an-automated-rapid-iterative-negative-geotaxis-assay-for-analyzing

An Automated Rapid Iterative Negative Geotaxis Assay for Analyzing Adult Climbing Behavior in a <em>Drosophila</em> Model of Neurodegeneration

Neurodegenerative diseases are frequently associated with a progressive loss of movement ability, reduced life span, and age-dependent neurodegeneration. ...

an-improved-method-for-collection-cerebrospinal-fluid-from

An Improved Method for Collection of Cerebrospinal Fluid from Anesthetized Mice

The cerebrospinal fluid (CSF) is a valuable body fluid for analysis in neuroscience research. It is one of the fluids in closest contact with the central ...

Chinese Academy of Sciences

University of Chinese Academy of Sciences, Chinese Academy of Science

Sino-Danish Center for Education and Research (SDC)

Le laminage panne (RBO) g&#xE8;ne code pour une int&#xE9;grale membrane plasmique lipase n&#xE9;cessaire pour phototransduction drosophile. Les photor&#xE9;cepteurs sont enrichis pour la prot&#xE9;ine RBO, et sensibles &#xE0; la temp&#xE9;rature mutants RBO montrent l&#x26;#39;&#xE9;limination r&#xE9;versible de phototransduction en quelques minutes, ce qui d&#xE9;montre un besoin aigu de la prot&#xE9;ine. Le bloc est une activit&#xE9; d&#xE9;pendante, ce qui indique que l&#x26;#39;action de RBO est d&#x26;#39;utiliser &#xE0; charge. Conditionnelles mutants RBO montrer d&#xE9;pendante de l&#x26;#39;activit&#xE9; appauvrissement de la couche d&#x26;#39;accumulation de diacylglyc&#xE9;rol et concomitante de phosphate de phosphatidylinositol et le phosphatidylinositol 4,5-bisphosphate dans les minutes suivant l&#x26;#39;induction, ce qui sugg&#xE8;re une r&#xE9;gulation n&#xE9;gative rapide de la phospholipase C (PLC) l&#x26;#39;activit&#xE9;. L&#x26;#39;exigence RBO identifie une &#xE9;tape essentielle dans la r&#xE9;glementation G-coupl&#xE9; &#xE0; la prot&#xE9;ine, PLC-signalisation d&#xE9;pendante des lipides inositol m&#xE9;diation activation de canaux TRP et TRPL pendant phototransduction.

Roulant Blackout, un nouvellement identifi&#xE9;s PIP2-DAG Pathway Lipase requis pour phototransduction Drosophila

W&#xE4;lzlager Blackout, ein neu identifiziertes PIP2-DAG-Lipase Weg f&#xFC;r Drosophila Phototransduktion erforderlich.

Il rolling blackout (rbo) gene codifica per una lipasi integrale di membrana plasmatica necessaria per phototransduction drosofila. Fotorecettori sono arricchiti per la proteina RBO, e mutanti termosensibili rbo Visualizza eliminazione reversibile di phototransduction in pochi minuti, dimostrando un&#39;esigenza acuta della proteina. Il blocco &egrave; attivit&agrave; dipendente, che indica che l&#39;azione di RBO &egrave; utilizzare dipendenti. Mutanti condizionali rbo Visualizza attivit&agrave; dipendente dalla deplezione di diacilglicerolo e concomitante accumulo di fosfatidilinositolo fosfati e fosfatidilinositolo 4,5-bisfosfato in pochi minuti di induzione, suggerendo rapida downregulation della fosfolipasi attivit&agrave; C (PLC). Il requisito di RBO identifica un passo normativo essenziale in lipidi G-proteina-accoppiato, PLC-dipendente inositolo segnalazione attivazione mediazione dei canali TRP e TRPL durante phototransduction.

Rolling blackout, una lipasi di pathway PIP2-DAG recentemente identificata necessaria per phototransduction drosofila.

&#x6EDA;&#x52A8;&#x505C;&#x7535; &#xFF08;rbo &#x90FD;) &#x57FA;&#x56E0;&#x8FDB;&#x884C;&#x7F16;&#x7801;&#x6240;&#x9700;&#x7684;&#x679C;&#x8747; phototransduction &#x79EF;&#x5206;&#x7EC6;&#x80DE;&#x819C;&#x8102;&#x80AA;&#x9176;&#x3002;Rbo &#x90FD;&#x86CB;&#x767D;&#xFF0C;&#x4E30;&#x5BCC;&#x4E86;&#x611F;&#x5149;&#x7EC6;&#x80DE;&#x548C;&#x7A81;&#x53D8;&#x4F53;&#x6E29;&#x5EA6;&#x654F;&#x611F; rbo &#x90FD;&#x663E;&#x793A;&#x53EF;&#x9006;&#x6D88;&#x9664;&#x7684; phototransduction &#x5728;&#x51E0;&#x5206;&#x949F;&#x4E4B;&#x5185;&#xFF0C;&#x8868;&#x660E;&#x86CB;&#x767D;&#x8D28;&#x6025;&#x6027;&#x8981;&#x6C42;&#x3002;&#x5757;&#x662F;&#x4F9D;&#x8D56;&#xFF0C;&#x6307;&#x793A;&#x884C;&#x52A8;&#x7684; rbo &#x90FD;&#x4F7F;&#x7528;&#x76F8;&#x5173;&#x7684;&#x6D3B;&#x52A8;&#x3002;&#x6761;&#x4EF6; rbo &#x90FD;&#x7A81;&#x53D8;&#x4F53;&#x663E;&#x793A;&#x6D3B;&#x6027;&#x4F9D;&#x8D56;&#x67AF;&#x7AED;&#x7684;&#x7518;&#x6CB9;&#x4E8C;&#x916F;&#x548C;&#x4F34;&#x968F;&#x79EF;&#x7D2F;&#x7684;&#x78F7;&#x9178;&#x808C;&#x9187;&#x548C;&#x78F7;&#x8102;&#x9170;&#x808C;&#x9187; 4&#xFF0C;5-&#x4E8C;&#x78F7;&#x9178;&#x7684;&#x611F;&#x5E94;&#xFF0C;&#x5206;&#x949F;&#x5185;&#x5EFA;&#x8BAE;&#x8FC5;&#x901F;&#x4E0B;&#x8C03;&#x7684;&#x78F7;&#x8102;&#x9176; C &#xFF08;PLC&#xFF09; &#x7684;&#x6D3B;&#x52A8;&#x3002;Rbo &#x90FD;&#x8981;&#x6C42;&#x6807;&#x8BC6; G &#x86CB;&#x767D;&#x5076;&#x8054;&#x3001; PLC &#x4F9D;&#x8D56;&#x808C;&#x9187;&#x8102;&#x8D28;&#x4FE1;&#x53F7;&#x8C03;&#x89E3;&#x6FC0;&#x6D3B;&#x8DE8;&#x56FD;&#x6FC0;&#x8FDB;&#x515A;&#x548C; TRPL &#x6E20;&#x9053;&#x5728; phototransduction &#x671F;&#x95F4;&#x76D1;&#x7BA1;&#x7684;&#x91CD;&#x8981;&#x4E00;&#x6B65;&#x3002;

&#x5206;&#x533A;&#x8F6E;&#x6D41;&#x505C;&#x7535;&#xFF0C;&#x6240;&#x9700;&#x7684;&#x679C;&#x8747; phototransduction &#x65B0;&#x53D1;&#x73B0; PIP2-&#x8FBE;&#x683C; &middot; &#x54C8;&#x901A;&#x8DEF;&#x8102;&#x80AA;&#x9176;&#x3002;

The rolling blackout (rbo) gene encodes an integral plasma membrane lipase required for Drosophila phototransduction. Photoreceptors are enriched for the RBO protein, and temperature-sensitive rbo mutants show reversible elimination of phototransduction within minutes, demonstrating an acute requirement for the protein. The block is activity dependent, indicating that the action of RBO is use dependent. Conditional rbo mutants show activity-dependent depletion of diacylglycerol and concomitant accumulation of phosphatidylinositol phosphate and phosphatidylinositol 4,5-bisphosphate within minutes of induction, suggesting rapid downregulation of phospholipase C (PLC) activity. The RBO requirement identifies an essential regulatory step in G-protein-coupled, PLC-dependent inositol lipid signaling mediating activation of TRP and TRPL channels during phototransduction.

