Tottering/leaner 생쥐 소뇌에서 조롱박세포의 가지돌기가시와 평형섬유염주사이의 신경연접 가소성

유임주(Im Joo Rhyu),박창현(Chang Hyun Park),전용혁(Yong Hyuck Chun),서영석(Young-Suk Suh) 대한해부학회 1999 Anatomy & Cell Biology Vol.32 No.2

Tottering 생쥐는 운동실조증, 소발작, 간헐성근경련과 유사한 운동장애등의 신경학적 증상을 보이는 변종생쥐이다. tottering (tg) 유전인자는 생쥐의 8번 염색체에 위치하는 것으로 알려져 있으며 보통염색체 열성으로 분류된다. 최근 tottering/leaner 생쥐의 전압의존성칼슘채널 α1A subunit 유전자 변이가 보고되었으며 사람의 보통염색체 우성 소뇌성운동실조증 (Autosomal dominant cerebellar ataxia: SCA6), 가족성반신불수성편두통 (Familial hemiplegic migraine) 및 제2 형간헐성운동실조증 (Episodic ataxia type-2) 등의 질환 모델로서 활용될 수 있게 되었다. 또한 이들 생쥐의 소뇌 분자층에서 과립세포 평행섬유와 조롱박세포 가지돌기가시 사이의 신경연접 가소성이 관찰되었다. 이에 본 연구는, tottering/leaner 생쥐에서 소뇌 분자층의 신경연접을 운동실조가 나타나는 시기에 따라 조롱박세포 가지돌기가 시극돌기와 과립세포 평형섬유팽대사이의 신경연접을 전자현미경을 이용하여 관찰하고, 전압의존성칼슘채널 α1A subunit 이상으로 유발된 세포내 전해질 변화가 조롱박세포의 형태에 미치는 영향을 이해하기 위해 성숙한 tottering/leaner 생쥐의 소뇌를 항 calbindin-28 kd 면역염색을 이용하여 관찰하였다. tottering/leaner 잡종생쥐의 조롱박세포에서 가지돌기가시는 정상대조군에 비하여 세포쪽부위에서 나오는 것이 많이 관찰 되었으며 축삭은 드물지 않게 축삭염주가 관찰되었다. 조롱박세포의 가지돌기가시와 과립세포의 평행섬유염주사이 신경연접에서 3개 이상의 신경연접은, tottering 생쥐에서는 30일군에서, tottering/leaner 잡종에서는 18일군에서, leaner 생쥐에서는 15일군에서 쉽게 관찰되었다. 이상에서 관찰된 형태학적 소견들은 이들 변종생쥐들의 운동실조증과 연관되어 있는 것으로 생각된다. The tottering (tg/tg) is neurologic mutant mouse exhibiting three neurological disorders: ataxia, petit mal-like absence seizures and myoclonic intermittent movement disorder. The tottering mouse carries an autosomal recessive single gene mutation on chromosome 8. The leaner (tgla) and Nagoya rolling (tgrol) are another two alleles of the tottering (tg). The combination of two mutant (tottering and leaner) produces compound heterozygous, tottering/leaner (tg/tgla) mouse. The genetic etilogy of the tottering and leaner was identified to be a mutation in voltage-dependent calcium channel α1A subunit. It made us link these animal model to human neurologic disease such as autosomal dominant cerebellar ataxia (SCA6), familial hemiplegic migraine and episodic ataxia type-2. The different onset and severity of neurological symptom of these three mutants (tg/tg, tg/tgla, tgla/tgla) offer good scale to analysis of pathophysiolgy of the neurologic disorder. Altered synapase between parallel fiber varicosity and dendritic spines of Purkinje cell was observed in adult tottering and leaner mice. Through the electron microscopic observation and anticalbindin-28 kd immunohistochemistry, we anaylzed not only the relationship between neurologic symptoms and synaptic plasticity around the ataxic onset of tottering, leaner and tottering leaner double mutation but also Purkinje cell morphology affected by voltage-sensitive calcium channel α1A subunit mutation in totterring mouse. Purkinje cell dendritic spines from proximal dendrites and axonal swellings of Purkine cell were observed frequently in wild type mice. The first apperance point of altered synapse based on semi-quantitative analysis was postnatal 15 days in leaner, postnatal 18 days in totering/leaner double mutation, and 30 days in tottering. These data suggest that altered synapse is associated with ataxia in tottering and leaner mice. Further study is needed to determine whether altered synapse is primary cause of ataxia.

