(3.236.214.19) 您好!臺灣時間:2021/05/09 21:20
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果

詳目顯示:::

: 
twitterline
研究生:林以文
研究生(外文):LIN YI-WEN
論文名稱:長期正腎上腺素性神經纖維去除對大白鼠CA1突觸可塑性的影響
論文名稱(外文):The Effect of Chronic Norepinephrine (NE) Fibers Depletion on Synaptic Plasticity in the Rat CA1 Synapses
指導教授:閔明源楊琇雯楊琇雯引用關係
指導教授(外文):MIN YUAN-MINGYANG HSIU-WEN
學位類別:碩士
校院名稱:中國醫藥學院
系所名稱:醫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2001
畢業學年度:89
語文別:中文
論文頁數:91
中文關鍵詞:海馬體長期記憶正腎上腺素長期增益
外文關鍵詞:HIPPOCAMPUSLONG TERM MEMORYNOREPINEPHRINE6OHDALTPLTD
相關次數:
  • 被引用被引用:0
  • 點閱點閱:111
  • 評分評分:系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔
  • 下載下載:17
  • 收藏至我的研究室書目清單書目收藏:0
於此研究中,我們的目的在於觀察對於大白鼠海馬體中長期的正腎上腺素性神經纖維的去除對於海馬迴CA1區域突觸可塑性的影響。大白鼠出生後連續給予皮下注射6OHDA (6-Hydroxydopamine) 100mg/Kg 或vehicle (0.1% ascorbic acid in saline) 四天,每隻動物每次注射量不超過0.1 ml,之後給予飼養於適當之環境。之後於第35-40天時,我們以DBH (dopamine-β-hydroxylase) 免疫組織染色的方式來觀察對照組與實驗組動物中正腎上腺素神經纖維的差異。在海馬體中,我們發現對照組動物的正腎上腺素神經纖維仍然完整,相對的於實驗組動物中其正腎上腺素神經纖維皆被6OHDA神經毒素給破壞了,可是在其他CNS的區域如subcortical,對照組和實驗組則沒有顯著的差異。
接著我們以離體海馬迴腦薄片及標準細胞外記錄的技術來觀察對照組和實驗組動物其海馬體CA1區域突觸可塑性的差異。我們以100 Hz頻率的θ波來引發long term potentiation (LTP)和用1Hz頻率900個刺激波來引發long term depression (LTD)。結果顯示於對照組中LTP及LTD皆能被引發,且能維持到實驗結束(40分鐘);於實驗組中LTD的引發和表現和對照組並無差異,然而LTP並無法被引發,我們由此得知正腎上腺素神經纖維的破壞會影響LTP的表現。
接著我們使用雙電極的方式來測試bath application正腎上腺素對於對照組及實驗組突觸可塑性的影響。於我們實驗中我們得知,於對照組的海馬迴薄片中加入正腎上腺素對於LTP並無影響,然而會抑制LTD。相對的在實驗組的實驗中我們得知,原本無法維持LTP的現象可以因外界加入正腎上腺素而恢復至和對照組一致。
進一步的,我們使用正腎上腺素各型接受器的致效劑來觀察NE對突觸可塑性的影響是經由何種接受器的作用,我們的結果顯示,是經由β型接受器而不是α型接受器。接著更進一步的探討β型接受器是透過何種次級傳導物質來發揮它的功能,於是我們加入各種次級傳導物質的活化劑,結果顯示正腎上腺素可經由活化β型接受器,進一步影響活化adenylyl cyclase,增加cAMP,並活化protein kinase A而可使得原本無法維持LTP的現象恢復至和對照組動物一致。
由本研究可以得知:
1.長期性的正腎上腺素神經纖維去除會使得突觸可塑性中的LTP表現異常,但是LTD並無差異。
2.長期正腎上腺素神經纖維去除後LTP表現異常的現象可以因bath application正腎上腺素而恢復至正常,而不是多巴胺。
3.正腎上腺素可以使得原本LTP表現異常的現象恢復是透過β型接受器,活化Gs蛋白質,因而活化了adenylyl cyclase,使得cAMP濃度上升,進一步活化protein kinase A使LTP得以表現。

In this study,we want to know the effect of chronic depletion of NE fibers on the bi-directional synaptic plasticity in rat hippocampus.Postnatal rat pups were given with 6-hydroxydopamine (100mg/Kg) or the vehicle (0.1% ascorbic acid in saline)for 4 days,starting at the day of birth.These procedures caused loss of NE fibers in the cortex and hippocampus,while spared those in subcortical areas,when estimated by dopamine-β- hydroxylase immunohisto- chemistry at postnatal day 35-40.Hippocampal slices taken from NE depleted or control animals aged from 25-40 days were subjected to standard electro- physiology techniques for induction of long term potentiation or depression (LTP or LTD) in CA1 synapses,using 100HZ theta burst or 900 pulse of 1 HZ stimulation.In control slices,both LTP (41.32±11.33%,N=8)and LTD(73.18±5.28%,N=8) were successfully induced;But in NE depleted slices,while the induction and expression of LTD (73.47±4.73%,N=8)seemed to be normal,LTP(2.84±5.27%,N=8) could not be induced successfully.Bath application of NE during the theta burst stimulation restored LTP(50.22± 8.50%,N=8) in NE depleted slices,it has no further effect of NE in control slices(58.04±14.81%,N=8). But bath application of NE not only in control(98.69±8.58%,N=8) but also NE depleted slices(102.92± 5.52% ,N=8)could inhibit the expression of LTD.While exchange the NE into dopamine into the bath,another depleted neuromodulator,it has no effect on bi-directional plasticity both in control and NE depleted slices.These results suggested that endogenous Ne but not dopamine is crucial for induction of LTP and inhibition of LTD.