Rolling blackout, a newly identified PIP2-DAG pathway lipase required for Drosophila phototransduction.

&#x8EE2;&#x304C;&#x308A;&#x505C;&#x96FB;&#xFF08;RBO&#xFF09;&#x907A;&#x4F1D;&#x5B50;&#x306F;&#x3001;&#x30B7;&#x30E7;&#x30A6;&#x30B8;&#x30E7;&#x30A6;&#x30D0;&#x30A8;&#x306E;&#x5149;&#x4F1D;&#x9054;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x5FC5;&#x8981;&#x4E0D;&#x53EF;&#x6B20;&#x306A;&#x7D30;&#x80DE;&#x819C;&#x30EA;&#x30D1;&#x30FC;&#x30BC;&#x3092;&#x30A8;&#x30F3;&#x30B3;&#x30FC;&#x30C9;&#x3057;&#x307E;&#x3059;&#x3002;&#x5149;&#x53D7;&#x5BB9;&#x4F53;&#x306F;&#x3001;RBO&#x306E;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;&#x3092;&#x6FC3;&#x7E2E;&#x3057;&#x3001;&#x6E29;&#x5EA6;&#x306B;&#x654F;&#x611F;&#x306A;RBO&#x5909;&#x7570;&#x4F53;&#x306F;&#x3001;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;&#x306E;&#x6025;&#x6027;&#x8981;&#x4EF6;&#x3092;&#x793A;&#x3057;&#x3066;&#x3001;&#x6570;&#x5206;&#x4EE5;&#x5185;&#x306B;&#x5149;&#x4F1D;&#x9054;&#x306E;&#x53EF;&#x9006;&#x7684;&#x306A;&#x9664;&#x53BB;&#x3092;&#x793A;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x30D6;&#x30ED;&#x30C3;&#x30AF;&#x306F;&#x3001;RBO&#x306E;&#x30A2;&#x30AF;&#x30B7;&#x30E7;&#x30F3;&#x306B;&#x4F9D;&#x5B58;&#x4F7F;&#x7528;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x3001;&#x6D3B;&#x52D5;&#x306B;&#x4F9D;&#x5B58;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x6761;&#x4EF6;RBO&#x5909;&#x7570;&#x4F53;&#x306F;&#x3001;&#x30DB;&#x30B9;&#x30DB;&#x30EA;&#x30D1;&#x30FC;&#x30BC;C&#xFF08;PLC&#xFF09;&#x6D3B;&#x6027;&#x306E;&#x6025;&#x901F;&#x306A;&#x30C0;&#x30A6;&#x30F3;&#x30EC;&#x30AE;&#x30E5;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x3092;&#x793A;&#x5506;&#x3057;&#x3001;&#x8A98;&#x5C0E;&#x6570;&#x5206;&#x4EE5;&#x5185;&#x306B;&#x30DB;&#x30B9;&#x30D5;&#x30A1;&#x30C1;&#x30B8;&#x30EB;&#x30A4;&#x30CE;&#x30B7;&#x30C8;&#x30FC;&#x30EB;&#x30EA;&#x30F3;&#x9178;&#x53CA;&#x3073;&#x30DB;&#x30B9;&#x30D5;&#x30A1;&#x30C1;&#x30B8;&#x30EB;&#x30A4;&#x30CE;&#x30B7;&#x30C8;&#x30FC;&#x30EB;4,5  - &#x30D3;&#x30B9;&#x30EA;&#x30F3;&#x9178;&#x306E;&#x30B8;&#x30A2;&#x30B7;&#x30EB;&#x30B0;&#x30EA;&#x30BB;&#x30ED;&#x30FC;&#x30EB;&#x3068;&#x4F75;&#x7528;&#x84C4;&#x7A4D;&#x306E;&#x6D3B;&#x52D5;&#x4F9D;&#x5B58;&#x7684;&#x306A;&#x67AF;&#x6E07;&#x3092;&#x793A;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002; RBO&#x306E;&#x8981;&#x4EF6;&#x306F;&#x3001;G&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;&#x5171;&#x5F79;&#x578B;&#x3001;TRP&#x3068;&#x5149;&#x60C5;&#x5831;&#x4F1D;&#x9054;&#x6642;&#x306E;TRPL&#x30C1;&#x30E3;&#x30CD;&#x30EB;&#x306E;&#x6D3B;&#x6027;&#x5316;&#x3092;&#x5A92;&#x4ECB;&#x3059;&#x308B;PLC-&#x4F9D;&#x5B58;&#x6027;&#x306E;&#x30A4;&#x30CE;&#x30B7;&#x30C8;&#x30FC;&#x30EB;&#x8102;&#x8CEA;&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x306B;&#x4E0D;&#x53EF;&#x6B20;&#x306A;&#x8ABF;&#x7BC0;&#x6BB5;&#x968E;&#x3092;&#x8B58;&#x5225;&#x3057;&#x307E;&#x3059;&#x3002;

&#x30ED;&#x30FC;&#x30EA;&#x30F3;&#x30B0;&#xB7;&#x30D6;&#x30E9;&#x30C3;&#x30AF;&#x3001;&#x30B7;&#x30E7;&#x30A6;&#x30B8;&#x30E7;&#x30A6;&#x30D0;&#x30A8;&#x306E;&#x5149;&#x60C5;&#x5831;&#x4F1D;&#x9054;&#x306B;&#x5FC5;&#x8981;&#x306A;&#x65B0;&#x305F;&#x306B;&#x540C;&#x5B9A;&#x3055;&#x308C;&#x305F;PIP2-DAG&#x7D4C;&#x8DEF;&#x30EA;&#x30D1;&#x30FC;&#x30BC;

&#xB864;&#xB9C1; &#xC815;&#xC804; (rbo) &#xC720;&#xC804;&#xC790;&#xB294; &#xCD08;&#xD30C;&#xB9AC; phototransduction&#xC5D0; &#xD544;&#xC694;&#xD55C; &#xD544;&#xC218;&#xC801;&#xC778; &#xD50C;&#xB77C;&#xC988;&#xB9C8; &#xBA64;&#xBE0C;&#xB808;&#xC778; lipase&#xB97C; &#xC778;&#xCF54;&#xB529;&#xD569;&#xB2C8;&#xB2E4;. &#xB300;&#xB1CC;&#xB294; RBO &#xB2E8;&#xBC31;&#xC9C8; &#xB18D;&#xCD95; &#xBC0F; &#xC628;&#xB3C4; &#xBBFC;&#xAC10;&#xD55C; rbo &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xB2E8;&#xBC31;&#xC9C8;&#xC5D0; &#xB300; &#xD55C; &#xAE09;&#xC131; &#xC694;&#xAD6C;&#xB97C; &#xBCF4;&#xC5EC;&#xC8FC;&#xB294; &#xBD84; &#xC774;&#xB0B4;&#xC5D0;, phototransduction&#xC758; &#xAC00;&#xC5ED; &#xC81C;&#xAC70; &#xD45C;&#xC2DC;. &#xBE14;&#xB85D;&#xC740; &#xC885;&#xC18D; RBO &#xC791;&#xC5C5; &#xC885;&#xC18D; &#xC0AC;&#xC6A9;&#xC740; &#xB098;&#xD0C0;&#xB0B4;&#xB294; &#xD65C;&#xB3D9;&#xC785;&#xB2C8;&#xB2E4;. &#xC870;&#xAC74;&#xBD80; rbo &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; phospholipase C (PLC) &#xD65C;&#xB3D9;&#xC758; &#xBE60;&#xB978; downregulation &#xC81C;&#xC548; &#xD65C;&#xB3D9; &#xC885;&#xC18D; &#xACE0;&#xAC08; &#xC720;&#xB3C4;, &#xBD84; &#xC774;&#xB0B4;&#xC5D0; phosphatidylinositol &#xC778;&#xC0B0; &#xC5FC; &#xBC0F; phosphatidylinositol 4, 5 bisphosphate&#xC758; diacylglycerol &#xBC0F; &#xC218; &#xBC18;&#xD558;&#xB294; &#xCD95;&#xC801;&#xC758; &#xD45C;&#xC2DC;. RBO &#xC694;&#xAD6C; G &#xB2E8;&#xBC31;&#xC9C8; &#xACB0;&#xD569;, PLC &#xC885;&#xC18D; inositol &#xC9C0;&#xC9C8; phototransduction &#xB3D9;&#xC548; &#xC911;&#xC7AC; &#xD65C;&#xC131;&#xD654; TRP TRPL &#xCC44;&#xB110;&#xC758; &#xC2E0;&#xD638;&#xC5D0; &#xD544;&#xC218;&#xC801;&#xC778; &#xADDC;&#xC81C; &#xB2E8;&#xACC4;&#xB97C; &#xC2DD;&#xBCC4; &#xD569;&#xB2C8;&#xB2E4;.

