two-algorithms-for-high-throughput-multi-parametric-quantification

Two Algorithms for High-throughput and Multi-parametric Quantification of <em>Drosophila</em> Neuromuscular Junction Morphology

Synaptic morphology is tightly related to synaptic efficacy, and in many cases morphological synapse defects ultimately lead to synaptic malfunction. The ...

Universidad Autónoma de Madrid

The LIM-HD protein Apterous has been shown to regulate expression of the FMRFamide neuropeptide in Drosophila neurons (Benveniste et al. [1998] Development 125:4757-4765). To test whether Apterous has a broader role in controlling neurosecretory identity, we analyzed the expression of several neuropeptides in apterous (ap) mutants. We show that Apterous is necessary for expression of the Leucokinin neuropeptide in a pair of brain neurons located in the lateral horn region of the protocerebrum (LHLK neurons). ap null mutants are depleted of Leucokinin in these cells, whereas hypomorphic mutants show reduced Leucokinin expression. Other Leucokinin-containing neurons are not affected by mutations in ap gene. Co-expression of apterous and Leucokinin is observed exclusively in the LHLK neurons, from larval stages to adulthood. Rescue assays performed in null ap mutants, by expressing Apterous protein under apGAL4 and elavGAL4 drivers, demonstrate the recovery of Leucokinin in the LHLK neurons. These results reinforce the emerging role of the LIM-HD proteins in determining neuronal identity. They also clarify the neuroendocrine phenotype of apterous mutants.

Neurosecretory identity conferred by the apterous gene: lateral horn leucokinin neurons in Drosophila.

&#x810A;&#x690E;&#x52A8;&#x7269;&#x8F7D;&#x8102;&#x86CB;&#x767D; D (ApoD) &#x662F;&#x5728;&#x795E;&#x7ECF;&#x53D1;&#x80B2;&#x548C;&#x8870;&#x8001;&#x8FC7;&#x7A0B;&#x4ECE;&#x5B50;&#x96C6;&#x7684;&#x795E;&#x7ECF;&#x5143;&#x548C;&#x80F6;&#x8D28;&#x7EC6;&#x80DE;&#x5206;&#x6CCC;&#x7684;&#x8F7D;&#x8102;&#x86CB;&#x767D;&#x3002;ApoD &#x9AD8;&#x6548;&#x8868;&#x8FBE;&#x548C;&#x8BB8;&#x591A;&#x795E;&#x7ECF;&#x7CFB;&#x7EDF;&#x75C5;&#x75C7;&#x548C;&#x80A5;&#x80D6;&#x3001; &#x7CD6;&#x5C3F;&#x75C5;&#x3001; &#x4EE5;&#x53CA;&#x8BB8;&#x591A;&#x5F62;&#x5F0F;&#x7684;&#x764C;&#x75C7;&#x4E4B;&#x95F4;&#x5B58;&#x5728;&#x5F3A;&#x5173;&#x8054;&#x3002;&#x7136;&#x800C;&#xFF0C;&#x4ECD;&#x672A;&#x77E5; ApoD &#x7684;&#x529F;&#x80FD;&#x4E0E;&#x8FD9;&#x4E9B;&#x75BE;&#x75C5;&#x7684;&#x75C5;&#x7406;&#x751F;&#x7406;&#x4E4B;&#x95F4;&#x7684;&#x786E;&#x5207;&#x5173;&#x7CFB;&#x3002;&#x6211;&#x4EEC;&#x5DF2;&#x751F;&#x6210;&#x529F;&#x80FD;&#x635F;&#x5931;&#x7684;&#x679C;&#x8747;&#x7A81;&#x53D8;&#x4F53;&#x4E3A;&#x80F6;&#x8D28; Lazarillo (&#x70ED;&#x5316;) &#x57FA;&#x56E0;&#xFF0C;&#x5728;&#x679C;&#x8747;&#xFF0C;&#x4E3B;&#x8981;&#x8868;&#x73B0;&#x5728;&#x6210;&#x4EBA;&#x795E;&#x7ECF;&#x80F6;&#x8D28;&#x7EC6;&#x80DE;&#x4E9A;&#x7FA4;&#x7684; ApoD &#x540C;&#x7CFB;&#x7269;&#x7684;&#x6C2F;&#x4EE3;&#x70C3;&#x3002;&#x70ED;&#x5316;&#x6CA1;&#x6709;&#x964D;&#x4F4E;&#x4E86;&#x673A;&#x4F53;&#x7684;&#x6297;&#x6C27;&#x5316;&#x5E94;&#x6FC0;&#x548C;&#x9965;&#x997F;&#xFF0C;&#x5E76;&#x7F29;&#x77ED;&#x4E86;&#x7537;&#x6027;&#x7684;&#x5BFF;&#x547D;&#x3002;&#x7A81;&#x53D8;&#x679C;&#x8747;&#x8868;&#x73B0;&#x51FA;&#x7531;&#x4E8E;&#x4E2D;&#x6027;&#x8102;&#x50A8;&#x5B58;&#x5728;&#x8102;&#x80AA;&#x4F53;&#x4F4E;&#x91CF;&#x8F83;&#x5C0F;&#x7684;&#x8EAB;&#x4F53;&#x8D28;&#x91CF;&#x3002;&#x795E;&#x7ECF;&#x7EC6;&#x80DE;&#x7684;&#x51CB;&#x4EA1;&#x589E;&#x52A0;&#x8001;&#x5E74;&#x82CD;&#x8747;&#x6216;&#x767E;&#x8349;&#x67AF;&#x7684;&#x5F85;&#x9047;&#xFF0C;&#x8FD8;&#x4F1A;&#x635F;&#x5BB3;&#x795E;&#x7ECF;&#x529F;&#x80FD;&#x8BC4;&#x5B9A;&#x7684;&#x884C;&#x4E3A;&#x6D4B;&#x8BD5;&#x4E4B;&#x4E0A;&#x3002;&#x7EFF;&#x8272;&#x8367;&#x5149;&#x86CB;&#x767D;&#x70ED;&#x5316;&#x878D;&#x5408;&#x86CB;&#x767D;&#x8868;&#x793A;&#x70ED;&#x5316;&#x5929;&#x7136;&#x542F;&#x52A8;&#x5B50;&#x63A7;&#x5236;&#x4E0B;&#x5BF9;&#x6C27;&#x5316;&#x5E94;&#x6FC0;&#x548C;&#x9965;&#x997F;&#xFF0C;&#x51CF;&#x5C11;&#x8102;&#x80AA;&#x50A8;&#x5B58;&#x7684;&#x9AD8;&#x654F;&#x611F;&#x5EA6;&#x8FD8;&#x539F;&#x4E3A;&#x63A7;&#x5236;&#x6C34;&#x5E73;&#x3002;&#x6700;&#x540E;&#xFF0C;&#x70ED;&#x5316;&#x7A81;&#x53D8;&#x4F53;&#x6709;&#x8F83;&#x5BC6;&#x96C6;&#x7684;&#x8102;&#x8D28;&#x8FC7;&#x6C27;&#x5316;&#x4F5C;&#x7528;&#x7684;&#x4EA7;&#x54C1;&#xFF0C;&#x6307;&#x5411;&#x4E00;&#x79CD;&#x8102;&#x8D28;&#x8FC7;&#x6C27;&#x5316;&#x4FDD;&#x62A4;&#x6216;&#x6E05;&#x7406;&#x4F5C;&#x4E3A;&#x6B64;&#x8F7D;&#x8102;&#x86CB;&#x767D;&#x7684;&#x884C;&#x52A8;&#x673A;&#x5236;&#x3002;&#x540C;&#x610F; walker &#x7B49; &#xFF08;&#xFF0C;&#x5728;&#x8FD9;&#x4E00;&#x95EE;&#x9898;&#x7684;&#x5F53;&#x4EE3;&#x751F;&#x7269;&#x5B66;&#xFF09;&#x3001; &#x8C01;&#x5206;&#x6790;&#x7684;&#x8D85;&#x91CF;&#x8868;&#x8FBE;&#x70ED;&#x5316;&#x5F71;&#x54CD;&#x3001; &#x70ED;&#x5316;&#x5728;&#x538B;&#x529B;&#x60C5;&#x51B5;&#x4E0B;&#x5177;&#x6709;&#x4FDD;&#x62A4;&#x4F5C;&#x7528;&#x548C;&#x5176;&#x7F3A;&#x5E2D;&#x5BFF;&#x547D;&#x51CF;&#x5C11;&#x4E86;&#x52A0;&#x901F;&#x795E;&#x7ECF;&#x53D8;&#x6027;&#xFF0C;&#x6211;&#x4EEC;&#x5F97;&#x51FA;&#x7684;&#x7ED3;&#x8BBA;&#x3002;

&#x80F6;&#x8D28;&#x7EC6;&#x80DE; lazarillo&#xFF0C;&#x540C;&#x7CFB;&#x7269;&#x7684;&#x8F7D;&#x8102;&#x86CB;&#x767D; D&#x3001; &#x635F;&#x5931;&#x51CF;&#x5C11;&#x4E86;&#x679C;&#x8747;&#x5BFF;&#x547D;&#x4E0E;&#x5E94;&#x6FC0;&#x62B5;&#x6297;&#x3002;

The vertebrate Apolipoprotein D (ApoD) is a lipocalin secreted from subsets of neurons and glia during neural development and aging . A strong correlation exists between ApoD overexpression and numerous nervous system pathologies as well as obesity, diabetes, and many forms of cancer . However, the exact relationship between the function of ApoD and the pathophysiology of these diseases is still unknown. We have generated loss-of-function Drosophila mutants for the Glial Lazarillo (GLaz) gene , a homolog of ApoD in the fruit fly, mainly expressed in subsets of adult glial cells. The absence of GLaz reduces the organism's resistance to oxidative stress and starvation and shortens male lifespan. The mutant flies exhibit a smaller body mass due to a lower amount of neutral lipids stored in the fat body. Apoptotic neural cell death increases in aged flies or upon paraquat treatment, which also impairs neural function as assessed by behavioral tests. The higher sensitivity to oxidative stress and starvation and the reduced fat storage revert to control levels when a GFP-GLaz fusion protein is expressed under the control of the GLaz natural promoter. Finally, GLaz mutants have a higher concentration of lipid peroxidation products, pointing to a lipid peroxidation protection or scavenging as the mechanism of action for this lipocalin. In agreement with Walker et al. (, in this issue of Current Biology), who analyze the effects of overexpressing GLaz, we conclude that GLaz has a protective role in stress situations and that its absence reduces lifespan and accelerates neurodegeneration.

