KISEP Experimental Research J Korean Neurosurg Soc , 1998 일과성뇌허혈후 Amyloid Precursor Protein mrna 와 Amyloid Precursor Protein 발현의변화 * 이상형 김

Similar documents
untitled

Chapter 26

γ

27 2, 1-16, * **,,,,. KS,,,., PC,.,,.,,. :,,, : 2009/08/12 : 2009/09/03 : 2009/09/30 * ** ( :

(Establishment and Management of Proteomics Core Facility)

환경중잔류의약물질대사체분석방법확립에 관한연구 (Ⅱ) - 테트라사이클린계항생제 - 환경건강연구부화학물질연구과,,,,,, Ⅱ 2010


Crt114( ).hwp

Lumbar spine


388 The Korean Journal of Hepatology : Vol. 6. No COMMENT 1. (dysplastic nodule) (adenomatous hyperplasia, AH), (macroregenerative nodule, MR

???춍??숏

Can032.hwp

Pharmacotherapeutics Application of New Pathogenesis on the Drug Treatment of Diabetes Young Seol Kim, M.D. Department of Endocrinology Kyung Hee Univ

Analyses the Contents of Points per a Game and the Difference among Weight Categories after the Revision of Greco-Roman Style Wrestling Rules Han-bong

DBPIA-NURIMEDIA

Dementia2

012임수진

<303920C3D6BFB5BCF728B1E8BBF3C8A3292E687770>

2009;21(1): (1777) 49 (1800 ),.,,.,, ( ) ( ) 1782., ( ). ( ) 1,... 2,3,4,5.,,, ( ), ( ),. 6,,, ( ), ( ),....,.. (, ) (, )

Kbcs002.hwp


패션 전문가 293명 대상 앙케트+전문기자단 선정 Fashionbiz CEO Managing Director Creative Director Independent Designer

54 한국교육문제연구제 27 권 2 호, I. 1.,,,,,,, (, 1998). 14.2% 16.2% (, ), OECD (, ) % (, )., 2, 3. 3

A 617

<35BFCFBCBA2E687770>

untitled

hwp

077~088 해계 유침과 전침이

(01) hwp

(

( )Kju269.hwp

( )Kjhps043.hwp

?

jaeryomading review.pdf

달생산이 초산모 분만시간에 미치는 영향 Ⅰ. 서 론 Ⅱ. 연구대상 및 방법 達 은 23) 의 丹 溪 에 최초로 기 재된 처방으로, 에 복용하면 한 다하여 난산의 예방과 및, 등에 널리 활용되어 왔다. 達 은 이 毒 하고 는 甘 苦 하여 氣, 氣 寬,, 結 의 효능이 있


16_이주용_155~163.hwp


재료및방법 1. 해마신경세포의분리와배양 전기생리학적측정 J Korean Neurosurg SocVolume 29November, 2000

저작자표시 - 비영리 - 변경금지 2.0 대한민국 이용자는아래의조건을따르는경우에한하여자유롭게 이저작물을복제, 배포, 전송, 전시, 공연및방송할수있습니다. 다음과같은조건을따라야합니다 : 저작자표시. 귀하는원저작자를표시하여야합니다. 비영리. 귀하는이저작물을영리목적으로이용할

Analysis of objective and error source of ski technical championship Jin Su Seok 1, Seoung ki Kang 1 *, Jae Hyung Lee 1, & Won Il Son 2 1 yong in Univ

α α α α α

< D B4D9C3CAC1A120BCD2C7C1C6AEC4DCC5C3C6AEB7BBC1EEC0C720B3EBBEC8C0C720BDC3B7C2BAB8C1A4BFA120B4EBC7D120C0AFBFEBBCBA20C6F2B0A E687770>


Alzheimer병과 Gangliosides 401 단세포군 항체 제조 재료와 방법 대뇌의 주요 gangliosides인 GM1, GD1a, GD1b, GT1a에 대한 4종류의 단세포군 항체를 하이브리도마법을 이용하여 제조 하여 각각을 GMB16, GMR17, GGR1