&#xC815;&#xC804;, &#xCD08;&#xD30C;&#xB9AC; phototransduction&#xC5D0; &#xD544;&#xC694;&#xD55C; &#xC0C8;&#xB85C; &#xD655;&#xC778;&#xB41C; &#xB41C; PIP2 DAG &#xD1B5;&#xB85C; lipase&#xB97C; &#xC555; &#xC5F0;.

El apag&#xF3;n de rodadura (RBO) codifica una parte integral de la membrana plasm&#xE1;tica lipasa necesaria para fototransducci&#xF3;n Drosophila. Los fotorreceptores son enriquecidos para la prote&#xED;na RBO, y sensibles a la temperatura mutantes RBO muestran eliminaci&#xF3;n reversible de fototransducci&#xF3;n en cuesti&#xF3;n de minutos, lo que demuestra un requisito aguda para la prote&#xED;na. El bloque es dependiente de la actividad, indicando que la acci&#xF3;n de RBO es utilizar dependiente. Mutantes condicionales RBO muestran una actividad dependiente de la disminuci&#xF3;n de la acumulaci&#xF3;n de diacilglicerol y concomitante de fosfato fosfatidilinositol bifosfato y fosfatidilinositol 4,5-a pocos minutos de la inducci&#xF3;n, lo que sugiere una r&#xE1;pida regulaci&#xF3;n a la baja de la fosfolipasa C (PLC) la actividad. El requisito de RBO identifica un paso esencial en la regulaci&#xF3;n de la prote&#xED;na G-acoplada, PLC-dependiente de se&#xF1;alizaci&#xF3;n de l&#xED;pidos de inositol mediar la activaci&#xF3;n de los canales TRP y TRPL durante fototransducci&#xF3;n.

De Rolling Blackout, una recientemente identificada PIP2-DAG lipasa Camino Necesario para Fototransducci&#xF3;n Drosophila

In the present study using extracellular electrophysiological recording techniques, we explored the temporal characteristics of hippocampal theta activation in relation to formalin nociception. Results indicate that, compared to hind paw injection of saline, formalin injection in behaving rat evoked biphasic increase in duration of dorsal CA1 theta. Such an increase broadly paralleled animal biphasic behavioral activation, especially lick and moment-to-moment agitated behaviors. Correspondingly, theta-modulated cell firing was observed following formalin injection in anesthetized rat. The formalin-induced theta activation in behaving rat was most marked during peak of theta activation in the 2nd theta state (11-40 min post-injection) comprising 73% of the time in the 5 min block. An increase in theta peak frequency was also observed with respect to pre-injection control. However, the peak of theta in the 2nd theta state mostly preceded the peak of lick and flinch of the affected paw. In the 41-60 min, following formalin injection while the animals displayed robust nociceptive flinching and lifting, the theta activity approached control levels. Furthermore, the theta peak frequency at peak of theta was higher than the corresponding values of sustained theta observed in correlation with the nociceptive behaviors; in contrast, high frequency theta rhythm was observed during formalin-induced other moment-to-moment agitated behaviors. These findings favor the notion that in the formalin model the theta state of the hippocampus reflects a neural drive that is dissociated from the duration of nociceptive experience and is not selective to the typical nociceptive indices of lick, flinch, and lift of the injured paw.

Hippocampal theta state in relation to formalin nociception.

&#x6EDA;&#x52A8;&#x505C;&#x7535; &#xFF08;rbo &#x90FD;&#xFF09; &#x662F;&#x5047;&#x5B9A;&#x7684;&#x8DE8;&#x819C;&#x8102;&#x80AA;&#x9176;&#x7518;&#x6CB9;&#x4E8C;&#x78F7;&#x8102;&#x9176; C &#x4F9D;&#x8D56;&#x78F7;&#x8102;&#x9170;&#x808C;&#x9187; 4&#xFF0C;5-&#x4E8C;&#x78F7;&#x9178;-&#x916F;&#x5728;&#x679C;&#x8747;&#x795E;&#x7ECF;&#x5143;&#x4FE1;&#x53F7;&#x6240;&#x9700;&#x3002;&#x6761;&#x4EF6;&#x6E29;&#x5EA6;&#x654F;&#x611F; (TS) rbo &#x90FD;&#x7A81;&#x53D8;&#x4F53;&#x5728;&#x5206;&#x949F;&#x5185;&#x663E;&#x793A;&#x5B8C;&#x6574;&#x7684;&#x53EF;&#x9006;&#x9EBB;&#x75F9;&#x8868;&#x660E; rbo &#x90FD;&#x5341;&#x5206;&#x9700;&#x8981;&#x7684;&#x8FD0;&#x52A8;&#x3002;Rbo &#x90FD;&#x86CB;&#x767D;&#x5728;&#x7A81;&#x89E6;&#x524D; boutons &#x5728;&#x795E;&#x7ECF;&#x808C;&#x8089;&#x63A5;&#x5934; &#xFF08;NMJ&#xFF09; &#x7A81;&#x89E6;&#x548C;&#x6574;&#x4E2A;&#x4E2D;&#x592E;&#x7684;&#x7A81;&#x89E6; neuropil&#xFF0C;&#x4E3B;&#x8981;&#x662F;&#x672C;&#x5730;&#x5316;&#x548C; rbo &#x90FD; TS &#x7A81;&#x53D8;&#x4F53;&#x663E;&#x793A;&#x7684;&#x4E2D;&#x67A2;&#x548C;&#x5916;&#x5468;&#x7A81;&#x89E6;&#x4F20;&#x9012;&#x5728;&#x51E0;&#x5206;&#x949F;&#x5185;&#x5B8C;&#x6210;&#x3001; &#x53EF;&#x9006;&#x5757;&#x3002;&#x6B64;&#x8868;&#x578B;&#x51FA;&#x73B0;&#x9650;&#x4E8E;&#x6210;&#x5E74;&#x4EBA;&#xFF0C;&#x56E0;&#x4E3A;&#x5E7C;&#x866B; NMJs &#x4E0D;&#x80FD;&#x8868;&#x73B0;&#x7684;&#x6025;&#x6027;&#x7684;&#x5C01;&#x9501;&#x3002;&#x7535;&#x955C;&#x89C2;&#x5BDF;&#x6210;&#x4EBA; rbo &#x90FD; TS &#x7A81;&#x53D8;&#x4F53; boutons &#x663E;&#x793A;&#x603B;&#x7A81;&#x89E6;&#x56CA;&#x6CE1; (SV) &#x7684;&#x5185;&#x5BB9;&#xFF0C;&#x4E0E;&#x7A81;&#x89E6;&#x524D; bouton &#x5927;&#x5C0F;&#x968F;&#x4E4B;&#x6536;&#x7F29;&#x548C;&#x79EF;&#x7D2F;&#x7684;&#x505C;&#x9760; SVs &#x5728;&#x7A81;&#x89E6;&#x524D;&#x6D3B;&#x52A8;&#x533A;&#x5728;&#x51E0;&#x5206;&#x949F;&#x5185;&#x589E;&#x52A0;&#x3002;&#x57FA;&#x56E0;&#x6D4B;&#x8BD5;&#x63ED;&#x793A; rbo &#x90FD;&#x548C; syntaxin1A TS &#x7A81;&#x53D8;&#xFF0C;&#x6697;&#x793A; rbo &#x90FD;&#x9700;&#x8981;&#x673A;&#x5236;&#x7684; N-ethylmaleimide-&#x654F;&#x611F;&#x56E0;&#x5B50;&#x9644;&#x4EF6;&#x86CB;&#x767D;&#x53D7;&#x4F53; &#xFF08;&#x9677;&#x9631;&#xFF09; &#x4E4B;&#x95F4;&#x7684;&#x534F;&#x540C;&#x4E92;&#x52A8;-&#x4ECB;&#x5BFC;&#x7684; SV &#x80DE;&#x6216;&#x5E76;&#x884C;&#x901A;&#x8DEF; SV &#x878D;&#x5408;&#x7684;&#x5FC5;&#x8981;&#x6761;&#x4EF6;&#x3002;Rbo &#x90FD; TS &#x7A81;&#x53D8;&#x5E76;&#x4E0D;&#x80FD;&#x591F;&#x68C0;&#x6D4B;&#x51FA;&#x6765;&#x6539;&#x53D8;&#x7F51;&#x7F57;&#x590D;&#x6742;&#x7EC4;&#x4EF6;&#xFF0C;&#x5EFA;&#x8BAE;&#x5728; SV &#x878D;&#x5408;&#x7684;&#x4E0B;&#x6E38;&#x9700;&#x6C42;&#x3002;&#x6211;&#x4EEC;&#x5F97;&#x51FA;&#x7684;&#x7ED3;&#x8BBA; rbo &#x90FD;&#x53D1;&#x6325;&#x91CD;&#x8981;&#x4F5C;&#x7528;&#xFF0C;&#x5728;&#x795E;&#x7ECF;&#x9012;&#x8D28;&#x91CA;&#x653E;&#xFF0C;&#x4E0B;&#x6E38;&#x7684; SV &#x5BF9;&#x63A5;&#xFF0C;&#x53EF;&#x80FD;&#x8C03;&#x89E3; SV &#x878D;&#x5408;&#x3002;

