patch-clamp-recordings-from-embryonic-zebrafish-mauthner-cells

Patch Clamp Recordings from Embryonic Zebrafish Mauthner Cells

Mauthner cells (M-cells) are large  reticulospinal neurons located in the hindbrain of teleost fish. They are key  neurons involved in a characteristic ...

University of Alberta

Alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate &#x53D7;&#x4F53; (AMPA-R) &#x52A8;&#x529B;&#x5B66;&#xFF0C;&#x901A;&#x8FC7;&#x53D7;&#x4F53;&#x4E9A;&#x578B;&#x4EA4;&#x6362;&#x673A;&#x53D1;&#x751F;&#x53D1;&#x5C55;&#x63D0;&#x901F;&#x7684;&#x8C37;&#x6C28;&#x9178;&#x795E;&#x7ECF;&#x7A81;&#x89E6;&#x6210;&#x719F;&#x7684;&#x5173;&#x952E;&#x4E00;&#x6B65;&#x3002;&#x4F46;&#x662F;&#xFF0C;&#x6240;&#x9700;&#x7684;&#x5F00;&#x5173;&#x5728;&#x53D7;&#x4F53;&#x7684;&#x5206;&#x5B50;&#x7EC4;&#x4EF6;&#x662F;&#x672A;&#x77E5;&#x7684;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#xFF0C;&#x6211;&#x4EEC;&#x4F7F;&#x7528;&#x6591;&#x9A6C;&#x9C7C;&#x51C6;&#x5907;&#x663E;&#x793A;&#x6FC0;&#x6D3B;&#x86CB;&#x767D;&#x6FC0;&#x9176; C (PKC) &#x4F3D;&#x739B;&#x5200;&#x662F;&#x5FC5;&#x8981;&#x7684; AMPA-R &#x52A8;&#x529B;&#x5B66;&#x53D1;&#x5C55;&#x52A0;&#x5FEB;&#x3002;PKCgamma &#x4E0E;&#x53CD;&#x4E49; morpholino &#x5BE1;&#x6838;&#x82F7;&#x9178; [PKCgamma-morpholino (PKCgamma-MO)] &#x7684;&#x76EE;&#x6807;&#x7684; &#x2477; &#x9632;&#x6B62;&#x6B63;&#x5E38;&#x8D85;&#x901F; AMPA-R Mauthner &#x7EC6;&#x80DE;&#x52A8;&#x529B;&#x5B66;&#x3002;PKCgamma MO &#x6CE8;&#x5165;&#x80DA;&#x80CE;&#x4E0D;&#x80FD;&#x8D29;&#x5356; AMPA Rs &#x540E;&#x4F5B;&#x6CE2; 12-&#x8089;&#x8C46;&#x853B;&#x9178;&#x916F; 13-&#x4E59;&#x9178;&#x916F;&#x6216; PKCgamma &#x7684;&#x5E94;&#x7528;&#x7A0B;&#x5E8F;&#x3002;PKCgamma MO &#x6CE8;&#x5165;&#x80DA;&#x80CE;&#x4E0D;&#x5B75;&#x5316;&#x6216;&#x5C55;&#x793A; C &#x5F00;&#x59CB;&#x9003;&#x8DD1;&#x53CD;&#x5E94;&#x3002;&#x7531;&#x9AD8;&#x67B6; K(+) &#x4ECB;&#x8D28;&#x7684;&#x5E94;&#x7528;&#x7A0B;&#x5E8F;&#x6216; N-&#x7532;&#x57FA;-D-&#x5929;&#x51AC;&#x6C28;&#x9178;&#x7684;&#x5E94;&#x7528;&#x589E;&#x52A0;&#x7A81;&#x89E6;&#x6D3B;&#x52A8; (33 h &#x68A8;&#x5B50;&#x623F;&#x80DA;&#x80CE;) &#x8BF1;&#x5BFC;&#x5FEB;&#x901F; PKCgamma &#x4F9D;&#x8D56;&#x8D29;&#x8FD0;&#x7684;&#x5FEB;&#x901F; AMPA Rs &#x5230;&#x795E;&#x7ECF;&#x7A81;&#x89E6;&#x3002;&#x6211;&#x4EEC;&#x7684;&#x7814;&#x7A76;&#x7ED3;&#x679C;&#x8868;&#x660E; PKCgamma &#x662F;&#x57FA;&#x672C;&#x7684; AMPA-Rs &#x6591;&#x9A6C;&#x9C7C; Mauthner &#x7EC6;&#x80DE;&#x53D1;&#x80B2;&#x52A0;&#x5FEB;&#x5206;&#x5B50;&#x94FE;&#x63A5;&#x3002;

