Intracavernosal Pressure Recording to Evaluate Erectile Function in Rodents

Erectile dysfunction (ED) is defined as the inability to attain or keep an erection of the penis, and this has become a prevalent male sexual disorder. ...

Nanjing University

&#x6211;&#x4EEC;&#x4EE5;&#x524D;&#x7684;&#x7814;&#x7A76;&#x8868;&#x660E;&#x7A33;&#x5B9A; Microrna (Mirna) &#x54FA;&#x4E73;&#x52A8;&#x7269;&#x8840;&#x6E05;&#x548C;&#x8840;&#x6D46;&#x4E2D;&#x79EF;&#x6781;&#x7684;&#x7EC4;&#x7EC7;&#x548C;&#x7EC6;&#x80DE;&#x5206;&#x6CCC;&#x548C;&#x53EF;&#x4EE5;&#x4F5C;&#x4E3A;&#x4E00;&#x7C7B;&#x65B0;&#x578B;&#x7684;&#x751F;&#x7269;&#x6807;&#x5FD7;&#x7269;&#x7684;&#x75BE;&#x75C5;&#xFF0C;&#x5E76;&#x4F5C;&#x4E3A;&#x4FE1;&#x53F7;&#x5206;&#x5B50;&#x5728;&#x7EC6;&#x80DE;&#x95F4;&#x901A;&#x8BAF;&#x4E2D;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#xFF0C;&#x6211;&#x4EEC;&#x62A5;&#x544A;&#x4EE4;&#x4EBA;&#x60CA;&#x8BB6;&#x7684;&#x53D1;&#x73B0;&#xFF0C;&#x5916;&#x6E90;&#x690D;&#x7269; Mirna &#x76EE;&#x524D;&#x5728;&#x8840;&#x6E05;&#x548C;&#x5404;&#x79CD;&#x52A8;&#x7269;&#x7684;&#x7EC4;&#x7EC7;&#x548C;&#x8FD9;&#x4E9B;&#x5916;&#x6E90;&#x690D;&#x7269; Mirna &#x4E3B;&#x8981;&#x53E3;&#x5934;&#x7684;&#x3001; &#x7ECF;&#x7531;&#x98DF;&#x7269;&#x6444;&#x5165;&#x91CF;&#x3002;MIR168a &#x76DB;&#x4EA7;&#x6C34;&#x7A3B;&#xFF0C;&#x662F;&#x5927;&#x591A;&#x6570;&#x7684;&#x9AD8;&#x6D53;&#x7F29;&#x5916;&#x6E90;&#x690D;&#x7269; Mirna &#x8840;&#x6E05;&#x4E2D;&#x6587;&#x79D1;&#x76EE;&#x4E4B;&#x4E00;&#x3002;&#x4F53;&#x5916;&#x548C;&#x4F53;&#x5185;&#x7684;&#x529F;&#x80FD;&#x7814;&#x7A76;&#x8868;&#x660E; MIR168a &#x53EF;&#x4EE5;&#x7ED1;&#x5B9A;&#x5230;&#x4EBA;&#x6743;/&#x9F20;&#x6807;&#x4F4E;&#x5BC6;&#x5EA6;&#x8102;&#x86CB;&#x767D;&#x53D7;&#x4F53;&#x9002;&#x914D;&#x5668;&#x86CB;&#x767D; 1 (LDLRAP1) mRNA&#xFF0C;&#x6291;&#x5236;&#x809D;&#x810F;&#xFF0C;LDLRAP1 &#x8868;&#x8FBE;&#x548C;&#x56E0;&#x6B64;&#x51CF;&#x5C11;&#x5C0F;&#x9F20;&#x8840;&#x6D46;&#x4F4E;&#x5BC6;&#x5EA6;&#x8102;&#x86CB;&#x767D;&#x53BB;&#x9664;&#x3002;&#x8FD9;&#x4E9B;&#x7814;&#x7A76;&#x7ED3;&#x679C;&#x8868;&#x660E;&#x5728;&#x98DF;&#x54C1;&#x4E2D;&#x7684;&#x5916;&#x6E90;&#x690D;&#x7269; Mirna &#x53EF;&#x4EE5;&#x8C03;&#x8282;&#x54FA;&#x4E73;&#x52A8;&#x7269;&#x4E2D;&#x76EE;&#x6807;&#x57FA;&#x56E0;&#x7684;&#x8868;&#x8FBE;&#x3002;

