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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.
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.
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.
Hajin Nam(남하진),In Koo Hwang(황인구),Harry Jung(정혜리),Seung-Hae Kwon(권승해),Ok Kyu Park(박옥규),Jun Gyo Suh(서준교) 한국생명과학회 2013 생명과학회지 Vol.23 No.9
인슐린은 근육세포 표면으로 포도당 수송체 4(glucose transporter 4, GLUT4)를 유도하여 혈액 속의 포도당을 세포 내로 유입시키도록 작용한다고 알려져 있다. Fagopyritol은 인슐린과 유사한 작용을 하는 것으로 알려져 있으므로, 본 연구에서는 혈당강하 효과가 있다고 알려진 fagopyritol을 랫드의 근육세포주(L6GLUT4myc 세포)에 처리하여, 아직 명확하게 밝혀지지 않은 fagopyritol의 혈당강하 기전을 규명하고자 수행하였다. Fagopyritol의 혈당강하 기전을 규명하기 위하여 근원세포(myoblast)와 근관세포(myotube)에 fagopyritol을 처리하여 액틴 필라멘트의 구조와 GLUT4에 미치는 영향을 분석하였다. Fagopyritol을 myoblast에 처리하였을 때, GLUT4가 처리군에서 대조군과 비교하여 유의 있게 원형질막 쪽으로 유도되는 것을 확인하였고, 액틴 필라멘트의 구조가 재조정 되면서 GLUT4의 이동을 돕는 것으로 생각된다. 또한 fagopyritol이 인슐린과 유사한 작용 경로를 가지는지 확인하기 위하여, 인슐린 작용 경로에서 중요한 역할을 하는 것으로 알려진 phosphatidylinositol 3-kinase (PI3K)의 억제제인 LY294002를 fagopyritol과 함께 처리하였을 때 GLUT4가 원형질막 쪽으로 유도되지 않는 것을 확인하였다. Fagopyritol을 myotube에 처리하였을 때, myoblast에 처리하였을 때와 유사한 결과를 나타내었다. 이러한 결과를 종합하면 fagopyritol이 인슐린과 유사한 작용을 하여 액틴 필라멘트의 구조 변경과 GLUT4의 이동을 촉진 시키는 것으로 사료된다. Insulin induces glucose transporter 4 (GLUT4) translocation to the muscle cell surface. As fagopyritol has insulin-like effects, the effects of fagopyritol on GLUT4 translocation and filamentous (F) actin remodeling in L6-GLUT4myc skeletal muscle cells were investigated. Fagopyritol significantly increased plasma membrane GLUT4 levels compared with the basal control in L6-GLUT4myc myoblast cells. Phosphatidylinositol (PI) 3-kinase inhibitor (LY294002) treatment prevented GLUT4 translocation to the plasma membrane in the myoblasts. Fagopyritol treatment apparently stimulates F-actin remodeling in myoblasts. In addition, fagopyritol treatment induced GLUT4 translocation and F-actin remodeling in myotubes. Taken together, these results suggest that fagopyritol promotes GLUT4 translocation and F-actin remodeling by activating the PI 3-kinase-dependent signaling pathway.
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.