모체 Thyroxine 투여가 태아알코올효과를 가진 흰쥐 뇌의 BDNF함유 신경세포 생후 발달에 미치는 영향

강양수(Yang Soo Kang),정윤영(Yoon Young Chung),박영란(Young Lan Park),현영식(Young Sig Hyun),김종중(Jong Joong Kim),문정석(Jeong Seok Moon),문영민(Young Min Mun),오재욱(Jae Wook Oh),신성희(Sung Heui Shin),배춘상(Choon Sang Bae) 대한해부학회 2006 Anatomy & Cell Biology Vol.39 No.4

임부의 알코올 남용은 태아 정신발육지연의 흔한 원인이며 특히 태아의 뇌 발생에 예민한 결정적 기간 (critical period) 동안에 지속적으로 음주하는 경우 태아알코올효과를 나타내기 쉽다. 본 연구에서는 임신 기간중 지속적으로 알코올을 섭취한 모체에 thyroxine을 투여하여 알코올의 유해한 영향으로 인한 태아알코올효과를 개선시킬 수 있는지를 관찰하기 위하여 흰쥐 뇌의 생후 연령에 따라 brain-derived neurotrophic factor(BDNF)함유 신경세포의 성숙 양상을 면역조직화학염색을 이용하여 관찰하였고 BDNF 함량을 측정하였다. 실험동물은 매일 35 칼로리 정도의 알코올을 섭취한 알코올군, 알코올 대신 dextrin이 첨가된 유동액을 섭취한 정상군 및 알코올을 섭취하고 thyroxine을 매일 5 μg/kg 피하 주사한 알코올+T4군으로 나누었으며 생후 0, 7, 14, 21, 28일에 희생시켰다. 본 연구 결과 BDNF 단백 양은 알코올+T4군에서 알코올군에 비해 생후 7, 14, 21일에 증가하였으며 특히 생후 7일의 알코올+T4군에서 가장 높았다. 알코올군은 모든 연령에서 정상군보다 감소하였다. 소뇌에서는 알코올+T4군에서 생후 14일부터 정상군과 유사한 BDNF함유 신경세포들이 조롱박세포층에 분포하였다. 알코올+T4군에서는 알코올군에 비해 대뇌겉질, 시상하부 및 해마에서 생후 7일에 성숙되고 두드러진 양성반응을 나타냈으며 해마에서는 생후 28일까지도 뚜렷한 양성반응을 나타냈다. 이상의 결과는 임신 중 알코올을 음용한 흰쥐 모체에 thyroxine을 투여하면 태아알코올효과를 가지고 태어날 수 있는 후손들의 뇌에 분포하는 BDNF함유 신경세포들이 생후 발달동안 정상군과 유사하거나 더 빠르게 발달시킬 수 있을 것으로 생각된다. 이는 모체에 투여하는 지속적인 thyroxine 처치가 출생 초기에 BDNF 합성을 증가시켜 모체의 알코올 남용으로 야기되는 태아알코올효과와 같은 출생결함을 개선시킬 수 있음을 시사한다. Maternal alcohol abuse is thought to be the common cause of mental retardation. Especially, continuous alcohol consumption during critical period of brain development induce fetal alcohol effects. In this study, the authors investigated the effects of maternal alcohol drinking on the postnatal changes of BDNF contents and patterns of BDNF-containing neuron in neonatal rat brain, and, the influence of maternal thyroxine treatment on the brain of pups of alcohol abused mother. Pregnant rats were divided into three groups. Alcohol-fed group (n=4) received 35 calories of liquid alcohol diet daily from gestation day 6; control pair-fed group (n=4) was fed a liquid diet in dextrin replaced alcohol isocalorically; alcohol+T4 group (n=4) received 35 calories liquid alcohol diet and exogenous thyroxine (5 μg/kg/day) subcutaneously. The amount of BDNF was significantly higher in the alcohol+T4 group as compared to the alcohol group at P7, P14 and P21, especially, alcohol+T4-exposed pups showed a significant increase of BDNF at P7. The decrease in BDNF was found in alcohol group compared to control pair-fed group at all ages. In alcohol+T4 group, BDNF-containing Purkinje cells exhibited mature pattern and monolayer arrangement at P14. Alcohol+T4 group showed mature pattern and numerical increase of BDNF-containing cells in cerebral cortex, hypothalamus and hippocampus at P7. The BDNF immunoreactivity of hippocampus continued to show prominent configuration in alcohol+T4 group at P28. These results indicate that the increase of the BDNF-containing neurons and BDNF amount in pups of thyroxinesupplemented alcohol-exposed dams as compared to control pair-fed and alcohol-exposed pups at P7, presumably suggest the early postnatal growth stimulatory effect of the exogenously supplemented thyroxine. Therefore, the increase of BDNF synthesis caused by maternal administration of exogenous thyroxine may ameliorate fetal alcohol effects, one of the ill effects as a result of the dysthyroid state following maternal alcohol abuse.

