Gerbil 삼차신경절에서 허혈-재관류 후 calbindin D-28k와 calretinin 면역반응의 경시적 변화

황인구(In Koo Hwang),박정훈(Jeong-Hoon Park),최우제(Woo-Je Choi),박노진(Noh-Jin Park),오해수(Hae-Soo Oh),안성진(Sung Jin An),박승국(Seung-Kook Park),윤대근(Dae-Kun Yoon),이원학(Won-Hak Lee),강태천(Tae-Cheon Kang),원무호(Moo Ho Won) 대한해부학회 2002 Anatomy & Cell Biology Vol.35 No.5

허혈에 대한 연구는 중추신경계통에서 대부분을 차지하고 있다. 그러나 허혈은 중추신경 뿐 아니라, 말초신경에도 많은 영향을 미칠 것으로 생각된다. 특히 머리 부분에 있는 대부분 장기들의 일반감각을 담당하는 삼차신경절은 매우 중요할 것으로 생각한다. 신경전달물질의 분비와 감각의 조절 등에 중요한 칼슘결합단백질은 세포질내 칼슘을 조절하여 감각의 전달에 매우 중요하다. 따라서 본 연구는 Mongolian gerbil의 삼차신경절에서 인위적으로 허혈-재관류를 유발하여 calbindin D-28k (CB)와 calretinin (CR)의 경시적인 변화와 그 의미를 연구해 보고자 하였다. 정상군에서 CB과 CR 면역반응은 큰 크기, 중간 크기, 작은 크기의 신경세포에서 관찰되었다. 허혈-재관류 후 12시간까 지는 면역반응세포 수의 변화에 큰 차이를 관찰할 수 없었다. 허혈-재관류 후 1일 경에는 CB과 CR을 함유한 큰 신경세 포의 수가 급격하게 증가하였으며, 그 이후부터 CB 면역반응세포는 감소하였고, CR 면역반응세포는 허혈-재관류 후 4일 경에 중간 크기 신경세포가 정상군보다 3배 가량 증가하였다. 이상의 연구 결과는 허혈-재관류 후 1일 경에 큰 신경세포에서 CB와 CR 면역반응의 증가가 있었는데, 큰 신경세포들 은 A fibers를 함유하고 있으므로, 이 시기에 CB와 CR이 기계적 자극의 전도에 영향을 줄 것으로 생각이 된다. 또한 허 혈-재관류 후 4일 이후에 중간 크기의 신경세포에서 CR 면역반응 세포의 증가는 이들이 Aδ 또는 C fiber를 함유하고 있 으므로, 이 시기에는 통증이나 온도 감각에 CR이 관여할 것으로 생각된다. Many researches have focused upon temporal changes of neurotransmitters and/or neuromodulators in the central nervous system after ischemic insult. In sensory neurons, the spatial and temporal alterations of neurotransmitters have been little studied. Calbindin D-28k (CB) and calretinin (CR) have been suggested to play a role in the transmission of neurotransmitters. Therefore, in the present study we investigated the chronological alteration of CB and CR immunoreactivity in the trigeminal ganglion cells of the Mongolian gerbil after ischemic insult. In the sham operated group, CB and CR immunoreactivities were found in small-, medium- and large-sized neurons. One and two days after ischemia-reperfusion, small and large-sized CB immunoreactive neurons increased significantly. Thereafter, number of the CB immunoreactive neurons decreased markedly. Furthermore, five days after ischemia-reperfusion, CB immunoreactivity was detected in a few neurons, and its immunoreactivity was also very weak in the cytoplasm. Number of the large-sized CR immunoreactive neurons increased significantly one day after ischemia-reperfusion. Thereafter, the number of the large-sized CR immunoreactive neurons decreased. Especially, the number of the medium-sized CR immunoreactive neurons increased dramatically 4 days after ischemia-reperfusion. These results suggest that an increase of CB and CR may play an important role in modulating the mechanoception 1 day after ischemia-reperfusion, because the immunoreactivities increased in large-sized neurons which have the myenlinated A fibers. These results also suggest that significant increase of CR expression in medium-sized neurons 4 and 5 days after ischemia-reperfusion may provoke CR in modulating the nociception or thermoception because the medium-sized neurons which have the myenlinated Aδ or C-fibers.