目 錄
封面
謝辭
目錄…………………………………………………………….I
圖目錄………………………………………………………...III
中文摘要……………………………………………………....V
第一章前言………………………………………………….1
第二章總論………………………………………………….3
第一節中樞神經的突觸傳導及突觸的可塑性………….3
第二節Long term Potentiation (LTP)及Long Term Depression (LTD)………………………………………………9
第三節海馬體在中樞神經學習及記憶中所扮演之角色………………………………………………...13
第四節中樞神經的正腎上腺素系統…………………...17
第五節此篇研究的目的………………………………...21
第三章實驗材料…………………………………………...23
第四章實驗方法…………………………………………...24
第一節實驗動物………………………………………..24
第二節 免疫組織化學染色法…………………………..24
第三節 電生理實驗…………………………………………27
第五章 結果………………………………………………………30
第六章 討論………………………………………………………64
第七章 參考文獻…………………………………………………75
英文摘要…………………………………………………………...81
圖目錄
Figure 1. DBH immunohistochemistry stain in the rat hippocampal
CA1 regions…………………………………………………….….43.
Figure 2. DBH immunohistochemistry stain in the rat hippocampal
CA3 regions………………………………………………………..44
Figure 3. DBH immunohistochemistry stain in the rat hippocampal
DG regions. ………………………………………………………..45
Figure 4. DBH immunohistochemistry stain in the rat subcortical
regions……… ………………………………….………………….46
Figure 5. Changes in synaptic efficancy upon conditioned stimulation
of different frequencyapplied to Schaffer collateral branch………..47
Figure 6. The relationship of conditioned stimulation frequency and fEPSP slope after stimulation in Con and 6OHDA slices………………………..48
Figure 7. The sample of two pathway stimulation of LTP……….…………...49
Figure 8 .The effect of Norepinephrine on LTP in control and 6OHDA slices………………………………………………………….…….50
Figure 9. The effect of Norepinephrine on LTD in control and 6OHDA slices. ………………………………………………………….…...51
Figure 10.The effect of dopamine on LTP in control and 6OHDA
Slices…………………………………………………..……….…..52
Figure 11.The effect of dopamine on LTD in control and 6OHDA
slices…………………………………………………………….….53
Figure 12.The effect of Phenylephrine on LTP in control and 6OHDA slices. ………………………………………………………….…...54
Figure 13.The effect of ISO on LTP in control and 6OHDA slices…………………………………………………………….….55
Figure 14. The effect of Phenylephrine on LTD in control and 6OHDA slices……….……………………………………………………….56
Figure 15.The effect of ISO on LTD in control and 6OHDA slices……….…57
Figure 16.The effect of Timolol on LTP in control slices………………….…58
Figure 17.LTP induced by TBS and ISO(or NE) with or without AP5 in 6OHDA slices………………………………………………………………..59
Figure 18.The effect of Forskolin in control slices…………………….……..60
Figure 19.Forskolin could restore the LTP in 6OHDA lesion slices…………61
Figure 20.Dibutyryl-cAMP and 8-bromo-cAMP have the same effect to restore the LTP in 6OHDA slices………………………………………..….62
Figure 21.LTP could not be maintained in PuTX and H89 pretreated slices…………………………………………………………..…….63

Alkadhi KA, AlHijailan RS, Malik K, Hogan YH. (2001) Retrograde carbon monoxide is required for induction of long-term potentiation in rat superior cervical ganglion. The Journal of Neuroscience, 21(10): 3515-3520.