&#x5916;&#x6E90;&#x690D;&#x7269; MIR168a &#x4E13;&#x95E8;&#x9488;&#x5BF9;&#x54FA;&#x4E73;&#x52A8;&#x7269; LDLRAP1&#xFF1A; microRNA &#x4EA4;&#x53C9;&#x738B;&#x56FD;&#x8C03;&#x8282;&#x7684;&#x8BC1;&#x636E;&#x3002;

Our previous studies have demonstrated that stable microRNAs (miRNAs) in mammalian serum and plasma are actively secreted from tissues and cells and can serve as a novel class of biomarkers for diseases, and act as signaling molecules in intercellular communication. Here, we report the surprising finding that exogenous plant miRNAs are present in the sera and tissues of various animals and that these exogenous plant miRNAs are primarily acquired orally, through food intake. MIR168a is abundant in rice and is one of the most highly enriched exogenous plant miRNAs in the sera of Chinese subjects. Functional studies in vitro and in vivo demonstrated that MIR168a could bind to the human/mouse low-density lipoprotein receptor adapter protein 1 (LDLRAP1) mRNA, inhibit LDLRAP1 expression in liver, and consequently decrease LDL removal from mouse plasma. These findings demonstrate that exogenous plant miRNAs in food can regulate the expression of target genes in mammals.

Exogenous plant MIR168a specifically targets mammalian LDLRAP1: evidence of cross-kingdom regulation by microRNA.

Nos &eacute;tudes ant&eacute;rieures ont d&eacute;montr&eacute; que stable microARN (miARN) dans le plasma et le s&eacute;rum chez les mammif&egrave;res sont activement s&eacute;cr&eacute;t&eacute;e par les cellules et les tissus et peut servir &agrave; une nouvelle classe de biomarqueurs des maladies et agissent comme des mol&eacute;cules de signalisation dans la communication intercellulaire. Nous rapportons ici le constat surprenant que plante exog&egrave;ne miARN est pr&eacute;sents dans le s&eacute;rum et les tissus de divers animaux et que ces miARN plante exog&egrave;ne est principalement acquis par voie orale, par le biais de la prise alimentaire. MIR168a est abondante dans le riz et est l&#39;un de la plupart miARN hautement enrichi plante exog&egrave;ne dans les s&eacute;rums de sujets chinois. Fonctionnelles &eacute;tudes in vitro et in vivo ont d&eacute;montr&eacute; que MIR168a pouvait lier &agrave; la prot&eacute;ine d&#39;adaptateur r&eacute;cepteur humain/souris lipoprot&eacute;ines de basse densit&eacute; 1 (LDLRAP1) mRNA, inhibent l&#39;expression de LDLRAP1 dans le foie et diminuent par cons&eacute;quent enl&egrave;vement de LDL du plasma de la souris. Ces r&eacute;sultats d&eacute;montrent que les miARN plante exog&egrave;ne dans les denr&eacute;es alimentaires peut r&eacute;glementer l&#39;expression de g&egrave;nes cibles chez les mammif&egrave;res.

Plante exog&egrave;ne MIR168a vise particuli&egrave;rement les mammif&egrave;res LDLRAP1 : preuve de r&egrave;glement de la Croix-Royaume de micro-ARN.

