric acid (GABA) immunoreactivities in the anterior horn of the spinal cord after ischemia-reperfusion. Materials and Methods: Spinal cord ischemia was induced by clamping the abdominal aorta for 15 minutes in New Zealand white rabbit, and then the spinal cord was reperfused. These animals were sacrificed at 0.5, 1, 3, 6, 12, 24 and 48 hours after ischemia-reperfusion. Spinal cord sections at the level of L7 were immunostained against glutamate and GABA. Results: Glutamate immunoreactive neurons and fibers were first detected in the lamina Ⅸ at 30 minutes, but at 1 hour, the immunoreactivity returned to the control level. At 6 hour, glutamate immunoreactivity was observed around the blood vessels and its immunoreactivity increased between 6 and 12 hour. Thereafter the immunoreactivity decreased and eventually disappeared at 48 hours. GABA immunoreactivity increased in the anterior horn from 6 to 12 hours. Thereafter, GABA immunoreactivity decreased and eventually disappeared at 48 hours. Conclusion: These results suggest that the alteration of the glutamate immunoreactivity may occur much rapidly than that of GABA immunoreactivity in spinal anterior horn after ischemia-reperfusion.

한국재래산양 뇌하수체의 성장자극세포와 젖샘자극세포에 관한 전자현미경적 연구

이인세,이흥식,원무호,서제훈,송승훈,남영삼,강태천,Lee, In-se,Lee, Heungshik S.,Won, Moo-ho,Seo, Jehoon,Song, Seung-hoon,Nam, Young-Sam,Kang, Tae-Cheon 대한수의학회 1998 大韓獸醫學會誌 Vol.38 No.3

Somatotropes, mammotropes and somatomammotropes of the Korean native goat hypophysis were studied by double immunoelectron microscopy using antisera to growth hormone(GH) and prolactin(PRL), and protein A-gold particles of different sizes. Mammotropes were round or oval in shape, and contained round and electron dense secretory granules. The size of secretory granules was variable from 460nm to 680nm in diameter. Somatotropes were elliptical or triangular in shape and the oval nucei were located eccentrically at the periphery of the cell. Secretory granules of the cell were oval in shape and clearly distinguished from round granules of mammotropes. The size of granules was 320~680nm in diameter, smaller than that of mammotropes. Somatomammotropes contained round or oval secretory granules. The granules had intermediate size between somatotropes and mammotropes. Some of granules contained both GH and PRL, while the others contained only one of them.

WKY 랫드 요수부분 척수신경절 내에서 Calcitonin gene-related peptide와 Substance P 면역반응세포의 분포

신재원,윤여성,원무호,오양석,Shin, Jae-won,Yoon, Yeo-sung,Won, Moo-ho,Oh, Yang-seok 대한수의학회 1997 大韓獸醫學會誌 Vol.37 No.4

Dorsal root ganglion(DRG) cells are primary sensory neurons which contain some biologically active neuropeptides which play a role as neurotransmitters or neuromodulators. This study was performed to observe normal distribution of calcitonin gene-related peptide (CGRP) and substance P (SP) immunoreactive cells and colocalization of CGRP and SP in a single DRG cell of the lumbar DRGs($L_1{\sim}L_6$) in the Wistar Kyoto(WKY) rat by immunohistochemistry. About 55.8% of DRG cells contained CGRP-immunoreactivity, while about 12.7% of DRG cells showed SP-immunoreactivity. There was no significant difference in percentage of each neuropeptied-immunoreactive cells between each neuropeptide-immunoreactive cells between each levels of DRGs ($L_1{\sim}L_6$) (p>0.01). In size distribution, CGRP-immunoreactive cells were identified below $1,500{\mu}m^2$; SP-immunoreactive cells below $600{\mu}m^2$. In serial sections, about 86.7% of the SP immunoreactive cells contained CGRP immunoreactivity.