&#x6EDA;&#x52A8;&#x505C;&#x7535;&#x662F;&#x5FC5;&#x9700;&#x7684;&#x7A81;&#x89E6;&#x56CA;&#x6CE1;&#x80DE;&#x3002;

Rolling blackout (RBO) is a putative transmembrane lipase required for phospholipase C-dependent phosphatidylinositol 4,5-bisphosphate-diacylglycerol signaling in Drosophila neurons. Conditional temperature-sensitive (TS) rbo mutants display complete, reversible paralysis within minutes, demonstrating that RBO is acutely required for movement. RBO protein is localized predominantly in presynaptic boutons at neuromuscular junction (NMJ) synapses and throughout central synaptic neuropil, and rbo TS mutants display a complete, reversible block of both central and peripheral synaptic transmission within minutes. This phenotype appears limited to adults, because larval NMJs do not manifest the acute blockade. Electron microscopy of adult rbo TS mutant boutons reveals an increase in total synaptic vesicle (SV) content, with a concomitant shrinkage of presynaptic bouton size and an accumulation of docked SVs at presynaptic active zones within minutes. Genetic tests reveal a synergistic interaction between rbo and syntaxin1A TS mutants, suggesting that RBO is required in the mechanism of N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE)-mediated SV exocytosis, or in a parallel pathway necessary for SV fusion. The rbo TS mutation does not detectably alter SNARE complex assembly, suggesting a downstream requirement in SV fusion. We conclude that RBO plays an essential role in neurotransmitter release, downstream of SV docking, likely mediating SV fusion.

Rolling blackout is required for synaptic vesicle exocytosis.

Roulant panne (RBO) est un transmembranaire putatif lipase n&#xE9;cessaire pour la phospholipase C-d&#xE9;pendante phosphatidylinositol 4,5-bisphosphate-diacylglyc&#xE9;rol signalisation dans les neurones de drosophile. Conditionnelle sensibles &#xE0; la temp&#xE9;rature (TS) mutants RBO afficher complet, paralysie r&#xE9;versible en quelques minutes, d&#xE9;montrant que RBO est vivement requise pour le mouvement. Prot&#xE9;ines RBO est localis&#xE9;e principalement dans boutons pr&#xE9;synaptiques &#xE0; la jonction neuromusculaire (JNM) synapses et tout au long neuropile synaptique centrale, et RBO mutants TS afficher une compl&#xE8;te, bloc r&#xE9;versible de la fois central et p&#xE9;riph&#xE9;rique de la transmission synaptique en quelques minutes. Ce ph&#xE9;notype semble limit&#xE9;e aux adultes, parce NMJs larvaires ne manifestent le blocus aigu&#xEB;. La microscopie &#xE9;lectronique de l&#x26;#39;adulte RBO TS boutons mutant r&#xE9;v&#xE8;le une augmentation de la v&#xE9;sicule synaptique total (SV) contenu, avec un retrait concomitant de la taille de bouton pr&#xE9;synaptique et une accumulation de amarr&#xE9; au SV zones actives pr&#xE9;synaptiques en quelques minutes. Les tests g&#xE9;n&#xE9;tiques r&#xE9;v&#xE8;lent une interaction synergique entre RBO et syntaxin1A mutants TS, ce qui sugg&#xE8;re que RBO est n&#xE9;cessaire dans le m&#xE9;canisme de la N-&#xE9;thylmal&#xE9;imide sensible r&#xE9;cepteur de prot&#xE9;ine facteur d&#x26;#39;attachement (SNARE) m&#xE9;di&#xE9;e par exocytose SV, ou dans une voie parall&#xE8;le n&#xE9;cessaire &#xE0; la fusion SV. La mutation RBO TS ne d&#xE9;tectable modifier SNARE assemblage complexe, ce qui sugg&#xE8;re une exigence en aval dans la fusion SV. Nous concluons que RBO joue un r&#xF4;le essentiel dans la lib&#xE9;ration de neurotransmetteurs, en aval de la SV d&#x26;#39;accueil, susceptibles de m&#xE9;diation de fusion SV.

Blackout roulant sont exig&#xE9;es pour l&#x26;#39;exocytose des v&#xE9;sicules synaptiques

Rolling Stromausfall (RBO) ist ein mutma&szlig;liches Transmembran-Lipase erforderlich f&uuml;r Phospholipase C-abh&auml;ngige Phosphatidylinositol 4, 5-bisphosphat-Diacylglycerol signalisieren in Drosophila Neuronen. Bedingte temperaturempfindliche (TS) Rbo Mutanten zeigen vollst&auml;ndige, reversible L&auml;hmung innerhalb von Minuten, demonstrieren, dass RBO f&uuml;r Bewegung akut erforderlich ist. RBO-Protein ist &uuml;berwiegend in pr&auml;synaptischen Bottoni in neuromuskul&auml;ren Synapse (NMJ) Synapsen und im gesamten zentralen synaptische Neuropil lokalisiert, und Rbo TS Mutanten zeigen einen vollst&auml;ndigen, reversiblen Block des zentralen und peripheren synaptischen &Uuml;bertragung innerhalb von Minuten. Dieser Ph&auml;notyp erscheint auf Erwachsene beschr&auml;nkt, da die larvalen NMJs nicht die akute Blockade offenbar. Elektronenmikroskopie von Erwachsenen Rbo TS mutant Bottoni zeigt eine Zunahme der gesamten synaptische Vesikel (SV) Inhalt, mit einer gleichzeitigen Schrumpfung des pr&auml;synaptischen Bouton Gr&ouml;&szlig;e und eine Ansammlung von angedockten SVs am pr&auml;synaptischen aktive Zonen innerhalb von Minuten. Genetische Tests zeigen eine synergistische Interaktion zwischen Rbo und syntaxin1A TS-Mutanten, die darauf hindeutet, dass die RBO in den Mechanismus der N-Ethylmaleimid-Sensitive Faktor Anlage Protein-Rezeptor (SNARE) erforderlich ist-vermittelte SV Exozytose, oder in einen parallelen Weg notwendig f&uuml;r SV-Fusion. Die Rbo TS-Mutation ver&auml;ndert nicht detectably SNARE komplexe Montage, eine nachgeschaltete Anforderung SV Fusion vorgeschlagen. Wir schlie&szlig;en, dass die RBO spielt eine wesentliche Rolle in der Neurotransmitter-Freisetzung, flussabw&auml;rts von SV Andocken, wahrscheinlich Vermittlung SV Verschmelzung.