Unsere fr&uuml;heren Studien haben bewiesen, dass stabile MicroRNAs (MiRNAs) in S&auml;ugetieren Serum und Plasma werden aktiv von Geweben und Zellen abgesondert und kann als eine neuartige Klasse von Biomarkern f&uuml;r Krankheiten dienen und als signalisierende Molek&uuml;le in interzellul&auml;re Kommunikation fungieren. Hier berichten wir &uuml;ber die &uuml;berraschend finden, dass exogene Pflanze MiRNAs im Serum und Gewebe verschiedener Tiere vorhanden sind und dass diese exogenen Pflanze MiRNAs haupts&auml;chlich m&uuml;ndlich, durch Nahrungsaufnahme erworben werden. MIR168a ist reichlich Reis und geh&ouml;rt zu den meisten hoch angereicherten exogene Pflanze MiRNAs in den Seren von chinesischen F&auml;cher. Funktionsstudien in vitro und in vivo gezeigt, dass MIR168a dem Mensch/Maus low density Lipoprotein Receptor Adapter Protein 1 (LDLRAP1)-mRNA binden k&ouml;nnte LDLRAP1 Ausdruck in der Leber zu hemmen, und folglich verringern LDL-Entfernung aus Maus-Plasma. Diese Ergebnisse zeigen, dass exogene Pflanze MiRNAs in Essen den Ausdruck der Gene gezielt in S&auml;ugetiere regulieren k&ouml;nnen.

Exogene Pflanze MIR168a richtet sich speziell an S&auml;ugetier-LDLRAP1: Beweise Kreuz-K&ouml;nigreich Verordnung durch MicroRNA.

Nostri studi precedenti hanno dimostrato che stabile microRNA (miRNAs) nel plasma e siero dei mammiferi sono attivamente secreti dalle cellule e tessuti e pu&ograve; servire come una nuova classe di marcatori biologici per le malattie e fungere da molecole di segnalazione in comunicazione intercellulare. Riportiamo il sorprendente trovare che pianta esogena miRNAs sono presenti nel siero e nei tessuti di vari animali e che questi miRNAs della pianta esogena principalmente sono acquisite per via orale, attraverso l&#39;assunzione di cibo. MIR168a &egrave; abbondante nel riso ed &egrave; uno del pi&ugrave; altamente arricchito pianta esogena miRNAs in sieri di soggetti cinesi. Studi funzionali in vitro e in vivo hanno dimostrato che MIR168a pu&ograve; legarsi alla proteina umana e mouse lipoproteine a bassa densit&agrave; del recettore adattatore 1 mRNA (LDLRAP1), inibiscono la LDLRAP1 espressione nel fegato e di conseguenza diminuiscono la rimozione di LDL dal plasma del mouse. Questi risultati dimostrano che pianta esogena miRNAs in alimento pu&ograve; regolare l&#39;espressione di geni bersaglio in mammiferi.

Pianta esogena MIR168a specificamente gli obiettivi LDLRAP1 mammiferi: prove di cross-Unito regolamento di microRNA.