Angulo MC, Lambolez B, Audinat E, Hestrin S, Rossier J.(1997) Subunit composition, kinetic, and permeation properties of AMPA receptors in single neocortical nonpyramidal cells.The Journal of Neuroscience, 17: 6685-6696.
Bartesaghi R and Ravasi L.(1999) Pyramidal neuron types in field CA2 of the guinea pig.Brain Research Bulletin, 50(4): 263-273.
Bear MF, Connors BW, Paradiso MA.(2001a).Synaptic transmission
in ”Neuroscience”. (Bear et al.,Ed). Chapter 5.
Bear MF, Connors BW, Paradiso MA.(2001b).Memory systems
In ”Neuroscience”. (Bear et al.,Ed). Chapter 23.
Bear MF, Connors BW, Paradiso MA.(2001c).Neurotransmitter systems.
in ”Neuroscience”. (Bear et al.,Ed). Chapter 6
Bear MF, Kirkwood A.(1996).Bidirectional plasticity of cortical synapses.
in”Cortacal Plasticity LTP and LTD”: pp.191-202.
Bear MF, Malenka RC (1994) Synaptic plasticity:LTP and LTD.Current Opinion
in Neurobiology, 4: 389-399
Bliss TVP, Lfmo T (1973): Long-lasting potentiation of synaptic transmission in
the Dentate area of the anaesthetized rabbit following stimulation of perforant
path. Journal of Physiology.(Lond), 232: 331-356
Bliss TVP, Collingridge GL.(1993).A synaptic model of memory: long-term
potentiation in the hippocampus. Nature, 361: 31-39
Blue ME,Parnavelas JG.(1982) The effect of neonatal 6-hydroxydopamine
treatment on synaptogenesis in the visual cortex of the rat. The Journal of
Comparative Neurology 205: 199-205.
Brown G P, Blitzer RD, Connor JH, Wong Tony, Shenolikar S, Iyengar R, and Landau EM(2000).Long-term potentiation induced by frequency stimulation is regulated by a protein phosphatase-1-operated gate. The Journal of Neuroscience, 20(21): 7880-7887
Calabresi P, Saiardi A, Pisani A, Baik JH, Centonze D, Mercuri NB, Bernardi G, Borrelli E.(1997) Abnormal synaptic plasticity in the striatum of mice lacking dopamine D2 receptors. The Journal of Neuroscience., 17 (12): 4536-4544
Chen WR, Lee S, Kato K, Spencer DD, Sheperd GM, Williamson A.(1996).Long-term modification of synaptic efficancy in the human inferior and middle temporal cortex. Proc.Natl Acad.Sci.USA 93: 8011-8015.
Cooper DMF, Mons N, Karpen JW.(1995).Adenylyl cyclase and the interaction between calcium and cAMP signaling. Nature, 374, 30.
Cooper LN.(1973) A possible organization for animal learning and
memory.In”Proceedings of the Nobel Symposium on collective Properties of
Physical Systems”. Academic Press, New York, pp.252-264.
Geiger JR, Melcher T, Koh DS, Sakmann B, Seeburg PH, Jonas P, Monyer H(1995).Relative abundance of subunit mRNAs determines gating and Ca2+ permeability of AMPA receptors in principal neurons and interneurons in rat CNS neuron.Neuron, 15, 193-204.
Hanson PI,Heuser JE,Jahn R.(1997) Neurotransmitter release-four years of
SNARE complexes. Current Opinion in Neurobiology 7:310-315.
