quantification-of-drosophila-grooming-behavior

Quantification of Drosophila Grooming Behavior

Drosophila grooming behavior is a complex multi-step locomotor program that requires coordinated movement of both forelegs and hindlegs. Here we present a ...

Williams College

We have cloned and characterized the first galectin to be identified in Drosophila melanogaster. The amino acid sequence of Drosophila galectin showed striking sequence similarity to invertebrate and vertebrate galectins and contained amino acids that are crucial for binding beta-galactoside sugars. Confirming its identity as a galectin family member, the Drosophila galectin bound beta-galactoside sugars. Structurally, the Drosophila galectin was a tandem repeat galectin containing two carbohydrate recognition domains connected by a unique peptide link. This divalent structure suggests that like mammalian galectins, Drosophila galectin may mediate cell-cell communication or facilitate cross-linking of receptors to trigger signal transduction events. The Drosophila galectin was very abundant in embryonic, larval, and adult Drosophila. During embryogenesis, Drosophila galectin had a unique and specific tissue distribution. Drosophila galectin expression was concentrated in somatic and visceral musculature and in the central nervous system. Similar to other insect lectins, Drosophila galectin may function in both embryogenesis and in host defense. Drosophila galectin was expressed by hemocytes, circulating phagocytic cells, suggesting a role for Drosophila galectin in the innate immune system.

Characterization of a novel Drosophila melanogaster galectin. Expression in developing immune, neural, and muscle tissues.

The differentiation of Drosophila blood cells relies on a functional hierarchy between the GATA protein, Serpent (Srp), and multiple lineage-specific transcription factors, such as the AML1-like protein, Lozenge (Lz). Two major branches of Drosophila hematopoiesis give rise to plasmatocytes/macrophages and crystal cells. Serrate signaling through the Notch pathway is critical in the regulation of Lz expression and the specification of crystal cell precursors, thus providing a key distinction between the two lineages. The expression of Serrate marks a discrete cluster of cells in the lymph gland, a signaling center, with functional similarities to stromal signaling in mammalian hematopoiesis.

A Serrate-expressing signaling center controls Drosophila hematopoiesis.

Arousal is fundamental to many behaviors, but whether it is unitary or whether there are different types of behavior-specific arousal has not been clear. In Drosophila, dopamine promotes sleep-wake arousal. However, there is conflicting evidence regarding its influence on environmentally stimulated arousal. Here we show that loss-of-function mutations in the D1 dopamine receptor DopR enhance repetitive startle-induced arousal while decreasing sleep-wake arousal (i.e., increasing sleep). These two types of arousal are also inversely influenced by cocaine, whose effects in each case are opposite to, and abrogated by, the DopR mutation. Selective restoration of DopR function in the central complex rescues the enhanced stimulated arousal but not the increased sleep phenotype of DopR mutants. These data provide evidence for at least two different forms of arousal, which are independently regulated by dopamine in opposite directions, via distinct neural circuits.

Two different forms of arousal in Drosophila are oppositely regulated by the dopamine D1 receptor ortholog DopR via distinct neural circuits.

Dopamine is a mediator of the stimulant properties of drugs of abuse, including ethanol, in mammals and in the fruit fly Drosophila. The neural substrates for the stimulant actions of ethanol in flies are not known. We show that a subset of dopamine neurons and their targets, through the action of the D1-like dopamine receptor DopR, promote locomotor activation in response to acute ethanol exposure. A bilateral pair of dopaminergic neurons in the fly brain mediates the enhanced locomotor activity induced by ethanol exposure, and promotes locomotion when directly activated. These neurons project to the central complex ellipsoid body, a structure implicated in regulating motor behaviors. Ellipsoid body neurons are required for ethanol-induced locomotor activity and they express DopR. Elimination of DopR blunts the locomotor activating effects of ethanol, and this behavior can be restored by selective expression of DopR in the ellipsoid body. These data tie the activity of defined dopamine neurons to D1-like DopR-expressing neurons to form a neural circuit that governs acute responding to ethanol.

A pair of dopamine neurons target the D1-like dopamine receptor DopR in the central complex to promote ethanol-stimulated locomotion in Drosophila.