&#x672C;&#x7814;&#x7A76;&#x306F;&#x3001;&#x54FA;&#x4E73;&#x985E;&#x306E;&#x8840;&#x6E05;&#x3068;&#x8840;&#x6F3F;&#x4E2D;&#x306E;&#x5B89;&#x5B9A;&#x3057;&#x305F;&#x30DE;&#x30A4;&#x30AF;&#x30ED; Rna &#xFF08;Mirna&#xFF09; &#x7A4D;&#x6975;&#x7684;&#x306B;&#x7D44;&#x7E54;&#x3084;&#x7D30;&#x80DE;&#x304B;&#x3089;&#x5206;&#x6CCC;&#x3055;&#x308C;&#x308B;&#x3068;&#x75C5;&#x306E;&#x30D0;&#x30A4;&#x30AA; &#x30DE;&#x30FC;&#x30AB;&#x30FC;&#x306E;&#x65B0;&#x898F;&#x30AF;&#x30E9;&#x30B9;&#x3068;&#x3057;&#x3066;&#x6A5F;&#x80FD;&#x3067;&#x304D;&#x7D30;&#x80DE;&#x9593;&#x30B3;&#x30DF;&#x30E5;&#x30CB;&#x30B1;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x306B;&#x304A;&#x3051;&#x308B;&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x5206;&#x5B50;&#x3068;&#x3057;&#x3066;&#x6A5F;&#x80FD;&#x3059;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;&#x5916;&#x56E0;&#x6027;&#x690D;&#x7269; Mirna &#x304C;&#x5404;&#x7A2E;&#x52D5;&#x7269;&#x306E;&#x7D44;&#x7E54;&#x304A;&#x3088;&#x3073;&#x8840;&#x6E05;&#x4E2D;&#x306B;&#x5B58;&#x5728;&#x3057;&#x3001;&#x98DF;&#x7269;&#x6442;&#x53D6;&#x3092;&#x901A;&#x3058;&#x3066;&#x3053;&#x308C;&#x3089;&#x306E;&#x5916;&#x56E0;&#x6027;&#x690D;&#x7269;&#x306E; miRNAs &#x4E3B;&#x306B;&#x53E3;&#x982D;&#x3067;&#x3001;&#x53D6;&#x5F97;&#x3057;&#x305F;&#x3044;&#x308B;&#x3053;&#x3068;&#x610F;&#x5916;&#x306A;&#x767A;&#x898B;&#x3092;&#x5831;&#x544A;&#x3057;&#x307E;&#x3059;&#x3002;MIR168a &#x30E9;&#x30A4;&#x30B9;&#x3067;&#x8C4A;&#x5BCC;&#x3067;&#x3042;&#x308B;&#x3001;&#x307B;&#x3068;&#x3093;&#x3069;&#x306E;&#x9AD8;&#x6FC3;&#x7E2E;&#x5916;&#x56E0;&#x6027;&#x690D;&#x7269; miRNAs &#x4E2D;&#x56FD;&#x88AB;&#x9A13;&#x8005;&#x306E;&#x8840;&#x6E05;&#x4E2D;&#x306E;&#x4E00;&#x3064;&#x3067;&#x3059;&#x3002;In vitro &#x304A;&#x3088;&#x3073; in vivo &#x3067;&#x306E;&#x6A5F;&#x80FD;&#x306E;&#x7814;&#x7A76; MIR168a &#x4EBA;&#x9593;/&#x30DE;&#x30A6;&#x30B9;&#x4F4E;&#x6BD4;&#x91CD;&#x30EA;&#x30DD;&#x86CB;&#x767D;&#x53D7;&#x5BB9;&#x4F53;&#x30A2;&#x30C0;&#x30D7;&#x30BF;&#x30FC;&#x86CB;&#x767D;&#x8CEA;&#x306B; 1 (LDLRAP1) mRNA &#x30D0;&#x30A4;&#x30F3;&#x30C9;&#x3067;&#x304D;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x305F;&#x809D; LDLRAP1 &#x306E;&#x767A;&#x73FE;&#x3092;&#x7981;&#x3058;&#x308B;&#x3001;&#x3057;&#x305F;&#x304C;&#x3063;&#x3066;&#x6E1B;&#x5C11; LDL &#x9664;&#x53BB;&#x30DE;&#x30A6;&#x30B9;&#x8840;&#x6F3F;&#x304B;&#x3089;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x8ABF;&#x67FB;&#x7D50;&#x679C;&#x306F;&#x3001;&#x5916;&#x56E0;&#x6027;&#x690D;&#x7269; miRNAs &#x98DF;&#x54C1;&#x3067;&#x54FA;&#x4E73;&#x985E;&#x6A19;&#x7684;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x767A;&#x73FE;&#x3092;&#x8ABF;&#x6574;&#x3067;&#x304D;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;

&#x5916;&#x56E0;&#x6027;&#x690D;&#x7269; MIR168a &#x5177;&#x4F53;&#x7684;&#x306B;&#x30BF;&#x30FC;&#x30B2;&#x30C3;&#x30C8;&#x54FA;&#x4E73;&#x985E; LDLRAP1: &#x30DE;&#x30A4;&#x30AF;&#x30ED; Rna &#x306B;&#x3088;&#x308B;&#x30AF;&#x30ED;&#x30B9;&#x738B;&#x56FD;&#x898F;&#x5236;&#x306E;&#x8A3C;&#x62E0;&#x3002;