척수를 손상시킨 후 꼬리정맥에 주입한 사람탯줄혈액세포가 뇌줄기에 미치는 영향

김종중,정윤영,박영란,문영민,현영식,정영욱,문정석 朝鮮大學校 附設 醫學硏究所 2007 The Medical Journal of Chosun University Vol.32 No.1

Background and Objectives: Stem cells are a valuable resource for treatment of many disease, but limited access to stem cells in some organs such as brain restricts their utility. Many approaches have been attempted to restore the function following brain stem injury (BSI) and spinal cord injury (SCI). The use of the human umbilical cord blood cells (hUCB) - a rich source of nonembryonic or adult stem cells - has recently been reported to ameliorate the behavioral consequences of stroke. Mateiials and Methods: Forty rats were divided into 8 groups: (1) SCI l+hUCB (infused 1 day post injury); (2) SCI 2+hUCB (infused 2 days post injury); (3) SCI 3+hUCB (infused 3 days post injury); (4) SCI 4+hUCB (infused 4 days post injury); (5) SCI 5+hUCB (mfusedt 5 days post injury); (6) SCI 6+hUCB (infused 6 days post injury); (7) LO+hUCB (laminectomy+hUCB); and (8) LO (laminectomy only). SCI was produced by compressing the spinal cord for one minute with an aneurysm clip calibrated to a closing pressure of 50 g. We report here that immunhistotochemical identification of fluorescent hUCB positive cells in the brain stem after compressed spinal cord injury using mouse anti-human mitochondria monoclonal antibody (MAB1273). Results: All SCI+hUCB (1-8) groups contained fluorescent hUCB positive cells in the all area of the brain stem. But especially a large number of fluorescent hUCB positive cells were observed in the whole area of the brain stem of the experimental 5 (SCI 5+hUCB) and 6 (SCI 6+hUCB)groups. No hUCB positive cells were found in the brain stem of group with non-injured spinal cord of these animals and group with laminectomy only. Conclusion: These results suggest that hUCB are potentially useful as a vector for treating a variety of the central nervous system disorders, and we are sure that continuous of stem cell study will give an best opportunity to treat the uncurable disorders in the future.

모체 Thyroxine 투여가 태아알코올효과를 가진 흰쥐 대뇌겉질 및 해마에서 NPY함유 신경세포의 생후 발달에 미치는 영향

김복,박상기,박영란,김종중,문정석,김주수,문영민,현영식,천관영,정윤영 朝鮮大學校 附設 醫學硏究所 2005 The Medical Journal of Chosun University Vol.30 No.2

Background and Objectives: Maternal alcohol abuse is considered to be one of the most prominent cause of neurobiological malformations in the postnatal and adult life of the offspring. In this study, we investigated the effects of maternal alcohol drinking on the postnatal development of NPY-containing neuron, and, the influence of thyroxine treatment on the cerebral cortex and hippocampus of pups of alcohol abused mother. Materials and Method: Time-pregnant rats were divided into three groups. Alcohol-fed group A received 35 calories of liquid alcohol diet daily from gestation day 6; control pair-fed group B was fed a liquid diet in dextrin replaced alcohol isocalorically: alcohol + T4 group C received 35 calories liquid alcohol diet and exogenous thyroxine subcutaneously. Results: Group C showed prominent NPY immunoreactivity in the cerebral cortex compared to group A and B at P7. In group C, NPY-containing neurons were widely distributed in the all layers of cerebral cortex after P14. Besides, numerical decrease of NPY-containing neuron as age increases was not found in group C. However, the decrease of NPY-containing neuron was clearly observed in group A compared to group C after P14. In hippocampus, group Band C were appeared similar patterns after P7. Additionally, in group C, NPY immunoreactivity was prominently appeared in CA2 and CA3 at P14 as compared to group B. Conclusions: The present results showed the increase of intensity and number of NPY-containing neurons in cerebral cortex and hippocampus of pups of exogenous T₄ supplemented alcohol-exposed dams as compared to control pair-fed and alcohol-exposed pups at P7. It presumably suggest that NPY-containing neurons might be regulated by the early postnatal growth stimulatory effect of the exogenously supplemented T₄. Therefore, the increase of NPY synthesis caused by maternal administration of exogenous thyroxine may ameliorate fetal alcohol effect, one of the ill effects as a result of the dysthyroid state following maternal alcohol abuse.