&#x8C03;&#x67E5;&#x7531;&#x751F;&#x7269;&#x80FA;&#x679C;&#x8747;&#x679C;&#x8747;&#x884C;&#x4E3A;&#x7684;&#x89C4;&#x7BA1;&#xFF0C;&#x6211;&#x4EEC;&#x5229;&#x7528;&#x5E7F;&#x6CDB;&#x7684;&#x56CA;&#x6CE1;&#x5355;&#x80FA;&#x8F6C;&#x8FD0;&#x4F53; (VMAT) &#x4E3A;&#x6C34;&#x75B1;&#x7684;&#x5B58;&#x50A8;&#x8981;&#x6C42;&#x548C;&#x9AD8;&#x5C14;&#x57FA;&#x6240;&#x6709;&#x5355;&#x80FA;&#x7C7B;&#x795E;&#x7ECF;&#x9012;&#x8D28;&#x7684;&#x91CA;&#x653E;&#x3002;&#x6211;&#x4EEC;&#x7528;&#x679C;&#x8747; VMAT (dVMAT) &#x7A7A;&#x7A81;&#x53D8;&#x6765;&#x5168;&#x7403; ablate &#x9AD8;&#x5C14;&#x57FA;&#x80FA;&#x91CA;&#x653E;&#x3001; &#x7136;&#x540E;&#x8FD8;&#x539F; DVMAT &#x6D3B;&#x52A8;&#x4E2D;&#x4E24;&#x4E2A;&#x4EBA;&#x6216;&#x591A;&#x4E2A;&#x4F7F;&#x7528;&#x8F6C;&#x57FA;&#x56E0;&#x7684;&#x7EC4;&#x7EC7;&#x80FA;&#x7C7B;&#x7CFB;&#x7EDF;&#x6551;&#x63F4;&#x6280;&#x672F;&#x3002;&#x6211;&#x4EEC;&#x53D1;&#x73B0;&#x662F;&#x4ED4;&#x9C7C;&#x5B58;&#x6D3B;&#x7387;&#x3001; &#x5E7C;&#x866B;&#x7684;&#x8FD0;&#x52A8;&#xFF0C;&#x4EE5;&#x53CA;&#x5973;&#x6027;&#x751F;&#x80B2;&#x80FD;&#x529B;&#x4E3B;&#x8981;&#x4F9D;&#x8D56;&#x4ECE;&#x6C34;&#x6CE1;&#x91CA;&#x653E; 5-&#x7F9F;&#x8272;&#x80FA;&#x6216;&#x591A;&#x5DF4;&#x80FA;&#x4E00;&#x70B9;&#x660E;&#x663E;&#x8F93;&#x5165; octopaminergic &#x7535;&#x8DEF;&#x3002;&#x76F8;&#x6BD4;&#x4E4B;&#x4E0B;&#xFF0C;&#x7537;&#x6027;&#x6C42;&#x5076;&#x548C;&#x80A5;&#x529B;&#x53EF;&#x4EE5;&#x7531;&#x8868;&#x793A; DVMAT octopaminergic &#x6216;&#x591A;&#x5DF4;&#x80FA;&#x80FD;&#x795E;&#x7ECF;&#x5143;&#xFF0C;&#x6697;&#x793A;&#x53EF;&#x80FD;&#x5197;&#x4F59;&#x7535;&#x8DEF;&#x4E2D;&#x6551;&#x51FA;&#x3002;&#x6210;&#x4EBA;&#x7684;&#x8FD0;&#x52A8;&#x548C;&#x60CA;&#x6115;&#x884C;&#x4E3A;&#x7684;&#x4E3B;&#x8981;&#x65B9;&#x9762;&#x7684;&#x6551;&#x63F4;&#x9700;&#x8981;&#x7AE0;&#x9C7C;&#x80FA;&#xFF0C;&#x4F46;&#x7684; 5-&#x7F9F;&#x8272;&#x80FA;&#x5F55;&#x5F97;&#x7684;&#x8865;&#x5145;&#x4F5C;&#x7528;&#x3002;&#x6709;&#x8DA3;&#x7684;&#x662F;&#xFF0C;&#x6210;&#x4EBA;&#x663C;&#x591C;&#x8282;&#x5F8B;&#x884C;&#x4E3A;&#x4E0D;&#x53EF;&#x80FD;&#x7531;&#x5355;&#x4E00;&#x7684;&#x5B50;&#x7C7B;&#x578B;&#x7EC4;&#x7EC7;&#x80FA;&#x7C7B;&#x795E;&#x7ECF;&#x5143;&#xFF0C;&#x4F46;&#x9700;&#x8981;&#x81F3;&#x5C11;&#x4E24;&#x4E2A;&#x7CFB;&#x7EDF;&#xFF0C;&#x8FD9;&#x8868;&#x660E;&#x610F;&#x5916;&#x5408;&#x4F5C;&#x4E92;&#x52A8;&#x7684;&#x53EF;&#x80FD;&#x6027;&#x5728; DVMAT &#x7684;&#x8868;&#x8FBE;&#x6551;&#x51FA;&#x3002;&#x4F7F;&#x7528;&#x6B64;&#x6A21;&#x578B;&#x7684;&#x8FDB;&#x4E00;&#x6B65;&#x5B9E;&#x9A8C;&#x5C06;&#x6709;&#x52A9;&#x4E8E;&#x786E;&#x5B9A;&#x5982;&#x4F55;&#x591A;&#x4E2A;&#x7EC4;&#x7EC7;&#x80FA;&#x7C7B;&#x7CFB;&#x7EDF;&#x53EF;&#x80FD;&#x6709;&#x52A9;&#x4E8E;&#x5176;&#x4ED6;&#x884C;&#x4E3A;&#x7684;&#x89C4;&#x7BA1;&#x3002;&#x6211;&#x4EEC;&#x7684;&#x6570;&#x636E;&#x8FD8;&#x7A81;&#x51FA;&#x4E0E;&#x5176;&#x4ED6;&#x673A;&#x5236;&#x7684;&#x6807;&#x51C6;&#x9AD8;&#x5C14;&#x57FA;&#x91CA;&#x653E;&#x53D7;&#x89C4;&#x7BA1;&#x7684;&#x884C;&#x4E3A;&#x4E4B;&#x95F4;&#x7684;&#x6F5C;&#x5728;&#x5DEE;&#x5F02;&#x3002;