Harik SI.(1984) Locus coeruleus lesion by local 6-hydroxydopamine infusion causes marked and specific destruction of noradrenergic neurons,long-term depletion of norepinephrine and the enzymes that synthesize it,and enhanced dopaminergic mechanisms in the ipsilateral cerebral cortex. The Journal of Neuroscience 4(3): 699-707
Heginbotham LR, Dunwiddie TV.(1991) Long-term increase in the evoked population spike in the CA1 region of rat hippocampus induced by β-adrenergic receptor activation. The Journal of Neuroscience , 11(8): 2519-2527
Hraetova S, Sacktor TC (1996).Bidirectional regulation of protein kinase Mζ
in the maintenance of long-term potentiation and long-term depression. The
Journal of Neuroscience, 16(17): 5324-5333
Huang Y-Y, Kandel ER (1994) Recruitment of long-lasting and protein kinase A-dependent long-term potentiation in the CA1 region of hippocampus requires repeated tetanization. Learn Mem 1: 74-82.
Huang YY, Kandel ER (1996).Modulation of both the early and the late
phase of mossy fiber LTP by the activation of b-adrenergic receptors
Neuron, 16: 611-617.
Hollmann M, Heinemann S(1994) Cloned glutamate receptors. Annual Review of Neuroscience.17: 31-108.
Jonas P, Bischofberger J, Sandkuhler J (1998) Corelease of two fast neurotransmitters at a central synaoses. Science 281: 419-424.
Jonas P, Racca C, Sakmann B, Seeburg PH, Monyer H(1994) Differences in Ca2+ permeability of AMPA-type glutamate receptor channels in neocortical neurons caused by differential GluR-B subunit expression. Neuron 12: 1281-1289
Johnston D and Amaral DG (1998) Hippocampus in “The synaptic organization of the brain”(GM Shepherd,Ed) chapter 11.
Katsuki H, Izumi Y, .Zorumski CF.(1997) Noradrenergic regulation of synaptic plasticity in the hippocampal CA1 region. The Journal of Neurophysiology 77: 3013-3020
Kirkwood A, Rozas C, Kirkwood J, Perez F, Bear MF.(1999) Modulation of long-term synaptic depression in visual cortex by acetylcholine and norepinephrine. The Journal of Neuroscience, 19 (5): 1599-1609
Kobayashi K, Noda Y, Matsushita N, Nishii K, Sawada H, Nagatsu T, Nakahara D, Fukabori R, Yasoshima Y, Yamamoto T, Miura M, Kano M, Mamiya T, Miyamoto Y, and Nabeshima T(2000) Modest neuropsychological deficits caused by reduced noradrenaline metabolism in mice heterozygous for a mutated tyrosine hydroxylase gene . The Journal of Neuroscience, 20: 2418-2426.
Larson J, Wong D, Lynch G (1986) Pattern stimulation at the theta frequency is optimal for the induction of hippocampal long-term potentiation. Brain Reasearch. 368: 347-350.
Maccaferri G, Toth K, McBain CJ.(1998).Target-specific expression of presynaptic Mossy fiber plasticity. Science. 279.27.
Makhinson M, Chotiner JK, Watson JB, O’Dell TJ.(1999) Adenylyl cyclase activation modulates activity-dependent change in synaptic strength and Ca2+
/Calmodulin-dependent kinase II autophosphorylation. The Journal of Neuroscience, 19 (12).
Milner B, Squire LR, Kandel ER.(1998) Congnitive neuroscience and the study of memory. Neuron 20: 445-468
Moore RY,Card JP (1984) Noradrenaline-containing neuron systems. Handbook
of chemical neuroanatomy, 2, 123-156.
Morris RGM, Anderson E, Lynch GS,Baudry M (1986).Selective impairment of learning and blockade of long-term potentiation by an N-methyl-D-aspartate receptor antagonist AP5. Nature 319: 774-776.
Mulkey RM, Endo S, Shenollkar S, Malenka RC.(1994).Involvement of a calcineurin/inhibitor-1 phosphatase cascade in hippocampal long-term depression. Nature, 369, 9 June.
Neher E.(1998).Vesicle pools and Ca2+ microdomains:new tools for
understandind their roles in neurotransmitter release. Neuron 20: 389-399.
Nicoll RA, Malenka RC, Kauer JA.(1990) Functional comparison of
neurotransmitter receptor subtypes in mammalian central nervous
system. Physiol.Rev.70: 513-565
Nicholls JG,Martin AR,Wallace BG(1992a) Principles of synaptic
transmission.in “FROM NEURON TO BRAIN”(J.G.Nicholls,Ed).Chapter 7.
Nicholls JG,Martin AR,Wallace BG(1992b) Cellular and molecular biochemistry of synaptic transmission.in “FROM NEURON TO BRAIN”(J.G.Nicholls,Ed).Chapter 9.