Loss of glial lazarillo, a homolog of apolipoprotein D, reduces lifespan and stress resistance in Drosophila.

Le vert&#xE9;br&#xE9; apolipoprot&#xE9;ine D (APOD) est un lipocaline s&#xE9;cr&#xE9;t&#xE9;e &#xE0; partir de sous-ensembles de neurones et cellules gliales au cours du d&#xE9;veloppement neuronal et le vieillissement. Une forte corr&#xE9;lation existe entre la surexpression ApoD et de nombreuses pathologies du syst&#xE8;me nerveux ainsi que l&#x26;#39;ob&#xE9;sit&#xE9;, le diab&#xE8;te, et de nombreuses formes de cancer. Cependant, la relation exacte entre la fonction de l&#x26;#39;ApoD et la physiopathologie de ces maladies est encore inconnue. Nous avons g&#xE9;n&#xE9;r&#xE9; une perte de fonction chez la drosophile mutants pour la Glial Lazarillo (Glaz) g&#xE8;ne, un homologue de l&#x26;#39;ApoD dans la mouche des fruits, principalement exprim&#xE9; dans les sous-ensembles de cellules gliales adultes. L&#x26;#39;absence de Glaz r&#xE9;duit la r&#xE9;sistance de l&#x26;#39;organisme au stress oxydatif et &#xE0; la famine et raccourcit la dur&#xE9;e de vie masculine. Les mouches mutantes pr&#xE9;sentent une masse corporelle plus faible en raison d&#x26;#39;une faible quantit&#xE9; de lipides neutres stock&#xE9;es dans la graisse corporelle. Apoptotiques neuronaux des augmentations de mort cellulaire chez les mouches &#xE2;g&#xE9;es ou lors d&#x26;#39;un traitement au paraquat, ce qui alt&#xE8;re &#xE9;galement la fonction neuronale &#xE9;valu&#xE9;e par des tests comportementaux. La plus grande sensibilit&#xE9; au stress oxydatif et famine et le stockage de graisse r&#xE9;duit en revenir &#xE0; contr&#xF4;ler les niveaux quand une prot&#xE9;ine de fusion GFP-Glaz est exprim&#xE9; sous le contr&#xF4;le de l&#x26;#39;Glaz promoteur naturel. Enfin, les mutants Glaz avoir une concentration plus &#xE9;lev&#xE9;e de produits de peroxydation lipidique, pointant vers une protection peroxydation lipidique ou de balayage que le m&#xE9;canisme d&#x26;#39;action de cette lipocaline. En accord avec Walker et al. (Dans ce num&#xE9;ro de la revue Current Biology), qui analysent les effets de la surexpression de Glaz, nous concluons que Glaz a un r&#xF4;le protecteur dans les situations de stress et que son absence r&#xE9;duit la dur&#xE9;e de vie et acc&#xE9;l&#xE8;re la neurod&#xE9;g&#xE9;n&#xE9;rescence.

Perte de Glial Lazarillo, un homologue de l&#x26;#39;apolipoprot&#xE9;ine D, r&#xE9;duit la r&#xE9;sistance Dur&#xE9;e de vie et le stress chez la drosophile

Die Wirbeltiere Apolipoprotein D (ApoD) ist ein Lipocalin abgesondert von Teilmengen von Neuronen und Glia w&auml;hrend der neuronalen Entwicklung und Alterung. Es besteht eine starke Korrelation zwischen ApoD-&Uuml;berexpression und zahlreiche Erkrankungen des Nervensystems wie Fettleibigkeit, Diabetes und vielen Formen von Krebs. Die genaue Beziehung zwischen der ApoD-Funktion und der Pathophysiologie dieser Krankheiten ist jedoch noch unbekannt. Wir haben die Verlustfunktion von Drosophila-Mutanten f&uuml;r den Gliazellen Lazarillo (GLaz) das Gen ein homologes von ApoD der Fruchtfliege, haupts&auml;chlich in Teilmengen von Erwachsenen Gliazellen ausgedr&uuml;ckt erzeugt. Das Fehlen von GLaz des Organismus gegen oxidativen Stress und Hunger reduziert und m&auml;nnliche Lebenserwartung verk&uuml;rzt. Die mutierten fliegen weisen einen kleineren K&ouml;rpermasse aufgrund geringerem Umfang neutralen Lipiden im K&ouml;rper Fett gespeichert. Apoptotic neuronalen Zelltod erh&ouml;ht in Alter fliegen oder bei Paraquat-Behandlung, die auch neuronale Funktion beeintr&auml;chtigt die verhaltensbasierte Pr&uuml;fungen zu bewerten. Die h&ouml;here Empfindlichkeit gegen&uuml;ber oxidativen Stress und Hunger und die reduzierten Fettspeicherung zu Steuerelement Ebenen zur&uuml;ckkehren, wenn ein GFP-GLaz-Fusionsprotein unter der Kontrolle des nat&uuml;rlichen Projekttr&auml;gers GLaz ausgedr&uuml;ckt wird. Schlie&szlig;lich haben die GLaz Mutanten eine h&ouml;here Konzentration von Lipid Peroxidation Produkte, zeigen ein Lipid-Peroxidation-Schutz oder als Mechanismus der Aktion f&uuml;r diese Lipocalin aufr&auml;umen. Im Einvernehmen mit Walker Et Al. (, in dieser Ausgabe von Current Biology), wer analysieren die Auswirkungen der &uuml;berexprimierenden GLaz, wir folgern, dass GLaz eine sch&uuml;tzende Rolle in Stresssituationen hat und seine Abwesenheit reduziert die Lebensdauer und beschleunigt die Neurodegeneration.

Verlust von Gliazellen Lazarillo, ein homologes von Apolipoprotein D, reduziert die Lebensdauer und Stress-Resistenz bei Drosophila.

Il vertebrato Apolipoprotein D (ApoD) &egrave; un Lipocalina secernuto da sottoinsiemi di neuroni e glia durante l&#39;invecchiamento e lo sviluppo neurale. Esiste una correlazione forte tra ApoD sovraespressione e numerose patologie del sistema nervoso cos&igrave; come obesit&agrave;, diabete e molte forme di cancro. Tuttavia, il rapporto esatto tra la funzione del ApoD e la fisiopatologia di queste malattie &egrave; ancora sconosciuto. Noi abbiamo generato mutanti Drosophila perdita di funzione del gene Lazarillo gliali (GLaz), un omologo del ApoD nel moscerino della frutta, principalmente espressa in sottoinsiemi di cellule gliale adulte. L&#39;assenza di GLaz riduce la resistenza dell&#39;organismo allo stress ossidativo e fame e accorcia la vita media maschile. Le mosche mutanti esibiscono una pi&ugrave; piccola massa di corpo a causa di una bassa quantit&agrave; di lipidi neutri memorizzati nel corpo grasso. Morte apoptotic delle cellule neurali aumenta in et&agrave; mosche o previo trattamento di paraquat, che altera anche funzione neurale, come valutato mediante prove comportamentali. La maggiore sensibilit&agrave; a stress ossidativo e fame e l&#39;accumulo di grasso ridotto tornare a livelli di controllo quando si esprime una proteina di fusione GFP-GLaz sotto il controllo del promotore naturale GLaz. Infine, GLaz mutanti hanno una maggiore concentrazione di prodotti di perossidazione lipidica, che punta a una protezione di perossidazione lipidica o lo scavenging come il meccanismo di azione per questo Lipocalina. D&#39;accordo con Walker et al (, in questo numero di Current Biology), che analizza gli effetti della con sovraesprimono relativa GLaz, concludiamo che GLaz ha un ruolo protettivo in situazioni di stress e che la sua assenza riduce la durata della vita e accelera la neurodegenerazione.

Perdita di glial lazarillo, un omologo del apolipoprotein D, riduce la resistenza durata della vita e dello stress in drosofila.