&#x591A;&#x5DF4;&#x80FA;&#x3001; &#x7AE0;&#x9C7C;&#x80FA;&#x548C; 5-&#x7F9F;&#x8272;&#x80FA;&#x5728;&#x679C;&#x8747;&#x4E2D;&#x7684;&#x53EF;&#x6709;&#x53EF;&#x65E0;&#x3001; &#x5197;&#x4F59;&#x3001; &#x4E92;&#x8865;&#x548C;&#x534F;&#x540C;&#x4F5C;&#x7528;&#x3002;

To investigate the regulation of Drosophila melanogaster behavior by biogenic amines, we have exploited the broad requirement of the vesicular monoamine transporter (VMAT) for the vesicular storage and exocytotic release of all monoamine neurotransmitters. We used the Drosophila VMAT (dVMAT) null mutant to globally ablate exocytotic amine release, and then restored DVMAT activity in either individual or multiple aminergic systems using transgenic rescue techniques. We find that larval survival, larval locomotion, and female fertility rely predominantly on octopaminergic circuits with little apparent input from the vesicular release of serotonin or dopamine. In contrast, male courtship and fertility can be rescued by expressing DVMAT in octopaminergic or dopaminergic neurons, suggesting potentially redundant circuits. Rescue of major aspects of adult locomotion and startle behavior required octopamine, but a complementary role was observed for serotonin. Interestingly, adult circadian behavior could not be rescued by expression of DVMAT in a single subtype of aminergic neurons, but required at least two systems, suggesting the possibility of unexpected cooperative interactions. Further experiments using this model will help determine how multiple aminergic systems may contribute to the regulation of other behaviors. Our data also highlight potential differences between behaviors regulated by standard exocytotic release versus other mechanisms.

Dispensable, redundant, complementary and cooperative roles of dopamine, octopamine and serotonin in Drosophila melanogaster.

Afin d&#39;&eacute;tudier la r&eacute;gulation du comportement de Drosophila melanogaster par les amines biog&egrave;nes, nous avons exploit&eacute; la condition large du transporteur de monoamine v&eacute;siculaire (VMAT) pour le stockage v&eacute;siculaire et la lib&eacute;ration de tous les neurotransmetteurs de monoamine exocytotique. Nous avons utilis&eacute; le mutant nul Drosophila VMAT (dVMAT) pour globalement l&#39;ablation lib&eacute;ration exocytotique amine et puis restaur&eacute; l&#39;activit&eacute; DVMAT en soit particulier ou plusieurs syst&egrave;mes aminergique utilisant transg&eacute;niques techniques de sauvetage. Nous trouvons que la survie des larves, larve locomotion et fertilit&eacute; femelle s&#39;appuient principalement sur des circuits d&#39;octopaminergique avec entr&eacute;e peu apparente de la lib&eacute;ration v&eacute;siculaire de s&eacute;rotonine ou de dopamine. En revanche, la fertilit&eacute; et les m&acirc;les peuvent &ecirc;tre sauv&eacute;es en exprimant DVMAT dans les neurones octopaminergique ou dopaminergiques, sugg&eacute;rant des circuits potentiellement redondants. Sauvetage des principaux aspects du comportement de locomotion et sursaut adult requise octopamine, mais un r&ocirc;le compl&eacute;mentaire a &eacute;t&eacute; observ&eacute; pour la s&eacute;rotonine. Fait int&eacute;ressant, comportement circadienne adulte ne pourrait pas &ecirc;tre sauv&eacute; par expression de DVMAT dans un sous-type unique de neurones aminergique, mais n&eacute;cessaires au moins deux syst&egrave;mes, sugg&eacute;rant la possibilit&eacute; d&#39;interactions coop&eacute;ratives inattendues. D&#39;autres exp&eacute;riences &agrave; l&#39;aide de ce mod&egrave;le permettra de d&eacute;terminer comment les divers syst&egrave;mes aminergique peuvent contribuer &agrave; la r&eacute;gulation d&#39;autres comportements. Nos donn&eacute;es soulignent &eacute;galement les diff&eacute;rences potentielles entre les comportements r&eacute;glement&eacute;s par la lib&eacute;ration exocytotique standard par rapport aux autres m&eacute;canismes.

R&ocirc;les dispensables, redondants, compl&eacute;mentaires et coop&eacute;ratifs de l&#39;octopamine, de dopamine et de s&eacute;rotonine chez Drosophila melanogaster.