<C1A63534C8B820BCBCB9CCB3AA2DC6EDC1FD2E687770>

스포츠과학 143호 내지.indd

03-서연옥.hwp

09È«¼®¿µ 5~152s

03-ÀÌÁ¦Çö

[ 영어영문학 ] 제 55 권 4 호 (2010) ( ) ( ) ( ) 1) Kyuchul Yoon, Ji-Yeon Oh & Sang-Cheol Ahn. Teaching English prosody through English poems with clon

example code are examined in this stage The low pressure pressurizer reactor trip module of the Plant Protection System was programmed as subject for

44-4대지.07이영희532~

김범수

서론 34 2

(Exposure) Exposure (Exposure Assesment) EMF Unknown to mechanism Health Effect (Effect) Unknown to mechanism Behavior pattern (Micro- Environment) Re



노영남

main.hwp

???? 1

09-감마선(dh)

목 차 회사현황 1. 회사개요 2. 회사연혁 3. 회사업무영역/업무현황 4. 등록면허보유현황 5. 상훈현황 6. 기술자보유현황 7. 시스템보유현황 주요기술자별 약력 1. 대표이사 2. 임원짂 조직 및 용도별 수행실적 1. 조직 2. 용도별 수행실적

<35335FBCDBC7D1C1A42DB8E2B8AEBDBAC5CDC0C720C0FCB1E2C0FB20C6AFBCBA20BAD0BCAE2E687770>

1..

<30372EC0CCC0AFC1F82E687770>

Jksvs019(8-15).hwp

19(1) 02.fm

32

Kaes010.hwp

간질과산화스트레스 - 발작으로인한산화스트레스 아코니타제 (Aconitase) 의비활성 (Inactivation) - 지방과산화 (Lipid peroxidation) DNA 산화 (DNA oxidation) SOD 변이생쥐 (SOD mutant mice)

7.ƯÁýb71ÎÀ¯È« š

step 1-1

264 축되어 있으나, 과거의 경우 결측치가 있거나 폐기물 발생 량 집계방법이 용적기준에서 중량기준으로 변경되어 자료 를 활용하는데 제한이 있었다. 또한 1995년부터 쓰레기 종 량제가 도입되어 생활폐기물 발생량이 이를 기점으로 크 게 줄어들었다. 그러므로 1996년부

139~144 ¿À°ø¾àħ

목원 한국화- 북경전을 준비하며 지난해부터 시작 된 한국의 목원대학교 한국화 전공의 해외미술체험은 제자와 스승의 동행 속에서 미술가로 성장하는 학생들의 지식에 샘을 채워주는 장학사업으로 진행되고 있으며, 한국의 우수한 창작인력 양성을 위해, 배움을 서로 나누는 스승들의

03이경미(237~248)ok

590호(01-11)

황지웅

untitled

한국전지학회 춘계학술대회 Contents 기조강연 LI GU 06 초강연 김동욱 09 안재평 10 정창훈 11 이규태 12 문준영 13 한병찬 14 최원창 15 박철호 16 안동준 17 최남순 18 김일태 19 포스터 강준섭 23 윤영준 24 도수정 25 강준희 26

[ReadyToCameral]RUF¹öÆÛ(CSTA02-29).hwp

Journal of Educational Innovation Research 2018, Vol. 28, No. 3, pp DOI: The Effect of Caree

(11+12.).pdf

충북의대학술지 Chungbuk Med. J. Vol. 27. No. 1. 1~ Charcot-Marie-Tooth Disease 환자의마취 : 증례보고 신일동 1, 이진희 1, 박상희 1,2 * 책임저자 : 박상희, 충북청주시서원구충대로 1 번지, 충북대학교

DBPIA-NURIMEDIA

뉴스래터수정12.20

09구자용(489~500)

<30352DB1E2C8B9C6AFC1FD2028C8ABB1E2C7F D36362E687770>


???? 1

노인정신의학회보14-1호

서강대학교 기초과학연구소대학중점연구소 심포지엄기초과학연구소

歯1.PDF

untitled

세포배양 방 TPH 효소활성측정 g 법 Korean J Psychopharmacol 2004;15(4):

DBPIA-NURIMEDIA

박선영무선충전-내지

KISEP Clinical Research J Korean Neurosurg Soc , 1999 척추동맥의박리성뇌동맥류의뇌혈관내수술 서범석 권양 권병덕 = Abstract = Endovascular Surgery of Vertebral Artery

Transcription:

KISEP Experimental Research J Korean Neurosurg Soc 271481-1489, 1998 일과성뇌허혈후 Amyloid Precursor Protein mrna 와 Amyloid Precursor Protein 발현의변화 * 이상형 김혜선 ** 정성진 ** 이영재 *** 서유헌 ** 양희진 정영섭 한대희 조병규 = Abstract = The Change of Expression in Amyloid Precursor Protein mrna and Amyloid Precursor Protein by Transient Ischemia Sang Hyung Lee, M.D., Hye Sun Kim, Ph.D.,** Sung Jin Jeong, BSc,** Young-Jae Lee, Ph.D.,*** Yoo Hun Suh, M.D.,** Hee Jin Yang, M.D., Young Seob Jung, M.D., Dae Hee Han, M.D., Byung-Kyu Cho, M.D. Departments of Neurosurgery and Pharmacology,** Seoul National University College of Medicine, Seoul, Korea Department of Veterinary Medicine,*** Cheju National University, Cheju, Korea KEY WORDS 서 론 J Korean Neurosurg SocVolume 27November, 1998 1481

일과성뇌허혈후 Amyloid Precursor Protein mrna 와 Amyloid Precursor Protein 발현의변화 연구범위및방법 1. 뇌허혈동물모델제조법 1482 2. 뇌조직에서 total RNA의분리 3. APP의 PCR 시발체 (Primer) 의설계 Table 1. Time schedule of experimental groups Group Occlusion time Reperfusion time A* B** 1 hr 0 hr C 1 hr 1 hr D 1 hr 3 hr E 1 hr 8 hr F 1 hr 1 day G 1 hr 3 day H 1 hr 7 day *normal control, **sham control, hrhour J Korean Neurosurg SocVolume 27 November, 1998

이상형 김혜선 정성진 이영재 서유헌 양희진 정영섭 한대희 조병규 4. 역전사중합효소연쇄반응 (RT-PCR(Reverse Transcription-Polymerase Chain Reaction)) 방법 Fig. 1. Amyloid Precursor Protein primer for PCR and their product size. 5. 단백질분리와 Western Blot 연구결과 1. 뇌허혈이유발된백서의대뇌피질에서의 APP mrna 의발현 J Korean Neurosurg SocVolume 27 November, 1998 1483

일과성 뇌허혈후 Amyloid Precursor Protein mrna와 Amyloid Precursor Protein 발현의 변화 는 더이상 증가하지 않아 30회로 고정하여 이후 실험을 시 후 실험에서는 DMSO를 첨가하지 않았다(Fig. 4 D-). 행하였다(Fig. 3). APP의 mrna의 발현은 뇌를 포함한 대 APP 유전자발현에 대한 일과성 허혈의 영향을 알아보기 부분의 조직에서 발현되는데 특히 APP695는 뇌에서 주로 위해 재관류후 1시간, 3시간, 8시간, 1일, 3일, 7일후에 백서의 나타났으며 APP770과 APP751는 매우 낮은 수준으로 발 대뇌피질을 적출하여 RNA를 추출한 후 RT-PCR을 시행한 현되었다. 반면, 신장과 간조직에서는 APP770과 APP751 결과, APP695의 mrna는 1일, 3일째에는 감소하는 것으로 이 주로 발현되었으며, 특히 신장에서는 APP751이 주로 보였으나 수술하기 전과 비교할 때 3일째에만 유의하게 감소 발현되는 것으로 나타났다(Fig. 4 D ). RT-PCR에 대한 하였다(p<0.05). APP751 및 APP770의 mrna는 1일 이 dimethyl sulfoxide(dmso)의 영향을 확인한 결과 DMSO 후에 증가하기 시작하여 3일째 최고 수준으로 증가하였으며, 를 첨가하지 않았을 때 PCR이 더 잘되는 것으로 나타나 이 7일째에는 다시 재관류하기 전으로 감소하였다. APP770의 mrna의 증가는 1일째에는 수술하기 전과 단순히 폐색시킨 것과 비교할 때 증가의 차이가 있었으며(p<0.01), 3일째에는 APP751 및 APP770의 mrna가 모두 통계적으로 유의한 증가를 보였다(p<0.01)(Fig. 5A, 5B, 7). 2. 뇌허혈이 유발된 백서의 대뇌피질에서의 APP단백질발현 허혈이 APP의 단백질의 발현에 대한 영향을 관찰하기 Fig. 2. Confirmation of the primer. PCR was performed using the synthesized primer with cdna. PCR products of 331, 274, 197bp equivalent to APP770, APP751, APP695 were obtained. Fig. 3. Amplication cycle-dependent PCR amplication of APP695 cdna. Fig. 4. The expression pattern of APP mrna isoforms in rat brain, liver and kidney(d Dimethylsulfoxide). 1484 Fig. 5. RT-PCR analysis of APP isoform mrna levels in cerebral cortex after I hour of ischemia. Lane M PCR marker, 1 group A(control), 2 group B(sham control), 3 group C, 4 group D, 5 group E, 6 group F, 7 group G, 8 group H. (A) Changes in the expression of APP 3 isoforms detected by RT-PCR. (B) Autoradiogram showing the changes in the expression of APP 3 isoforms, using 32P-α dctp β-actin band was weakened by reducing the exposure time for exact comparison with 3 APP isoform bands. Fig. 6. Immunoblotting analysis of the cortex homogenates after ischemia using 22C11. Lane 1 group A(control) 2 group B(sham control), 3 group C, 4 group D, 5 group E, 6 group F, 7 group G, 8 group H. J Korean Neurosurg Soc/ Volume 27 / November, 1998