흰쥐 소뇌 발달과정에서 발현의 변화를 보이는 glutamate_aspartate transporter (GLAST) 유전자 클로닝 및 발현양상 규명

장윤정(Yoon-Jeong Chang),심재호(Jae Ho Shim),김창미(Changmee Kim),김 현(Hyun Kim),유임주(Im Joo Rhyu),서영석(Young-Suk Suh),전용혁(Yong-Hyuck Chun) 대한해부학회 2000 Anatomy & Cell Biology Vol.33 No.4

Ordered differential display PCR로 흰쥐 뇌 발달과정에서 발현의 변화를 보이는 유전자를 분리하여 GLAST(L -glutamate/L-aspartate transporter) 유전자인 S59158과 동일한 유전자임을 확인하였으며, 이 유전자의 특성 규명을 위해 흰쥐 뇌 발달과정에서의 발현양상 및 glutamate 아나로그인 kainic acid를 처리한 흰쥐 뇌에서의 유전자 발현양상을 in situ hybridization 조직화학법으로 확인하였다. GLAST S59158은 신경조직 특이적인 발현양상을 보였으며, 특히 출생 후 소뇌 발달과정에 따라 현저한 발현의 변화를 보였다. S59158은 신경아교세포 생성이 왕성하게 일어나는 시기인 배자기 14일에서 20일 사이에 신경아교세포 생성이 일 어나는 부위인 뇌실밑부분 (subventricular zone)에서 높게 발현하였다. 출생 후에는 뇌실밑부분에서의 발현이 현저히 감소한 반면, 소뇌 조롱박세포층 (purkinje cell layer)의 Bergmann 세포에서 매우 강한 발현을 보였으며, 해마 (hippocampus), 치아이랑 (dentate gyrus) 및 후각망울 (olfactory bulb)에서는 중등도의 발현을 보였다. 이상의 결과에서 GLAST(S59158) 가 흰쥐 신경계의 발달 과정에서 신경아교세포의 생성과 관계가 있음을 생각할 수 있으며, 세포 외 glutamate 농도 증가에 의해 발현이 증가하는 것을 예상할 수 있다. In this study, S59158, a gene of GLAST(L -Glutamate/L-Aspartate transporter), was cloned by ordered differential display PCR with developing rat brains. The mRNA expression of this gene in the developing rat brain and the effect of kainic acid (KA), glutamate analogue, on this gene were investigated with in situ hybridization histochemistry. The expression of S59158 was restricted to nervous system and observed from E12 (embryonic day 12), peaked at E20, and gradually decreased to adult level. In embryos, S59158 was prominently expressed in the subventricular zones throughout the brain. After birth, strong expression was observed in the purkinje cell layer of cerebellum and moderate level of expression was observed in the subventricular zone, olfactory bulb, hippocampal formation, and cerebral cortex. In the KA treated rat brains, the expression of S59158 was significantly increased in dentate gyrus, hippocampus, and cerebral cortex. From these results, it may be suspected that S59158 is related to the development of the brain and is induced by increased extracellular glutamate level.

흰쥐 신경계 발달에 따른 전압의존성칼슘채널 α1 subunits (α1D, α1B, α1A, α1E) 유전자들의 발현에 관한 연구

김창미(Chang-Mee Kim),한승훈(Seung Hun Han),한상준(Sang Jun Han),김동연(Dong Yeon Kim),이경상(Kyung Sang Lee),전용혁(Yong-Hyuck Chun) 대한해부학회 1998 Anatomy & Cell Biology Vol.31 No.5