SD 랫드와 WKY 랫드의 허리 및 엉치부분 척수신경절 내에서 CGRP 면역반응세포의 분포

신재원(Jae-Won Shin),윤여성(Yeo-Sung Yoon),강태천(Tae-Cheon Kang),오양석(Yang-Seok Oh),원무호(Moo Ho Won) 한국실험동물학회 1999 Laboratory Animal Research Vol.15 No.3

WKY 랫드 요수부분 척수신경절 내에서 Substance P와 Somatostatin 면역반응세포의 분포

신재원(Jae-Won Shin),윤여성(Yeo-Sung Yoon),원무호(Moo-Ho Won),성제경(Je-Kyung Seong),서준교(Jun-Gyo Suh),오양석(Yang-Seok OH) 한국실험동물학회 1997 Laboratory Animal Research Vol.13 No.1

청서 시상하부의 growth hormone-releasing factor 및 somatostatin 면역반응신경세포의 분포

정영길,손화영,윤원기,김길수,원무호,류시윤,조성환,김무강,Jeong, Young-gil,Son, Hwa-young,Yoon, Won-kee,Kim, Kil-soo,Won, Moo-ho,Ryu, Si-yun,Cho, Sung-whan,Kim, Moo-kang 대한수의학회 1995 大韓獸醫學會誌 Vol.35 No.4

This study was attempted to investigate the topographical distribution, shape and immunoreactivity of growth hormone-releasing factor(GRF)- and somatostatin(SOM)-immunoreactive neurons in the hypothalamus of the Korean squirrels(Sciurus vulgalis coreae). For the light microscopical examination of immunohistochemistry, the brains were fixed with 4% paraformaldehyde solution by means of intracardiac perfusion. And the frozen sections($40{\mu}m$ thick) were stained immunohistochemically by ABC method. Distribution of GRF immunoreactive neurons($12-17{\mu}m$) was highest in the paraventricular nucleus, moderate in the periventricular and supraoptic nuclei, and low in the arcuate nucleus and lateral hypothalamic area. Their immunoreactive fibers were found very high in the median eminence, moderately in the supraoptic, paraventricular and periventricular nuclei, and low in the arcuate nucleus and lateral hypothalamic area. SOM immunoreactive perikarya($14-18{\mu}m$) were found moderately in the periventricular nucleus near the subependymal layer of the third ventricle, and low in the arcuate and suprachiasmatic nuclei. SOM immunoreactive fibers were found high in the median eminence, and moderately or low in the arcuate and periventricular nuclei.

BB랫드의 췌장 샘포세포의 미세구조적 특징

정영길,김길수,이철호,원무호,조성환,김무강,Jeong, Young-gil,Kim, Kil-soo,Lee, Chul-ho,Won, Moo-ho,Cho, Sung-whan,Kim, Moo-kang 대한수의학회 1996 大韓獸醫學會誌 Vol.36 No.1

Functional and morphological characteristics of the exocrine pancreas in genetic model BB rat of insulin dependent diabetes medllitus(IDDM) were carried out. Wistar rat was used as control animal. Flow rate of pancreatic juice, output of amylase and protein, and plasma glucose and insulin levess were examined. Also light and ultrastructural characteristics of the exocrine pancreas were observed. Pancreatic flow rate, output of amylase and protein, and insulin level were lower;glucose level was higher comparing with those of the control Wistar rat. In Wistar rat, exocrine pancreas was typical light microscopically. Zymogen granules and cell organelles were well developed in fine structure. Cell size of the periinsular acini was larger, and number of zymogen granules were more than those of the teleinsular acini. Most acinar cells were dark cells which containe well-developed RER in their cytoplasm. On the other hand, some light cells which have the dilated RER cisterns were found. In BB rat exocrine pancreas, cell size of per-and tele-insular acini similar to that of Wistar rat. The number of light cells occupied 40-50% compairing with that of Wistar rat. Zymogen granules were lower in number than that of Wistar rat and divied into three types in morphological characteristics ; type I showing normal structure, type II showing the wide hallo and small electron dense core in center of the zymogen granule and type III not having the electron dense core in the zymogen granule. The present ratio of type I, type II and type III are less than 5%, 30-40% and more than 50%, respectively.