한국재래산양 십이지장의 장관신경계통에 분포하는 Substance P, CGRP 및 칼슘결합단백질 반응세포에 대한 면역조직화학적 연구

이인세,이흥식,송승훈,윤성태,황인구,강태천,원무호,서제훈,Lee, In-se,Lee, Heungshik S.,Song, Seung-hoon,Yoon, Sung-tae,Hwang, In-koo,Kang, Tae-cheon,Won, Moo-ho,Seo, Je-hoon 대한수의학회 1999 大韓獸醫學會誌 Vol.39 No.3

The localization of substance P(SP), calcitonin gene-related peptide(CGRP) and three calcium binding proteins, calbindin D-28k(CB), calretinin(CR) and parvalbumin(PA) was immunohistochemically examined in the myenteric and submucous plexuses of Korean native goat duodenum. In the neurons of myenteric and submucous plexuses of duodenum, immunoreactivities of SP, CGRP and CB were confirmed in both nerve cell bodies and fibers. In contrast, CR immunoreactivity was found only in nerve fibers of myenteric plexuses, while PA immunoreactivity was found only in nerve cell bodies of submucous plexuses. In the inner circular muscle layer, dense SP-like immunoreactive fibers were prominent but only a few CGRP-like immunoreactivities were observed. Most of SP- and CGRP-like immunoreactive neurons of both plexuses colocalized with CB. This result showed that SP and CGRP may have a important role for the movement of small intestine. The colocalizations of CB with SP or CGRP in myenteric and submucous plexuses suggest that CB may serve neuromodulatory role for SP- and CGRP-immunoreacted neurons on the movement of intestinal wall.

한국재래산양 송과체와 앞쪽목신경절의 관계규명을 위한 면역조직화학적 연구

이흥식,이인세,송승훈,윤성태,황인구,이충현,Lee, Heungshik S.,Lee, In-Se,Song, Seung-hoon,Yoon, Sung-tae,Hwang, In-koo,Lee, Choong-hyun 대한수의학회 2000 大韓獸醫學會誌 Vol.40 No.2

The pineal body have been known to be affected by superior cervical ganglia, and most of its nerve fibers containing peptidergic neurotransmitters have been considered to be originated from this ganglia. To confirm this relationships, some peptidergic neurotransmitters were identified in both of pineal body and superior cervical ganglia of the Korean native goat, which were divided into two group; breeding season and non-breeding season. The localizations of two catecholamine-synthesizing enzymes; tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH), were investigated by immunohistochemistry in the superior cervical ganglia and the pineal body of adult Korean native goats. Substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY) and galanin (GAL) were also identified in these organs by immunohistochemical and double immunofluorescent methods. In superior cervical ganglia, immunoreactivities for TH and DBH were confirmed in the same ganglion cells. The immunoreactivites for SP, VIP(only in male), NPY and GAL were identified in both of ganglion cell bodies and nerve fibers in the ganglia. CGRP immunoreactivity, however, was observed only in nerve fibers. Most NPY- and VIP-immunoreactive(IR) ganglion cells also contained TH. SP and TH were colocalized in the cell bodies, but not in the nerve fibers. TH immunoreactivity was shown in almost all of ganglion cells in the superior cervical ganglia. The immunoreactivity for NPY had some seasonal variation and was stronger in breeding season than in non-breeding season. In pineal body, lots of TH-IR fibers were observed throughout the parenchyma including the pineal stalk and most of them also contained DBH. SP- and NPY-IR fibers were also immunostained with TH or DBH. But a few SP- and NPY-IR fibers were not colocalized with TH or DBH. Exceptionally, a bipolar neuron-like cell was observed to be immunostained with NPY in the pineal body. A few CGRP and GAL-IR fibers were observed, while VIP-IR fibers were not present. It is concluded that most TH- and DBH-IR fibers as well as the peptidergic immunoreactive fibers of the pineal body might be originated from the superior cervical ganglia. Some peptidergic immunoreactive fibers, however, might be come from other regions of brain. We also suggest that NPY in pineal body plays a important role for pineal function. The seasonal variation of NPY immunoreactivity indicates that the synthesis and use of NPY may be different between in breeding and non-breeding seasons.