&#x810A;&#x690E;&#x52D5;&#x7269;&#x306E;&#x30A2;&#x30DD;&#x30EA;&#x30DD;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;D&#xFF08;APOD&#xFF09;&#x306F;&#x3001;&#x795E;&#x7D4C;&#x767A;&#x9054;&#x3068;&#x8001;&#x5316;&#x306E;&#x9593;&#x306B;&#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3;&#x3068;&#x30B0;&#x30EA;&#x30A2;&#x306E;&#x30B5;&#x30D6;&#x30BB;&#x30C3;&#x30C8;&#x304B;&#x3089;&#x5206;&#x6CCC;&#x3055;&#x308C;&#x308B;&#x30EA;&#x30DD;&#x30AB;&#x30EA;&#x30F3;&#x3067;&#x3059;&#x3002;&#x5F37;&#x3044;&#x76F8;&#x95A2;&#x304C;APOD&#x306E;&#x904E;&#x5270;&#x767A;&#x73FE;&#x3068;&#x591A;&#x6570;&#x306E;&#x4E2D;&#x67A2;&#x795E;&#x7D4C;&#x7CFB;&#x306E;&#x75C5;&#x614B;&#x3060;&#x3051;&#x3067;&#x306A;&#x304F;&#x3001;&#x80A5;&#x6E80;&#x3001;&#x7CD6;&#x5C3F;&#x75C5;&#x3001;&#x764C;&#x306E;&#x591A;&#x304F;&#x306E;&#x5F62;&#x614B;&#x306E;&#x9593;&#x306B;&#x5B58;&#x5728;&#x3057;&#x307E;&#x3059;&#x3002;&#x3057;&#x304B;&#x3057;&#x3001;APOD&#x306E;&#x6A5F;&#x80FD;&#x3068;&#x3053;&#x308C;&#x3089;&#x306E;&#x75BE;&#x60A3;&#x306E;&#x75C5;&#x614B;&#x751F;&#x7406;&#x306E;&#x9593;&#x306E;&#x6B63;&#x78BA;&#x306A;&#x95A2;&#x4FC2;&#x306F;&#x307E;&#x3060;&#x4E0D;&#x660E;&#x3067;&#x3059;&#x3002;&#x79C1;&#x305F;&#x3061;&#x306F;&#x3001;&#x4E3B;&#x306B;&#x5927;&#x4EBA;&#x306E;&#x30B0;&#x30EA;&#x30A2;&#x7D30;&#x80DE;&#x306E;&#x30B5;&#x30D6;&#x30BB;&#x30C3;&#x30C8;&#x306B;&#x767A;&#x73FE;&#x3057;&#x30B0;&#x30EA;&#x30A2;Lazarillo&#xFF08;&#x30B0;&#x30E9;&#x30FC;&#x30C4;&#xFF09;&#x907A;&#x4F1D;&#x5B50;&#x3001;&#x30B7;&#x30E7;&#x30A6;&#x30B8;&#x30E7;&#x30A6;&#x30D0;&#x30A8;&#x306E;APOD&#x306E;&#x30DB;&#x30E2;&#x30ED;&#x30B0;&#x306E;&#x305F;&#x3081;&#x306E;&#x6A5F;&#x80FD;&#x55AA;&#x5931;&#x30B7;&#x30E7;&#x30A6;&#x30B8;&#x30E7;&#x30A6;&#x30D0;&#x30A8;&#x306E;&#x5909;&#x7570;&#x4F53;&#x3092;&#x751F;&#x6210;&#x3057;&#x3066;&#x3044;&#x308B;&#x3002;&#x30B0;&#x30E9;&#x30FC;&#x30C4;&#x306E;&#x4E0D;&#x5728;&#x306F;&#x3001;&#x9178;&#x5316;&#x30B9;&#x30C8;&#x30EC;&#x30B9;&#x3068;&#x98E2;&#x9913;&#x306B;&#x751F;&#x7269;&#x306E;&#x62B5;&#x6297;&#x3092;&#x4F4E;&#x6E1B;&#x3057;&#x3001;&#x7537;&#x6027;&#x306E;&#x5BFF;&#x547D;&#x304C;&#x77ED;&#x304F;&#x306A;&#x308A;&#x307E;&#x3059;&#x3002;&#x5909;&#x7570;&#x30CF;&#x30A8;&#x306F;&#x3001;&#x8102;&#x80AA;&#x4F53;&#x306B;&#x683C;&#x7D0D;&#x3055;&#x308C;&#x305F;&#x4E2D;&#x6027;&#x8102;&#x8CEA;&#x306E;&#x4E0B;&#x9650;&#x91CF;&#x306E;&#x305F;&#x3081;&#x306B;&#x5C0F;&#x3055;&#x306A;&#x30DC;&#x30C7;&#x30A3;&#x306E;&#x8CEA;&#x91CF;&#x3092;&#x793A;&#x3059;&#x3002;&#x9AD8;&#x9F62;&#x30CF;&#x30A8;&#x3084;&#x884C;&#x52D5;&#x30C6;&#x30B9;&#x30C8;&#x306B;&#x3088;&#x3063;&#x3066;&#x8A55;&#x4FA1;&#x3055;&#x308C;&#x308B;&#x3088;&#x3046;&#x306B;&#x3082;&#x795E;&#x7D4C;&#x6A5F;&#x80FD;&#x3092;&#x640D;&#x306A;&#x3046;&#x30D1;&#x30E9;&#x30B3;&#x30FC;&#x30C8;&#x51E6;&#x7406;&#x3001;&#x6642;&#x306E;&#x30A2;&#x30DD;&#x30C8;&#x30FC;&#x30B7;&#x30B9;&#x795E;&#x7D4C;&#x7D30;&#x80DE;&#x6B7B;&#x304C;&#x5897;&#x52A0;&#x3057;&#x307E;&#x3059;&#x3002;&#x9178;&#x5316;&#x30B9;&#x30C8;&#x30EC;&#x30B9;&#x3068;&#x98E2;&#x9913;&#x3068;&#x4F4E;&#x8102;&#x80AA;&#x306E;&#x84C4;&#x7A4D;&#x306B;&#x9AD8;&#x3044;&#x611F;&#x5EA6;&#x306F;&#x3001;GFP-&#x30B0;&#x30E9;&#x30FC;&#x30C4;&#x878D;&#x5408;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;&#x306F;&#x30B0;&#x30E9;&#x30FC;&#x30C4;&#x306E;&#x5929;&#x7136;&#x30D7;&#x30ED;&#x30E2;&#x30FC;&#x30BF;&#x30FC;&#x306E;&#x5236;&#x5FA1;&#x4E0B;&#x3067;&#x767A;&#x73FE;&#x3055;&#x308C;&#x305F;&#x3068;&#x304D;&#x306E;&#x30EC;&#x30D9;&#x30EB;&#x3092;&#x5236;&#x5FA1;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x623B;&#x308A;&#x307E;&#x3059;&#x3002;&#x6700;&#x5F8C;&#x306B;&#x3001;&#x30B0;&#x30E9;&#x30FC;&#x30C4;&#x306E;&#x5909;&#x7570;&#x4F53;&#x306F;&#x3001;&#x8102;&#x8CEA;&#x904E;&#x9178;&#x5316;&#x4FDD;&#x8B77;&#x3092;&#x6307;&#x3057;&#x3066;&#x3044;&#x308B;&#x3001;&#x307E;&#x305F;&#x306F;&#x3053;&#x306E;&#x30EA;&#x30DD;&#x30AB;&#x30EA;&#x30F3;&#x306E;&#x305F;&#x3081;&#x306E;&#x884C;&#x52D5;&#x306E;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x3068;&#x3057;&#x3066;&#x3001;&#x6E05;&#x6383;&#x3001;&#x8102;&#x8CEA;&#x904E;&#x9178;&#x5316;&#x751F;&#x6210;&#x7269;&#x306E;&#x9AD8;&#x3044;&#x6FC3;&#x5EA6;&#x3092;&#x6301;&#x3063;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002; Walker&#x3089;&#x3068;&#x306E;&#x5408;&#x610F;&#x3067;&#x3042;&#x308B;&#x3002; &#xFF08;&#x73FE;&#x5728;&#x306E;&#x751F;&#x7269;&#x5B66;&#x306F;&#x3001;&#x3053;&#x306E;&#x554F;&#x984C;&#x3067;&#xFF09;&#x904E;&#x5270;&#x767A;&#x73FE;&#x30B0;&#x30E9;&#x30FC;&#x30C4;&#x306E;&#x52B9;&#x679C;&#x3092;&#x5206;&#x6790;&#x3059;&#x308B;&#x4EBA;&#x306F;&#x3001;&#x6211;&#x3005;&#x306F;&#x3001;&#x30B0;&#x30E9;&#x30FC;&#x30C4;&#x306F;&#x3001;&#x30B9;&#x30C8;&#x30EC;&#x30B9;&#x72B6;&#x6CC1;&#x3067;&#x306E;&#x4FDD;&#x8B77;&#x306E;&#x5F79;&#x5272;&#x3092;&#x6301;&#x3063;&#x3066;&#x3044;&#x308B;&#x3053;&#x3068;&#x3068;&#x3001;&#x305D;&#x306E;&#x4E0D;&#x5728;&#x304C;&#x5BFF;&#x547D;&#x3092;&#x6E1B;&#x3089;&#x3057;&#x3001;&#x795E;&#x7D4C;&#x5909;&#x6027;&#x3092;&#x4FC3;&#x9032;&#x3059;&#x308B;&#x3068;&#x7D50;&#x8AD6;&#x4ED8;&#x3051;&#x3066;&#x3044;&#x308B;&#x3002;

&#x30B0;&#x30EA;&#x30A2;Lazarillo&#x306E;&#x640D;&#x5931;&#x3001;&#x30A2;&#x30DD;&#x30EA;&#x30DD;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;D&#x306E;&#x30DB;&#x30E2;&#x30ED;&#x30B0;&#x306F;&#x3001;&#x30B7;&#x30E7;&#x30A6;&#x30B8;&#x30E7;&#x30A6;&#x30D0;&#x30A8;&#x306E;&#x5BFF;&#x547D;&#x3068;&#x30B9;&#x30C8;&#x30EC;&#x30B9;&#x62B5;&#x6297;&#x3092;&#x4F4E;&#x6E1B;