Zinc 대사와 관련된 Paneth 세포활성의 변화에 관한 조직화학적 연구

조승묵,김성준,박승국,강태천,원무호,Jo, Seung-Mook,Kim, Sung-Jun,Park, Seung-Kook,Kang, Tae-Cheon,Won, Moo-Ho 한국현미경학회 2000 Applied microscopy Vol.30 No.4

포유류 장생 상피세포의 하나인 파네스세포(Paneth cell)는 여러 가지 금속류를 함유하고 있으며, 생체가 과잉의 이들 금속류에 노출되거나 부족할 때에 장 내강을 통해 제거하거나 흡수함으로써 금속류의 항상성에 기여한다고 알려져 있다. 이번 연구는 이러한 연구보고에 근거하여 금속류중에서 zinc를 실험적으로 과량투여한 후 파네스세포내 zinc의 분포에 어떤 변화가 초래되는 지를 광학 및 전자현미경적 autometallography(AMG)로 관찰함으로써 파네스세포의 zinc와 관련된 세포생물학적 기능을 규명하고자 하였다. Wistar 랫드에 체중 당 20mg의 zinc chloride를 생리식염수 5ml에 녹여 복강주사한 후 2시간에 이르러 0.1 M phosphate buffer(PB)에 녹인 0.5% sodium sulphide-3% glutaraldehyde 흔합액으로 관류고정하였다. 회장(ileum)의 일부를 Dry Ice나 $CO_2$ gas로 얼린 후 cryostat을 이용하여 $20{\mu}m$ 두께의 조직절편을 만들어 및 단계의 AMG법을 시행하였다. 이와는 별도로 EM용으로 선택된 회장절편은 일련의 전자현미경 표본제작과정을 거쳐, 100nm두께의 thin section을 만들어 uranyl-lead 이중염 색 추 전자현미경으로 관찰하였다. Zinc를 투여한 동물에서 관찰된 소견은 생리식염수만을 투여한 대조군의 것과 비교할 때 큰 차이를 보였다. 우선 대조군에서는 파네스세포의 꼭대기(apex)부위에서 낱알모양을 띤 AMG양성반응 구조물(AMG grain)들이 분비과립과 사이토졸(cytosol)에서 관찰되었고, 세포사이공간에서도 적은 양의 grain이 분포하고 있었다. 반면 zinc를 투여한 랫드의 파네스세포에서는 AMG 입자가 이상의 부위에서 훨씬 많은 양의 grain이 분포해 있었으며, 특히 분비과립에서는 가장 높은 농도로 관찰되었다. 더욱이 대조군에서 관찰되지 많았던 장샘의 분비관(Lieberkuhn crypt)및 고유판 혈관의 내에서도 많은 grain이 관찰되었다. 이상의 결과를 요약하면 과량의 zinc에 노출된 생체는 혈관을 경유하여, 파네스세포의 분비과립을 매개로 하여, 장관의 내강쪽으로 과잉의 zinc 를 피내고 있는 모습을 보인다. 따라서 연구자는 파네스세포가 생체내 zinc의 양을 조절하는데 중요한 역할이 있을 것으로 본다. Paneth cells have been suggested to contribute to the elimination of excess metals into the intestinal lumen. The purpose of this study wat to investigate the changes of the zinc pools in rats subjected to functional loading with zinc salt by mean of both light and electron microscopical autometallography (AMG). Wistar rats 4 were administrated with zinc chloride (20 mg/kg body weight) intraperitoneally dissolved in 1 ml distilled water. The control group received 1 ml saline IP. After further one hour the animals were transcardially perfused with 0.4% sodium sulphide dissolved in 0.1 M PB fellowed by 3% glutaraldehyde solution for 10 minutes. Pieces of ileum were frozen with solid $CO_2$ and sectioned on a cryostat. The sections $(20{\mu}m)$ were autometallographically developed. Sections selected for EM were reembedded on top of a blank Epon block, from which ultrathin sections (100 nm) were cut. The ultrathin sections were double stained with uranyl acetate (30 min) and lead citrate (5 min), then examined under electron microscope. Studies of comparable sections from control and zinc loaded animals with the AMG selenium method gave quite different results. The control animals demonstrated a weakly positive staining in the cytoplasm of the Paneth cells. In the electron microscope the AMG silver grains were found to be located in the cytoplasm, while the electron dense secretary granules and other cell organelles were void of staining. Few AMG grains were located at the apical surface of the Paneth cells. In sections from zinc loaded rats, the AMG grains were seen in abundance in the lumen of the Lieberkuhn crypts at light microscopic levels. At EM levels the zinc revealing silver grains were located in the cytoplasm as in the controls, but much more AMG grains were shifted into the secretary granules. Furthermore, profound AMG grains were found in the lumen of the crypts and surrounding vessels. And a few grains were seen in the endothelium. The AMG technique demonstrated a pattern of AMG grains in the Paneth cells that strongly suggests a transport of zinc ions through these cells.