전압의존성 칼슘채널 (voltage dependent calcium channel, VDCC)의 구성요소 중 칼슘이온통로로 작용하는 α1 subunit 중에서 신경계에 발현하는 것으로 알려진 α1A, α1B, α1D 및 α1E subunit의 유전자 발현양상을 출생 전 태자기와 출생 후 성장기의 흰쥐 신경계에서 in situ hybridization 조직화학법으로 관찰하였다. 성숙한 흰쥐 뇌에서 각각의 유전자는 광범위하고 특이한 발현양상을 보였다. VDCC α1A 유전자는 해마의 CA3 부위와 소 뇌에서 높게 발현하였으며, VDCC α1B 유전자는 뇌 전체에서 광범위하고 낮게 발현하였다. VDCC α1D 유전자는 후각망울의 승모세포층, 시각교차위핵, 치아이랑 등에서 높게 발현하였으며, VDCC α1E 유전자는 후각망울, 해마와 치아이랑, 안쪽고삐핵소뇌에서 높은 발현을 보였다. 태자기 각종 조직에서 VDCC α1D 유전자는 신경조직에서 뿐만 아니라 허파, 창자, 콩팥, 간, 뼈대근육 등의 비신경계조직에서도 발현이 관찰되었으나, VDCC α1A, VDCC α1B 및 VDCC α1E 유전자는 신경조직 이외의 부위에서는 거의 발현을 보이지 않는 신경조직 특이성을 보였다. 흰쥐 뇌 발달과정에서 VDCC α1A, VDCC α1D 및 VDCC α1E 유전자 발현은 14일된 태자에서 관찰되기 시작하여 출생 후 14일까지 발현부위 및 정도가 증가되었으며 그 이후 점차 감소하였다. 한편 VDCC α1B 유전자는 14일된 태자에서 발현하기 시작하여 출생 직전과 직후에 가장 높은 발현을 보였으며 그 이후 점차 감소되는 양상을 보였다. 이들 유전자들은 공통적으로 뇌실층 (ventricular layer)에 비해 외투층(mantle layer)에서 높게 발현하였다. 성숙한 흰쥐 신경계에서의 특이한 발현양상으로부터 각각의 전압의존성 칼슘채널이 고유한 역할을 하는 것을 예측할 수 있으며, 발달과정에서 신경세포의 분화가 일어나는 부위에서 높게 발현하며 분화가 왕성하게 일어나는 시기 이후에는 발현이 감소하는 양상에서 신경계 분화에 관여하는 것을 예측할 수 있다. Voltage dependent calcium channels (VDCCs) mediate Ca++ influx into cells and are responsible for regulation of a variety of physiological effects. The key functional property of VDCCs are attributed to the calcium-pore forming α1 subunit. In this study, distribution pattern of α1 subunit (α1D, α1B, α1A, α1E) mRNA of VDCCs in developing and adult rat brain was investigated by in situ hybridization histochemistry. In the adult rat brain, each α1 subunit mRNA displayed a specific and distinct distribution pattern. α1D was highly expressed in the olfactory bulb, dentate gyrus, pituitary gland, pineal gland, hypothalamus, superior colliculus and cerebellum. Relatively low level of α1B was expressed throughout the whole brain and strong expression of α1A was observed in CA3 area of Ammon's horn, medial geniculate body, inferior colliculus and cerebellum. High level of α1E was found in the olfactory bulb, hippocampus, dentate gyrus, medial habenular nucleus and cerebellum. Moreover, α1B, α1A and α1E were expressed only in the nervous system but α1D was expressed not only in the nervous system but also in other tissues including liver, heart, lung and skeletal muscle. Generally the expression of α1D, α1A, and α1E subunit was observed from E14 and thereafter the intensity of labeling was gradually increased to P14 and then decreased to the adult level. But the expression of α1B subunit was observed from E14 and gradually increased to E20 and P0 and then decresaed. From the differential expressions of VDCC α1 subunits in developing and adult rat brain, it is suggested that each type of VDCCs may play a distinct roles in neural and nonneural tissues, and the VDCCs may be related with development of nervous system.