&#xCC99;&#xCD94; Apolipoprotein D (ApoD) &#xC2E0;&#xACBD; &#xBC1C;&#xB2EC; &#xBC0F; &#xB178;&#xD654; &#xC911; &#xC2E0;&#xACBD; &#xC138;&#xD3EC;&#xC640; &#xBA85;&#xACFC;&#xC758; &#xD558;&#xC704; &#xC9D1;&#xD569;&#xC5D0;&#xC11C; &#xBD84; &#xBE44; &#xD558;&#xB294; lipocalin&#xC785;&#xB2C8;&#xB2E4;. ApoD overexpression &#xBC0F; &#xC218;&#xB9CE;&#xC740; &#xC2E0; &#xACBD;&#xACC4; &#xBCD1; &#xB9AC;&#xB85C; &#xBE44;&#xB9CC;, &#xB2F9;&#xB1E8;&#xBCD1;, &#xADF8;&#xB9AC;&#xACE0; &#xB2E4;&#xC591; &#xD55C; &#xD615;&#xD0DC;&#xC758; &#xC554; &#xC0AC;&#xC774; &#xAC15;&#xD55C; &#xC0C1;&#xAD00; &#xAD00;&#xACC4; &#xC874;&#xC7AC;. &#xADF8;&#xB7EC;&#xB098;, Apod&#xC758; &#xAE30;&#xB2A5;&#xACFC; &#xC774;&#xB7EC;&#xD55C; &#xC9C8;&#xBCD1;&#xC758; &#xC774;&#xC0C1; &#xC0AC;&#xC774;&#xC758; &#xC815;&#xD655;&#xD55C; &#xAD00;&#xACC4;&#xB294; &#xC544;&#xC9C1; &#xC54C;. &#xC6B0;&#xB9AC; &#xAD50; Lazarillo (GLaz) &#xC720;&#xC804;&#xC790;, &#xC8FC;&#xB85C; &#xC131;&#xC778; glial &#xC138;&#xD3EC;&#xC758; &#xD558;&#xC704; &#xC9D1;&#xD569;&#xC73C;&#xB85C; &#xD45C;&#xD604; &#xD558;&#xB294; &#xACFC;&#xC77C; &#xD30C;&#xB9AC;&#xC5D0;&#xC11C; Apod&#xC758; &#xCCB4;&#xC5D0; &#xB300; &#xD55C; &#xC190;&#xC2E4;&#xC758; &#xAE30;&#xB2A5;&#xC758; &#xCD08;&#xD30C;&#xB9AC; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xC0DD;&#xC131;. GLaz &#xBD80;&#xC7AC; &#xC0B0;&#xD654; &#xC2A4;&#xD2B8;&#xB808;&#xC2A4;&#xC640; &#xAE30;&#xC544;&#xC5D0; &#xB300; &#xD55C; &#xC720;&#xAE30; &#xCCB4;&#xC758; &#xC800;&#xD56D;&#xC744; &#xAC10;&#xC18C; &#xD558; &#xACE0; &#xB0A8;&#xC131; &#xC218;&#xBA85; &#xB2E8;&#xCD95;. &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xD30C;&#xB9AC; &#xC804;&#xC2DC; &#xC911;&#xB9BD; &#xC9C0;&#xC9C8; &#xC9C0;&#xBC29;&#xC774; &#xC2E0;&#xCCB4;&#xC5D0; &#xC800;&#xC7A5; &#xB41C;&#xC758; &#xB0AE;&#xC740; &#xAE08;&#xC561;&#xC73C;&#xB85C; &#xC778;&#xD574; &#xC791;&#xC740; &#xC2E0;&#xCCB4; &#xC9C8;&#xB7C9;. Apoptotic &#xC2E0;&#xACBD; &#xC138;&#xD3EC; &#xC8FD;&#xC74C; &#xC138; &#xD30C;&#xB9AC; &#xB610;&#xB294; &#xD30C;&#xB77C;&#xCF70;&#xD2B8; &#xCE58;&#xB8CC;, &#xD589;&#xB3D9; &#xD14C;&#xC2A4;&#xD2B8;&#xB85C; &#xD3C9;&#xAC00; &#xB610;&#xD55C; &#xC2E0;&#xACBD; &#xAE30;&#xB2A5;&#xC744; &#xC190;&#xC0C1; &#xC2DC; &#xC99D;&#xAC00; &#xD569;&#xB2C8;&#xB2E4;. &#xC0B0;&#xD654; &#xC2A4;&#xD2B8;&#xB808;&#xC2A4;&#xC640; &#xAE30;&#xC544; &#xAC10;&#xC18C; &#xC9C0;&#xBC29; &#xC2A4;&#xD1A0;&#xB9AC;&#xC9C0;&#xB97C; &#xB192;&#xC740; &#xAC10;&#xB3C4; GLaz GFP &#xC735;&#xD574; &#xB2E8;&#xBC31;&#xC9C8; GLaz &#xC790;&#xC5F0; &#xBC1C;&#xAE30;&#xC778; &#xD1B5;&#xC81C; &#xD45C;&#xD604; &#xC81C;&#xC5B4; &#xC218;&#xC900;&#xC73C;&#xB85C; &#xB3CC;&#xC544;&#xAC11;&#xB2C8;&#xB2E4;. &#xB9C8;&#xC9C0;&#xB9C9;&#xC73C;&#xB85C;, GLaz &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xC9C0;&#xC9C8; &#xACFC;&#xC0B0;&#xD654; &#xBCF4;&#xD638;&#xB97C; &#xAC00;&#xB9AC;&#xD0A4;&#xB294; &#xB610;&#xB294;&#xC774; lipocalin&#xC5D0; &#xB300; &#xD55C; &#xD589;&#xB3D9;&#xC758; &#xBA54;&#xCEE4;&#xB2C8;&#xC998;&#xC73C;&#xB85C; &#xCCAD;&#xC18C; &#xC9C0;&#xC9C8; &#xACFC;&#xC0B0;&#xD654; &#xC81C;&#xD488;&#xC758; &#xB192;&#xC740; &#xB18D;&#xB3C4;&#xAC00; &#xC788;&#xB2E4;. &#xC6CC;&#xCEE4; &#xC678;. &#xB3D9;&#xC758; (, &#xD604;&#xC7AC; &#xC0DD;&#xBB3C;&#xD559;&#xC758;&#xC774; &#xBB38;&#xC81C;&#xC5D0;), &#xB204;&#xAC00; GLaz overexpressing&#xC758; &#xC601;&#xD5A5;&#xC744; &#xBD84;&#xC11D;, &#xC6B0;&#xB9AC;&#xAC00; &#xACB0;&#xB860;&#xC744; GLaz &#xC2A4;&#xD2B8;&#xB808;&#xC2A4; &#xC0C1;&#xD669;&#xC5D0;&#xC11C; &#xBCF4;&#xD638; &#xC5ED;&#xD560;&#xC744; &#xD558;&#xACE0;&#xC788;&#xB2E4; &#xADF8;&#xB9AC;&#xACE0; &#xADF8;&#xAC83;&#xC758; &#xD734;&#xBB34; &#xC218;&#xBA85;&#xC774; &#xC904;&#xC5B4;&#xB4E4;&#xACE0; neurodegeneration&#xC744; &#xAC00;&#xC18D;.

&#xAD50; lazarillo, apolipoprotein D &#xCCB4;&#xC758; &#xC190;&#xC2E4; &#xCD08;&#xD30C;&#xB9AC;&#xC5D0;&#xC11C; &#xC218;&#xBA85; &#xBC0F; &#xC2A4;&#xD2B8;&#xB808;&#xC2A4; &#xC800;&#xD56D;&#xC744; &#xAC10;&#xC18C; &#xC2DC;&#xD0A8;&#xB2E4;.

El vertebrado apolipoprote&#xED;na D (APOD) es un lipocalina secreta de los subconjuntos de neuronas y c&#xE9;lulas gliales durante el desarrollo neuronal y el envejecimiento. Existe una fuerte correlaci&#xF3;n entre la sobreexpresi&#xF3;n de Apod y numerosas patolog&#xED;as del sistema nervioso, as&#xED; como la obesidad, la diabetes, y muchas formas de c&#xE1;ncer. Sin embargo, la relaci&#xF3;n exacta entre la funci&#xF3;n de Apod y la fisiopatolog&#xED;a de estas enfermedades sigue siendo desconocido. Hemos generado la p&#xE9;rdida de funci&#xF3;n de mutantes de Drosophila para el Lazarillo glial (Glaz) de genes, un hom&#xF3;logo de Apod en la mosca de la fruta, expresada principalmente en los subgrupos de adultos c&#xE9;lulas gliales. La ausencia de Glaz reduce la resistencia del organismo al estr&#xE9;s oxidativo y el hambre y acorta la vida &#xFA;til masculina. Las moscas mutantes presentan una masa corporal menor como consecuencia de una menor cantidad de l&#xED;pidos neutros almacena en la grasa corporal. Apoptosis neuronal aumenta la muerte celular en moscas de la edad o tras el tratamiento el paraquat, que tambi&#xE9;n altera la funci&#xF3;n neuronal seg&#xFA;n la evaluaci&#xF3;n de las pruebas de comportamiento. La mayor sensibilidad al estr&#xE9;s oxidativo y la inanici&#xF3;n y el almacenamiento de grasa reducida volver a controlar los niveles de cuando una prote&#xED;na de fusi&#xF3;n GFP-Glaz se expresa bajo el control del promotor Glaz natural. Por &#xFA;ltimo, los mutantes Glaz tienen una mayor concentraci&#xF3;n de productos de peroxidaci&#xF3;n lip&#xED;dica, que apunta a una protecci&#xF3;n de la peroxidaci&#xF3;n de l&#xED;pidos o de recolecci&#xF3;n de residuos, como el mecanismo de acci&#xF3;n para este lipocalina. De acuerdo con Walker et al. (En este n&#xFA;mero de la revista Current Biology), que analizan los efectos de la sobreexpresi&#xF3;n de Glaz, llegamos a la conclusi&#xF3;n de que Glaz tiene un papel protector en situaciones de estr&#xE9;s y que su ausencia reduce la vida &#xFA;til y acelera la neurodegeneraci&#xF3;n.

La p&#xE9;rdida de glial Lazarillo, un hom&#xF3;logo de la apolipoprote&#xED;na D, reduce la resistencia de Longevidad y el estr&#xE9;s en Drosophila

One of the most widely studied phenomena in the establishment of neuronal identity is the determination of neurosecretory phenotype, in which cell-type-specific combinatorial codes direct distinct neurotransmitter or neuropeptide selection. However, neuronal types from divergent lineages may adopt the same neurosecretory phenotype, and it is unclear whether different classes of neurons use different or similar components to regulate shared features of neuronal identity. We have addressed this question by analyzing how differentiation of the Drosophila larval leucokinergic system, which is comprised of only four types of neurons, is regulated by factors known to affect expression of the FMRFamide neuropeptide. We show that all leucokinergic cells express the transcription factor Squeeze (Sqz). However, based on the effect on LK expression of loss- and gain-of-function mutations, we can describe three types of Lk regulation. In the brain LHLK cells, both Sqz and Apterous (Ap) are required for LK expression, but surprisingly, high levels of either Sqz or Ap alone are sufficient to restore LK expression in these neurons. In the suboesophageal SELK cells, Sqz, but not Ap, is required for LK expression. In the abdominal ABLK neurons, inhibition of retrograde axonal transport reduces LK expression, and although sqz is dispensable for LK expression in these cells, it can induce ectopic leucokinergic ABLK-like cells when over-expressed. Thus, Sqz appears to be a regulatory factor for neuropeptidergic identity common to all leucokinergic cells, whose function in different cell types is regulated by cell-specific factors.

Squeeze involvement in the specification of Drosophila leucokinergic neurons: Different regulatory mechanisms endow the same neuropeptide selection.