Rolling Blackout ist f&uuml;r synaptische Vesikel-Exozytose erforderlich.

Rolling blackout (RBO) &egrave; una lipasi putativa transmembrana necessaria per la fosfolipasi C-dipendente fosfatidilinositolo 4,5-bisfosfato-diacilglicerolo segnalazione in drosofila neuroni. Condizionale mutanti di rbo di (TS) di sensibili alla temperatura visualizzano paralisi completa, reversibile in pochi minuti, dimostrando che RBO &egrave; acutamente necessaria per il movimento. RBO proteina &egrave; localizzata prevalentemente nella presinaptica boutons alla giunzione neuromuscolare (NMJ) sinapsi e tutta neuropilo sinaptica centrale e mutanti rbo TS visualizzare un blocco reversibile, completo di trasmissione sinaptica sia centrale che periferico, in pochi minuti. Questo fenotipo appare limitato agli adulti, perch&eacute; NMJs larvale non manifestano il blocco acuto. Microscopia elettronica di boutons mutante rbo adulti TS rivela un aumento del contenuto totale della vescicola sinaptica (SV), con una concomitante contrazione della dimensione bouton presinaptica e un accumulo di SVs ancorata presso presinaptiche zone attive in pochi minuti. Test genetici rivelano un&#39;interazione sinergica tra mutanti TS rbo e syntaxin1A, suggerendo che RBO &egrave; necessaria nel meccanismo di N-ethylmaleimide-sensibili al fattore attachment protein receptor (SNARE)-mediata esocitosi SV, o in un percorso parallelo necessario per la fusione di SV. La mutazione rbo TS non altera distintamente assemblaggio complesso SNARE, suggerendo un requisito a valle in fusione di SV. Possiamo concludere che la filiale regionale svolge un ruolo essenziale nel rilascio di neurotrasmettitore, a valle della SV docking, probabilmente mediando fusione SV.

Rolling blackout &egrave; richiesto per esocitosi della vescicola sinaptica.

&#x8EE2;&#x304C;&#x308A;&#x505C;&#x96FB;&#xFF08;RBO&#xFF09;&#x306F;&#x30B7;&#x30E7;&#x30A6;&#x30B8;&#x30E7;&#x30A6;&#x30D0;&#x30A8;&#x306E;&#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3;&#x306B;&#x304A;&#x3051;&#x308B;&#x30DB;&#x30B9;&#x30DB;&#x30EA;&#x30D1;&#x30FC;&#x30BC;C&#x4F9D;&#x5B58;&#x6027;&#x30DB;&#x30B9;&#x30D5;&#x30A1;&#x30C1;&#x30B8;&#x30EB;&#x30A4;&#x30CE;&#x30B7;&#x30C8;&#x30FC;&#x30EB;4,5  - &#x30D3;&#x30B9;&#x30EA;&#x30F3;&#x9178; - &#x30B8;&#x30A2;&#x30B7;&#x30EB;&#x30B0;&#x30EA;&#x30BB;&#x30ED;&#x30FC;&#x30EB;&#x306E;&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x306B;&#x5FC5;&#x8981;&#x306A;&#x63A8;&#x5B9A;&#x819C;&#x8CAB;&#x901A;&#x30EA;&#x30D1;&#x30FC;&#x30BC;&#x3067;&#x3042;&#x308B;&#x3002;&#x6761;&#x4EF6;&#x4ED8;&#x304D;&#x306E;&#x6E29;&#x5EA6;&#x611F;&#x53D7;&#x6027;&#xFF08;TS&#xFF09;RBO&#x5909;&#x7570;&#x4F53;&#x306F;&#x3001;RBO&#x304C;&#x6025;&#x6027;&#x904B;&#x52D5;&#x304C;&#x5FC5;&#x8981;&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x3092;&#x5B9F;&#x8A3C;&#x3057;&#x3001;&#x6570;&#x5206;&#x4EE5;&#x5185;&#x306B;&#x5B8C;&#x4E86;&#x3059;&#x308B;&#x3068;&#x3001;&#x53EF;&#x9006;&#x7684;&#x306A;&#x9EBB;&#x75FA;&#x3092;&#x8868;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002; RBO&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;&#x306F;&#x3001;&#x795E;&#x7D4C;&#x7B4B;&#x63A5;&#x5408;&#x90E8;&#xFF08;NMJ&#xFF09;&#x306E;&#x30B7;&#x30CA;&#x30D7;&#x30B9;&#x3067;&#x3001;&#x4E2D;&#x592E;&#x306E;&#x30B7;&#x30CA;&#x30D7;&#x30B9;&#x306E;&#x795E;&#x7D4C;&#x7DB2;&#x3092;&#x901A;&#x3058;&#x3066;&#x30B7;&#x30CA;&#x30D7;&#x30B9;&#x524D;&#x7D42;&#x672B;&#x306B;&#x4E3B;&#x306B;&#x5C40;&#x5728;&#x3057;&#x3001;RBO TS&#x5909;&#x7570;&#x4F53;&#x306F;&#x3001;&#x6570;&#x5206;&#x4EE5;&#x5185;&#x306B;&#x3001;&#x4E2D;&#x67A2;&#x304A;&#x3088;&#x3073;&#x672B;&#x68A2;&#x30B7;&#x30CA;&#x30D7;&#x30B9;&#x4F1D;&#x9054;&#x306E;&#x4E21;&#x65B9;&#x3092;&#x5B8C;&#x5168;&#x306B;&#x3001;&#x53EF;&#x9006;&#x30D6;&#x30ED;&#x30C3;&#x30AF;&#x3092;&#x8868;&#x793A;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x5E7C;&#x866B;NMJs&#x306F;&#x6025;&#x6027;&#x5C01;&#x9396;&#x3092;&#x660E;&#x3089;&#x304B;&#x306B;&#x3057;&#x307E;&#x305B;&#x3093;&#x306E;&#x3067;&#x3001;&#x3053;&#x306E;&#x8868;&#x73FE;&#x578B;&#x306F;&#x3001;&#x6210;&#x4EBA;&#x306B;&#x9650;&#x5B9A;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x5927;&#x4EBA;&#x306E;RBO TS&#x5909;&#x7570;&#x4F53;&#x7D42;&#x672B;&#x306E;&#x96FB;&#x5B50;&#x9855;&#x5FAE;&#x93E1;&#x306F;&#x3001;&#x30B7;&#x30CA;&#x30D7;&#x30B9;&#x30D6;&#x30FC;&#x30C8;&#x30F3;&#x6A19;&#x672C;&#x306E;&#x5927;&#x304D;&#x3055;&#x3068;&#x6570;&#x5206;&#x4EE5;&#x5185;&#x306B;&#x524D;&#x30B7;&#x30CA;&#x30D7;&#x30B9;&#x306E;&#x30A2;&#x30AF;&#x30C6;&#x30A3;&#x30D6;&#x30BE;&#x30FC;&#x30F3;&#x306B;&#x30C9;&#x30C3;&#x30AD;&#x30F3;&#x30B0;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;SV&#x306E;&#x84C4;&#x7A4D;&#x306E;&#x540C;&#x6642;&#x53CE;&#x7E2E;&#x3067;&#x3001;&#x5408;&#x8A08;&#x30B7;&#x30CA;&#x30D7;&#x30B9;&#x5C0F;&#x80DE;&#xFF08;SV&#xFF09;&#x542B;&#x91CF;&#x306E;&#x5897;&#x52A0;&#x3092;&#x660E;&#x3089;&#x304B;&#x306B;&#x3059;&#x308B;&#x3002;&#x907A;&#x4F1D;&#x5B50;&#x691C;&#x67FB;&#x306F;&#x3001;RBO&#x306F;&#x3001;N-&#x30A8;&#x30C1;&#x30EB;&#x30DE;&#x30EC;&#x30A4;&#x30DF;&#x30C9;&#x611F;&#x53D7;&#x6027;&#x56E0;&#x5B50;&#x4ED8;&#x7740;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;&#x53D7;&#x5BB9;&#x4F53;&#xFF08;SNARE&#xFF09;&#x5A92;&#x4ECB;SV&#x30A8;&#x30AD;&#x30BD;&#x30B5;&#x30A4;&#x30C8;&#x30FC;&#x30B7;&#x30B9;&#x3001;&#x307E;&#x305F;&#x306F;SV&#x306E;&#x878D;&#x5408;&#x306E;&#x305F;&#x3081;&#x306B;&#x5FC5;&#x8981;&#x306A;&#x4E26;&#x5217;&#x7D4C;&#x8DEF;&#x3067;&#x306E;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x304C;&#x5FC5;&#x8981;&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x5506;&#x3057;&#x3001;RBO&#x3068;syntaxin1A TS&#x5909;&#x7570;&#x4F53;&#x3068;&#x306E;&#x9593;&#x306E;&#x76F8;&#x4E57;&#x7684;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x3092;&#x660E;&#x3089;&#x304B;&#x306B;&#x3057;&#x305F;&#x3002; RBO TS&#x5909;&#x7570;&#x306F;&#x691C;&#x51FA;&#x53EF;&#x80FD;&#x306A;SV&#x878D;&#x5408;&#x306E;&#x4E0B;&#x6D41;&#x8981;&#x4EF6;&#x3092;&#x793A;&#x5506;&#x3057;&#x3001;SNARE&#x8907;&#x96D1;&#x306A;&#x30A2;&#x30BB;&#x30F3;&#x30D6;&#x30EA;&#x3092;&#x5909;&#x66F4;&#x3059;&#x308B;&#x3053;&#x3068;&#x306F;&#x3042;&#x308A;&#x307E;&#x305B;&#x3093;&#x3002;&#x6211;&#x3005;&#x306F;&#x3001;RBO&#x53EF;&#x80FD;&#x6027;&#x304C;&#x9AD8;&#x3044;SV&#x306E;&#x878D;&#x5408;&#x3092;&#x5A92;&#x4ECB;&#x3059;&#x308B;&#x3001;&#x4E0B;&#x6D41;SV&#x30C9;&#x30C3;&#x30AD;&#x30F3;&#x30B0;&#x306E;&#x3001;&#x795E;&#x7D4C;&#x4F1D;&#x9054;&#x7269;&#x8CEA;&#x306E;&#x653E;&#x51FA;&#x306B;&#x91CD;&#x8981;&#x306A;&#x5F79;&#x5272;&#x3092;&#x679C;&#x305F;&#x3057;&#x3066;&#x3044;&#x308B;&#x3068;&#x7D50;&#x8AD6;&#x4ED8;&#x3051;&#x3066;&#x3044;&#x308B;&#x3002;