&#xC6B0;&#xB9AC;&#xC758; &#xC774;&#xC804; &#xC5F0;&#xAD6C; &#xD3EC;&#xC720;&#xB958; &#xD608; &#xCCAD; &#xBC0F; &#xD50C;&#xB77C;&#xC988;&#xB9C8;&#xC5D0; &#xC548;&#xC815;&#xC801;&#xC778; microRNAs (miRNAs) &#xC801;&#xADF9;&#xC801;&#xC73C;&#xB85C; &#xC870;&#xC9C1;&#xACFC; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xBD84; &#xBE44;, &#xC218; &#xC9C8;&#xBCD1;, &#xBC14;&#xC774;&#xC624; &#xB9C8;&#xCEE4;&#xC758; &#xC18C;&#xC124; &#xD074;&#xB798;&#xC2A4;&#xB85C; &#xC0AC;&#xC6A9;&#xD560;&#xC744; &#xC138;&#xD3EC; &#xD1B5;&#xC2E0; &#xC2E0;&#xD638; &#xBD84;&#xC790; &#xD589;&#xC704;&#xB85C; &#xC785;&#xC99D;. &#xC5EC;&#xAE30;, &#xC6B0;&#xB9AC;&#xAC00; exogenous &#xACF5;&#xC7A5; miRNAs &#xC138;&#xB77C;&#xC640; &#xB2E4;&#xC591; &#xD55C; &#xB3D9;&#xBB3C;&#xC758; &#xC870;&#xC9C1;&#xC5D0; &#xC874;&#xC7AC; &#xD558;&#xBA70; &#xC774;&#xB7EC;&#xD55C; exogenous &#xACF5;&#xC7A5; miRNAs &#xC74C;&#xC2DD; &#xC12D;&#xCDE8;&#xB97C; &#xD1B5;&#xD574; &#xAD6C;&#xB450;&#xB85C; &#xC778;&#xC218; &#xC8FC;&#xB85C; &#xB180;&#xB77C;&#xC6B4; &#xC77C;&#xC774; &#xCC3E;&#xB294; &#xBCF4;&#xACE0;. MIR168a &#xD48D;&#xBD80;&#xD55C; &#xC300;&#xC5D0;&#xC11C; &#xC774;&#xBA70; &#xC911;&#xAD6D; &#xACFC;&#xBAA9;&#xC758; &#xC138;&#xB77C;&#xC5D0;&#xC11C; &#xAC00;&#xC7A5; &#xACE0;&#xB3C4;&#xB85C; &#xB18D;&#xCD95;&#xB41C; exogenous &#xACF5;&#xC7A5; Mirnas&#xC758; &#xD558;&#xB098;&#xC785;&#xB2C8;&#xB2E4;. &#xC0DD;&#xCCB4; &#xC678; &#xBC0F; &#xC0DD;&#xCCB4; &#xC870;&#xAC74; &#xAE30;&#xB2A5; &#xC5F0;&#xAD6C; MIR168a 1 (LDLRAP1) mRNA &#xC778;&#xAC04;/&#xB9C8;&#xC6B0;&#xC2A4; &#xC800;&#xBC00;&#xB3C4; &#xC9C0; &#xB2E8;&#xBC31;&#xC9C8; &#xC218;&#xC6A9; &#xCCB4; &#xC5B4;&#xB311;&#xD130; &#xB2E8;&#xBC31;&#xC9C8;&#xC5D0; &#xBC14;&#xC778;&#xB529;&#xD560; &#xC218; &#xC2DC;&#xC5F0; LDLRAP1 &#xC2DD; &#xAC04;, &#xC5B5;&#xC81C; &#xADF8;&#xB9AC;&#xACE0; &#xACB0;&#xACFC;&#xC801;&#xC73C;&#xB85C; LDL &#xC81C;&#xAC70; &#xB9C8;&#xC6B0;&#xC2A4; &#xD50C;&#xB77C;&#xC988;&#xB9C8;&#xC5D0;&#xC11C; &#xAC10;&#xC18C;. &#xC774;&#xB7EC;&#xD55C; &#xC5F0;&#xAD6C; &#xACB0;&#xACFC; &#xC74C;&#xC2DD;&#xC5D0; exogenous &#xACF5;&#xC7A5; miRNAs &#xD3EC;&#xC720;&#xB958;&#xC5D0;&#xC11C; &#xB300;&#xC0C1; &#xC720;&#xC804;&#xC790;&#xC758; &#xD45C;&#xC815;&#xC744; &#xC870;&#xC808;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4; &#xBCF4;&#xC5EC; &#xC90D;&#xB2C8;&#xB2E4;.