Chinchilla 달팽이핵에서 Calbindin D-28ka 면역반응세포의 미세구조에 관한 연구

문영민,박영란,정윤영,김종증 朝鮮大學校 附設 醫學硏究所 2008 The Medical Journal of Chosun University Vol.33 No.3

Chinchilla 달팽이핵의 신경표지물질의 하나인 calbindin D-28ka은 칼슘결합단백질의 일종으로 여러 포유동물의 신경계통에 널리 분포하고 있는 것으로 알려지고 있다. Chinchilla 달팽이핵에서 CB에 대한 고도의 특이성을 지닌 단세포군항체를 이용하여 광학 및 전자현미경적 면역화학염색방법을 이용하여 CB양성세포의 분포상과 세포의 형태 및 크기와 CB세포의 미세구조에 대하여 관찰한 결과 다음과 같은 결론을 얻었다. 1. Calbindin D-28ka 양성반응세포는 chinchilla 달팽이핵의 전 분야에서 관찰할 수 있었으나 특히 등쪽달팽이핵에서 더 많이 관찰할 수 있었으며 CB양성세포의 형태는 방추형, 타원형, 이극 및 뭇극형이었고 세포의 크기는 16-30?μm 였다. 2. 수많은 미토콘도리아와 축삭세포체 연접, 미세섬유등을 관찰할 수 있으며 축삭을 둘러싸고 있는 말이집과 calbindin D-28ka 양성반응세포의 침착물과 모세혈관을 관찰할 수 있었고 커다란 핵과 축삭의 주변에 작은 슈반핵과 말이집으로 둘러싸인 가로 단면의 수많은 축삭들을 관찰할 수 있었다. 이상의 결과로 보아 calbindin D-28ka 양성반응세포는 청각기능세포를 보호할 뿐만 아니라 청각기능을 수행하는데 매우 중요한 역할을 하는 것으로 판단된다. A unique class cells, strongly immunopositive for anti-calbindin D-28ka were observed in and near the cochlear nucleus of the chinchilla (chinchilla laniger). They were preferentially located in the dorsal cochlear nucleus, as well as in the adjacent superior olivary nucleus. The calbindin D-28ka positive neurons had relatively large cell bodies with thick, heavy spinal dendrites, and were typically situated in the immediate subpial position. The calbindin D-28ka immunoreactive neurons were very varied morphologically; bitufted, bipolar, multipolar and horizontal cells in shape, and the diameters of the neurons are 16-30 μm have been described in the cochlear nucleus of the chinchilla. Also, the calbindin D-28ka positive dendrites were considerably more densely arrayed than those of other cochlear nuclear cells, including the cartwheel cells of the dorsal cochlear nucleus. The fine structure of the cells in the cochlear nucleus in the chinchilla(chinchilla laniger) was studied by transmission electron microscopy. The neurons had a large, round, centrally located nucleus, and their cytoplasm contained numerous mitochondria, myelinated axon, dense granular endoplasmic reticulum, abundant Golgi apparatus, groups of free ribosomes, some heterogeneous particles and neurofilaments. Cell surfaces were studied irregularly with small spinous processes, and they contained a few fine irregularly arranged neurofilaments and some granular endoplasmic reticulum. Boutons contacting the soma and dendrites of the cochlear nucleus neurons were defined as the axosomatic synapses. Based on differences in the bouton and vesicle morphology, the four synaptic bouton types were identified; 1. Asymmetrical and symmetrical synapses with small rounded vesicles. 2. Asymmerical synapses with pleomorphic admixture contained predominantly spherical vesicles. 3. Symmerical synapses with pleomorphic vesicles of flattened, spherical and dense-core vesicles. 4. Asymmetrical as well as symmetrical synapses with heterogeneous and large dense-core vesicles. Synaptic boutons containing the rounded vesicles and the large densecore vesicles were most frequently observed.

한국인 태아 및 사산아에서 얼굴신경에 대한 형태학적 연구

강양수,김종중,김복,정상봉,문영민,설경수,정윤영 朝鮮大學校 附設 醫學硏究所 2005 The Medical Journal of Chosun University Vol.30 No.3

It is critical that the accurate anatomy of the facial nerve IS known for uncomplicated successful facial surgery. Such an operation is hazardous because of the intimate relationship between the facial nerve and the expressional muscles. It is hoped that such basic information will be helpful to those who treat conditions involving the facial region. In order to define the detail of this relationship, many studies have been undertaken on dissected adult specimens. This study was based on Korean fetus and stillborn infant cadaveric dissections, and describes anatomical variations of the peripheral branches of the facial nerve that pose a potential danger in a number of surgical procedure on the face. The branching patterns were classified into six types: the frequencies of occurrence were: type I, 4.9%; type II, 24.4%; type III, 34.1%; type IV, 19.5%; type V, 12.2%; and type VI, 4.9%. Types II, III and IV together accounted for almost 80% of the specimens. In the discussion we compare the results of the present study with those of others and try to clarify the anatomical characteristics of the facial nerve in infants, which demand that special attention be paid to the avoidance of injuries during facial surgery.