이상형 김혜선 정성진 이영재 서유헌 양희진 정영섭 한대희 조병규 Fig. 7. The changes of ratio of mrna of three isoforms. APP770, APP751 and APP695 in cerebral cortex. Each point represent s the means.e.m.n5 percentage of the total of three isoforms. The intensity of the bands of five autoradiograms were quantitated by densitometer and statistical analysis was made by ANOVA**p0.01. Fig. 8. The effect of transient ischemia on the level of amyloid precursor protein in rat cerebral cortex. There was no significant difference by ANOVA. 고찰 J Korean Neurosurg SocVolume 27 November, 1998 1485

일과성뇌허혈후 Amyloid Precursor Protein mrna 와 Amyloid Precursor Protein 발현의변화 1486 J Korean Neurosurg SocVolume 27 November, 1998

이상형 김혜선 정성진 이영재 서유헌 양희진 정영섭 한대희 조병규 결론 References 1) Abe K, Tanzi RE, Kogure KInduction of HSP70 mrna after transient ischemia in gerbil brain. Neurosci Lett 125 166-168, 1991a 2) Abe K, Tanzi RE, Kogure KSelective induction of Kunitztype protease inhibitor domain-containing amyloid precursor protein mrna after persistent focal ischemia in rat cerebral cortex. Neurosci Lett 125172-174, 1991b 3) Araki W, Kitaguchi N, Tokushima Y, et altrophic effct of -amyloid precursor protein on cerebral cortical neurons in culture. Biochem Biophys Res Commun 181265-271, 1991 4) Busciglio J, Lorenzo A, Yankner BA, et almethodological variables in the assessment of beta amyloid neurotoxicity. Neurobiol Aging 13609-612, 1992 5) Card JP, Meade RP, Davis LGImmunocytochemic localization of the precursor protein for -amyloid in the central nervous system. Neuron 1835-846, 1988 6) Checler FProcessing of the beta-amyloid precursor protein and its regulation in Alzheimer s disease. J Neurochem 65 1431-1444, 1995 7) Chen ST, Hsu CY, Hogan EL, et ala model of focal ischemic strokereprducible extensive cortical infarction. Stroke 17 738-743, 1996 J Korean Neurosurg SocVolume 27 November, 1998 1487