The Drosophila melanogaster gene diskette (also known as dik or dAda3) encodes a protein 29% identical to human ADA3, a subunit of GCN5-containing histone acetyltransferase (HAT) complexes. The fly dADA3 is a major contributor to oogenesis, and it is also required for somatic cell viability. dADA3 localizes to chromosomes, and it is significantly reduced in dGcn5 and dAda2a, but not in dAda2b, mutant backgrounds. In dAda3 mutants, acetylation at histone H3 K9 and K14, but not K18, and at histone H4 K12, but not K5, K8, and K16, is significantly reduced. Also, phosphorylation at H3 S10 is reduced in dAda3 and dGcn5 mutants. Variegation for white (w(m4)) and scute (Hw(v)) genes, caused by rearrangements of X chromosome heterochromatin, is modified in a dAda3(+) gene-dosage-dependent manner. The effect is not observed with rearrangements involving Y heterochromatin (bw(D)), euchromatin (Scutoid), or transvection effects on chromosomal pairing (white and zeste interaction). Activity of scute gene enhancers, targets for Iroquoi transcription factors, is abolished in dAda3 mutants. Also, Iroquoi-associated phenotypes are sensitive to dAda3(+) gene dosage. We conclude that dADA3 plays a role in HAT complexes which acetylate H3 and H4 at specific residues. In turn, this acetylation results in chromatin structure effects of certain rearrangements and transcription of specific genes.

Transcriptional adaptor ADA3 of Drosophila melanogaster is required for histone modification, position effect variegation, and transcription.

&#x666E;&#x6D1B;&#x65AF;&#x662F;&#x6240;&#x9700;&#x671F;&#x95F4;&#x8F74;&#x7A81;&#x6307;&#x5BFC; &#xFF1B; &#x5212;&#x5206;&#x7EB5;&#x5411;&#x80F6;&#x8D28;&#x7EC6;&#x80DE; (LG)&#x6700;&#x521D;&#xFF0C;&#x4F7F;&#x80F6;&#x8D28;&#x53F8;&#x56DE;&#x5E94;&#x795E;&#x7ECF;&#x5143;&#x7684;&#x8054;&#x7CFB;&#x4EBA;&#x4E2D;&#xFF0C;&#x5E76;&#x968F;&#x540E;&#x4FDD;&#x6301;&#x80F6;&#x8D28;&#x7EC6;&#x80DE;&#x524D;&#x4F53;&#x5728;&#x9759;&#x6B62;&#x72B6;&#x6001;&#x4E0E;&#x6709;&#x4E1D;&#x5206;&#x88C2;&#x7684;&#x6F5C;&#x529B;&#x3002;&#x53EA;&#x662F;&#x666E;&#x7F57;&#x65AF;&#x4F69;&#x7F57;&#x9633;&#x6027; LG &#x54CD;&#x5E94;&#x795E;&#x7ECF;&#x5143;&#x6D88;&#x878D;&#x7531; over-proliferating&#xFF0C;&#x6A21;&#x4EFF;&#x4E00;&#x79CD;&#x80F6;&#x8D28;&#x7EC6;&#x80DE;&#x4FEE;&#x590D;&#x53CD;&#x5E94;&#x3002;&#x666E;&#x6D1B;&#x65AF;&#x662F;&#x901A;&#x8FC7;&#x7EB5;&#x5411; glioblast &#x6CBF;&#x88AD;&#x7684;&#x5B50;&#x4EE3;&#x7EC6;&#x80DE;&#x5F97;&#x5230;&#x516C;&#x5E73;&#x5206;&#x914D;&#x3002;&#x53EA;&#x662F;&#x4E4B;&#x524D;&#x8F74;&#x7A81;&#x63A5;&#x89E6;&#x666E;&#x6D1B;&#x65AF;&#x5F7C;&#x7F57;&#x7684;&#x6D53;&#x5EA6;&#x8F83;&#x9AD8;&#x4E2D;&#x7684;&#x4E24;&#x4E2A;&#x56DB;&#x4E2A;&#x5B50;&#x4EE3;&#x7EC6;&#x80DE;&#xFF0C;&#x548C;&#x540E;&#x8F74;&#x7A81;&#x6307;&#x5BFC;&#x666E;&#x6D1B;&#x65AF;&#x76EE;&#x524D;&#x4EC5;&#x5728;&#x516D;&#x51FA;&#x7684;&#x5341;&#x4E2A;&#x540E;&#x4EE3; LG.&#x5728;&#x8FD9;&#x91CC;&#x6211;&#x4EEC;&#x95EE;&#x666E;&#x6D1B;&#x65AF;&#x5982;&#x4F55;&#x5206;&#x914D;&#x4E0D;&#x5E73;&#x7B49;&#x5728;&#x8FD9;&#x4E9B;&#x4E24;&#x4E2A;&#x4E0D;&#x540C;&#x7684;&#x9636;&#x6BB5;&#x3002;&#x6211;&#x4EEC;&#x7684;&#x524D;&#x663E;&#x793A;&#x795E;&#x7ECF;&#x5143;&#x7684;&#x8054;&#x7CFB;&#x4EBA;&#x4E2D;&#xFF0C;&#x7EB5;&#x5411; glioblasts &#x63A5;&#x53D7; invaginating &#x7684;&#x5206;&#x88C2;&#xFF0C;&#x5782;&#x76F4;&#x4E8E;&#x5916;&#x80DA;&#x5C42;&#x5C42;&#x3002;&#x7C73;&#x5170;&#x8FBE;&#x88AB;&#x9700;&#x8981;&#x9694;&#x79BB;&#x666E;&#x6D1B;&#x65AF;&#x5F7C;&#x7F57;&#x975E;&#x5BF9;&#x79F0;&#x76F4;&#x5230;&#x80F6;&#x8D28;&#x7EC6;&#x80DE;&#x540E;&#x4EE3;&#x56DB;&#x4E2A;&#x9636;&#x6BB5;&#x3002;&#x795E;&#x7ECF;&#x5143;&#x63A5;&#x89E6;&#x540E;&#x666E;&#x6D1B;&#x65AF;&#x76EE;&#x524D;&#x5728;&#x53EA;&#x6709;&#x6FC0;&#x6D3B; Notch &#x4FE1;&#x53F7;&#x5728;&#x952F;&#x9F7F;&#x72B6;&#x9AD3;&#x540E;&#x8FDE;&#x5408;&#x8F74;&#x7A81;&#xFF0C;&#x6240;&#x63D0;&#x4F9B;&#x7684;&#x54CD;&#x5E94;&#x7684; LG &#x548C; Numb &#x4EC5;&#x9650;&#x4E8E;&#x4E0D;&#x5305;&#x542B;&#x666E;&#x6D1B;&#x65AF;&#x5F7C;&#x7F57;&#x7684;&#x80F6;&#x8D28;&#x7EC6;&#x80DE;&#x3002;&#x666E;&#x6D1B;&#x65AF;&#x5F7C;&#x7F57;&#x90E8;&#x7F72;&#x6B64;&#x53CC;&#x91CD;&#x76D1;&#x7BA1;&#xFF0C;&#x7531;&#x4E8E;&#x80F6;&#x8D28;&#xFF0C;&#x52A0;&#x4E0A;&#x7684;&#x5F62;&#x6210;&#x548C;&#x7EF4;&#x62A4;&#x7684;&#x5F25;&#x6F2B;&#x6027;&#x8F74;&#x7D22;&#x8F68;&#x8FF9;&#x3002;

&#x4E24;&#x79CD;&#x4E0D;&#x540C;&#x673A;&#x5236;&#x5728;&#x7EB5;&#x5411;&#x80F6;&#x8D28;&#x7EC6;&#x80DE;&#x4E0E;&#x795E;&#x7ECF;&#x8F74;&#x7A81;&#x7684;&#x4EA4;&#x4E92;&#x7684;&#x65F6;&#x95F4;&#x5B89;&#x6392;&#x7684;&#x57FA;&#x672C;&#x9694;&#x79BB;&#x666E;&#x6D1B;&#x65AF;&#x5F7C;&#x7F57;&#x3002;

Prospero is required in dividing longitudinal glia (LG) during axon guidance; initially to enable glial division in response to neuronal contact, and subsequently to maintain glial precursors in a quiescent state with mitotic potential. Only Prospero-positive LG respond to neuronal ablation by over-proliferating, mimicking a glial-repair response. Prospero is distributed unequally through the progeny cells of the longitudinal glioblast lineage. Just before axon contact the concentration of Prospero is higher in two of the four progeny cells, and after axon guidance Prospero is present only in six out of ten progeny LG. Here we ask how Prospero is distributed unequally in these two distinct phases. We show that before neuronal contact, longitudinal glioblasts undergo invaginating divisions, perpendicular to the ectodermal layer. Miranda is required to segregate Prospero asymmetrically up to the four glial-progeny stage. After neuronal contact, Prospero is present in only the LG that activate Notch signalling in response to Serrate provided by commissural axons, and Numb is restricted to the glia that do not contain Prospero. As a result of this dual regulation of Prospero deployment, glia are coupled to the formation and maintenance of axonal trajectories.

Two distinct mechanisms segregate Prospero in the longitudinal glia underlying the timing of interactions with axons.

Prospero est requis en divisant glia longitudinale (LG) au cours de guidage axonal ; initialement pour activer gliale division en r&eacute;ponse &agrave; un contact neuronal et par la suite maintenir gliales pr&eacute;curseurs dans un &eacute;tat de repos avec potentiel mitotique. Seulement Prospero positifs LG r&eacute;pondre &agrave; ablation neuronale en over-proliferating, mimant une r&eacute;action gliale-r&eacute;paration. Prospero est in&eacute;galement r&eacute;partie &agrave; travers les cellules de la descendance de la lign&eacute;e glioblast longitudinale. Juste avant l&#39;arriv&eacute;e de l&#39;axone, la concentration de Prospero est plus &eacute;lev&eacute;e dans deux des quatre cellules prog&eacute;niture, et apr&egrave;s que le guidage axonal Prospero est pr&eacute;sent uniquement chez les descendants de six sur dix LG. Ici, nous demandons comment Prospero est r&eacute;partie in&eacute;galement dans ces deux phases distinctes. Nous montrons qu&#39;avant contact neuronale, glioblasts longitudinales subissent une augmentation de divisions, perpendiculaires &agrave; la couche ectodermique. Miranda est n&eacute;cessaire pour s&eacute;parer les Prospero asym&eacute;triquement jusqu&#39;au stade de glial-prog&eacute;niture quatre. Apr&egrave;s contact neuronale, Prospero est pr&eacute;sente dans seulement le LG qui activent l&#39;encoche signalling in response to Serrate fournie par les axones commissures et Numb est restreinte &agrave; la glie qui ne contiennent pas de Prospero. &Agrave; la suite de ce r&egrave;glement double de d&eacute;ploiement de Prospero, cellules gliales sont coupl&eacute;s &agrave; la formation et le maintien des trajectoires axonales.