Exogenous &#xACF5;&#xC7A5; MIR168a &#xD2B9;&#xD788; &#xD45C;&#xC801;&#xC73C;&#xB85C; &#xD3EC;&#xC720;&#xB958; LDLRAP1: Microrna&#xC5D0; &#xC758;&#xD574; &#xD06C;&#xB85C;&#xC2A4;-&#xB098;&#xB77C; &#xB808; &#xADE4; &#xB808;&#xC774; &#xC158;&#xC758; &#xC99D;&#xAC70;.

Nuestros estudios anteriores han demostrado que estable microRNAs (miRNAs) en mam&iacute;fero suero y plasma se secretan activamente de tejidos y c&eacute;lulas y puede servir como una nueva clase de biomarcadores de enfermedades y act&uacute;an como mol&eacute;culas de se&ntilde;alizaci&oacute;n en la comunicaci&oacute;n intercelular. Aqu&iacute;, Divulgamos el sorprendente hallazgo que miRNAs ex&oacute;genos de la planta est&aacute;n presentes en el suero y tejidos de diversos animales y que estos miRNAs planta ex&oacute;geno se adquieren principalmente por v&iacute;a oral, a trav&eacute;s de la ingesta de alimentos. MIR168a es abundante en arroz y es uno de la mayor&iacute;a miRNAs altamente enriquecido planta ex&oacute;geno en el suero de sujetos chinos. Estudios funcionales in vitro e in vivo demostraron que MIR168a podr&iacute;a unirse a la prote&iacute;na de adaptador de receptor de lipoprote&iacute;na de baja densidad de humanos/rat&oacute;n 1 (LDLRAP1) mRNA, inhiben la expresi&oacute;n de LDLRAP1 en el h&iacute;gado y en consecuencia disminuyen la eliminaci&oacute;n de LDL del plasma de rat&oacute;n. Estos resultados demuestran que planta ex&oacute;geno miRNAs en alimentos puede regular la expresi&oacute;n de genes diana en mam&iacute;feros.

Planta ex&oacute;geno MIR168a dirigido espec&iacute;ficamente LDLRAP1 mam&iacute;feros: evidencia de la regulaci&oacute;n de la Cruz-Unido por microARN.

Autophagy is activated in cancer cells during chemotherapy and often contributes to tumor chemotherapy resistance. In this study, we characterized the role of microRNA-30a (miR-30a) in the coordination of cancer cell apoptosis and autophagy, which determines the sensitivity of cancer cells to chemotherapy. First, the autophagy activity in cancer cells increased after cis-dichloro-diamine platinum (cis-DDP) or Taxol treatment, as indicated by the enhanced expression of beclin 1, a key regulator of autophagy, and increased number of LC3-positive autophagosomes. Second, miRNA screening using a TaqMan probe-based quantitative RT-PCR assay identified that miR-30a, a miRNA that targets beclin 1, was significantly reduced in tumor cells by cis-DDP treatment. Forced expression of miR-30a significantly reduced beclin 1 and the autophagy activity of tumor cells induced by cis-DDP. Third, the blockade of tumor cell autophagy activity by miR-30a expression or 3-methyladenine significantly increased tumor cell apoptosis induced by cis-DDP treatment. Finally, an in vivo tumor implantation mouse model clearly showed that elevation of miR-30a in implanted tumor cells by administration of the recombinant lentivirus expressing miR-30a strongly enhanced cis-DDP-induced apoptosis of tumor cells. In conclusion, our results demonstrate for the first time that miR-30a can sensitize tumor cells to cis-DDP via reducing beclin 1-mediated autophagy and that increasing miR-30a level in tumor cells represents a novel approach to enhance the efficacy of chemotherapy during cancer treatment.

MicroRNA-30a sensitizes tumor cells to cis-platinum via suppressing beclin 1-mediated autophagy.