Vitamin E 投與마우스 飢餓에 對한 抵抗과 胃腸管粘膜에 미치는 影響에 關한 組織學的 硏究

全容赫 고려대학교 의과대학 1975 고려대 의대 잡지 Vol.12 No.3

To clarify the one of the physiological effects of vitamin E, 1976 adult albino mice were used in present study. Vitamin E was injected on a daily dose of 5㎎; 10 days for the adequate dose group and 20 days for the overdose group. After completion of injections, animals were put into the starvation-absolute and free access of water groups. Change of body weight, survival time withstanding to the starvation, histochemical observations of the gastrointestinal mucosa were calculated at the periods of starvation; 24, 48 hours and at the time of death. The results of the study are summarized as follows: 1. Administration of adequate dose of vitamin E resulted prolongation of survival time against starvation, but excess amount resulted shortening tendency in survival time. 2. Vitamine E administration accelerated the rate of body weight increment and the loss of body weight due to starvation was rather slow and moderate. 3. There was not much difference in histometrical feature of the gastrointestinal mucosa in adequate vitamin E administration group, but in histochemical finding, amount of cellular synthetic material was increased. However, overdose vitamin E group showed edematous mucous membrane and decreased stainability because of the reduction of the number of mucous secreting cells and cellular synthetic material. 4. The changes of histological finding due to starvation were more prominent in duodenal mucosa compared to that of stomach, but decrease in mucous material was more prominant in stomach. The neutral mucous substance in gastric mucosa, the acid mucous substance in duodenal crypt and the neutral mucus in duodenal villi were mainly decreased respectively. 5. Decrease of intensity and stain reactivity in secretory granules of Paneth cell due to starvation was suppressed by the vitamin E. It seems that vitamin E causes acceleration of the secretory granule synthesis. 6. Reduction in the number of epithelial lining cell of duodenum following starvation was more apparent in the villi than the crypt, while goblet cells decreased generally less grade. The tendency showed that vitamin E injected groups were suppressed in the cell reduction. 7. In general, free access of water groups survived longer than absolute starvation groups and showed mild reduction whenever fasting proceed. However, any noticeable histological difference was not found between two groups.

正常韓國人의 血淸 Isocitric Dehydrogenase 活性値

全容赫 우석대학교 의과대학 1970 우석의대잡지 Vol.7 No.1

To evaluate normal value and range of serum isocitric dehydrogenase activity in Koreans, sera of the 50 apparently healthy Korean male adults were analysed and the results were as follows: Mean: 105±32 Sigma unit/ml with range of 55~175 Sigma units/ml which were not significantly different from those of Europeans.

Vitamin E 投與가 마우스 諸組織에 미치는 影響에 關한 實驗的 硏究 : Ⅱ. 腎臟의 死後組織溶解에 關한 觀察

全容赫,李福周,洪貞姬 최신의학사 1974 最新醫學 Vol.17 No.5

Histopathological observation was made to compare the post-mortem renal autolysis between vitamin E injected group and control mice. Animals were received 5mg of vitamin E injection as a single dose for 10 days and 20 days intramuscularly everyday. One slice was fixed in the Zenker's solution promptly after death and the others were allowed to remain carcass in the normal saline solution for 24 hours at roomtemperature. The result indicated that the post-mortem autolysis was apparently prevented by the administration of adequate dose of vitamin E which would seemed 10-day injection. In general, histolysis affected more intensely in the cortex than medulla and renal corpuscles showed extensive changes.

四肢發生 過程에 關한 觀察

全容赫,朴善華 고려대학교 의과대학 1992 고려대 의대 잡지 Vol.29 No.2

Developmental sequence of events of fore-limb in DDY mouse embryo were observed to obtain the data on shape ; on histology with special reference to the apical ectodermal ridge(AER), programmed cell death(PCD) and chondro-osteogenesis and ; on changes of glycoconjugates. Fore-limb buds appeared between the day 9.5 and 10 of gestation(plug day=0) and distal ends grew into paddle-like plates from day 11, plates condensed to form digital rays on day 12, webbed fingers on day 13 and fingers were separated on day 14. Well developed AER appeared with the beginning of elevation of buds and decreased in size from day 11 and remained remnants on day 12.5 of gestation. Proximal core mesenchyme began to condense form day 11 and transformed to precartilaginous zone 12 hours later chondrified from day 12.5 and ossified from day 15 and these phenomena proceeded proximo-distal sequence. Programmed cell death happened only between the day 10.5 to 11.5 and were confined to the proximal and middle zone of mesenchyme, which comes into question how PCD In motioned area and time correlate with subsequent morphogenses. PAS positive reactivity was gradually increased in the epithelium. AER and cartilaginous zone with the development proceed but that of mesenchyme was weak throughout the period. and in comparison with PAS reactivity, PAPS reactivity to the epithelium was always less but no difference in cartilaginous zone which suggest that amount and component of glycoeonjugates are variable and tissue/stage-specific in the developing limb.