&#x86CB;&#x767D;&#x6FC0;&#x9176; Cgamma &#x662F;&#x4E00;&#x4E2A;&#x4FE1;&#x53F7;&#x5206;&#x5B50;&#x6240;&#x9700;&#x7684; alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate &#x53D7;&#x4F53;&#x52A8;&#x529B;&#x5B66;&#x7684;&#x53D1;&#x5C55;&#x52A0;&#x5FEB;&#x3002;

A key step in the maturation of glutamate synapses is the developmental speeding of alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate receptor (AMPA-R) kinetics, which occurs via a switch in receptor subtypes. However, the molecular components required for the switch in receptors are unknown. Here, we used the zebrafish preparation to show that activation of protein kinase C (PKC)gamma is necessary for the developmental speeding of AMPA-R kinetics. Targeted knockdown of PKCgamma with an antisense morpholino oligonucleotide [PKCgamma-morpholino (PKCgamma-MO)], prevents the normal speeding up of AMPA-R kinetics in Mauthner cells. PKCgamma-MO-injected embryos are incapable of trafficking AMPA-Rs following application of phorbol 12-myristate 13-acetate or PKCgamma. PKCgamma-MO-injected embryos do not hatch or exhibit the C-start escape response. Increasing synaptic activity (33 h post-fertilization embryos) by application of an elevated K(+) medium or by application of N-methyl-D-aspartate induces rapid PKCgamma-dependent trafficking of fast AMPA-Rs to synapses. Our findings reveal that PKCgamma is a molecular link underlying the developmental speeding of AMPA-Rs in zebrafish Mauthner cells.

Protein kinase Cgamma is a signaling molecule required for the developmental speeding of alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate receptor kinetics.

Une &eacute;tape cl&eacute; dans la maturation des synapses de glutamate est l&#39;acc&eacute;l&eacute;ration du d&eacute;veloppement de la cin&eacute;tique des r&eacute;cepteurs alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate (AMPA-R), qui s&#39;effectue via un commutateur dans les sous-types de r&eacute;cepteurs. Cependant, les composants mol&eacute;culaires requis pour le commutateur dans les r&eacute;cepteurs sont inconnus. Ici, nous avons utilis&eacute; la pr&eacute;paration de poisson z&egrave;bre pour montrer que l&#39;activation de gamma de prot&eacute;ine kinase C (PKC) est n&eacute;cessaire pour l&#39;acc&eacute;l&eacute;ration du d&eacute;veloppement de la cin&eacute;tique de l&#39;AMPA-R. Pr&eacute;cipitation cibl&eacute;e de PKCgamma avec un oligonucl&eacute;otide morpholino antisens [PKCgamma-morpholino (PKCgamma-MO)], emp&ecirc;che l&#39;acc&eacute;l&eacute;ration normale de la cin&eacute;tique de la LPCA-R dans les cellules de Mauthner. PKCgamma-MO-inject&eacute; des embryons sont incapables de trafic AMPA-Rs apr&egrave;s application de phorbol 12-myristate 13-ac&eacute;tate ou PKCgamma. Embryons PKCgamma-MO-inject&eacute; n&#39;&eacute;closent ou pi&egrave;ce de la r&eacute;action de fuite C d&eacute;marrage. Augmenter l&#39;activit&eacute; synaptique (embryons auteur h 33) par application d&#39;un milieu k &eacute;lev&eacute; ou par application de N-m&eacute;thyl-D-aspartate induit rapide d&eacute;pendant PKCgamma traite des rapides AMPA-Rs de synapses. Nos r&eacute;sultats r&eacute;v&egrave;lent que le PKCgamma est un lien mol&eacute;culaire qui sous-tendent l&#39;acc&eacute;l&eacute;ration du d&eacute;veloppement de AMPA-r dans les cellules de Mauthner zebrafish.