In this study, we first characterized synaptosome microRNA (miRNA) profiles using microarray and qRT-PCR. MicroRNAs were detected in isolated synaptic vesicles, and Ago2 immunoprecipitation studies revealed an association between miRNAs and Ago2. Second, we found that miR-29a, miR-99a, and miR-125a were significantly elevated in synaptosome supernatants after depolarization. MiRNA secretion by the synaptosome was Ca(2+)-dependent and was inhibited by the exocytosis inhibitor, okadaic acid. Furthermore, application of nerve growth factor increased miRNA secretion without altering the spontaneous release of miRNAs. Conversely, kainic acid decreased miRNA secretion and enhanced the spontaneous release of miRNAs. These results indicate that synaptosomes could secrete miRNAs. Finally, synthesized miRNAs were taken up by synaptosomes, and the endocytosis inhibitor Dynasore blocked this process. After incubation with miR-125a, additional miR-125a was bound to Ago2 in the synaptosome, and expression of the miR-125a target gene (PSD95 mRNA) was decreased; these findings suggest that the ingested miRNAs were assembled in the RNA-induced silencing complex, resulting in the degradation of target mRNAs. To our knowledge, this is the first study that demonstrates the secretion of miRNAs by synaptosomes under physiological stimulation and demonstrates that secreted miRNAs might be functionally active after being taken up by the synaptic fraction via the endocytic pathway.

Synaptosomes secrete and uptake functionally active microRNAs via exocytosis and endocytosis pathways.

Selective secretion of microRNA in CNS system.

Evidence of a causal link between male obesity and subfertility or infertility has been demonstrated previously. However, the mechanism underlying this link is incompletely understood. Here, we report that sustained high protein-tyrosine phosphatase 1B (PTP1B) activity in sperm of obese donors plays an essential role in coupling male obesity and subfertility or infertility. First, PTP1B level and activity were significantly higher in sperm from ob/ob mice than in wild-type littermates. High PTP1B level and activity in sperm was also observed in obese patients compared with non-obese donors. The enhanced sperm PTP1B level and activity in ob/ob mice and obese patients correlated with a defect of the sperm acrosome reaction (AR). Second, treating sperm from male ob/ob mice or obese men with a specific PTP1B inhibitor largely restored the sperm AR. Finally, blockade of sperm AR by enhanced PTP1B activity in male ob/ob mice or obese men was due to prolonged dephosphorylation of N-ethylmaleimide-sensitive factor by PTP1B, leading to the inability to reassemble the trans-SNARE complexes, which is a critical step in sperm acrosomal exocytosis. In summary, our study demonstrates for the first time that a sustained high PTP1B level or activity in the sperm of obese donors causes a defect of sperm AR and that PTP1B is a novel potential therapeutic target for male infertility treatment.

Sustained high protein-tyrosine phosphatase 1B activity in the sperm of obese males impairs the sperm acrosome reaction.

Aging-related erectile dysfunction (A-ED) is a neurovascular and refractory disorder with complicated pathophysiological mechanisms and a high prevalence. MicroRNAs (miRNAs), which modulate a variety of cell functions, may be involved in the pathophysiological processes of this disorder.

Identification and characterization of the MicroRNA profile in aging rats with erectile dysfunction.

Small RNA existed in commercial reverse transcriptase: primary evidence of functional small RNAs.

Previous studies reported that the expression of miR-23b-27b cluster was downregulated in embryonic brain cortices during hypoxia-induced neuronal apoptosis. However, the mechanism underlying this downregulation is not completely understood. Here, we report that the transcription factor c-Myc plays an important role in regulating the expression of miR-23b-27b cluster during hypoxia. First, the c-Myc protein level was significantly elevated in embryonic brain cortices in a mouse model of fetal distress. Second, forced overexpression or knockdown of c-Myc could suppress or increase the expression of miR-23b-27b cluster polynucleotides. Third, we identified 2 conserved c-Myc binding sites (E-boxes) in the enhancer and promoter regions of miR-23b-27b cluster in the mouse genome. Finally, we showed that elevated c-Myc expression led to an increase in the Apaf-1 level by suppressing miR-23b-27b cluster expression and that this enhanced neuronal sensitivity to apoptosis. In summary, our study demonstrates that c-Myc may suppress the expression of the miR-23b-27b cluster, resulting in additional neuronal apoptosis during hypoxia. 

