1、正常及MPTP帕金森病模型小鼠蒼白球orexin-A的電生理效應(yīng)及其受體機(jī)制 研究生：陳安琪 導(dǎo) 師：陳 蕾 教授 青島大學(xué)醫(yī)學(xué)院神經(jīng)生物學(xué)專業(yè)1基底神經(jīng)節(jié)2基底神經(jīng)節(jié)直接和間接通路示意圖直接通路：提高皮層的興奮性間接通路：抑制皮層的興奮性3帕金森病(PD)4OrexinOrexin是下丘腦神經(jīng)肽家族中的一員，可直接參與調(diào)節(jié)中樞運(yùn)動(dòng)功能。長(zhǎng)度為130個(gè)氨基酸的前orexin原經(jīng)水解后可生成orexin-A和orexin-B5Orexin的主要作用參與睡眠/覺醒狀態(tài)的調(diào)控參與能量代謝過程調(diào)控?cái)z食行為影響學(xué)習(xí)記憶活動(dòng)神經(jīng)興奮作用運(yùn)動(dòng)功能調(diào)控67Orexin與PD帕金森病晚期病人下丘腦orexin能

2、神經(jīng)元缺失約3845%，前額葉皮質(zhì)orexin-A水平減少40%帕金森病晚期患者腦脊液orexin水平降低，且orexin水平與帕金森病嚴(yán)重程度呈負(fù)相關(guān)性，即orexin水平越低，病情越重8蒼白球與PD蒼白球神經(jīng)元放電頻率降低，通過間接通路增強(qiáng)對(duì)基底神經(jīng)節(jié)所支配區(qū)域的抑制效應(yīng)，從而導(dǎo)致帕金森病的運(yùn)動(dòng)減少等癥狀蒼白球神經(jīng)元簇狀放電的出現(xiàn)直接導(dǎo)致帕金森病的靜止性震顫癥狀，并且其電活動(dòng)的改變程度與帕金森病震顫及運(yùn)動(dòng)減少等癥狀的嚴(yán)重性直接相關(guān)9Orexin與蒼白球 已有文獻(xiàn)報(bào)道，蒼白球接受下丘腦orexin能神經(jīng)元的直接投射，并且orexin受體在蒼白球內(nèi)有表達(dá) Orexin對(duì)大鼠蒼白球神經(jīng)元具有興奮

3、作用，并且該興奮性作用是由OX1R和OX2R共同介導(dǎo)的10問題 Orexin-A對(duì)正常小鼠蒼白球神經(jīng)元自發(fā)放電的影響 Orexin-A對(duì)蒼白球神經(jīng)元電生理效應(yīng)的受體及信號(hào)轉(zhuǎn)導(dǎo)機(jī)制 Orexin-A對(duì)PD模型小鼠蒼白球的電生理效應(yīng)是否改變 Orexin-A是否參與調(diào)控PD模型小鼠的運(yùn)動(dòng)行為異常11內(nèi)容觀察orexin-A對(duì)正常及PD模型小鼠蒼白球神經(jīng)元自發(fā)放電頻率及模式的影響觀察orexin-A對(duì)蒼白球神經(jīng)元放電影響的受體機(jī)制觀察orexin-A電生理效應(yīng)的信號(hào)轉(zhuǎn)導(dǎo)途徑觀察內(nèi)源性orexins對(duì)正常及PD模型小鼠蒼白球神經(jīng)元放電的影響觀察正常與PD模型小鼠蒼白球OX1受體和OX2受體的表達(dá)觀察o

4、rexin-A對(duì)PD模型小鼠僵直和運(yùn)動(dòng)減少等行為的影響12實(shí)驗(yàn)技術(shù)與方法在體細(xì)胞外電生理記錄MPTP帕金森病模型小鼠制備免疫組織化學(xué)技術(shù)帕金森病模型小鼠行為學(xué)測(cè)試13實(shí)驗(yàn)結(jié)果1415小鼠蒼白球神經(jīng)元規(guī)則放電ISI histogramAutocorrelogram16小鼠蒼白球神經(jīng)元不規(guī)則放電ISI histogramAutocorrelogram17小鼠蒼白球神經(jīng)元簇狀放電ISI histogramAutocorrelogram組別規(guī)則放電不規(guī)則放電簇狀放電細(xì)胞總數(shù)n (%)n (%)n (%)n正常小鼠29 (49.15%)22 (37.29%)8 (13.56%)59PD小鼠12 (42.

5、86%)16 (57.14%)*0 (0%)28正常及PD模型小鼠蒼白球三種放電模式神經(jīng)元的比較18*P0.0519正常及PD模型小鼠蒼白球神經(jīng)元基礎(chǔ)放電頻率無差異(n=39)(n=23)nsns: not significantOrexin-A對(duì)正常小鼠蒼白球神經(jīng)元自發(fā)放電的影響0.01mmolOrexin-A20Orexin-A對(duì)正常小鼠蒼白球神經(jīng)元放電頻率的影響GroupNFiring rate before drugapplication(Hz)Firing rate before drugapplication(Hz)Excitatory neurons218.870.9611.10

6、1.23*Inhibitory neurons37.820.304.551.33*P0.0012122Orexin-A對(duì)PD模型小鼠蒼白球神經(jīng)元自發(fā)放電的影響0.01mmolOrexin-A23Orexin-A對(duì)PD模型小鼠蒼白球神經(jīng)元放電頻率的影響GroupNFiring rate before drugapplication(Hz)Firing rate before drugapplication(Hz)Excitatory neurons147.061.2810.641.61*Inhibitory neurons25.242.373.121.02*P0.0010.01mmolSB334

7、867SB334867對(duì)正常小鼠蒼白球神經(jīng)元自發(fā)放電的影響2425SB334867對(duì)正常小鼠蒼白球神經(jīng)元放電頻率的影響GroupNFiring rate before drugapplication(Hz)Firing rate before drugapplication(Hz)Inhibitory neurons236.260.793.470.65*Excitatory neurons12.673.9*P0.001 26GroupNFiring rate before drugapplication(Hz)Firing rate before drugapplication(Hz)Inhi

8、bitory neurons36.772.293.872.19*SB334867對(duì)PD模型小鼠蒼白球神經(jīng)元放電頻率的影響*P0.05Orexin-A對(duì)正常和PD模型小鼠蒼白球神經(jīng)元放電頻率的影響27正常PDSB334867對(duì)正常和PD模型小鼠蒼白球神經(jīng)元放電頻率的影響28正常PD29GroupNCV before drug applicationCV after drug applicationFF before drug applicationFF after drug applicationOrexin-A210.500.090.630.09*0.120.040.110.04SB33486

9、7230.890.121.170.12*0.210.070.700.16*Orexin-A和SB334867對(duì)正常小鼠蒼白球神經(jīng)元放電模式的影響*P0.05, *P0.01 CV (Coefficient of Variance); FF (Fano Factor)30GroupNCV before drug applicationCV after drug applicationFF before drug applicationFF after drug applicationOrexin-A140.490.090.60.110.160.080.130.07SB33486741.090.181.500.16*0.180.070.670.19*Orexin-A和SB334867對(duì)PD小鼠蒼白球神經(jīng)元放電模式的影響*P0.05 CV (Coefficient of Variance); FF (Fano Factor)31200m100m50m正常小鼠蒼白球OX-1受體的表達(dá)32正常PD正常及PD模型小鼠下丘腦外側(cè)區(qū)orexi