La prot&eacute;ine kinase Cgamma est une mol&eacute;cule de signalisation n&eacute;cessaire &agrave; l&#39;acc&eacute;l&eacute;ration du d&eacute;veloppement de la cin&eacute;tique du RECEPTEUR alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate.

Ein wichtiger Schritt bei der Reifung von Glutamat Synapsen ist die Entwicklungsbiologie Beschleunigung der alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate-Rezeptor (AMPA-R) Kinetik, die &uuml;ber einen Schalter im Rezeptor-Subtypen auftritt. Molekularen Komponenten f&uuml;r den Switch-Rezeptoren sind jedoch unbekannt. Hier haben wir die Zebrafisch-Vorbereitung verwendet, um zu zeigen, dass die Aktivierung der Protein-Kinase C (PKC) Gamma f&uuml;r die Entwicklungsbiologie Beschleunigung der AMPA-R Kinetik notwendig. Gezielte Niederschlag des PKCgamma mit einem Morpholino antisense-Oligonukleotid [PKCgamma-Morpholino (PKCgamma-MO)], verhindert die normale Beschleunigung der AMPA-R Kinetik in Mauthner Zellen. PKCgamma-MO-injiziert Embryonen Menschenhandel AMPA-Rs unf&auml;hig sind, nach Anwendung der Phorbol 12-Isopropylmyristat 13-Acetat oder PKCgamma. PKCgamma-MO-injiziert Embryonen nicht schl&uuml;pfen oder die C-Start-Flucht-Reaktion aufweisen. Synaptische Aktivit&auml;t (33 h post-fertilization Embryonen) erh&ouml;hen, durch Anwendung eines erh&ouml;hten K(+) Mediums oder durch Anwendung von N-Methyl-D-Aspartat induziert rapid PKCgamma-abh&auml;ngige Menschenhandel schnell AMPA-Rs zu Synapsen. Unsere Ergebnisse zeigen, dass PKCgamma eine molekulare Verbindung die Entwicklungsbiologie Beschleunigung von AMPA-Rs im Zebrafisch Mauthner Zellen zugrunde.

Proteinkinase Cgamma ist eine signalisierende Molek&uuml;l ben&ouml;tigt f&uuml;r die Entwicklungsbiologie Beschleunigung der alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate-Rezeptor-Kinetik.

Un passo fondamentale nella maturazione delle sinapsi glutammato &egrave; accelerare lo sviluppo della cinetica di alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate del recettore (AMPA-R), che avviene tramite un interruttore in sottotipi del recettore. Tuttavia, le componenti molecolari necessarie per il passaggio nei recettori sono sconosciute. Qui, abbiamo utilizzato la preparazione di zebrafish per mostrare che l&#39;attivazione della gamma di proteina chinasi C (PKC) &egrave; necessaria per l&#39;accelerazione dello sviluppo della cinetica di AMPA-R. Atterramento mirato di PKCgamma con un oligonucleotide antisenso oligonucleotides [PKCgamma-oligonucleotides (PKCgamma-MO)], impedisce il normale accelerazione della cinetica di AMPA-R in cellule del Mauthner. PKCgamma-MO-iniettato embrioni sono incapaci di traffico AMPA-Rs dopo l&#39;applicazione del forbolo miristato 12 13-acetato o PKCgamma. PKCgamma-MO-iniettato embrioni non schiudono o esporre la risposta di fuga C-start. Aumento dell&#39;attivit&agrave; sinaptica (embrioni post-fertilization 33 h) dall&#39;applicazione di un&#39;elevata K(+) medio o dall&#39;applicazione di N-metil-D-aspartato induce rapida PKCgamma-dipendente dal traffico di veloce AMPA-Rs di sinapsi. I nostri risultati rivelano che PKCgamma &egrave; un legame molecolare sottostante accelerare lo sviluppo di AMPA-Rs in cellule del Mauthner zebrafish.