Um die Regulierung von Drosophila Melanogaster Verhalten von biogenen Aminen zu untersuchen, haben wir das breite Erfordernis der vesikul&auml;ren Monoamin-Transporter (VMAT) f&uuml;r die vesikul&auml;re Speicherung und exocytotic Freisetzung von allen Monoamin-Neurotransmittern ausgenutzt. Wir der Taufliege VMAT (dVMAT)-null-Mutant zum Global exocytotic Amin Freigabe entfernen verwendet und dann wiederhergestellt DVMAT Aktivit&auml;t in jedem einzelnen oder mehrerer Aminergic-Systeme, die mit transgenen retten Techniken. Wir finden, dass Larven &uuml;berleben, larval Fortbewegung und weibliche Fruchtbarkeit &uuml;berwiegend auf Octopaminergic Schaltungen mit wenig offensichtliche Input aus der vesikul&auml;ren Freisetzung von Serotonin oder Dopamin. Im Gegensatz dazu k&ouml;nnen m&auml;nnliche Verlobungszeit und Fruchtbarkeit gerettet werden, zum Ausdruck bringen, DVMAT in Octopaminergic oder dopaminergen Neuronen, die potenziell redundante Schaltungen vorgeschlagen. Rettung der Hauptaspekte der Erwachsenen Fortbewegung und Startle Verhalten erforderlich Octopamine, aber eine erg&auml;nzende Rolle wurde f&uuml;r Serotonin beobachtet. Interessanterweise konnten Erwachsene circadian Verhalten nicht Ausdruck der DVMAT in einen einzigen Untertyp der Aminergic Neuronen, sondern ben&ouml;tigt mindestens zwei Systeme, die M&ouml;glichkeit der unerwarteten kooperative Wechselwirkungen gerettet werden. Weitere Experimente mit diesem Modell werden helfen, festzustellen, wie mehrere Aminergic Systeme zur Regulierung der anderen Verhaltensweisen beitragen k&ouml;nnen. Unsere Daten zeigen auch m&ouml;gliche Unterschiede zwischen Verhaltensweisen von standard exocytotic-Release im Vergleich zu anderen Mechanismen reguliert.

Entbehrlich, redundante, komplement&auml;re und kooperative Funktionen Octopamine, Dopamin und Serotonin in Drosophila Melanogaster.

Per studiare la regolazione del comportamento di Drosophila melanogaster da ammine biogene, noi abbiamo sfruttato il requisito del trasportatore vescicolare di monoamine (VMAT) per l&#39;immagazzinamento vescicolare ampia e questo rilascio di tutti i neurotrasmettitori di monoamine. Abbiamo usato il mutante null drosofila VMAT (dVMAT) per l&#39;ablazione globalmente rilascio questo ammina e poi ripristinati attivit&agrave; DVMAT in ogni individuo o pi&ugrave; sistemi di aminergic usando transgenici tecniche di salvataggio. Troviamo che sopravvivenza larvale, locomozione delle larve e fertilit&agrave; femminile si affidano prevalentemente octopaminergic circuiti con poco apparente input dal rilascio vescicolare di serotonina o dopamina. Al contrario, fertilit&agrave; e corteggiamento maschile pu&ograve; essere salvati dalla esprimendo DVMAT in octopaminergic o dopaminergico neuroni, suggerendo circuiti potenzialmente ridondanti. Salvataggio dei principali aspetti del comportamento degli adulti di locomozione e startle necessaria Octopamina, ma &egrave; stato osservato un ruolo complementare per la serotonina. &Egrave; interessante notare che, comportamento circadiano degli adulti non poteva essere salvato dalla espressione di DVMAT in un singolo sottotipo di aminergic neuroni, ma richiesti almeno due sistemi, suggerendo la possibilit&agrave; di interazioni impreviste cooperative. Ulteriori esperimenti utilizzando questo modello aiuter&agrave; a determinare quanto pi&ugrave; aminergic sistemi possono contribuire alla regolazione di altri comportamenti. I nostri dati, inoltre, evidenziare potenziali differenze tra comportamenti regolati da standard questo rilascio contro altri meccanismi.

Superflui, ridondanti, complementari e cooperativi ruoli di dopamina, octopamina e serotonina nel melanogaster della drosofila.