Deux m&eacute;canismes distincts s&eacute;parer Prospero dans la glie longitudinale qui sous-tendent le timing des interactions avec les axones.

Prospero &egrave; necessaria nella divisione longitudinale glia (LG) durante la guida dell&#39;assone; inizialmente per consentire la divisione gliali in risposta al contatto neuronale e successivamente per mantenere i precursori gliali in uno stato quiescente con potenziale mitotica. Prospero-positivo LG rispondere solo per ablazione neuronale da over-proliferating, mimando una risposta gliale-riparazione. Prospero &egrave; inegualmente distribuita attraverso le cellule della progenie della stirpe glioblast longitudinale. Appena prima del contatto assone la concentrazione di Prospero &egrave; superiore in due delle quattro cellule progenie, e dopo guida assone Prospero &egrave; presente solo in sei su dieci progenie LG. Qui chiediamo come Prospero &egrave; distribuito diversamente in queste due fasi distinte. Mostriamo che prima del contatto neuronale, glioblasts longitudinale subiscono divisioni invaginating, perpendicolare allo strato ectodermica. Miranda &egrave; necessaria per segregare asimmetricamente Prospero fino alla fase quattro gliali progenie. Dopo contatto neuronale, Prospero &egrave; presente solo la LG che attivare la segnalazione in risposta a dentellato fornito da assoni commissural Notch e Numb &egrave; limitato per l&#39;itaGlia che non contengono Prospero. A seguito di questo regolamento dual della distribuzione di Prospero, glia sono accoppiati alla formazione e alla manutenzione delle traiettorie assonale.

Due meccanismi distinti segregano Prospero nell&#39;Itaglia longitudinale sottostante la tempistica delle interazioni con assoni.

&#x30D7;&#x30ED;&#x30B9;&#x30DA;&#x30ED;&#x30A6;&#x306F;&#x8EF8;&#x7D22;&#x30AC;&#x30A4;&#x30C0;&#x30F3;&#x30B9;&#x4E2D;&#x7E26;&#x30B0;&#x30EA;&#x30A2; &#xFF08;LG&#xFF09; &#x306E;&#x5272;&#x308A;&#x306B;&#x306F;&#x5FC5;&#x8981;&#x3067;&#x3059;&#x3002;&#x5F53;&#x521D;&#x30B0;&#x30EA;&#x30A2;&#x90E8;&#x9580;&#x795E;&#x7D4C;&#x304A;&#x554F;&#x3044;&#x5408;&#x308F;&#x305B;&#x3078;&#x306E;&#x5FDC;&#x7B54;&#x3092;&#x6709;&#x52B9;&#x306B;&#x3057;&#x3066;&#x3001;&#x305D;&#x306E;&#x5F8C;&#x30B0;&#x30EA;&#x30A2;&#x524D;&#x99C6;&#x7D30;&#x80DE;&#x306F;&#x7D30;&#x80DE;&#x5206;&#x88C2;&#x306E;&#x53EF;&#x80FD;&#x6027;&#x3068;&#x9759;&#x6B62;&#x72B6;&#x614B;&#x3067;&#x7DAD;&#x6301;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x3002;&#x30D7;&#x30ED;&#x30B9;&#x30DA;&#x30ED;&#x30A6;&#x6B63; LG &#x5FDC;&#x7B54;&#x306E;&#x307F;&#x795E;&#x7D4C;&#x30A2;&#x30D6;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3; over-proliferating &#x306B;&#x3088;&#x3063;&#x3066;&#x30B0;&#x30EA;&#x30A2;&#x4FEE;&#x5FA9;&#x5FDC;&#x7B54;&#x3092;&#x6A21;&#x5023;&#x3057;&#x307E;&#x3059;&#x3002;&#x30D7;&#x30ED;&#x30B9;&#x30DA;&#x30ED;&#x30A6;&#x7E26; glioblast &#x7CFB;&#x7D71;&#x306E;&#x5B50;&#x5B6B;&#x7D30;&#x80DE;&#x3092;&#x5747;&#x7B49;&#x306B;&#x5206;&#x6563;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x8EF8;&#x7D22;&#x9023;&#x7D61;&#x3059;&#x308B;&#x524D;&#x306B;&#x3061;&#x3087;&#x3046;&#x3069;&#x30D7;&#x30ED;&#x30B9;&#x30DA;&#x30ED;&#x6FC3;&#x5EA6; 2 &#x3064; 4 &#x3064;&#x306E;&#x5B50;&#x5B6B;&#x7D30;&#x80DE;&#x306E;&#x8EF8;&#x7D22;&#x30AC;&#x30A4;&#x30C0;&#x30F3;&#x30B9; &#x30D7;&#x30ED;&#x30B9;&#x30DA;&#x30ED;&#x30A6;&#x304C; 6 &#x5272;&#x5B50;&#x5B6B; lg &#x96FB;&#x5B50;&#x3067;&#x306E;&#x307F;&#x5B58;&#x5728;&#x306E;&#x5F8C;&#x3067;&#x9AD8;&#x304F;&#x306A;&#x3063;&#x3066;&#x3044;&#x307E;&#x3059;&#x3053;&#x3053;&#x3067;&#x6211;&#x3005; &#x306F;&#x8CEA;&#x554F;&#x3092;&#x3069;&#x306E;&#x3088;&#x3046;&#x306B;&#x30D7;&#x30ED;&#x30B9;&#x30DA;&#x30ED;&#x5747;&#x7B49;&#x306B;&#x3053;&#x308C;&#x3089; 2 &#x3064;&#x306E;&#x30D5;&#x30A7;&#x30FC;&#x30BA;&#x3067;&#x914D;&#x7F6E;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x79C1;&#x9054;&#x306F;&#x795E;&#x7D4C;&#x9023;&#x7D61;&#x3059;&#x308B;&#x524D;&#x306B;&#x3001;invaginating &#x90E8;&#x9580;&#x3001;&#x5916;&#x80DA;&#x8449;&#x5C64;&#x306B;&#x5782;&#x76F4;&#x7E26; glioblasts &#x3092;&#x53D7;&#x3051;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;&#x30DF;&#x30E9;&#x30F3;&#x30C0; &#x30FB; &#x30D7;&#x30ED;&#x30B9;&#x30DA;&#x30ED;&#x975E;&#x5BFE;&#x79F0; 4 &#x30B0;&#x30EA;&#x30A2;&#x5B50;&#x5B6B;&#x6BB5;&#x968E;&#x307E;&#x3067;&#x3092;&#x5206;&#x96E2;&#x3059;&#x308B;&#x5FC5;&#x8981;&#x304C;&#x3042;&#x308A;&#x307E;&#x3059;&#x3002;&#x795E;&#x7D4C;&#x63A5;&#x89E6;&#x306E;&#x5F8C;&#x3001;&#x30D7;&#x30ED;&#x30B9;&#x30DA;&#x30ED; &#x30CE;&#x30C3;&#x30C1;&#x4EA4;&#x9023;&#x8EF8;&#x7D22;&#x306B;&#x3088;&#x3063;&#x3066;&#x63D0;&#x4F9B;&#x3055;&#x308C;&#x308B; Serrate &#x3078;&#x306E;&#x5FDC;&#x7B54;&#x306E;&#x4FE1;&#x53F7;&#x3092;&#x30A2;&#x30AF;&#x30C6;&#x30A3;&#x30D6;&#x306B;&#x3059;&#x308B; lg &#x96FB;&#x5B50;&#x3060;&#x3051;&#x306E;&#x5B58;&#x5728;&#x3067;&#x3042;&#x308A;&#x30D5;&#x30C3;&#x30AF;&#x30E1;&#x30BD;&#x30C3;&#x30C9; &#x30D7;&#x30ED;&#x30B9;&#x30DA;&#x30ED;&#x3092;&#x542B;&#x307E;&#x306A;&#x3044;&#x30B0;&#x30EA;&#x30A2;&#x306B;&#x5236;&#x9650;&#x3002;&#x30D7;&#x30ED;&#x30B9;&#x30DA;&#x30ED;&#x30A6;&#x5C55;&#x958B;&#x306F;&#x3053;&#x306E;&#x4E8C;&#x91CD;&#x306E;&#x5236;&#x5FA1;&#x7D50;&#x679C;&#x3068;&#x3057;&#x3066;&#x30B0;&#x30EA;&#x30A2;&#x7D30;&#x80DE;&#x306F;&#x7D50;&#x5408;&#x5F62;&#x6210;&#x3068;&#x8EF8;&#x7D22;&#x306E;&#x8ECC;&#x9053;&#x306E;&#x30E1;&#x30F3;&#x30C6;&#x30CA;&#x30F3;&#x30B9;&#x306B;&#x3002;

2 &#x3064;&#x306E;&#x7570;&#x306A;&#x308B;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0; &#x30D7;&#x30ED;&#x30B9;&#x30DA;&#x30ED;&#x30A6;&#x306F;&#x8EF8;&#x7D22;&#x3068;&#x306E;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x306E;&#x30BF;&#x30A4;&#x30DF;&#x30F3;&#x30B0;&#x306E;&#x57FA;&#x790E;&#x3068;&#x306A;&#x308B;&#x7E26;&#x306E;&#x30B0;&#x30EA;&#x30A2;&#x7D30;&#x80DE;&#x3067;&#x5206;&#x96E2;&#x3057;&#x307E;&#x3059;&#x3002;