Chinasi di proteina Cgamma &egrave; una molecola di segnalazione necessaria per accelerare lo sviluppo della cinetica del recettore alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate.

&#x30B0;&#x30EB;&#x30BF;&#x30DF;&#x30F3;&#x9178;&#x30B7;&#x30CA;&#x30D7;&#x30B9;&#x306E;&#x6210;&#x719F;&#x306E;&#x91CD;&#x8981;&#x306A;&#x30B9;&#x30C6;&#x30C3;&#x30D7;&#x306F;&#x3001;&#x53D7;&#x5BB9;&#x4F53;&#x30B5;&#x30D6;&#x30BF;&#x30A4;&#x30D7;&#x306E;&#x30B9;&#x30A4;&#x30C3;&#x30C1;&#x3092;&#x4ECB;&#x3057;&#x3066;&#x767A;&#x751F;&#x3057;&#x307E;&#x3059; alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate &#x53D7;&#x5BB9;&#x4F53; (AMPA-R&#xFF09; &#x306E;&#x53CD;&#x5FDC;&#x901F;&#x5EA6;&#x8AD6;&#x306E;&#x767A;&#x9054;&#x30B9;&#x30D4;&#x30FC;&#x30C9;&#x9055;&#x53CD;&#x3067;&#x3059;&#x3002;&#x3057;&#x304B;&#x3057;&#x3001;&#x53D7;&#x5BB9;&#x4F53;&#x3067;&#x30B9;&#x30A4;&#x30C3;&#x30C1;&#x306B;&#x5FC5;&#x8981;&#x306A;&#x5206;&#x5B50;&#x8981;&#x7D20;&#x306F;&#x77E5;&#x3089;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;&#x6D3B;&#x6027;&#x5316;&#x86CB;&#x767D;&#x8CEA;&#x30AD;&#x30CA;&#x30FC;&#x30BC; C (PKC) &#x30AC;&#x30F3;&#x30DE;&#x304C; AMPA R &#x30AB;&#x30A4;&#x30CD;&#x30C6;&#x30A3;&#x30AF;&#x30B9;&#x306E;&#x767A;&#x9054;&#x306E;&#x30B9;&#x30D4;&#x30FC;&#x30C9;&#x9055;&#x53CD;&#x306E;&#x305F;&#x3081;&#x306B;&#x5FC5;&#x8981;&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3059;&#x306B;&#x30BC;&#x30D6;&#x30E9;&#x30D5;&#x30A3;&#x30C3;&#x30B7;&#x30E5;&#x6E96;&#x5099;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x307E;&#x3059;&#x3002;PKCgamma &#x3068; morpholino &#x306E;&#x30A2;&#x30F3;&#x30C1;&#x30BB;&#x30F3;&#x30B9; &#x30AA;&#x30EA;&#x30B4;&#x30CC;&#x30AF;&#x30EC;&#x30AA;&#x30C1;&#x30C9; [PKCgamma-morpholino &#xFF08;PKCgamma-&#x30DF;&#x30BA;&#x30FC;&#x30EA;&#xFF09;]&#x3001;&#x30BF;&#x30FC;&#x30B2;&#x30C3;&#x30C8;&#x3092;&#x7D5E;&#x3063;&#x305F;&#x30CE;&#x30C3;&#x30AF;&#x30C0;&#x30A6;&#x30F3;&#x3092;&#x9632;&#x304E;&#x307E;&#x3059; AMPA