&#x751F;&#x4F53;&#x30A2;&#x30DF;&#x30F3;&#x306B;&#x3088;&#x308B;&#x30B7;&#x30E7;&#x30A6;&#x30B8;&#x30E7;&#x30A6;&#x30D0;&#x30A8; &#x30AD;&#x30A4;&#x30ED;&#x30B7;&#x30E7;&#x30A6;&#x30B8;&#x30E7;&#x30A6;&#x30D0;&#x30A8;&#x884C;&#x52D5;&#x306E;&#x5236;&#x5FA1;&#x3092;&#x8ABF;&#x67FB;&#x3059;&#x308B;&#x306B;&#x306F;&#x3001;&#x5E45;&#x306E;&#x5E83;&#x3044;&#x8981;&#x4EF6;&#x5C0F;&#x80DE;&#x306E;&#x30E2;&#x30CE;&#x30A2;&#x30DF;&#x30F3;&#x8F38;&#x9001;&#x4F53;&#x306E;&#x5C0F;&#x80DE;&#x306E;&#x4FDD;&#x7BA1; (VMAT) &#x3068;&#x3059;&#x3079;&#x3066;&#x306E;&#x30E2;&#x30CE;&#x30A2;&#x30DF;&#x30F3;&#x795E;&#x7D4C;&#x4F1D;&#x9054;&#x7269;&#x8CEA;&#x306E;&#x5206;&#x6CCC;&#x306E;&#x30EA;&#x30EA;&#x30FC;&#x30B9;&#x3092;&#x60AA;&#x7528;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x79C1;&#x305F;&#x3061;&#x306F;&#x30B0;&#x30ED;&#x30FC;&#x30D0;&#x30EB;&#x306B;&#x5206;&#x6CCC;&#x30A2;&#x30DF;&#x30F3; &#x30EA;&#x30EA;&#x30FC;&#x30B9;&#x3092;&#x713C;&#x707C;&#x3059;&#x308B;&#x30B7;&#x30E7;&#x30A6;&#x30B8;&#x30E7;&#x30A6;&#x30D0;&#x30A8; VMAT (dVMAT) null &#x5909;&#x7570;&#x4F53;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3001;DVMAT &#x306E;&#x6D3B;&#x52D5;&#x306E;&#x3044;&#x305A;&#x308C;&#x304B;&#x306E;&#x500B;&#x3005; &#x3092;&#x5FA9;&#x5143;&#x307E;&#x305F;&#x306F;&#x30C8;&#x30E9;&#x30F3;&#x30B9;&#x30B8;&#x30A7;&#x30CB;&#x30C3;&#x30AF;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3066;&#x8907;&#x6570;&#x306E;&#x30A2;&#x30DF;&#x30F3; &#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x6551;&#x52A9;&#x6280;&#x8853;&#x3002;&#x5E7C;&#x866B;&#x306E;&#x751F;&#x5B58;&#x3001;&#x5E7C;&#x866B;&#x306E;&#x6B69;&#x884C;&#x3001;&#x5973;&#x6027;&#x4E0D;&#x598A;&#x4E3B;&#x30AA;&#x30AF;&#x30C8;&#x30D1;&#x30DF;&#x30F3;&#x30A2;&#x30B4;&#x30CB;&#x30B9;&#x30C8;&#x6D3B;&#x6027;&#x56DE;&#x8DEF;&#x30BB;&#x30ED;&#x30C8;&#x30CB;&#x30F3;&#x3084;&#x30C9;&#x30FC;&#x30D1;&#x30DF;&#x30F3;&#x306E;&#x5C0F;&#x80DE;&#x306E;&#x30EA;&#x30EA;&#x30FC;&#x30B9;&#x304B;&#x3089;&#x5C11;&#x306A;&#x3044;&#x306E;&#x898B;&#x304B;&#x3051;&#x4E0A;&#x306E;&#x5165;&#x529B;&#x3092;&#x983C;&#x308B;&#x3053;&#x3068;&#x3092;&#x898B;&#x3064;&#x3051;&#x307E;&#x3059;&#x3002;&#x5BFE;&#x7167;&#x7684;&#x306B;&#x3001;&#x6F5C;&#x5728;&#x7684;&#x5197;&#x9577;&#x56DE;&#x8DEF;&#x3092;&#x793A;&#x5506;&#x3001;&#x30AA;&#x30AF;&#x30C8;&#x30D1;&#x30DF;&#x30F3;&#x30A2;&#x30B4;&#x30CB;&#x30B9;&#x30C8;&#x6D3B;&#x6027;&#x307E;&#x305F;&#x306F;&#x30C9;&#x30D1;&#x30DF;&#x30F3;&#x306E;&#x795E;&#x7D4C;&#x7D30;&#x80DE;&#x3067;&#x306E; DVMAT &#x306E;&#x8868;&#x73FE;&#x306B;&#x3088;&#x3063;&#x3066;&#x96C4;&#x306E;&#x6C42;&#x611B;&#x3068;&#x4E0D;&#x598A;&#x6CBB;&#x7642;&#x3092;&#x6551;&#x51FA;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;&#x6210;&#x4EBA;&#x306E;&#x6B69;&#x884C;&#x3068;&#x9A5A;&#x6115;&#x306E;&#x884C;&#x52D5;&#x306E;&#x4E3B;&#x8981;&#x306A;&#x5074;&#x9762;&#x306E;&#x6551;&#x52A9;&#x30AA;&#x30AF;&#x30C8;&#x30D1;&#x30DF;&#x30F3;&#x3001;&#x5FC5;&#x8981;&#x304C;&#x3001;&#x88DC;&#x5B8C;&#x7684;&#x306A;&#x5F79;&#x5272;&#x306F;&#x3001;&#x30BB;&#x30ED;&#x30C8;&#x30CB;&#x30F3;&#x306E;&#x89B3;&#x5BDF;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x8208;&#x5473;&#x6DF1;&#x3044;&#x3053;&#x3068;&#x306B;&#x3001;&#x6210;&#x4EBA;&#x306E;&#x6982;&#x65E5;&#x884C;&#x52D5;&#x306F;&#x4E88;&#x671F;&#x305B;&#x306C;&#x5354;&#x529B;&#x7684;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x306E;&#x53EF;&#x80FD;&#x6027;&#x3092;&#x793A;&#x5506;&#x3057;&#x3066;&#x5358;&#x4E00;&#x306E;&#x30B5;&#x30D6;&#x30BF;&#x30A4;&#x30D7;&#x306E;&#x30A2;&#x30DF;&#x30F3; &#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3;&#x304C;&#x5FC5;&#x8981;&#x5C11;&#x306A;&#x304F;&#x3068;&#x3082; 2 &#x3064;&#x306E;&#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x306B;&#x304A;&#x3051;&#x308B; DVMAT &#x306E;&#x767A;&#x73FE;&#x306B;&#x3088;&#x3063;&#x3066;&#x6551;&#x51FA;&#x3055;&#x308C;&#x308B;&#x53EF;&#x80FD;&#x6027;&#x306F;&#x3042;&#x308A;&#x307E;&#x305B;&#x3093;&#x3002;&#x3053;&#x306E;&#x30E2;&#x30C7;&#x30EB;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3066;&#x306E;&#x3055;&#x3089;&#x306A;&#x308B;&#x5B9F;&#x9A13;&#x65B9;&#x6CD5;&#x8907;&#x6570;&#x306E;&#x30A2;&#x30DF;&#x30F3; &#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x4ED6;&#x306E;&#x884C;&#x52D5;&#x306E;&#x8ABF;&#x7BC0;&#x306B;&#x5BC4;&#x4E0E;&#x3057;&#x3066;&#x53EF;&#x80FD;&#x6027;&#x304C;&#x3042;&#x308A;&#x307E;&#x3059;&#x6C7A;&#x5B9A;&#x306B;&#x5F79;&#x7ACB;&#x3061;&#x307E;&#x3059;&#x3002;&#x6211;&#x3005; &#x306E;&#x30C7;&#x30FC;&#x30BF;&#x306F;&#x307E;&#x305F;&#x6A19;&#x6E96;&#x306E;&#x5206;&#x6CCC;&#x30EA;&#x30EA;&#x30FC;&#x30B9;&#x5BFE;&#x4ED6;&#x306E;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x306B;&#x3088;&#x3063;&#x3066;&#x898F;&#x5236;&#x884C;&#x52D5;&#x306E;&#x6F5C;&#x5728;&#x7684;&#x306A;&#x9055;&#x3044;&#x3092;&#x5F37;&#x8ABF;&#x8868;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;