畸形原的 危害性 判別을 위한 Hydra Attenuata System의 評價

全容赫 고려대학교 의과대학 1983 고려대 의대 잡지 Vol.20 No.2

The multiplicity of potential teratogenic mechanisms requires that a rapid and practical screening system for adverse effects on development must be able to assess not just one or two developmental events but as much of the entire phenomena as possible because it is not yet possible to explain the mechanism on triggering event for and teratogenic outcome. Artificial hydra "embryo" is obviously quite different from a real embryo on first inspection. On closer examination, however, it seems to share a great many developmental capabilities with any ontogenic system. Sequence of events the "embryo" undergoes during reaggregation and regeneration, such as spatical orientation, cell sorting, recognition of neighbors and formation of cell junction, formationa of extracellular material, tissue organization, response to induction and formation of organ fields all appear to be similar to those events required during mammalian embryogenesis. In addition, the "embryo" 1) has an obvious and complex ontogeny, 2) reacts to a teratogen in a dose=related manner, 3) has an evident threshold effect to the drug, 4) illustrate that developmental critical periods exist and that the developmental stage at the time of insult is more important perhaps than the severity or nature of the insult, 5) reacts in a somewhat stereotyped manner and 6) the system itself, also can provide the two relevant dose levels based on toxicity as adequately as standard laboratory rodents but at a significantly lower cost in time and effort.

Vitamin E 投與가 마우스 諸組織에 미치는 影響에 關한 實驗的 硏究 : Ⅰ. 骨格筋纖維에 關한 組織學的 觀察

全容赫 고려대학교 의과대학 1973 고려대 의대 잡지 Vol.10 No.2

Changes of muscle fiber size that follow vitamin E administration and vitamin E deficiency in the mouse were observed. Mean diameter and percentile size distribution were calculated in the sartorius and back muscles. Of sartorius muscle, the mean diameter was measured as 36.0 microns and their size distribution extends from 10 microns to 70 microns in normal control group. Of back muscle, mean diameter was 35.5 microns and range also between 10 to 70 microns. Vitamin E administration resulted slight increase in the mean diameter of fibers, while a deficiency caused positive decrease and irregularity in size distribution in both muscles.

白鼠胎仔 골격발생과정에 미치는 Chlorambucil과 Tocopherol의 相互作用에 關한 實驗的 硏究

宋志南,全容赫 고려대학교 의과대학 1982 고려대 의대 잡지 Vol.19 No.3

The effect of tocopherol to the teratogenecity due to chlorambucil have been studied on the developing skeleton in prenatal offsprings from 27 mothers of albino rat through macroscopic and microscopic observations. 1. None of the significant abnormality has been observed on either macroscopic or microscopic features in the fetal group treated with single dose of tocopherol administration the mother, except, the group treated with continuous 3 doses of it to the mother, of which the measures of the weight and stature of them are generally decreased than those of the normal contrast. 2. Though the degree of teratogenecity on the developing fetal skeletal frame due to the chlorambucil is mainly depend upon the amount of the drug and the time of administration during pregnancy of mother, the highest to teratogenetic effect could be elicitable on the fetuses at the day 12th, while, none of significant effect could be detectable in the fetuses after the time of day 14th of pregnancy on the administration of the chlorambucil. 3. The protective effectiveness of the tocopherol from the teratogenecity on the fetal skeletogenesis due to the chlorambucil appears not to be efficient. On the contrary, the concomitant use of tocopheral with chlorarmbucil tends to be facilitate the teratogenetic effect of the chlorambucil, furthermore, the rate is higher against with increase of the date of pregnancy.