The transcription factor c-Myc suppresses MiR-23b and MiR-27b transcription during fetal distress and increases the sensitivity of neurons to hypoxia-induced apoptosis.

BRCA1-associated protein-1 (BAP1) is an important nuclear-localized deubiquitinating enzyme that serves as a tumor suppressor in lung cancer; however, its function and its regulation are largely unknown. In this study, we found that BAP1 protein levels were dramatically diminished in lung cancer tissues while its mRNA levels did not differ significantly, suggesting that a post-transcriptional mechanism was involved in BAP1 regulation. Because microRNAs (miRNAs) are powerful post-transcriptional regulators of gene expression, we used bioinformatic analyses to search for miRNAs that could potentially bind BAP1. We predicted and experimentally validated miR-31 as a direct regulator of BAP1. Moreover, we showed that miR-31 promoted proliferation and suppressed apoptosis in lung cancer cells and accelerated the development of tumor growth in xenograft mice by inhibiting BAP1. Taken together, this study highlights an important role for miR-31 in the suppression of BAP1 in lung cancer cells and may provide insights into the molecular mechanisms of lung carcinogenesis. 

BAP1 suppresses lung cancer progression and is inhibited by miR-31.

To better understand the molecular aetiology of type 2 diabetes mellitus-associated erectile dysfunction (T2DMED) and to provide candidates for further study of its diagnosis and treatment, this study was designed to investigate differentially expressed microRNAs (miRNAs) in the corpus cavernosum (CC) of mice with T2DMED using GeneChip array techniques (Affymetrix miRNA 4.0 Array) and to predict target genes and signalling pathways regulated by these miRNAs based on bioinformatic analysis using TargetScan, the DAIAN web platform and DAVID. In the initial screening, 21 miRNAs appeared distinctly expressed in the T2DMED group (fold change ≥3, p ≤ 0.01). Among them, the differential expression of miR-18a, miR-206, miR-122, and miR-133 were confirmed by qRT-PCR (p < 0.05 and FDR <5 %). According to bioinformatic analysis, the four miRNAs were speculated to play potential roles in the mechanisms of T2DMED via regulating 28 different genes and several pathways, including apoptosis, fibrosis, eNOS/cGMP/PKG, and vascular smooth muscle contraction processes, which mainly focused on influencing the functions of the endothelium and smooth muscle in the CC. IGF-1, as one of the target genes, was verified to decrease in the CCs of T2DMED animals via ELISA and was confirmed as the target of miR-18a or miR-206 via luciferase assay. Finally, these four miRNAs deserve further confirmation as biomarkers of T2DMED in larger studies. Additionally, miR-18a and/or miR-206 may provide new preventive/therapeutic targets for ED management by targeting IGF-1.

Differentially expressed microRNAs in the corpus cavernosum from a murine model with type 2 diabetes mellitus-associated erectile dysfunction.

Zika virus (ZIKV) can lead to severe birth defects especially microcephaly in newborns by infecting human neural progenitors and impairing brain development. However, as the resident immune cells in the brain, the role of microglia in the ZIKV pathology is not clearly defined. To understand the interplay between immune response and neural cells, we investigate the interaction between microglia and NPCs during ZIKV infection. Our results demonstrate that primary microglia infected with ZIKV induces an inflammatory response similar to that in human, producing high level of tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), interleukin 1β (IL-1β) and inducible nitric oxide synthase (iNOS). Furthermore, conditional medium (CM) of ZIKV infected microglia showed inhibitory effects on cell proliferation and neuronal differentiation of neural precursor cells (NPCs) derived from E14 mice brain. Blocking cytokines in the CM remarkably improved neurogenesis and decreased astrocytic differentiation of NPCs. Together, our results suggest that microglia mediated neuroinflammation plays an important role in neuropathogenesis during ZIKV infection.

Qipeng Zhang

Nanjing University

Intracavernosal Pressure Recording to Evaluate Erectile Function in Rodents