한국재래산양 시각교차위핵에 관한 형태학적 연구

송승훈,이흥식,이인세,황인구,이충현,Song, Seung-hoon,Lee, Heungshik S.,Lee, In-se,Hwang, In-koo,Lee, Choong-hyun 대한수의학회 2000 大韓獸醫學會誌 Vol.40 No.4

The present study was undertaken to investigate the morphological characteristics of the suprachiasmatic nucleus (SCN) of the Korean native goat by cresyl violet stain. The SCN was located ventrolateral to the third ventricle and dorsal to the optic chiasm. This nucleus was showed carrot form in longitudinal section. Its size was 1.8 mm in length, 0.9 mm in maximum height and 1.6 mm in maximum width. In coronal sections, the SCN shaped very thin plate in rostral part, but it was changed to sweet-potato form in middle part, and ornamental jade form in caudal part. The size of SCN was larger in caudal part than in rostral part. The size of the neuron of SCN was about $10{\mu}m$ in diameter with round or oval shape. The boundary of SCN was not firmly defined in caudal part because the neurons were dispersed into the hypothalamic region. It is concluded that the SCN of the Korean native goat has a morphological characteristics.

Immunogold법에 의한 한국재래산양 샘뇌하수체의 성샘자극세포에 관한 연구

이인세,이흥식,송승훈,윤성태,황인구,서제훈,강태천,원무호,Lee, In-se,Lee, Heungshik S.,Song, Seung-hoon,Yoon, Sung-tae,Hwang, In-koo,Seo, Je-hoon,Kang, Tae-cheon,Won, Moo-ho 대한수의학회 1999 大韓獸醫學會誌 Vol.39 No.5

There have been a number of studies of gonadotropes secreting LH and FSH in the adenohypophysis, but the pattern of hormone storage and secretion of these cells still remains a controversial matter. In this study, we examined whether gonadotropes contained both of LH and FSH, and if so, how these hormones were distributed within the secretory granules. Hypophyseal sections of Korean native goat were simultaneously immunostained for LH and FSH antisera by protein A-gold technique. It was found that most gonadotropes contained both FSH and LH, but hormone storages in the secretory granules were some different among cells. Three types of gonadotropes were identified by the shape and size of the secretory granules and their hormone storage patterns. One type(I) of gonadotropes contained oval secretory granules, which immunoreactivity for FSH and LH were very weak. The size of secretory granules ranged from 160 to 310nm in diameter. Most granules contained both FSH and LH, but some contained only one of them. In another type(II) of gonadotropes, the immunreactivity and hormone storage patterns of the secretory granules were similar to those of type I cells. However, the secretory granules were round in shape and larger in size than those of type I. The other gonadotropes(type III) were distinctly distinguished by plenty of hormones in their secretory granules which were densely packed with numerous immunolabelled gold particles. These data are some inconsistent with other results that have been obtained in other ruminants like as cattle and sheep.

Wistar 랫드와 자연발증 고혈압 랫드(SHR)의 주후각망울에서 노화에 따른 catalase 면역반응의 변화

김덕수(Duk Soo Kim),유기연(Ki Yeon Yoo),안성진(Sung-Jin An),박승국(Seung-Kook Park),황인구(In Koo Hwang),정주영(Ju-Young Jung),이경우(Kyoung Woo Lee),조준휘(Jun Hwi Cho),강태천(Tae-Cheon Kang),원무호(Moo Ho Won) 대한해부학회 2003 Anatomy & Cell Biology Vol.36 No.5