일과성뇌허혈후 Amyloid Precursor Protein mrna 와 Amyloid Precursor Protein 발현의변화 8) Choi DWThe role of glutamate neurotoxicity in hypoxicischemic neuronal death. Ann Rev Neurosci 13171-182, 1990 9) Cochran E, Bacci B, Chen Y, et alamyloid precursor protein and ubiquitin immunoreactivity in dystrophic axons is not unique to Alzheimer s disease. Am J Pathol 139485-489, 1991 10) Coombs DJ, Dempsey RJ, Donaldson DHyperglycemia supresses c-fos mrna expression following transient cerebral ischemia in gerbil. J Cereb Flow Metabol 12169-172, 1992 11) Curran T, Franza BR JrFos and JunThe AP-1 connection. Cell 55395-397, 1988 12) Estus S, Golde TE, Kunishita T, et alpotentially amyloidogenic carboxyl-terminal derivatives of amyloid precursor protein. Science 255726-728, 1992 13) Frautschy SA, Baird A, Cole GMEffects of injected Alzheimer beta-amyloid cores in rat brain. Proc Natl Acad SciUSA 888362-8366, 1991 14) Glenner GAlzheimer s diseasethe commonest form of amyloidosis. Arch Path Lab Med 107281-282, 1983 15) Glenner GG, Wong CWAlzhemier s diseaseinitial report of the purification and characterization of a novel cerebrovascular amyloid protein. Biochem Biophys Res 120885-890, 1984 16) Goldgaber D, Lerman MI, Mcbride W, et alcharacterization and chromosomal localization of a cdna encoding brain amyloid of Alzheimer s disease. Science 235877-880, 1987 17) Hoffmann MC, Nitsch C, Scotti AL, et althe prolonged sequence of glia-derived nexin, an endogenous protease inhibitor, in the hippocampus after ischemia-induced delayed neuronal death. Neuroscience 49397-408, 1992 18) Jendroska K, Poewe W, Daniel SE, et alischemic stress induces deposition amyloid beta immunoreactiviy in human brain. Acta Neuropathol 90461-466, 1995 19) Jorgensen MB, Deckert J, Wright DC, et aldelayed C-fos protooncogene expression in the rat hippocampus induced by transient global ischemia. Brain Res 484393-398, 1989 20) Kang J, Lemaire H-G, Unterbeck A, et althe precursor of Alzheimer s disease amyloid A4 protein resembles a cell surface receptor. Nature 325733-736, 1987 21) Kawabata S, Higgins GA, Gordon JWAmyloid plaques, neurofibrillary tangles and neuronal loss in brains of transgenic mice overexpressing a C-terminal fragment of human amyloid precursor protein. Nature 334476-478, 1991 22) Kiessling M, Dienel GA, Jacewicz M, et alprotein synthesis in postischemic rat braina two dimensional electrophoretic analysis. J Cereb Blood Flow Metab 6642-649, 1986 23) Kitaguchi N, Takahashi Y, Tokushima Y, et alnovel precursor of Alzheimer s disease amyloid protein shows protease inhibitory activity. Nature 331530-532, 1988 24) Kogure K, Kato HAltered gene expression in cerebral ischemia. Stroke 242121-2127, 1993 25) Kowall NW, Beal NF, Busciglio J, et alan in vivo model for the neurodegenerative effects of beta amyloid and protection by substance P. Proc Natl cad SciUSA 887247-7251, 1991 26) Longa ZE, Weinstein PR, Carlson S, et alreversible middle cerebral artery occlusion without craniectomy in rats. Stroke 2084-91, 1989 27) Mattson MP, Cheng B, Culwell AR, et alevidence for excitoprotective and intraneuronal calcium-regulating roles of secreted forms of -amyloid precursor protein. Neuron 10243-254, 1993a 28) Mattson MP, Tomas KJ, Rydel RECalcium-destabilizing and neurodegenerative effects of aggravated beta-amyloid peptide are attenuated by basic FGF. Brain Res 62135-49, 1993b 29) Morgan JI, Curran TStimulus-transcription coupling in the nervous systeminvolvement of the inducible proto-oncogenes fos and jun. Ann Rev Neurosci 14421-451, 1991 30) Neve RL, Finch EA, Dawes LRExpression of the Alzheimer s amyloid precursor gene transcripts in the human brain. Neuron 1669-677, 1988 31) Nishimoto I, Okamoto T, Matsuura T, et alalzheimer amyloid protein precursor complexes with brain GTP-binding protein G. Nature 36275-79, 1993 32) Nowak Ts Jr, Bond U, Schesinger MJHeat shock RNA levels in brain and other tissues after hyperthermia and transient ischemia. J Neurochem 54451-458, 1990 33) Palpacios G, Mengod G, Tortosa A, et alincreased -amyloid precursor protein expression in astrocyte in the gerbil hippocampus follwoing ischemiaassociation with prolifeation of astrocytes. Euro J Neurosci 7501-510, 1995 34) Pike CJ, Burdick D, Walencewicz AJ, et alneurodegeneration induced by beta-amyloid peptide in vitrothe role of peptide assembly state. J Neurosci 131676-1687, 1993 35) Ponte P, Gonzalez-Dewhitt P, Schilling J, et ala new A4 amyloid mrna contains domain homologous to serine protease inhibitors. Nature 331525-527, 1988 36) Quon D, Wang Y, Catalano R, et alformation of -amyloid protein deposits in brains of transgenic mice. Nature 352 239-241, 1991 37) Robakis NK, Ramakrishna N, Walfe G, et almolecular cloning and characterization of a cdna encoding the cerebrovascular and the neurite plaque amyloid peptides. Proc. Natl Acad SciUSA 844190-4194, 1987 38) Robison PM, Clemens JA, Stephenson D, et aldecrease of amyloid precursor protein precedes hippocampal degenearation in rat brain following transient ischemia. Brain Res 608 334-337, 1993 39) Rush DK, Aschmies S, Merriman MCIntracerebral betaamyloid2535 produces tissue damageis it neurotoxic? Neurobiol Aging 13591-594, 1992 40) Saitoh T, Sundsmo M, Roch J, et alsecreted form of amyloid beta protein precursor is involved in the growth regulation of fibroblasts. Cell 58615-622, 1989 41) Salbaum JM, Weidermann A, Lemaire HG, et alethe promoter of Alzheimer s disease amyloid A4 precursor gene. EM- 1488 J Korean Neurosurg SocVolume 27 November, 1998