&#x30ED;&#x30FC;&#x30EA;&#x30F3;&#x30B0;&#xB7;&#x30D6;&#x30E9;&#x30C3;&#x30AF;&#x306F;&#x3001;&#x30B7;&#x30CA;&#x30D7;&#x30B9;&#x5C0F;&#x80DE;&#x30A8;&#x30AD;&#x30BD;&#x30B5;&#x30A4;&#x30C8;&#x30FC;&#x30B7;&#x30B9;&#x304C;&#x5FC5;&#x8981;&#x3067;&#x3059;

&#xC815;&#xC804; (RBO) &#xB864;&#xB9C1; phospholipase C &#xC885;&#xC18D; phosphatidylinositol 4, 5-bisphosphate-diacylglycerol Drosophila &#xC2E0;&#xACBD; &#xC138;&#xD3EC;&#xC5D0; &#xC2E0;&#xD638;&#xC5D0; &#xD544;&#xC694;&#xD55C; &#xC0C1; &#xB9C9; &#xD6A1;&#xB2E8; lipase&#xB294;. &#xC870;&#xAC74;&#xBD80; &#xC628;&#xB3C4; &#xBBFC;&#xAC10;&#xD55C; (TS) rbo &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; RBO &#xC2EC;&#xD558;&#xAC8C; &#xC6C0;&#xC9C1;&#xC784;&#xC744; &#xC704;&#xD574; &#xD544;&#xC694; &#xD558;&#xB2E4;&#xB294; &#xAC83;&#xC744; &#xBCF4;&#xC5EC;&#xC8FC;&#xB294; &#xBA87; &#xBD84; &#xC548;&#xC5D0;, &#xC804;&#xCCB4;, &#xAC00;&#xC5ED; &#xB9C8;&#xBE44;&#xB97C; &#xD45C;&#xC2DC; &#xD569;&#xB2C8;&#xB2E4;. RBO &#xB2E8;&#xBC31;&#xC9C8;&#xC740; &#xC8FC;&#xB85C; presynaptic boutons &#xC2E0;&#xACBD; &#xADFC;&#xC721; &#xD559; &#xAD50;&#xCC28;&#xB85C; (NMJ) &#xC2DC; &#xB0C5; &#xC2A4; &#xBC0F; &#xC911;&#xC559; &#xC2DC; &#xB0C5; &#xC2A4; neuropil&#xC5D0; &#xAC78;&#xCCD0; &#xBC0F; rbo TS &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xBD84; &#xC774;&#xB0B4; &#xC911;&#xC559;&#xACFC; &#xC8FC;&#xBCC0; &#xC2DC; &#xB0C5; &#xC2A4; &#xC804;&#xC1A1;&#xC758; &#xC644;&#xB8CC;, &#xAC00;&#xC5ED; &#xBE14;&#xB85D; &#xD45C;&#xC2DC;. &#xC560;&#xBC8C;&#xB808; NMJs &#xAE09;&#xC131; &#xBD09;&#xC1C4;&#xB97C; &#xCC38;&#xACE0; &#xD558;&#xC9C0; &#xC54A;&#xC2B5;&#xB2C8;&#xB2E4; &#xB54C;&#xBB38;&#xC5D0;&#xC774; &#xD615; &#xC131;&#xC778;, &#xC81C;&#xD55C; &#xB41C; &#xB098;&#xD0C0;&#xB0A9;&#xB2C8;&#xB2E4;. &#xC131;&#xC778; rbo TS &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; boutons&#xC758; &#xC804;&#xC790; &#xD604;&#xBBF8;&#xACBD; presynaptic bouton &#xD06C;&#xAE30;&#xC758; &#xBD80;&#xC218;&#xC801;&#xC778; &#xC218;&#xCD95; &#xBC0F; &#xBA87; &#xBD84; &#xC548;&#xC5D0; presynaptic &#xD65C;&#xC131; &#xC601;&#xC5ED;&#xC5D0; &#xB3C4;&#xD0B9; &#xB41C; Svs&#xC758; &#xCD95;&#xC801; &#xCD1D; &#xC2DC; &#xB0C5; &#xC2A4; &#xC18C;&#xD3EC; (SV) &#xCF58;&#xD150;&#xCE20; &#xC99D;&#xAC00; &#xBCF4;&#xC5EC;&#xC900;&#xB2E4;. &#xC720;&#xC804;&#xC790; &#xAC80;&#xC0AC; &#xACF5;&#xAC1C; RBO N ethylmaleimide &#xBBFC;&#xAC10;&#xD55C; &#xC694;&#xC18C; &#xCCA8;&#xBD80; &#xB2E8;&#xBC31;&#xC9C8; &#xC218;&#xC6A9; &#xCCB4; (&#xC62C; &#xBB34;)&#xC758; &#xBA54;&#xCEE4;&#xB2C8;&#xC998;&#xC5D0; &#xD544;&#xC694;&#xD55C; &#xC81C;&#xC548; rbo &#xBC0F; syntaxin1A TS &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xAC04;&#xC758; &#xC2DC;&#xB108;&#xC9C0; &#xC791;&#xC6A9;-SV exocytosis &#xC911;&#xC7AC; &#xB610;&#xB294; &#xBCD1;&#xB82C; &#xD1B5;&#xB85C; SV &#xC735;&#xD569;&#xC5D0; &#xD544;&#xC694;&#xD55C;. Rbo TS &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; detectably SV &#xD4E8;&#xC804;&#xC5D0;&#xC11C; &#xB2E4;&#xC6B4;&#xC2A4;&#xD2B8;&#xB9BC; &#xC694;&#xAD6C; &#xC0AC;&#xD56D;&#xC744; &#xC81C;&#xC548; &#xC62C; &#xBB34; &#xBCF5;&#xC7A1; &#xD55C; &#xC5B4;&#xC148;&#xBE14;&#xB9AC;&#xB97C; &#xBCC0;&#xACBD; &#xD558;&#xC9C0; &#xC54A;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xB294; RBO &#xC2E0;&#xACBD; &#xC804;&#xB2EC; &#xBB3C;&#xC9C8; &#xBC29;&#xCD9C;&#xC5D0; &#xD544;&#xC218;&#xC801;&#xC778; &#xC5ED;&#xD560;&#xC744; &#xD55C;&#xB2E4; &#xACB0;&#xB860; SV &#xB3C4;&#xD0B9;, &#xC544;&#xB9C8; SV &#xD4E8;&#xC804; &#xC911;&#xC7AC;&#xC758; &#xB2E4;&#xC6B4;&#xC2A4;&#xD2B8;&#xB9BC;.

