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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 閔明源(Ming-Yuan Min) | |
| dc.contributor.author | Hsin-wei Lu | en |
| dc.contributor.author | 呂昕煒 | zh_TW |
| dc.date.accessioned | 2021-06-13T15:19:16Z | - |
| dc.date.available | 2009-07-26 | |
| dc.date.copyright | 2008-07-26 | |
| dc.date.issued | 2008 | |
| dc.date.submitted | 2008-07-24 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/37104 | - |
| dc.description.abstract | 食慾蛋白(Orexin)為大鼠下視丘一群神經元所分泌的多肽神經傳導物質，它的神經末稍遍布全腦，調控了許多生理上功能，包括痛覺的傳遞。而許多研究指出，A7兒茶酚胺細胞群投射軸突至脊髓背角處，並可產生止痛的效果。為了解食慾蛋白是否在腦幹中調控A7兒茶酚胺神經細胞群的活動，本實驗進行了型態上與電生理上的研究。染色結果顯示，A7核區中有很豐富的食慾蛋白神經末稍分布。從腦薄片上的全細胞電生理紀錄則可發現，給予食慾蛋白後，這些神經元的自發性動作電位頻率有顯著的提升，而如果將膜電位箝制在-70mV，則可觀察到食慾蛋白-A引發了一往細胞內流的電流。此種興奮性的效果會被食慾蛋白受體1的阻斷劑—SB-334867所抑制。除此之外，該電流的反轉電位接近-34mV，且若將細胞外大部分鈉離子取代為NMDG，便偵測不到此食慾蛋白引起的電流，暗示著食慾蛋白可能開啟了一種無選擇性的陽離子通道，如Transient Receptor Potential(TRP)通道。事實上，這股電流在一些非專一性TRP通道的抑制劑(如2-APB, SKF-96365, Ruthenium red)下均有效被抑制；另外，capsazepine(一種專一性的TRPV1通道的拮抗劑)也能顯著減少此食慾蛋白引發的電流。免疫染色結果指出在這些神經元上的確有TRPV1通道的分布，而且此通道的活化劑Capsaicin也能夠引發內流電流，其反轉電位亦接近於-34mV，種種電生理特性皆與食慾蛋白-A引起之反應相符。本研究顯示，食慾蛋白-A會作用在A7兒茶酚胺細胞群的食慾蛋白受體1上，並可能打開了TRPV1通道，進而提升這些神經元的膜興奮性。 | zh_TW |
| dc.description.abstract | Orexin is a neuropeptide that involves in multiple brain functions, including pain modulation. Therefore we investigated the effect of orexin-A on noradrenergic (NAergic) neurons of A7 catecholamine cell group, which projects NAergic fibers to the dorsal spinal cord and plays a role in spinal antinociception. From whole cell recordings of NAergic neurons in the rat brain slices, bath application of orexin-A increases spontaneous firing rate under current clamp mode and activates an inward current(IOXA) under voltage clamp mode with holding voltage -70mV. SB-334867, an orexin receptor 1 antagonist, blocked this response. In addition, IOXA reversed at -33.9mV and was almost eliminated when extracellular Na+ was substituted by N-methyl-D-glucamine(NMDG), suggesting IOXA was mediated by a nonselective cation conductance, presumably by the transient receptor potential(TRP) channels. Indeed, IOXA was significantly blocked by several antagonists of TRP channels, such as 2-APB, SKF-96365, ruthenium red, and a specific TRPV1 antagonist capsazepine. We then tested whether TRPV1 expresses on A7 NAergic neurons. Application of specific TRPV1 agonist capsaicin produced an inward current, which reversed at -33.4mV and was sensitive to both NMDG and capsazepine. Rich expression of TRPV1 proteins was also found on these NAergic neurons using immunohistochemistry techniques. Taken together, we conclude that orexin-A acting on OXR1 excites NAergic neurons in A7 catecholamine cell group through opening TRPV1 channels. Other TRP-like channels may also mediate the response. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T15:19:16Z (GMT). No. of bitstreams: 1 ntu-97-R95b41009-1.pdf: 1669061 bytes, checksum: 896c19e24bed22851d7f0e3cc8be4ff1 (MD5) Previous issue date: 2008 | en |
| dc.description.tableofcontents | Introduction………………………………………………………………………....11
A7 catecholamine cell group: anatomy and physiological function....................11 Orexins and orexin receptors...............................................................................14 Distribution of orexinergic neurons, fibers, and orexin receptors in rat CNS.....15 Physiolgical effects of orexins..............................................................................16 Cellular responses to orexins...............................................................................18 Aim of this study...................................................................................................19 Materials and Methods……………………………………………………………..21 Preparation of brain stem slices...........................................................................21 Electrophysiology.................................................................................................21 Measurements of liquid Junction potentials.........................................................23 Data analysis.......................................................................................................24 Drugs ...................................................................................................................25 Filling recorded neurons with biocytin and immunocytochemistry.....................26 Immunohistochemistry of TRPV1 and orexin-A fibers.........................................27 Results...……………………………………………………………………………..29 Identification of NAergic neurons of A7 catecholamine cell group in rat brain stem slices.............................................................................................................29 Rich innervations of Orexinergic fibers in the A7 catecholamine cell group.....................................................................................................................29 Orexin-A increases spontaneous firing rates of NAergic neurons in A7 cell group.....................................................................................................................30 Orexin-A activates an inward current through OXR1..........................................31 Ionic basis of the orexin A evoked inward current...............................................32 Orexin-A may activate TRP-like channels but not Na+/Ca2+ exchangers............35 Expression of TRPV1 channels in A7 NAergic neurons.......................................38 Possible cellular mechanisms underlying orexin-A-induced response................39 Discussion……………………………………………………………………………41 Na+/Ca2+ exchanger and K+ channels are less likely mediate orexin A-induced current..................................................................................................................42 Involvement of Nonselective Cation conductances..............................................45 TRPV1 is one of the target nonselective cation channels of orexin-A..................46 Other possible nonselective cation channels activated by orexin-A....................50 Possible cellular signaling pathway for orexin-A to excite A7 NAergic neurons.................................................................................................................51 Physiological implications...................................................................................53 References…………………………………………………………………………...54 Figures……………………………………………………………………………….65 | |
| dc.language.iso | en | |
| dc.subject | 全細胞電生理記錄 | zh_TW |
| dc.subject | TRPV1通道 | zh_TW |
| dc.subject | 痛覺傳導 | zh_TW |
| dc.subject | 腦幹 | zh_TW |
| dc.subject | A7核區 | zh_TW |
| dc.subject | 食慾蛋白 | zh_TW |
| dc.subject | pain transmission | en |
| dc.subject | brainstem | en |
| dc.subject | A7 cell group | en |
| dc.subject | orexin | en |
| dc.subject | whole-cell recording | en |
| dc.subject | TRPV1 channels | en |
| dc.title | 食慾蛋白-A對大鼠A7核區正腎上腺素神經元細胞膜興奮性之影響 | zh_TW |
| dc.title | Effects of Orexin-A on Membrane Excitabilities of Noradrenergic Neurons of A7 Cell Group in Rats | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 96-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 楊琇雯,潘建源,陳志成 | |
| dc.subject.keyword | 腦幹,A7核區,食慾蛋白,全細胞電生理記錄,TRPV1通道,痛覺傳導, | zh_TW |
| dc.subject.keyword | brainstem,A7 cell group,orexin,whole-cell recording,TRPV1 channels,pain transmission, | en |
| dc.relation.page | 77 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2008-07-24 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 動物學研究所 | zh_TW |
| 顯示於系所單位： | 動物學研究所 | |
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