R &#x30DE;&#x30A6;&#x30B9;&#x30CA;&#x30FC;&#x7D30;&#x80DE;&#x52D5;&#x614B;&#x306E;&#x901A;&#x5E38;&#x9AD8;&#x901F;&#x5316;&#x3002;PKCgamma MO &#x6CE8;&#x5165;&#x80DA;&#x306F;&#x4EBA;&#x8EAB;&#x58F2;&#x8CB7;&#x306E; AMPA Rs &#x306E;&#x53EF;&#x80FD;&#x306A;&#x6B21;&#x306E; 12-13-&#x30D5;&#x30A9;&#x30EB;&#x30DC;&#x30FC;&#x30EB;&#x30A8;&#x30B9;&#x30C6;&#x30EB;&#x307E;&#x305F;&#x306F; PKCgamma &#x306E;&#x9069;&#x7528;&#x3002;PKCgamma MO &#x6CE8;&#x5165;&#x80DA;&#x5B75;&#x5316;&#x3084;&#x5C55;&#x793A; C &#x30B9;&#x30BF;&#x30FC;&#x30C8; &#x30A8;&#x30B9;&#x30B1;&#x30FC;&#x30D7;&#x5FDC;&#x7B54;&#x3092;&#x3057;&#x306A;&#x3044;&#x3067;&#x304F;&#x3060;&#x3055;&#x3044;&#x3002;&#x30B7;&#x30CA;&#x30D7;&#x30B9;&#x6D3B;&#x52D5; (33 h post-fertilization &#x80DA;&#xFF09; &#x306F;&#x9AD8;&#x67B6;&#x306B;&#x65BC;&#x3044;&#x3066;&#x306E;&#x5A92;&#x4F53;&#x306E;&#x30A2;&#x30D7;&#x30EA;&#x30B1;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x3001;&#x307E;&#x305F;&#x306F;&#x30A2;&#x30D7;&#x30EA;&#x30B1;&#x30FC;&#x30B7;&#x30E7;&#x30F3; N &#x30E1;&#x30C1;&#x30EB;-D-&#x30A2;&#x30B9;&#x30D1;&#x30E9;&#x30AE;&#x30F3;&#x9178;&#x306E;&#x5897;&#x52A0;&#x306F;&#x6025;&#x901F;&#x306A; PKCgamma &#x4F9D;&#x5B58;&#x6027;&#x30B7;&#x30CA;&#x30D7;&#x30B9;&#x306B;&#x5BFE;&#x3059;&#x308B;&#x9AD8;&#x901F; AMPA Rs &#x306E;&#x4EBA;&#x8EAB;&#x58F2;&#x8CB7;&#x3092;&#x8A98;&#x5C0E;&#x3057;&#x307E;&#x3059;&#x3002;&#x6211;&#x3005; &#x306E;&#x8ABF;&#x67FB;&#x7D50;&#x679C; PKCgamma AMPA-Rs &#x30BC;&#x30D6;&#x30E9;&#x30D5;&#x30A3;&#x30C3;&#x30B7;&#x30E5; &#x30FB; &#x30DE;&#x30A6;&#x30B9;&#x30CA;&#x30FC;&#x7D30;&#x80DE;&#x306E;&#x767A;&#x9054;&#x306E;&#x30B9;&#x30D4;&#x30FC;&#x30C9;&#x9055;&#x53CD;&#x306E;&#x57FA;&#x306B;&#x306A;&#x308B;&#x5206;&#x5B50;&#x30EA;&#x30F3;&#x30AF;&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x3092;&#x660E;&#x3089;&#x304B;&#x306B;&#x3057;&#x307E;&#x3059;&#x3002;