Catalase (CAT)는 신진대사의 과정동안 Cu, Zn-superoxide dismutase (SOD)나 Mn-SOD에 의해서 oxygen radical (O2-)이나 hydroxyl radical (OH-)이 H2O2로 분해되고, 이렇게 분해된 H2O2를 H2O와 O2로 변환시키는 작용을 하기 때문에 reactive oxygen species (ROS)에 대항하는 중요한 내인성 항산화효소중의 하나이다. 따라서 본 연구에서는 6개월령에서 24개월령까지의 Wistar 랫드와 유전적으로 자연발증 고혈압 랫드인 spontaneous hypertensive rat (SHR)의 주후각망울에서 CAT의 경시적인 변화를 관찰하여 다음과 같은 결과를 얻었다. Wistar 랫드에서의 CAT 면역반응은 glomerular layer (GL), external plexiform layer (EPL), internal plexiform layer (IPL) 그리고 granule cell layer (GCL)에서 나타났다. CAT 면역반응은 노화와 함께 서서히 감소하였으며, 24개월령에서는 거의 면역반응이 사라지는 것을 관찰할 수 있었다. 6개월군과 12개월군의 EPL에서 나타난 CAT 면역반응 신경세포는 형태학적으로 granule cell과 유사하였다. 그러나, 18개월군과 24개월군의 EPL과 mitral cell layer (MCL)에서 나타난 CAT 면역반응 신경세포는 각각 tufted cell과 형태학적으로 유사하였다. SHR에서의 CAT 면역반응은 주후각망울의 전 층에서 관찰되었으며, 노화와 함께 서서히 감소하는 것을 관찰할 수 있었다. 특히 24개월군의 GL과 EPL에서 면역반응이 현저하게 감소하는 것을 알 수 있었다. 이러한 결과는 CAT 면역반응의 감소가 산화적 손상 (oxidative stress)의 증가에 대한 원인 중의 하나로 작용하고, 이러한손상이 주후각망울의 노화에 따른 후각 저하와 밀접한 연관성이 있음을 시사한다. 더욱이 Wistar 랫드보다 SHR에서 CAT 면역반응이 상대적으로 빠르게 감소하는 것은 고혈압과 연관된 신경손상의 원인과 관련이 있는 것으로 추측된다. The reactive oxygen species (ROS) is well-known for the causative factors inducing ischemia, Parkinson’s disease, Alzheimer, amylotrophic lateral sclerosis, hypertension and aging. Catalase (CAT) is an important endogenous antioxidant enzyme against ROS because it removes H2O2 during metabolic processes. Hence, we examined the age-related changes of CAT-immunoreactivity in the main olfactory bulbs (MOB) of the Wistar and spontaneous hypertensive rat (SHR) at various aging stages over 2 years periods; postnatal month 6 (PM 6), PM 12, PM 18 and PM 24. CATimmunoreactive (IR) neurons in Wistar rats were located in the glomerular layer (GL), external plexiform layer (EPL), internal plexiform layer (IPL) and granule cell layer (GCL). The number of CAT-IR neurons slightly decreased agedependently and nearly disappeared at PM 24. At PM 6 and PM 12, the CAT-IR neurons located in the EPL were morphologically identified as granule cells. However, at PM 18 and PM 24, CAT-IR neurons located in the EPL and mitral cell layer (MCL) were morphologically identified as tufted and mitral cells, respectively. CAT-IR neurons in the SHR were located in all layers of the MOB. The number of CAT-IR neurons and CAT immunoreactivity decreased agedependently and nearly disappeared especially in the GL and EPL at PM 24. These findings indicate that the decrease of CAT immunoreactivity may be one of the causative factors for increase of oxidative stress, and these damages may underlie age-related changes in the olfactory process. The early decrease of CAT immunoreactivity in the SHR than in the Wistar rat suggests that the early decreae of CAT may be associated with the cause of hypertensive neuronal damage

간질에 의한 Mongolian gerbil 줄무늬체에서의 미세관 관련 단백질들의 변화

안성진(Sung-Jin An),박승국(Seung-Kook Park),황인구(In-Koo Hwang),원무호(Moo Ho Won),강태천(Tae-Cheon Kang) 대한해부학회 2002 Anatomy & Cell Biology Vol.35 No.5