&#xC815;&#xC804;&#xC744; &#xC555; &#xC5F0; &#xD558;&#xB294; &#xAC83;&#xC740; &#xC2DC; &#xB0C5; &#xC2A4; &#xC18C;&#xD3EC; exocytosis &#xD544;&#xC694; &#xD569;&#xB2C8;&#xB2E4;.

Rodamientos apag&#xF3;n (RBO) es un supuesto necesario para transmembrana lipasa dependiente de la fosfolipasa C fosfatidilinositol 4,5-bifosfato-diacilglicerol se&#xF1;alizaci&#xF3;n en las neuronas de Drosophila. Condicional sensibles a la temperatura (TS) RBO mutantes muestran una par&#xE1;lisis completa y reversible en cuesti&#xF3;n de minutos, lo que demuestra que es sumamente RBO requerida para el movimiento. RBO prote&#xED;nas se localiza predominantemente en boutons presin&#xE1;ptica en la uni&#xF3;n neuromuscular (UNM) y en las sinapsis neuropilo sin&#xE1;ptica central y RBO mutantes TS mostrar un bloque completo, reversible de la central y la transmisi&#xF3;n sin&#xE1;ptica perif&#xE9;rica en cuesti&#xF3;n de minutos. Este fenotipo parece limitada a los adultos, porque NMJs larvas no manifiestan el bloqueo agudo. La microscop&#xED;a electr&#xF3;nica de los adultos RBO TS mutante boutons revela un aumento de la ves&#xED;cula sin&#xE1;ptica total de (SV) de contenido, con una contracci&#xF3;n concomitante de tama&#xF1;o bouton presin&#xE1;ptica y una acumulaci&#xF3;n de SV atracado en las zonas activas presin&#xE1;pticas en cuesti&#xF3;n de minutos. Las pruebas gen&#xE9;ticas revelan una interacci&#xF3;n sin&#xE9;rgica entre RBO y mutantes syntaxin1A TS, lo que sugiere que RBO se requiere en el mecanismo de N-etilmaleimida sensible receptor del factor de prote&#xED;na de fijaci&#xF3;n (SNARE)-mediada exocitosis SV, o en una v&#xED;a paralela necesario para la fusi&#xF3;n SV. La mutaci&#xF3;n RBO TS no detectable alterar conjunto complejo SNARE, lo que sugiere un requisito posterior a la fusi&#xF3;n SV. Llegamos a la conclusi&#xF3;n de que RBO juega un papel esencial en la liberaci&#xF3;n de neurotransmisores, aguas abajo de la SV de acoplamiento, probablemente la mediaci&#xF3;n de la fusi&#xF3;n SV.

Apag&#xF3;n del balanceo se requiere para la exocitosis de ves&#xED;culas sin&#xE1;pticas

Alzheimer's disease (AD) is attributable to synapse dysfunction and loss, but the nature and progression of the presynaptic structural and functional changes in AD are essentially unknown. We expressed wild-type or arctic form of beta amyloid(1-42) (Abeta) in a small group of neurons in the adult fly and performed extensive time course analysis of the function and structure of both axon and presynaptic terminals at the identified single-neuron level. Abeta accumulated intracellularly and induced a range of age-dependent changes, including depletion of presynaptic mitochondria, slowdown of bi-directional transports of axonal mitochondria, decreased synaptic vesicles, increased large vacuoles, and elevated synaptic fatigue. These structural and functional synaptic changes correlated with age-dependent deficit in motor behavior. All these alterations were accelerated in flies expressing the arctic form of Abeta. The depletion of presynaptic mitochondria was the earliest detected phenotype and was not caused by the change in axonal transport of mitochondria. Moreover, axonal mitochondria exhibited a dramatic reduction in number but a significant increase in size in aged Abeta-expressing flies, indicating a global depletion of mitochondria in the neuron and an impairment of mitochondria fission. These results suggest that Abeta accumulation depletes presynaptic and axonal mitochondria, leading to other presynaptic deficits.

Expression of beta-amyloid induced age-dependent presynaptic and axonal changes in Drosophila.

Multidrug resistance-associated protein 1 (MRP1), an efflux multidrug transporter, was shown to be elevated in both glia and neurons in seizure focus in refractory epilepsy patients. Up-regulation of MRP1 and other multidrug transporters in perivascular astrocytes was suggested to cause resistance to antiepileptic drugs (AEDs) by reducing the concentration of AEDs at the epileptogenic areas. However, it is not known whether the up-regulation of MRP1 in neurons can cause resistance to AEDs, such as sodium phenytoin (PHT) and valproic acid (VPA). PHT inhibits voltage-gated sodium channel (VGSC) by occluding it, but whether PHT enters the channel through its inner or outer pore is not known. The authors overexpressed human MRP1 protein only in neurons in a Drosophila genetic seizure model, bang senseless (bss) mutants. The authors found that overexpression of MRP1 blocked the attenuation of the seizure behavior of bss mutants by acute and chronic application of PHT, and by chronic application of VPA. Conversely, overexpression of MRP1 in neurons increased the tolerance of bss flies to high-dosage PHT and VPA. Thus, up-regulation of MRP1 expression only in neurons causes resistance to AED in seizure flies. Moreover, the current data suggest that PHT enters VGSC through its inner pore.

Overexpression of human MRP1 in neurons causes resistance to antiepileptic drugs in Drosophila seizure mutants.