&#xC9C0;&#xC8FC; &#xB2D8; &#xCD95; &#xC0AD; &#xC9C0;&#xB3C4;; &#xACBD;&#xB3C4; &#xBA85;&#xACFC; (lg &#xC804;&#xC790;) &#xBD84;&#xD560; &#xD544;&#xC694; &#xCC98;&#xC74C; &#xC2E0;&#xACBD; &#xC811;&#xCD09;&#xC5D0; &#xB300; &#xD55C; &#xC751;&#xB2F5;&#xC5D0;&#xC11C; &#xD3D0;&#xD574; &#xBD80;&#xBB38; &#xD65C;&#xC131;&#xD654; &#xBC0F; &#xC774;&#xD6C4; &#xC720;&#xC9C0; &#xAD00;&#xB9AC;&#xAC00; mitotic &#xC7A0;&#xC7AC;&#xB825; &#xBB34;&#xBD80;&#xD558; &#xC0C1;&#xD0DC;&#xC5D0;&#xC11C; &#xD3D0;&#xD574; &#xC120;&#xAD6C;&#xC790;. &#xB9CC; &#xC9C0;&#xC8FC; &#xB2D8; &#xAE0D;&#xC815;&#xC801;&#xC778; LG &#xC2E0;&#xACBD; &#xC808;&#xC81C; &#xD558; &#xC5EC; &#xC751;&#xB2F5; over-proliferating, &#xD3D0;&#xD574; &#xBCF5;&#xAD6C; &#xC751;&#xB2F5;&#xC744; &#xD749;&#xB0B4; &#xB0B8;. &#xC9C0;&#xC8FC; &#xB2D8;&#xC740; &#xACBD;&#xB3C4; glioblast &#xD608;&#xD1B5;&#xC758; &#xC790;&#xC190; &#xC138;&#xD3EC;&#xB97C; &#xD1B5;&#xD574; &#xADE0;&#xB4F1; &#xD558; &#xAC8C; &#xBD84;&#xC0B0; &#xB429;&#xB2C8;&#xB2E4;. &#xCD95; &#xC0AD; &#xC811;&#xCD09; &#xC9C1;&#xC804; &#xC9C0;&#xC8FC; &#xB2D8;&#xC758; &#xB18D;&#xB3C4; 4 &#xAC1C;&#xC758; &#xC790;&#xC190; &#xC138;&#xD3EC;&#xC640; &#xCD95; &#xC0AD; &#xC9C0;&#xB3C4; &#xC9C0;&#xC8FC; &#xB2D8; lg. 6 10 &#xC790;&#xC190;&#xC5D0;&#xB9CC; &#xD6C4; &#xB450;&#xC5D0;&#xC11C; &#xB192;&#xC740; &#xC5EC;&#xAE30; &#xC5B4;&#xB5BB;&#xAC8C; &#xC9C0;&#xC8FC; &#xB2D8;&#xC740; &#xADE0;&#xB4F1; &#xD558;&#xC9C0; &#xC774;&#xB7EC;&#xD55C; &#xB450; &#xAC00;&#xC9C0; &#xB2E8;&#xACC4;&#xB85C; &#xBD80;&#xD0C1; &#xB4DC;&#xB9BD;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xAC00; &#xBCF4;&#xC5EC; &#xC2E0;&#xACBD; &#xC811;&#xCD09; &#xC804;&#xC5D0; &#xACBD;&#xB3C4; glioblasts invaginating &#xBD80;&#xBB38;, ectodermal &#xCE35;&#xC5D0; &#xC218;&#xC9C1;&#xC73C;&#xB85C; &#xBC1B; &#xB2E4;. &#xBBF8; &#xB780;&#xB2E4;&#xB294; 4 &#xAC1C;&#xC758; &#xD3D0;&#xD574; &#xC790;&#xC190; &#xBB34;&#xB300; &#xC9C0;&#xC8FC; &#xB2D8;&#xC744; &#xBE44;&#xB300;&#xCE6D;&#xC801;&#xC73C;&#xB85C; &#xBD84;&#xB9AC; &#xD574;&#xC57C; &#xD569;&#xB2C8;&#xB2E4;. &#xC2E0;&#xACBD; &#xC811;&#xCD09; &#xD6C4; &#xC9C0;&#xC8FC; &#xB2D8;&#xB9CC; &#xB178;&#xCE58; Serrate commissural &#xCD95; &#xC0AD;&#xC5D0; &#xC758;&#xD574; &#xC81C;&#xACF5;&#xC5D0; &#xB300; &#xD55C; &#xC751;&#xB2F5; &#xC2E0;&#xD638;&#xB97C; &#xD65C;&#xC131;&#xD654; &#xD558;&#xB294; lg &#xC804;&#xC790;&#xC5D0; &#xC874;&#xC7AC; &#xD558;&#xB294; &#xBC0F; &#xB9C8;&#xBE44; &#xBA85;&#xACFC; &#xC9C0;&#xC8FC; &#xB2D8;&#xC744; &#xD3EC;&#xD568; &#xD558;&#xC9C0; &#xC54A;&#xB294; &#xC81C;&#xD55C; &#xB429;&#xB2C8;&#xB2E4;. &#xC9C0;&#xC8FC; &#xB2D8; &#xBC30;&#xD3EC; &#xC774;&#xC911; &#xADDC;&#xC81C;&#xC758; &#xACB0;&#xACFC;&#xB85C; &#xBA85;&#xACFC; &#xD615;&#xC131; &#xBC0F; axonal &#xADA4;&#xB3C4;&#xC758; &#xC720;&#xC9C0; &#xBCF4;&#xC218;&#xC5D0; &#xACB0;&#xD569; &#xB41C;&#xB2E4;.

&#xB450; &#xAC00;&#xC9C0; &#xBA54;&#xCEE4;&#xB2C8;&#xC998; &#xACBD;&#xB3C4; &#xBA85;&#xACFC; &#xAE30;&#xBCF8; &#xC0C1;&#xD638;&#xC791;&#xC6A9; &#xCD95; &#xC0AD;&#xC758; &#xD0C0;&#xC774;&#xBC0D;&#xC5D0;&#xC5D0;&#xC11C; &#xC9C0;&#xC8FC; &#xB2D8;&#xC744; &#xBD84;&#xB9AC; &#xD569;&#xB2C8;&#xB2E4;.

Prospero es necesaria en la divisi&oacute;n longitudinal glia (LG) durante la orientaci&oacute;n del ax&oacute;n; inicialmente para permitir la divisi&oacute;n glial en respuesta al contacto neuronal y posteriormente mantener precursores gliales en un estado quieto con potencial mit&oacute;tico. S&oacute;lo positivo Prospero LG responden a ablaci&oacute;n neuronal por over-proliferating, imitando una respuesta de reparaci&oacute;n glial. Prospero se distribuye desigualmente a trav&eacute;s de las c&eacute;lulas de la progenie del linaje glioblast longitudinal. Justo antes del contacto del ax&oacute;n, la concentraci&oacute;n de Prospero es mayor en dos de las cuatro c&eacute;lulas de la progenie, y despu&eacute;s direcci&oacute;n axon Prospero est&aacute; presente s&oacute;lo en seis de cada diez progenie LG. Aqu&iacute; preguntamos c&oacute;mo Prospero es distribuido de forma desigual en estas dos fases distintas. Demostramos que antes del contacto neuronal, glioblasts longitudinales sufren divisiones invaginante, perpendiculares a la capa ectod&eacute;rmica. Miranda es necesaria para segregar Prospero asim&eacute;trico hasta la fase cuatro de progenie glial. Despu&eacute;s del contacto neuronal, Prospero est&aacute; presente en s&oacute;lo el LG que activan la muesca se&ntilde;alizaci&oacute;n en respuesta a serrado de axones commissural y Numb est&aacute; restringida a la glia que no contengan Prospero. Como resultado de este doble regulaci&oacute;n de despliegue de Prospero, glia est&aacute;n acoplados a la formaci&oacute;n y mantenimiento de trayectorias axonal.

Dos mecanismos distintos segregan Prospero en la glia longitudinal que subyace a la sincronizaci&oacute;n de las interacciones con los axones.

It is becoming increasingly clear that the activation of specific terminal differentiation genes during neural development is critically dependent upon the establishment of unique combinatorial transcription factor codes within distinct neural cell subtypes. However, it is still unclear to which extent these codes are shared by lineage-unrelated neurons expressing the same terminal differentiation genes. Additionally, it is not known if the activation of a specific terminal differentiation gene is restricted to cells born at a particular developmental time point. Here, we utilize the terminal differentiation gene FMRFa which is expressed by the Ap4 and SE2 neurons in the Drosophila ventral nerve cord, to explore these issues in depth. We find that the Ap4 and SE2 neurons are generated by different neural progenitors and use different combinatorial codes to activate FMRFa expression. Additionally, we find that the Ap4 and SE2 neurons are generated in different temporal gene expression windows. Extending the investigation to include a second Drosophila terminal differentiation gene, Leucokinin, we find similar results, suggesting that neurons generated by different progenitors might commonly use different transcription factor codes to activate the same terminal differentiation gene. Furthermore, these results imply that the activation of a particular terminal differentiation gene in temporally unrestricted.

Lineage-unrelated neurons generated in different temporal windows and expressing different combinatorial codes can converge in the activation of the same terminal differentiation gene.

The central nervous system contains a wide variety of neuronal subclasses generated by neural progenitors. The achievement of a unique neural fate is the consequence of a sequence of early and increasingly restricted regulatory events, which culminates in the expression of a specific genetic combinatorial code that confers individual characteristics to the differentiated cell. How the earlier regulatory events influence post-mitotic cell fate decisions is beginning to be understood in the Drosophila NB 5-6 lineage. However, it remains unknown to what extent these events operate in other lineages. To better understand this issue, we have used a very highly specific marker that identifies a small subset of abdominal cells expressing the Drosophila neuropeptide Capa: the ABCA neurons. Our data support the birth of the ABCA neurons from NB 5-3 in a cas temporal window in the abdominal segments A2-A4. Moreover, we show that the ABCA neuron has an ABCA-sibling cell which dies by apoptosis. Surprisingly, both cells are also generated in the abdominal segments A5-A7, although they undergo apoptosis before expressing Capa. In addition, we have performed a targeted genetic screen to identify players involved in ABCA specification. We have found that the ABCA fate requires zfh2, grain, Grunge and hedgehog genes. Finally, we show that the NB 5-3 generates other subtype of Capa-expressing cells (SECAs) in the third suboesophageal segment, which are born during a pdm/cas temporal window, and have different genetic requirements for their specification.

A targeted genetic screen identifies crucial players in the specification of the Drosophila abdominal Capaergic neurons.