&#x86CB;&#x767D;&#x8CEA;&#x30AD;&#x30CA;&#x30FC;&#x30BC; Cgamma alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate &#x53D7;&#x5BB9;&#x4F53;&#x306E;&#x52D5;&#x614B;&#x306E;&#x767A;&#x9054;&#x306E;&#x30B9;&#x30D4;&#x30FC;&#x30C9;&#x9055;&#x53CD;&#x306E;&#x305F;&#x3081;&#x306B;&#x5FC5;&#x8981;&#x306A;&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x5206;&#x5B50;&#x3067;&#x3042;&#x308A;&#x307E;&#x3059;&#x3002;

&#xC870;&#xBBF8;&#xB8CC; &#xB0C5;&#xC758; &#xC131;&#xC219;&#xC5D0; &#xC911;&#xC694; &#xD55C; &#xB2E8;&#xACC4;&#xB294; alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate &#xC218;&#xC6A9; &#xCCB4; (AMPA-R) &#xC18D;&#xB3C4; &#xB860; &#xC218;&#xC6A9; &#xCCB4;&#xC758; &#xD558;&#xC704;&#xC5D0; &#xC788;&#xB294; &#xC2A4;&#xC704;&#xCE58;&#xB97C; &#xD1B5;&#xD574; &#xBC1C;&#xC0DD; &#xD558;&#xB294;&#xC758; &#xBC1C;&#xB2EC; &#xACFC;&#xC18D;. &#xADF8;&#xB7EC;&#xB098; &#xC218;&#xC6A9; &#xCCB4;&#xC758; &#xC2A4;&#xC704;&#xCE58;&#xC5D0; &#xD544;&#xC694;&#xD55C; &#xBD84;&#xC790; &#xAD6C;&#xC131; &#xC54C; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC5EC;&#xAE30;, zebrafish &#xC900;&#xBE44; &#xC0AC;&#xC6A9; &#xD558;&#xB294; &#xB2E8;&#xBC31;&#xC9C8; &#xD0A4; &#xB2C8; &#xC544; &#xC81C; C (PKC) &#xAC10;&#xB9C8;&#xC758; &#xD65C;&#xC131;&#xD654; &#xD544;&#xC694; AMPA-R &#xC18D;&#xB3C4; &#xB860; &#xBC1C;&#xB2EC; &#xACFC;&#xC18D;&#xC5D0; &#xB300; &#xD55C; &#xD45C;&#xC2DC;. &#xC13C;&#xC2A4; morpholino oligonucleotide [PKCgamma-morpholino (PKCgamma-&#xBAA8;)]&#xC640; Pkcgamma&#xC758; &#xD0C0;&#xAC9F;&#xB41C; &#xCD5C;&#xC800;&#xC758; AMPA-R &#xC18D;&#xB3C4; &#xB860; Mauthner &#xC138;&#xD3EC;&#xC5D0;&#xC11C;&#xC758; &#xC815;&#xC0C1;&#xC801;&#xC778; &#xC18D;&#xB3C4;&#xB97C; &#xBC29;&#xC9C0;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. PKCgamma &#xBAA8; &#xC8FC;&#xC785; &#xBC30;&#xC544;&#xB294; &#xBC00;&#xB9E4; AMPA-Rs&#xC758; &#xB2A5;&#xB825;&#xC774; phorbol 12-myristate 13-&#xC544;&#xC138;&#xD14C;&#xC774;&#xD2B8; &#xB610;&#xB294; PKCgamma &#xC751;&#xC6A9; &#xD504;&#xB85C;&#xADF8;&#xB7A8;&#xC744; &#xB2E4;&#xC74C;&#xACFC; &#xAC19;&#xC740;. PKCgamma &#xBAA8; &#xC8FC;&#xC785; &#xBC30;&#xC544; &#xBD80; &#xD654; &#xB418;&#xC9C0; &#xC54A;&#xAC70;&#xB098; C &#xC2DC;&#xC791; &#xD0C8;&#xCD9C; &#xC751;&#xB2F5; &#xC804;&#xC2DC;. &#xB192;&#xC740; K(+) &#xB9E4;&#xCCB4;&#xC758; &#xC751;&#xC6A9; &#xD504;&#xB85C;&#xADF8;&#xB7A8; &#xB610;&#xB294; &#xC751;&#xC6A9; &#xD504;&#xB85C;&#xADF8;&#xB7A8; N-&#xBA54; &#xD2F8;-D-aspartate&#xC758; &#xC2DC; &#xB0C5; &#xC2A4; &#xD65C;&#xB3D9; (33 h post-fertilization &#xBC30;&#xC544;)&#xC744; &#xC99D;&#xAC00; &#xAE09;&#xC18D; &#xD55C; PKCgamma &#xC885;&#xC18D; &#xB0C5;&#xC5D0; &#xBE60;&#xB978; AMPA Rs&#xC758; &#xC778;&#xC2E0; &#xB9E4;&#xB9E4; &#xC720;&#xB3C4; &#xD569;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xC758; &#xC5F0;&#xAD6C; &#xACB0;&#xACFC; &#xACF5;&#xAC1C; PKCgamma &#xBD84;&#xC790; &#xB9C1;&#xD06C; &#xBC1C;&#xB2EC; &#xACFC;&#xC18D; zebrafish Mauthner &#xC138;&#xD3EC;&#xC5D0;&#xC11C; AMPA-Rs&#xC758; &#xAE30;&#xBCF8; &#xC774;&#xB2E4;.

