KISEP Original Articles J Korean Neuropsychiatr Assoc Vol 38, No 2 내측전전두피질에 6-hydroxydopamine 주입후 흰쥐신선조체도파민수용체의변화에관한 자가방사기록법적연구 권준수 * 이동영 *

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1 KISEP Original Articles J Korean Neuropsychiatr Assoc Vol 38, No 2 내측전전두피질에 6-hydroxydopamine 주입후 흰쥐신선조체도파민수용체의변화에관한 자가방사기록법적연구 권준수 * 이동영 * 우종인 * An Autoradiographic Study on the Rat Neostriatal Dopamine Receptor Changes after 6-hydroxydopamine Injection into the Medial Prefrontal Cortex Jun Soo Kwon, M.D.,* Dong Young Lee, M.D.*, Jong Inn Woo, M.D.* 국문초록 중심단어 서 Department of Psychiatry & Institute for Neuroscience, Seoul National University, College of Medicine, Seoul Corresponding author 론 408

2 409

3 1. 실험동물및재료 연구대상및방법 2. 입체외과수술 (Stereotaxic surgery) Fig. 1. Cursor box placements in brain regions of coronal sections collected 9.2 to 10.2mm anterior to the interaural plane are represented here at anterior. Left hemisphere labels apply to neuroanatomical names according to Paxinos and Watson1986. ac, anterior commissurecc, corpus calosumclau, claustrumcpu, caudate-putamenfrpam, frontal parietal motor parietal motor cortexfrpass, frontal parietal somatosensory cortexlot, lateral olfactory tractna, nucleus accumbensot, olfactory tubercle. 3. 조직준비및보온 (Incubation) 410

4 Fig. 2. Saturation curve for 3 H-SCH bindingd1 receptor binding in caudate-putamen sections of rat. Specific and non-specific binding of 3 H- SCH to striatal sections. Fig. 4. Saturation curve for 3 H-spiperone bindingd2 receptor binding in caudate-putamen sections of rat. Specific and non-specific binding of 3 H- spiperone to striatal sections. Fig. 3. Representive scatchard plot of specific 3 H-SCH bindingd1 receptor binding in caudateputamen sections of rat. Fig. 5. Representive scatchard plot of specific 3 H-spiperone bindingd2 receptor binding in caudateputamen sections of rat. 411

Fig. 6. Autoradiographic distribution of [ 3 H]SCH 23390 binding in caudate-putamen and nucleus accumbens.

Total binding(left side) and nonspecific binding(right part) are shown. cold buffer로 5분간 씻은 후 0 의 증류수에 담근 후 실온에서 건조시켰다.

0nM의 6가지 농도를 사용하 였고 역시 5-HT2 수용체 및 α1 아드레날린 수용체를 차단하기 위하여 mianserin 100nM을 넣었다.

5 Fig. 6. Autoradiographic distribution of [ 3 H]SCH binding in caudate-putamen and nucleus accumbens. Illustrated are a 6-hydroxydopamine injected rat(upper part) and a shamlesioned rat(lower part). Total binding(left side) and nonspecific binding(right part) are shown. cold buffer로 5분간 씻은 후 0 의 증류수에 담근 후 실온에서 건조시켰다. 비특이적 결합을 측정하기 위하 여 특이결합에 사용된 바로 옆 절편을 각각 butaclamol 10μM을 넣고 보온하였다. D2 수용체 결합을 측정하기 위하여 [3H]spiperone 0.3, 0.5, 0.7, 1.0, 1.5, 2.0nM의 6가지 농도를 사용하 였고 역시 5-HT2 수용체 및 α1 아드레날린 수용체를 차단하기 위하여 mianserin 100nM을 넣었다. 같은 방 법으로 ice cold buffer로 5분간 씻은 후 0 의 증류수 에 담근 후 실온에서 건조시켰으며, 비특이적 결합을 측 정하기 위하여 바로 옆 절편을 각각 butaclamol 2μM 을 넣고 보온하였다. 4. 자가방사기록법 및 컴퓨터 영상분석 Fig. 7. Comparison of Bmax and Kd of D1 receptor in caudate-putamen and nucleus accumbens between control and 6-OHDA group. *p<0.05, as compared with control group(by Mann-Whitney U-test) 뇌조직이 있는 유리슬라이드를 빛을 차단하는 X-ray cassette에서 Hyperfilm-3H 위에 순서대로 나란히 배 이 놓은 후 실온의 암실에서 3주([3H]SCH23390), 또 열한다. 광학도를 fmol/mg tissue weight 값으로 전환 는 4주([3H]spiperone)간 보관한 후 Kodak D19로 3 하기 위해 표준 [3H]-microscale(Amersham)도 같 분간 현상한 후 1분 간 증류수에 씻는다. 다시 Kodak 412

Fig. 8. Autoradiographic distribution of [ 3 H]spiperone binding in caudate-putamen and nucleus accumbens.