Drosophila embryonic neuroblasts generate different cell types at different time points. This is controlled by a temporal cascade of Hb→Kr→Pdm→Cas→Grh, which acts to dictate distinct competence windows sequentially. In addition, Seven up (Svp), a member of the nuclear hormone receptor family, acts early in the temporal cascade, to ensure the transition from Hb to Kr, and has been referred to as a 'switching factor'. However, Svp is also expressed in a second wave within the developing CNS, but here, the possible role of Svp has not been previously addressed. In a genetic screen for mutants affecting the last-born cell in the embryonic NB5-6T lineage, the Ap4/FMRFamide neuron, we have isolated a novel allele of svp. Expression analysis shows that Svp is expressed in two distinct pulses in NB5-6T, and mutant analysis reveals that svp plays two distinct roles. In the first pulse, svp acts to ensure proper downregulation of Hb. In the second pulse, which occurs in a Cas/Grh double-positive window, svp acts to ensure proper sub-division of this window. These studies show that a temporal factor may play dual roles, acting at two different stages during the development of one neural lineage.

Seven up acts as a temporal factor during two different stages of neuroblast 5-6 development.

HCN channels are becoming pharmacological targets mainly in cardiac diseases. But apart from their well-known role in heart pacemaking, these channels are widely expressed in the nervous system where they contribute to the neuron firing pattern. Consequently, abolishing Ih current might have detrimental consequences in a big repertoire of behavioral traits. Several studies in mammals have identified the Ih current as an important determinant of the firing activity of dopaminergic neurons, and recent evidences link alterations in this current to various dopamine-related disorders. We used the model organism Drosophila melanogaster to investigate how lack of Ih current affects dopamine levels and the behavioral consequences in the sleep:activity pattern. Unlike mammals, in Drosophila there is only one gene encoding HCN channels. We generated a deficiency of the DmIh core gene region and measured, by HPLC, levels of dopamine. Our data demonstrate daily variations of dopamine in wild-type fly heads. Lack of Ih current dramatically alters dopamine pattern, but different mechanisms seem to operate during light and dark conditions. Behaviorally, DmIh mutant flies display alterations in the rest:activity pattern, and altered circadian rhythms. Our data strongly suggest that Ih current is necessary to prevent dopamine overproduction at dark, while light input allows cycling of dopamine in an Ih current dependent manner. Moreover, lack of Ih current results in behavioral defects that are consistent with altered dopamine levels.

Ih current is necessary to maintain normal dopamine fluctuations and sleep consolidation in Drosophila.

A number of transcription factors that are expressed within most, if not all, embryonic neuroblast (NB) lineages participate in neural subtype specification. Some have been extensively studied in several NB lineages (e.g. components of the temporal gene cascade) whereas others only within specific NB lineages. To what extent they function in other lineages remains unknown. Klumpfuss (Klu), the Drosophila ortholog of the mammalian Wilms tumor 1 (WT1) protein, is one such transcription factor. Studies in the NB4-2 lineage have suggested that Klu functions to ensure that the two ganglion mother cells (GMCs) in this embryonic NB lineage acquire different fates. Owing to limited lineage marker availability, these observations were made only for the NB4-2 lineage. Recent findings reveal that Klu is necessary for larval neuroblast growth and self-renewal. We have extended the study of Klu to the well-known embryonic NB5-6T lineage and describe a novel role for Klu in the Drosophila embryonic CNS. Our results demonstrate that Klu is expressed specifically in the postmitotic Ap4/FMRFa neuron, promoting its differentiation through the initiation of BMP signaling. Our findings indicate a pleiotropic function of Klu in Ap cluster specification in general and particularly in Ap4 neuron differentiation, indicating that Klu is a multitasking transcription factor. Finally, our studies indicate that a transitory downregulation of klu is crucial for the specification of the Ap4/FMRFa neuron. Similar to WT1, klu seems to have either self-renewal or differentiation-promoting functions, depending on the developmental context.

Klumpfuss controls FMRFamide expression by enabling BMP signaling within the NB5-6 lineage.

Sensory cilia are often encapsulated by an extracellular matrix (ECM). In Caenorhabditis elegans, Drosophila melanogaster, and vertebrates, this ECM is thought to be directly involved in ciliary mechanosensing by coupling external forces to the ciliary membrane. Drosophila mechano- and chemosensory cilia are both associated with an ECM, indicating that the ECM may have additional roles that go beyond mechanosensory cilium function. Here, we identify Artichoke (ATK), an evolutionarily conserved leucine-rich repeat ECM protein that is required for normal morphogenesis and function of ciliated sensilla in Drosophila. atk is transiently expressed in accessory cells in all ciliated sensory organs during their late embryonic development. Antibody stainings show ATK protein in the ECM that surrounds sensory cilia. Loss of ATK protein in atk null mutants leads to cilium deformation and disorientation in chordotonal organs, apparently without uncoupling the cilia from the ECM, and consequently to locomotion defects. Moreover, impaired chemotaxis in atk mutant larvae suggests that, based on ATK protein localization, the ECM is also crucial for the correct assembly of chemosensory receptors. In addition to defining a novel ECM component, our findings show the importance of ECM integrity for the proper morphogenesis of ciliated organs in different sensory modalities.

The extracellular matrix protein artichoke is required for integrity of ciliated mechanosensory and chemosensory organs in Drosophila embryos.

Studies in the Drosophila embryonic NB4-2 lineage have suggested that the transcription factor Klumpfuss (Klu) functions within embryonic neuroblast lineages to differentiate between the identities of two adjacent ganglion mother cells (GMCs). However, because of the limited lineage markers available, these observations have been made only for the NB4-2 lineage. Recent findings have placed this transcription factor in the vanguard of Drosophila neural stem cell biology by demonstrating that Klu is necessary for larval neuroblast growth and self-renewal. Here, we have studied the role of klu in an incipient model in order to address basic mechanisms of neural specification: the Va system. None of the previously reported roles of Klu satisfactorily explain our observations. Unexpectedly, in this lineage, klu is necessary for differentiating between the fates of the two neurons born from a unique GMC; klu mutants produce two B-type cells, rather than one B-type (Notch-OFF) and one A-type (Notch-ON) cell. Additionally, our results demonstrate that Klu operates in the GMC and/or in the newly born neuron, but not in the neuroblast. Unlike in larval neuroblasts in which Klu is an executor of Notch signaling, we have found that Klu does not lie downstream of the Notch pathway in this cell division context.

A new role of Klumpfuss in establishing cell fate during the GMC asymmetric cell division.

The morphology of synapses is of central interest in neuroscience because of the intimate relation with synaptic efficacy. Two decades of gene manipulation studies in different animal models have revealed a repertoire of molecules that contribute to synapse development. However, since such studies often assessed only one, or at best a few, morphological features at a given synapse, it remained unaddressed how different structural aspects relate to one another. Furthermore, such focused and sometimes only qualitative approaches likely left many of the more subtle players unnoticed. Here, we present the image analysis algorithm 'Drosophila_NMJ_Morphometrics', available as a Fiji-compatible macro, for quantitative, accurate and objective synapse morphometry of the Drosophila larval neuromuscular junction (NMJ), a well-established glutamatergic model synapse. We developed this methodology for semi-automated multiparametric analyses of NMJ terminals immunolabeled for the commonly used markers Dlg1 and Brp and showed that it also works for Hrp, Csp and Syt. We demonstrate that gender, genetic background and identity of abdominal body segment consistently and significantly contribute to variability in our data, suggesting that controlling for these parameters is important to minimize variability in quantitative analyses. Correlation and principal component analyses (PCA) were performed to investigate which morphometric parameters are inter-dependent and which ones are regulated rather independently. Based on nine acquired parameters, we identified five morphometric groups: NMJ size, geometry, muscle size, number of NMJ islands and number of active zones. Based on our finding that the parameters of the first two principal components hardly correlated with each other, we suggest that different molecular processes underlie these two morphometric groups. Our study sets the stage for systems morphometry approaches at the well-studied Drosophila NMJ.

Article	Year
Neurosecretory identity conferred by the apterous gene: lateral horn leucokinin neurons in Drosophila. The Journal of comparative neurology\| PubMed ID: 12541314	2003
Loss of glial lazarillo, a homolog of apolipoprotein D, reduces lifespan and stress resistance in Drosophila. Current biology : CB\| PubMed ID: 16581513	2006
Squeeze involvement in the specification of Drosophila leucokinergic neurons: Different regulatory mechanisms endow the same neuropeptide selection. Mechanisms of development\| PubMed ID: 17442544	2007
Transcriptional adaptor ADA3 of Drosophila melanogaster is required for histone modification, position effect variegation, and transcription. Molecular and cellular biology\| PubMed ID: 17967867	2008
Two distinct mechanisms segregate Prospero in the longitudinal glia underlying the timing of interactions with axons. Neuron glia biology\| PubMed ID: 18634579	2007
Lineage-unrelated neurons generated in different temporal windows and expressing different combinatorial codes can converge in the activation of the same terminal differentiation gene. Mechanisms of development\| PubMed ID: 20732418	2010
A targeted genetic screen identifies crucial players in the specification of the Drosophila abdominal Capaergic neurons. Mechanisms of development\| PubMed ID: 21236339	2011
Seven up acts as a temporal factor during two different stages of neuroblast 5-6 development. Development (Cambridge, England)\| PubMed ID: 22071101	2011
Ih current is necessary to maintain normal dopamine fluctuations and sleep consolidation in Drosophila. PloS one\| PubMed ID: 22574167	2012
Klumpfuss controls FMRFamide expression by enabling BMP signaling within the NB5-6 lineage. Development (Cambridge, England)\| PubMed ID: 23633512	2013
The extracellular matrix protein artichoke is required for integrity of ciliated mechanosensory and chemosensory organs in Drosophila embryos. Genetics\| PubMed ID: 24496014	2014
A new role of Klumpfuss in establishing cell fate during the GMC asymmetric cell division. Cell and tissue research\| PubMed ID: 25129108	2014
A New Fiji-Based Algorithm That Systematically Quantifies Nine Synaptic Parameters Provides Insights into Drosophila NMJ Morphometry. PLoS computational biology\| PubMed ID: 26998933	2016

Laura Torroja

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