&#xB2E8;&#xBC31;&#xC9C8; &#xD0A4; &#xB2C8; &#xC544; &#xC81C; Cgamma&#xC740; &#xC18D;&#xB3C4; &#xB860; alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate &#xC218;&#xC6A9; &#xCCB4;&#xC758; &#xBC1C;&#xB2EC; &#xC18D;&#xB3C4;&#xC5D0; &#xD544;&#xC694;&#xD55C; &#xC2E0;&#xD638; &#xBD84;&#xC790; &#xC774;&#xB2E4;.

Un paso clave en la maduraci&oacute;n de las sinapsis de glutamato es la aceleraci&oacute;n del desarrollo de la cin&eacute;tica de la alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate del receptor (AMPA-R), que se produce a trav&eacute;s de un interruptor en subtipos del receptor. Sin embargo, los componentes moleculares necesarios para el interruptor en receptores son desconocidos. Aqu&iacute;, utilizamos la preparaci&oacute;n del pez cebra para mostrar que la activaci&oacute;n de la gamma de la prote&iacute;na quinasa C (PKC) es necesaria para la aceleraci&oacute;n del desarrollo de la cin&eacute;tica de AMPA-R. Espec&iacute;fica ca&iacute;da de PKCgamma con un oligonucle&oacute;tido antisentido Morfolino [PKCgamma-Morfolino (PKCgamma-MO)], previene la aceleraci&oacute;n normal de cin&eacute;tica de AMPA-R en c&eacute;lulas de Mauthner. Inyecci&oacute;n de PKCgamma-MO embriones son incapaces de AMPA-Rs tr&aacute;fico despu&eacute;s de la aplicaci&oacute;n de forbol miristato 12 13-acetato o PKCgamma. PKCgamma-MO-inyectado embriones no eclosionan o exhiben la respuesta de escape de C-Inicio. Aumento de la actividad sin&aacute;ptica (embriones poste-fertilizaci&oacute;n de 33 h) por el uso de un medio de capacidad elevado o por aplicaci&oacute;n de N-metil-D-aspartato induce r&aacute;pida PKCgamma-dependiente trata de AMPA-Rs r&aacute;pidos a las sinapsis. Nuestros resultados revelan que el PKCgamma es un enlace molecular subyacente a la aceleraci&oacute;n del desarrollo de AMPA-Rs en pez cebra Mauthner c&eacute;lulas.