유전적인 간질 모델 중에 하나인 gerbil의 줄무늬체에서 미세관 관련 단백질 1A (microtubule-associated protein 1A, MAP1A)와 미세관 관련 단백질 2 (microtubule-associated protein 2, MAP2) 면역반응의 변화를 관찰한 결과 다음과 같은 결과를 얻었다. 정상군의 경우, MAP1A 면역반응성은 줄무늬체의 꼬리-조가비핵의 신경세포체에서 미약하게 관찰된 반면, 간질전 군 (pre-seizure group)에서는 신경세포체 및 신경가지에서 강하게 관찰되었다. 그러나, 간질발작 30분 후에는 신경세포체의 MAP1A 면역반응이 감소하기 시작하여, 간질발작 3시간 후에는 정상군과 유사한 반응성을 나타내었다. MAP2 면역반응성은, MAP1A의 경우와 유사하여, 정상군에 비해 간질전군에서 강한 염색성을 나타내었다. 간질발작 30 분 후에는 신경세포체 및 신경망에서의 MAP2 면역반응성이 감소하기 시작하여, 간질발작 3시간 후에는 매우 미약한 MAP2 면역반응성이 관찰되었다. 그러나 이러한 MAP1A 및 MAP2 면역반응성의 감소는 간질발작 12시간 후에 모두 간질전군 수준으로 회복되었다. 이러한 결과는 간질군에서 MAPs 면역반응성의 증가 및 간질발작에 따른 MAPs 면역반응성의 감소는 줄무늬체에서의 MAPs 발현의 변화는 간질발작과 매우 밀접한 연관성이 있음을 시사한다. 따라서, 간질군에서의 MAPs 면역반응성의 변화는 간질발작에 의한 신경세포손상을 복구하기 위한 일종의 보상작용으로 추측된다. The present study involves a chronological and comparative analysis of both microtubule-associated protein 1A (MAP1A) and microtubule-associated protein 2 (MAP2) immunoreactivities in the striatum of both seizure resistant (SR) and seizure sensitive (SS) gerbil. The MAP1A immunoreactivity is weakly detected in perikarya of SR gerbils. However, MAP1A immunoreactivity is more accumulated in perikarya and dendrites in the pre-seizure group. At 30 min postictal, MAP1A immunoreactivity in the perikarya is decreased. At 3 hr postictal, MAP1A immunoreactivity in perikarya and dendrites is similarly decreased to the level of SR gerbils. The MAP2 immunoreactivity is weakly detected in the perikarya and dendrites of SR gerbils. However, MAP2 immunoreactivity is more accumulated in perikarya and dendrites. In particular, the neuropil between unstained fiber tracts obviously contains strong MAP2 immunoreactivity. At 30 min postictal, MAP2 immunoreactivity isn’t almost observed in striatum. At 3 hr postictal, the MAP2 immunoreactivity is not different in the 30 min post-seizure groups but is only observed in the neuropil. However, at 12 hr postictal, the decrease of both MAP1A and MAP2 immunoreactivities had recovered to the pre-seizure level of SS gerbils. These results suggest that MAPs immunoreactivity in the striatum is different in SR and SS gerbils, and that this difference may be the results of seizure activity in this animal.

Pregnancy Inhibits Neuroblast Differentiation without Neuronal Damage in the Hippocampal Dentate Gyrus of C57BL/6 Mice

In Koo Hwang(황인구),Dae Young Yoo,Ki-Yeon Yoo,Choong Hyun Lee,Jung Hoon Choi,Ji Won Choi,Je Kyung Seong,Yeo Sung Yoon,Moo-Ho Won 한국실험동물학회 2009 한국실험동물학회 학술발표대회 논문집 Vol.2009 No.2

Effects of Adrenalectomy on the Cell Proliferation and Neuroblasts in the Dentate Gyrus

In Koo Hwang(황인구),Sun Shin Yi,Dae Young Yoo,Ki-Yeon Yoo,Choong Hyun Lee,Jung Hoon CHoi,Moo-Ho Won,Je Kyung Seong,Yeo Sung Yoon 한국실험동물학회 2009 한국실험동물학회 학술발표대회 논문집 Vol.2009 No.-

불확실한 연속형 및 이산형 시스템에서의 이상검출법

황인구,권오규,Hwang, In-Koo,Kwon, Oh-Kyu 대한전자공학회 1990 전자공학회논문지 Vol.27 No.10

이 논문은 비선혀성, 모델링오차 그리고 잡음입력이 존재하는 선형 및 비선형시스템에서의 모델에 근거한 이상검출방법을 제시한다. 대상 시스템은 연속형이나 이산형 모두에 적용할 수 있도록 통합연산자$(unified operator)^[5]$로써 표시한다. 이 논문에서 제시되는 이상검출법은 잡음과 모델의 부정합과 비선형성을 고려한 것이다. 모델링 오차는 더하기꼴로 나타내며 계수추정에서 불확실성의 한계를 정량화시키기 위해 공칭모델 분모는 사건실험을 통해 고정시키는 것으로 한다. 공칭모델의 분자 계수들은 최소자승법으로 추정한다. 컴퓨터 모의실험을 추정하여 이 논문에서 제시한 방법이 기존의 방법보다 우수한 성능을 지니고 있음을 보인다. This paper proposes a model-based fault detection method for linear/nonlinear system having modelling errors, nonlinearities and measurement noise. The system model is represented by the unified operator [5] in order to apply to both the continuous-time and discrete-time problems. The fault detection method suggested here accounts for the effects of noise, model mismatch and nonlinearities. Modelling errors are depicted by additive forms and the nominal model denominator is fixed via prior experiments in order to quantify the nucertainty bound on the parameter estima-tion. The least square method is used to estimate the numerator parameters of the nominal model. performance than traditional methods.