Oleskevich S, Descarries L, Lacaille JC (1989) Quantified distribution of the noradrenaline innervation in the hippocampus of adult rat. The Journal of Neuroscience, 9(11): 3803-3814
Otmakhova NA, Lisman JE.(1996) D1/D5 dopamine receptor activation
increases the magnitude of early long-term potentiation at CA1 hippocampal
synapses The Journal of Neuroscience, 16(23): 7478-7485
Otmakhova NA, Otmakhov N, Mortenson LH,Lisman JE.(2000) Inhibition of the cAMP pathway decrease early long-term potentiation at CA1 hippocampal synapses. The Journal of Neuroscience, 20(12), 4446-4451
Pockett S, Slack JR, Peacock S.(1993) Cyclic AMP and long-term potentiation in the CA1 region of rat hippocampus.Neuroscience. 52(2): 229-236.
Ravi lyengar.(1996) Gating by cyclic AMP:Expanded role for an old signaling pathway. SCIENCE. 271, 26 Jan.
Scanziani M, Gahwiler BH., Thompson SM. (1993) Presynaptic inhibition of excitatory synaptic transmission mediated by α-adrenregic receptors in area CA3 oh the rat hippocampus in vitro. The Journal of Neuroscience.13(12): 5393-5401
SharmaVK, Harik SI, Busto R, Banerjee SP.(1981) Effect of noradrenaline depletion on adrenergic and muscarinic cholinergic receptors in the cerebral cortex,hippocampus and cerebellum. Exp.Neurol.72: 179-194
Shepherd GM and Koch C.(1998a) Introduction to synaptic circuits.in”The synaptic organization of the brain”(G.M. Shepherd,Ed.), chapter1.
Sherrington C.(1906).Integrative action of the nervous system.New Haven:Yale
University Press
Trimarchi JR and Murphey RK.(1997) The shaking-B2 mutation disrupts
electrical synapses in a flight circuit in adult drosophila.The Journal of
Neuroscience, 17(12): 4700-4710.
Wang SJ, Chemg LL, Gean PW. 1999) Cross-modulation of synaptic plasticity by β-adrenergic and 5-HT1A receptors in the rat basolateral Amygdala. The Journal of Neuroscience, 19(2): 570-577
Winder DG, Martin KC., Muzzio IA, Rohrer D, Chruscinski A, Kobilka B, Kandel ER.(1999) ERK plays a regulatory role on induction of LTP by theta frequency stimulation and its modulation by β-adrenergic Receptor. Neuron 24: 715-726

QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
1. 楊清江,《警察業務概說》,警政學報(8),中央警官學校,民國74年。
2. 李湧清,《論當代民主社會中警察的角色與功能》,警學叢刊,30卷6期,民國89年。
3. 楊永年,《警察組織績效評估-就李總統六個月改善治安之承諾論述》,警學叢刊,28卷1期,民國86年7月。
4. 蔡進閱,《激勵管理與我國警察人員升遷體系之研究》,警學叢刊,24卷3期,民國83年3月。
5. 張金鑑,《人群關係的理論與方法》,人事行政季刊,第45期,民國82年。
6. 章光明,《從行政改革論我國警察業務功能之演變》,中央警察大學學報(34),民國88年。
7. 張世賢,《政策執行資源投入度之分析》,公共行政學報,第1期,民國74年。
8. 官政哲,《探討提升警政組織層級之政策思考與效益》,警光第五二二期,民國88年1月。
9. 李湧清,《論警大警專兩校合併-一個應重開的辯論與議題》,警學叢刊,29卷5期,民國88年3月。
10. 李湧清,《企業管理與警政經營-我們可以學到什麼》,警學叢刊,27卷3期,民國85年。
11. 吳堯峰,《臺灣省人事機構組織再造工程芻議》,人事月刊,21卷3期,民國84年。
12. 吳定,《再造工程方法應用於工作簡化之探討》,人事管理月刊,第34卷1期,民國85年。
13. 江明修,《非營利組織領導行為之研究》,問題與研究34卷10期,民國84年。
14. 朱楠賢,《談官僚機構的政治面》,人事月刊,10卷2期,民國79年。
15. 朱金池,《各國警察組織類型之研究:以美國日本及中國大陸為例》,警學叢刊,29卷2期,民國87年9月。
 
系統版面圖檔 系統版面圖檔