Prote&iacute;na quinasa Cgamma es una mol&eacute;cula de se&ntilde;alizaci&oacute;n necesaria para la aceleraci&oacute;n del desarrollo de la cin&eacute;tica del receptor alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate.

Encoding new information requires dynamic changes in synaptic strength. The brain can boost synaptic plasticity through the secretion of neuromodulatory substances, including acetylcholine and noradrenaline. Considerable effort has focused on elucidating how neuromodulatory substances alter synaptic properties. However, determination of the potential synergistic interactions between different neuromodulatory systems remains incomplete. Previous results indicate that coactivation of Î²-adrenergic and cholinergic receptors facilitated the conversion of STP to LTP through an extracellular signal-regulated kinase (ERK)-dependent mechanism. ERK signaling has been linked to synaptically localized translation regulation. Thus, we hypothesized that costimulation of noradrenergic and cholinergic receptors could initiate the transformation of STP to LTP through up-regulation of protein synthesis. Our results indicate that a protocol which yields STP (5 Hz, 5 sec) when paired with coapplication of the Î²-adrenergic agonist, isoproterenol (ISO), and the cholinergic agonist, carbachol (CCh), induces translation-dependent LTP in mouse CA1. This form of LTP requires both Î²1-adrenergic and M1 muscarinic receptor activation, as blocking either receptor subtype prevented LTP induction. Blocking ERK, mTOR, or translation reduced the expression of LTP induced with ISO + CCh. Taken together, our data demonstrate that coactivation of Î²-adrenergic and muscarinic receptors facilitates the conversion of STP to LTP through a mechanism requiring translation initiation.

Conversion of short-term potentiation to long-term potentiation in mouse CA1 by coactivation of Î²-adrenergic and muscarinic receptors.

The function of the cellular prion protein (PrP(C)) in healthy brains remains poorly understood, in part because Prnp knockout mice are viable. On the other hand, transient knockdown of Prnp homologs in zebrafish (including two paralogs, prp1 and prp2) has suggested that PrP(C) is required for CNS development, cell adhesion, and neuroprotection. It has been argued that zebrafish Prp2 is most similar to mammalian PrP(C), yet it has remained intransigent to the most thorough confirmations of reagent specificity during knockdown. Thus we investigated the role of prp2 using targeted gene disruption via zinc finger nucleases. Prp2(-/-) zebrafish were viable and did not display overt developmental phenotypes. Back-crossing female prp2(-/-) fish ruled out a role for maternal mRNA contributions. Prp2(-/-) larvae were found to have increased seizure-like behavior following exposure to the convulsant pentylenetetrazol (PTZ), as compared to wild type fish. In situ recordings from intact hindbrains demonstrated that prp2 regulates closing of N-Methyl-d-aspartate (NMDA) receptors, concomitant with neuroprotection during glutamate excitotoxicity. Overall, the knockout of Prp2 function in zebrafish independently confirmed hypothesized roles for PrP, identifying deeply conserved functions in post-developmental regulation of neuron excitability that are consequential to the etiology of prion and Alzheimer diseases.

Article	Year
Protein kinase Cgamma is a signaling molecule required for the developmental speeding of alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate receptor kinetics. The European journal of neuroscience\| PubMed ID: 20525069	2010
Conversion of short-term potentiation to long-term potentiation in mouse CA1 by coactivation of Î²-adrenergic and muscarinic receptors. Learning & memory (Cold Spring Harbor, N.Y.)\| PubMed ID: 23077334	2012
Targeted mutation of the gene encoding prion protein in zebrafish reveals a conserved role in neuron excitability. Neurobiology of disease\| PubMed ID: 23523635	2013

Birbickram Roy

Biography

JoVE Journal Publications

Other Publications