&#x30C9;&#x30FC;&#x30D1;&#x30DF;&#x30F3;, &#x30AA;&#x30AF;&#x30C8;&#x30D1;&#x30DF;&#x30F3;&#x3068;&#x30AD;&#x30A4;&#x30ED;&#x30B7;&#x30E7;&#x30A6;&#x30B8;&#x30E7;&#x30A6;&#x30D0;&#x30A8;&#x306B;&#x304A;&#x3051;&#x308B;&#x30BB;&#x30ED;&#x30C8;&#x30CB;&#x30F3;&#x306E;&#x4E0D;&#x53EF;&#x6B20;&#x3067;&#x3001;&#x5197;&#x9577;&#x3001;&#x88DC;&#x5B8C;&#x3001;&#x5354;&#x8ABF;&#x306E;&#x5F79;&#x5272;&#x3002;

&#xC0DD;&#xBB3C; &#xC544;&#xBBFC;&#xC5D0; &#xC758;&#xD574; Drosophila melanogaster &#xB3D9;&#xC791;&#xC758; &#xADDC;&#xC81C; &#xC870;&#xC0AC;, &#xC6B0;&#xB9AC;&#xB294; &#xAE30;&#xACF5;&#xC744; &#xBAA8;&#xB178; &#xC6B4;&#xC1A1;&#xC5C5; &#xC790; &#xAE30;&#xACF5;&#xC744; &#xC800;&#xC7A5;&#xC6A9; (VMAT)&#xC758; &#xAD11;&#xBC94;&#xC704; &#xD55C; &#xC694;&#xAD6C; &#xC0AC;&#xD56D; &#xBC0F; &#xBAA8;&#xB4E0; &#xBAA8;&#xB178; &#xC2E0;&#xACBD; &#xC804;&#xB2EC; &#xBB3C;&#xC9C8;&#xC758; exocytotic &#xCD9C;&#xC2DC; &#xC545;&#xC6A9;. &#xC6B0;&#xB9AC;&#xB294; null &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xCD08;&#xD30C;&#xB9AC; VMAT (dVMAT)&#xB97C; &#xC0AC;&#xC6A9; &#xD558;&#xB294; &#xC138;&#xACC4;&#xC801;&#xC73C;&#xB85C; exocytotic &#xC544;&#xBBFC; &#xB9B4;&#xB9AC;&#xC2A4; &#xC808;&#xC81C; &#xC5B4;&#xB290; &#xAC1C;&#xC778;&#xC5D0; DVMAT &#xD65C;&#xB3D9;&#xC744; &#xBCF5;&#xC6D0; &#xB610;&#xB294; &#xC720;&#xC804;&#xC790; &#xBCC0;&#xD615; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xC5EC;&#xB7EC; &#xAC1C;&#xC758; aminergic &#xC2DC;&#xC2A4;&#xD15C; &#xAD6C;&#xC870; &#xAE30;&#xC220;. &#xC560;&#xBC8C;&#xB808;&#xC758; &#xC0DD;&#xC874;, &#xC560;&#xBC8C;&#xB808; &#xC6B4;&#xB3D9; &#xBC0F; &#xC5EC;&#xC131; &#xBD88; &#xC784; &#xC138;&#xB85C;&#xD1A0;&#xB2CC; &#xC774;&#xB098; &#xB3C4;&#xD30C;&#xBBFC;&#xC758; &#xAE30;&#xACF5;&#xC744; &#xB9B4;&#xB9AC;&#xC2A4;&#xC5D0;&#xC11C; &#xC57D;&#xAC04; &#xBA85;&#xBC31;&#xD55C; &#xC785;&#xB825; octopaminergic &#xD68C;&#xB85C;&#xC5D0; &#xC8FC;&#xB85C; &#xC758;&#xC874;&#xC744; &#xCC3E;&#xC744; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xBC18;&#xBA74;, &#xB0A8;&#xC131; &#xAD6C;&#xC560;&#xC640; &#xBD88; &#xC784; &#xC7A0;&#xC7AC;&#xC801; &#xC911;&#xBCF5; &#xD68C;&#xB85C; &#xC81C;&#xC548; octopaminergic &#xB610;&#xB294; dopaminergic &#xC2E0;&#xACBD; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; DVMAT&#xC744; &#xD45C;&#xD604; &#xD558; &#xC5EC; &#xAD6C;&#xCD9C; &#xB420; &#xC218; &#xC788;&#xB2E4;. &#xC131;&#xC778; &#xC6B4;&#xB3D9; &#xBC0F; startle &#xB3D9;&#xC791;&#xC758; &#xC8FC;&#xC694; &#xCE21;&#xBA74;&#xC758; &#xAD6C;&#xC870; octopamine, &#xD558;&#xC9C0;&#xB9CC; &#xC0C1;&#xD638; &#xBCF4;&#xC644;&#xC801;&#xC778; &#xC5ED;&#xD560;&#xC744; &#xC138;&#xB85C;&#xD1A0;&#xB2CC;&#xC5D0; &#xB300; &#xD55C; &#xAD00;&#xCC30; &#xB418;&#xC5C8;&#xB2E4;. &#xD765;&#xBBF8;&#xB86D;&#xAC8C;&#xB3C4;, &#xC131;&#xC778; circadian &#xB3D9;&#xC791;&#xC774; &#xC608;&#xAE30;&#xCE58; &#xC54A;&#xC740; &#xD611;&#xB825; &#xC0C1;&#xD638; &#xC791;&#xC6A9;&#xC758; &#xAC00;&#xB2A5;&#xC131;&#xC744; &#xC81C;&#xC548; aminergic &#xC2E0;&#xACBD;, &#xD558;&#xC9C0;&#xB9CC; &#xD544;&#xC694;&#xD55C; &#xB450; &#xAC1C; &#xC774;&#xC0C1;&#xC758; &#xC2DC;&#xC2A4;&#xD15C;&#xC758; &#xB2E8;&#xC77C; &#xD558;&#xC704; &#xD615;&#xC2DD;&#xC5D0; DVMAT&#xC758; &#xC2DD;&#xC5D0; &#xC758;&#xD574; &#xAD6C;&#xCD9C; &#xB418; &#xACE0; &#xD558;&#xC9C0;. &#xC774; &#xBAA8;&#xB378;&#xC744; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xCD94;&#xAC00; &#xC2E4;&#xD5D8; &#xC5B4;&#xB5BB;&#xAC8C; &#xC5EC;&#xB7EC; aminergic &#xC2DC;&#xC2A4;&#xD15C; &#xB2E4;&#xB978; &#xD589;&#xB3D9;&#xC758; &#xC870;&#xC808;&#xC5D0; &#xAE30;&#xC5EC;&#xD560; &#xC218;&#xB97C; &#xACB0;&#xC815; &#xD558;&#xB294; &#xB370; &#xB3C4;&#xC6C0;&#xC774; &#xB429;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xC758; &#xB370;&#xC774;&#xD130;&#xB294; &#xB610;&#xD55C; &#xB3D9;&#xC791; &#xD45C;&#xC900; exocytotic &#xB9B4;&#xB9AC;&#xC2A4; &#xB300; &#xB2E4;&#xB978; &#xBA54;&#xCEE4;&#xB2C8;&#xC998;&#xC5D0; &#xC758;&#xD574; &#xADDC;&#xC81C;&#xC758; &#xC7A0;&#xC7AC;&#xC801;&#xC778; &#xCC28;&#xC774;&#xC810;&#xC744; &#xAC15;&#xC870; &#xD45C;&#xC2DC; &#xD569;&#xB2C8;&#xB2E4;.

&#xB3C4;&#xD30C;&#xBBFC;, octopamine &#xBC0F; &#xC138;&#xB85C;&#xD1A0;&#xB2CC; Drosophila melanogaster&#xC5D0; &#xBD88;&#xAC00;&#xACB0;, &#xC911;&#xBCF5;, &#xBCF4;&#xC644; &#xBC0F; &#xD611;&#xB825; &#xC5ED;&#xD560;.