The accumulation of beta amyloid (Aβ) can cause synaptic impairments, but the characteristics and mechanisms of the synaptic impairment induced by the accumulation of Aβ in Alzheimer's disease (AD) remain unclear. In identified single neurons in a newly developed Drosophila AD model, in which Aβ accumulates intraneuronally, we found an age-dependent reduction in the synaptic vesicle release probability that was associated with a decrease in the density of presynaptic calcium channel clusters and an increase in the presynaptic and postsynaptic contact length. Moreover, these alterations occurred in the absence of presynaptic bouton loss. In addition, we found that Aβ expression also produced an age-dependent decrease in the amount of Bruchpilot (Brp), which plays an important role in controlling Ca(2+) channel clustering and synaptic vesicle release in the presynaptic active zone. Our study indicates that the chronic accumulation of intraneuronal Aβ can induce functional and structural changes in the presynaptic active zone prior to a loss of presynaptic buttons in the same neuron.

Age-dependent alterations in the presynaptic active zone in a Drosophila model of Alzheimer's disease.

Beta amyloid (Aβ42)-induced dysfunction and loss of synapses are believed to be major underlying mechanisms for the progressive loss of learning and memory abilities in Alzheimer's disease (AD). The vast majority of investigations on AD-related synaptic impairment focus on synaptic plasticity, especially the decline of long-term potentiation of synaptic transmission caused by extracellular Aβ42. Changes in other aspects of synaptic and neuronal functions are less studied or undiscovered. Here, we report that intraneuronal accumulation of Aβ42 induced an age-dependent slowing of neuronal transmission along pathways involving multiple synapses.

Intraneuronal accumulation of Aβ42 induces age-dependent slowing of neuronal transmission in Drosophila.

The Mini-Mental State Examination (MMSE) and the Montreal Cognitive Assessment (MoCA) are brief cognitive screening tools that have been developed for the screening of patients with Mild Cognitive Impairment.

The use of MMSE and MoCA in patients with acute ischemic stroke in clinical.

The negative-geotaxis climbing assay is used to efficiently study aging and neurodegeneration in Drosophila. To make it suitable for large-scale study, a method called the rapid iterative negative geotaxis (RING) assay has been established by simultaneously photographing the climbing of multiple groups of flies when they are manually tapped down in test tubes. Here, we automated the assay by using a well-controlled electric motor to drive the tapping, and a homemade program to analyze the climbing height of flies. Using the automated RING (aRING) assay, we found that the climbing ability of a strain of wild-type flies, males in particular, declined rapidly before day 21 after eclosion, but slowly from day 21 to 35. We also found that the expression of arctic mutant Aβ42 accelerated the age-dependent decline in the climbing ability of flies. Moreover, using aRING, we examined the effect of third chromosome deficiencies on the accelerated locomotor decline in Aβ42-expressing flies, and isolated 7 suppressors and 15 enhancers.

Automated rapid iterative negative geotaxis assay and its use in a genetic screen for modifiers of Aβ(42)-induced locomotor decline in Drosophila.

Phosphoinositides and their metabolizing enzymes are involved in Aβ42 metabolism and Alzheimer's disease pathogenesis. In yeast and mammals, Eighty-five requiring 3 (EFR3), whose Drosophila homolog is Rolling Blackout (RBO), forms a plasma membrane-localized protein complex with phosphatidylinositol-4-kinase Type IIIα (PI4KIIIα) and a scaffold protein to tightly control the level of plasmalemmal phosphatidylinositol-4-phosphate (PI4P). Here, we report that RBO binds to Drosophila PI4KIIIα, and that in an Aβ42-expressing Drosophila model, separate genetic reduction of PI4KIIIα and RBO, or pharmacological inhibition of PI4KIIIα ameliorated synaptic transmission deficit, climbing ability decline, premature death, and reduced neuronal accumulation of Aβ42 Moreover, we found that RBO-PI4KIIIa downregulation increased neuronal Aβ42 release and that PI4P facilitated the assembly or oligomerization of Aβ42 in/on liposomes. These results indicate that RBO-PI4KIIIa downregulation facilitates neuronal Aβ42 release and consequently reduces neuronal Aβ42 accumulation likely via decreasing Aβ42 assembly in/on plasma membrane. This study suggests the RBO-PI4KIIIα complex as a potential therapeutic target and PI4KIIIα inhibitors as drug candidates for Alzheimer's disease treatment.SIGNIFICANCE STATEMENT Phosphoinositides and their metabolizing enzymes are involved in Aβ42 metabolism and Alzheimer's disease pathogenesis. Here, in an Aβ42-expressing Drosophila model, we discovered and studied the beneficial role of downregulating RBO or its interacting protein PI4KIIIα-a protein that tightly controls the plasmalemmal level of PI4P-against the defects caused by Aβ42 expression. Mechanistically, RBO-PI4KIIIα downregulation reduced neuronal Aβ42 accumulation, and interestingly increased neuronal Aβ42 release. This study suggests the RBO-PI4KIIIα complex as a novel therapeutic target, and PI4KIIIα inhibitors as new drug candidates.

Downregulation of RBO-PI4KIIIα Facilitates Aβ42 Secretion and Ameliorates Neural Deficits in Aβ42-Expressing Drosophila.

Stroke is a leading cause of death worldwide. Up to one thousand potential drugs or interventions have been developed to treat stroke, out of which ~160 have gone on to clinical trials. However, none of them has been successful. New insights into the molecular and cellular mechanisms of ischemia-induced injury are needed for discovering new therapeutic targets. Recently, Drosophila has been used to uncover new hypoxia-related genes. In this study, we describe an efficient and reliable assay with a sophisticated apparatus for studying the effects of oxygen deprivation on flies. Using this assay, wild-type flies were exposed to an anoxic environment for varying lengths of time, then the cumulative death rate and mobility recovery were systematically analyzed. We found that anoxia for over one hour caused lethality. The cumulative death rate on day 5 after anoxia was linearly and positively correlated with the duration of anoxia, and reached 50% when the duration was 2.5 h-3 h. We also found that the mobility recovery in normoxia was slow, as the climbing ability remained largely unchanged 4 h-6 h after 2.5-h of anoxia. We suggest that 2.5 h-3 h of anoxia and 4 h-6 h of recovery before mobility analysis are appropriate for future use of the anoxia assay.

ArticleTotal : 2	Year
Publication title Cited by 28	2017
Publication title Cited by 80	2018

Article	Year
Rolling blackout, a newly identified PIP2-DAG pathway lipase required for Drosophila phototransduction. Nature neuroscience\| PubMed ID: 15361878	2004
Hippocampal theta state in relation to formalin nociception. Pain\| PubMed ID: 16480829	2006
Rolling blackout is required for synaptic vesicle exocytosis. The Journal of neuroscience : the official journal of the Society for Neuroscience\| PubMed ID: 16510714	2006
Expression of beta-amyloid induced age-dependent presynaptic and axonal changes in Drosophila. The Journal of neuroscience : the official journal of the Society for Neuroscience\| PubMed ID: 20107079	2010
Overexpression of human MRP1 in neurons causes resistance to antiepileptic drugs in Drosophila seizure mutants. Journal of neurogenetics\| PubMed ID: 22026728	2011
Age-dependent alterations in the presynaptic active zone in a Drosophila model of Alzheimer's disease. Neurobiology of disease\| PubMed ID: 23149068	2013
Intraneuronal accumulation of Aβ42 induces age-dependent slowing of neuronal transmission in Drosophila. Neuroscience bulletin\| PubMed ID: 24733651	2014
The use of MMSE and MoCA in patients with acute ischemic stroke in clinical. The International journal of neuroscience\| PubMed ID: 26000804	2016
Automated rapid iterative negative geotaxis assay and its use in a genetic screen for modifiers of Aβ(42)-induced locomotor decline in Drosophila. Neuroscience bulletin\| PubMed ID: 26077703	2015
Downregulation of RBO-PI4KIIIα Facilitates Aβ42 Secretion and Ameliorates Neural Deficits in Aβ42-Expressing Drosophila. The Journal of neuroscience : the official journal of the Society for Neuroscience\| PubMed ID: 28424219	2017
An Efficient and Reliable Assay for Investigating the Effects of Hypoxia/Anoxia on Drosophila. Neuroscience bulletin\| PubMed ID: 28866769	2017

Fu-de Huang

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