Total binding(left side) and nonspecific binding(right part) are shown. rapid fixer에 약 4분 간 고정한 후 깨끗이 씻고 건조 D2 수용체의 결합도를 보기 위해 미상핵-피각, 측좌 시킨다.

총결합에서 바로 옆 뇌절편의 영상인 비특이성결 analysis system을 이용하여 분석을 하였다. Video 합을 뺀 것이 특이성결합인데 fmol/mg tissue weight camera(dage-mti series 68 NEWVICON)에 의해 로 계산하였다.

Video signal은 512X512 pixel에 chard 분석을 시행하여 최대 결합력(maximum number digitize되고 각 pixel의 밝기는 256 gray level로 바뀌 of binding site)인 Bmax과 해리 상수(dissociation 게 된다.

6 Fig. 8. Autoradiographic distribution of [ 3 H]spiperone binding in caudate-putamen and nucleus accumbens. Illustrated are a 6-hydroxydopamine injected rat(upper part) and a sham-lesioned rat(lower part). Total binding(left side) and nonspecific binding(right part) are shown. rapid fixer에 약 4분 간 고정한 후 깨끗이 씻고 건조 D2 수용체의 결합도를 보기 위해 미상핵-피각, 측좌 시킨다. 핵 부위를 분석하였고(그림 1) 정확한 지형학적 위치 감광된 필름의 영상을 분석시스템인 digital autoradio- 는 Paxinos와 Watson의 Atlas(1986)를 참조하였 graphic system인 Amersham RAS-R1000 receptor 다. 총결합에서 바로 옆 뇌절편의 영상인 비특이성결 analysis system을 이용하여 분석을 하였다. Video 합을 뺀 것이 특이성결합인데 fmol/mg tissue weight camera(dage-mti series 68 NEWVICON)에 의해 로 계산하였다. 이와 같이 얻은 결과들을 포화곡선(그 필름을 통과한 빛을 기록한 후 color TV monitor에 영 림 2 for D1 그림 4 for D2)을 산출하였고, Scat- 상화시키게 된다. Video signal은 512X512 pixel에 chard 분석을 시행하여 최대 결합력(maximum number digitize되고 각 pixel의 밝기는 256 gray level로 바뀌 of binding site)인 Bmax과 해리 상수(dissociation 게 된다. constant)인 Kd를 구하였다(그림 3 for D1 그림 5 먼저 거리를 검도(calibration)한 후 microscale이 감 광된 부위의 평균광학도(optical density)를 이용하여 표준곡선을 구한다. 이 표준곡선에 의해 측정하고자 하 는 부위의 감광정도에 따라 광학도를 구한다. 정량분석은 관심영역(area of interest)을 설정하고 관심영역부위에서 마우스(mouse)로 직사각형 부위를 만들고 그 부위의 광학도의 평균을 측정하였다. D1 및 for D2). 5. 통계처리 실험군인 6-OHDA 투여군과 대조군의 Bmax와 Kd 를 비교하기 위하여 SPSS 프로그램(version 7.01)을 이용하여 비모수 통계방법인 Mann-Whitney test를 사용하였다 413

7 Fig. 9. Comparison of Bmax and Kd of D2 receptor in caudate-putamen and nucleus accumbens between control and 6-OHDA group. 결 1. 내측전전두엽 (MPFC) 에 6-OHDA 투여후흰쥐뇌의 D 1 수용체의변화 2. 내측전전두엽 (MPFC) 에 6-OHDA 투여후흰쥐뇌의 D 2 수용체의변화 과 고찰 414

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서론 50 Korean J Psychopharmacol 1998;91:49-57

서론 50 Korean J Psychopharmacol 1998;91:49-57 KISEP Original Articles 9 1 1998 Haloperidol, Clozapine 및 Haloperidol과 Serotonin 성제제병합약에의한흰쥐뇌 FOS 면역반응성 ABSTRACT 김광수 박원명 서상원 FOS-like Immunoreactivity in Rat Brain Induced by Haloperidol, Clozapine, and