Para investigar la regulaci&oacute;n del comportamiento de Drosophila melanogaster por aminas bi&oacute;genas, nos hemos explotado el requisito amplio del transportador monoamino vesicular (VMAT) para el almacenamiento vesicular y exocit&oacute;tico liberaci&oacute;n de neurotransmisores de monoamino todos. Se utiliz&oacute; al mutante nulo de Drosophila VMAT (dVMAT) a ablar globalmente liberaci&oacute;n exocit&oacute;tico amina y luego hab&iacute;a restaurada actividad DVMAT en cualquier individuo o t&eacute;cnicas de rescate de m&uacute;ltiples sistemas de amin&eacute;rgico que utilizan transg&eacute;nicos. Encontramos que supervivencia larvaria, locomoci&oacute;n larva y la fertilidad de la mujer dependen predominantemente octopaminergic circuitos con entrada poco evidente de la vesicular liberaci&oacute;n de serotonina o dopamina. En cambio, fertilidad y cortejo masculino pueden ser rescatados expresando DVMAT en las neuronas octopaminergic o dopamin&eacute;rgicos, sugiriendo circuitos potencialmente redundantes. Rescate de los principales aspectos del comportamiento adulto de locomoci&oacute;n y susto necesaria Octopamina, pero se observ&oacute; un papel complementario para la serotonina. Curiosamente, comportamiento circadiano adultos no podr&iacute;a ser rescatada por la expresi&oacute;n de DVMAT en un &uacute;nico subtipo de amin&eacute;rgico neuronas, pero requiere al menos dos sistemas, sugiriendo la posibilidad de interacciones cooperativas inesperadas. Otros experimentos utilizando este modelo ayudar&aacute; a determinar c&oacute;mo m&uacute;ltiples sistemas amin&eacute;rgico pueden contribuir a la regulaci&oacute;n de otros comportamientos. Nuestros datos tambi&eacute;n resaltar las diferencias potenciales entre comportamientos regulados por liberaci&oacute;n exocit&oacute;tico est&aacute;ndar frente a otros mecanismos.

Funciones prescindibles, redundantes, complementarias y cooperativas de Octopamina, dopamina y serotonina en Drosophila melanogaster.

Sleep is an essential behavioral state of rest that is regulated by homeostatic drives to ensure a balance of sleep and activity, as well as independent arousal mechanisms in the central brain. Dopamine has been identified as a critical regulator of both sleep behavior and arousal. Here, we present results of a genetic screen that selectively restored the Dopamine Receptor (DopR/DopR1/dumb) to specific neuroanatomical regions of the adult Drosophila brain to assess requirements for DopR in sleep behavior. We have identified subsets of the mushroom body that utilizes DopR in daytime sleep regulation. These data are supported by multiple examples of spatially restricted genetic rescue data in discrete circuits of the mushroom body, as well as immunohistochemistry that corroborates the localization of DopR protein within mushroom body circuits. Independent loss of function data using an inducible RNAi construct in the same specific circuits also supports a requirement for DopR in daytime sleep. Additional circuit activation of discrete DopR(+) mushroom body neurons also suggests roles for these subpopulations in sleep behavior. These conclusions support a new separable function for DopR in daytime sleep regulation within the mushroom body. This daytime regulation is independent of the known role of DopR in nighttime sleep, which is regulated within the Fan-Shaped Body (FSB). This study provides new neuroanatomical loci for exploration of dopaminergic sleep functions in Drosophila, and expands our understanding of sleep regulation during the day vs. night.

A Genetic Screen To Assess Dopamine Receptor (DopR1) Dependent Sleep Regulation in Drosophila.

Optomotor behavior represents a stereotyped locomotor response to visual motion that is found in both vertebrate and invertebrate models. The Fly Stampede assay was developed to study an optomotor response in freely walking populations of Drosophila. Here we share optimized assay designs and software for production of a modified stampede assay that can be used for genetic screens, and improved tracking outputs for understanding behavioral parameters of visual-motion responses and arousal state of individual animals. Arousal state influences behavioral performance in the stampede assay. As proof of principle experiments we show parametric modulation of visual stimuli and startle stimuli in both wildtype and mutant flies for the type I family dopamine receptor Dop1R1 (DopR). DopR mutants are hyperactive and perform poorly in the stampede assay, suggesting a potential role in visual perception and/or arousal. The stampede assay creates an efficient platform for rapid screening of mutant animals or circuit manipulations for investigating attentional processes in Drosophila.

Article	Year
Characterization of a novel Drosophila melanogaster galectin. Expression in developing immune, neural, and muscle tissues. The Journal of biological chemistry\| PubMed ID: 11809773	2002
A Serrate-expressing signaling center controls Drosophila hematopoiesis. Genes & development\| PubMed ID: 12569125	2003
Two different forms of arousal in Drosophila are oppositely regulated by the dopamine D1 receptor ortholog DopR via distinct neural circuits. Neuron\| PubMed ID: 19945394	2009
A pair of dopamine neurons target the D1-like dopamine receptor DopR in the central complex to promote ethanol-stimulated locomotion in Drosophila. PloS one\| PubMed ID: 20376353	2010
Dispensable, redundant, complementary and cooperative roles of dopamine, octopamine and serotonin in Drosophila melanogaster. Genetics\| PubMed ID: 23086220	2012
A Genetic Screen To Assess Dopamine Receptor (DopR1) Dependent Sleep Regulation in Drosophila. G3 (Bethesda, Md.)\| PubMed ID: 27760793	2016
Fly Stampede 2.0: A Next Generation Optomotor Assay for Walking Behavior in Drosophila Melanogaster. Frontiers in molecular neuroscience\| PubMed ID: 28105003	2016

Tim Lebestky

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