이상형 김혜선 정성진 이영재 서유헌 양희진 정영섭 한대희 조병규 BO J 72807-2819, 1988 42) Seubert D, Cole G, Saitoh T, et alamyloid beta protein is a mitogern. Biochem Biophys Res Commun 16283-88, 1989 43) Smith-Swintosky VL, Pettigrew LC, Craddok SD, et alsecreted forms of -amyloid precursor protein protect against ischemic brain injury. J Neurochem 63781-784, 1994 44) Stephenson DT, Rash K, Clemens JAAmyloid precursor protein accumulates in regions fo neurodegeneration following focal cerebral ischemia in the rat. Brain Res 593128-135, 1992 45) Tanaka S, Shiojiri S, Takahashi Y, et altissue-specific expression of three types of a protein precusor mrnaenhancement of protease inhibitor-harboring types in Alzheimer s disease. Biochem Biophys Res Commun 1651406-1414, 1989 46) Tanzi R, Mcclalatchey AL, Lamperti ED, et alprotease inhibitor domain encoded by an amyloid protein precusor mrna associated with Alzheimer s disease. Nature 331528-530, 1988 47) Tomimoto H, Akiguchi I, Wakita H, et alultrastuructural localization of amyloid precursor protein in the normal and post ischemic brain. Brain Res 672187-195, 1995 48) Uemura Y, Kowall NW, Beal MFGlobal ischemia induces NMDA recepter mediated c-fos expression in neurons resistant to injury in gerbil hippocampus. Brain Res 542343-347, 1991 49) Vass K, Welch WJ, Nowak TS JrLocalization of 70-kDa stress protein induction in gerbil brain after ischemia. Acta NeuropatholBerl 77128-135, 1988 50) Waite J, Cole GM, Frautsch SA, et alsolvent effects on beta protein toxicity in vivo. Neurobiol Aging 13595-599, 1992 51) Wakita H, Tomimoto H, Akiguchi I, et alregional accumulation of amyloid precursor protein in the gerbil brain following transient ischemia. Neuroscience Lett 146135-138, 1992 52) Yankner BA, Dawes LR, Fischer S, et alneurotoxity of a fragment of the amyloid precursor associated with Alzheimer s disease. Science 245417-420, 1989 53) Yankner BA, Duffy LK Kirssehner DANeurotrophic and neurotoxic effects of amyloid proteinreversal of by tachykinin neropeptides. Science 250279-282, 1990 54) Yoneda K, Okada YEffects of anoxia and recovery on the neurotransmission and level of high energy phosphates in thin hippocampal slices from the guinea-pig. Neuroscience 28 401-407, 1989 J Korean Neurosurg SocVolume 27 November, 1998 1489