Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae

Vector mosquitoes inflict more human suffering than any other organism&#8212;and kill more than one million people each year. The mosquito genome projects ...

Indiana University School of Medicine

University of Notre Dame

We use monoclonal antibodies against synaptic proteins and anterograde tracing with neurobiotin to describe the architecture of the antennal lobes in different castes of two ant species -Camponotus sericeus and Camponotus compressus. The reproductives and worker classes are readily categorized based on size and external morphology. The overall organization of brain neuropile is comparable between castes with differences only in the visual ganglia. Males have a larger fraction of neuropile occupied by the medulla and lobula than females. In the diurnal species, C. sericeus these regions are more highly represented, than in the nocturnal species C. compressus. The most striking differences are in the antennal lobe where males possess a macroglomerulus, which is about ten times larger in volume than the other glomeruli; such a specialization is absent in females. Minor workers possess a significantly larger number of glomeruli than the majors despite the smaller overall volume of the lobe. These caste-specific differences occur mainly within glomerular clusters that receive input from sensory neurons that project in tracts - T4 and T5 - within the antennal nerve. The comparative anatomy of different castes of ants provides an entry point into a future systematic analysis of how divergent brain architectures can arise within a single species.

Caste and sex specific olfactory glomerular organization and brain architecture in two sympatric ant species Camponotus sericeus and Camponotus compressus (Fabricius, 1798).

The antennal lobes of different castes of the ant species Camponotus compressus show a marked diversity in the organization of their olfactory glomeruli. Notably, there is a significant difference in the number and size of glomeruli between the reproductives and the workers and among the different worker castes. In this report, we investigate the notion that these caste-specific differences in glomerular number might be accounted for, at least in part, by the differences in numbers of olfactory sensilla that target the antennal lobe. For this, we examine the number of sensilla on the antennal flagella of all the individual castes of C. compressus. This analysis reveals a striking correlation between sensillar number and the number of antennal glomeruli in a given caste. As a first step in investigating the causal mechanisms that might give raise to this correlation, we carry out an initial characterization of olfactory system development in the minor workers of C. compressus. We analyze the temporal pattern of innervations of the developing antennal lobe by olfactory sensory neuron axons. We document the development of the olfactory glomeruli in the antennal lobe during this process, which occurs during early pupal stages. Our findings provide the basis for future manipulative developmental studies on the role of sensory afferent number in glomerular development of different castes within the same species.

Morphological and developmental analysis of peripheral antennal chemosensory sensilla and central olfactory glomeruli in worker castes of Camponotus compressus (Fabricius, 1787).

Un esfuerzo considerable ha sido dirigido hacia la comprensi&oacute;n de la organizaci&oacute;n y la funci&oacute;n de perif&eacute;rico y el sistema nervioso central de mosquitos vectores de enfermedades como el Aedes aegypti. Hasta la fecha, todas estas investigaciones se han realizado en adultos, pero ninguno de los estudios dirigi&oacute; el desarrollo del sistema nervioso durante las etapas de larvas y pupas en mosquitos. Aqu&iacute;, nosotros primero pantalla un conjunto de 30 anticuerpos, que se han utilizado para estudiar el desarrollo del cerebro en Drosophila e identificar a 13 de ellos antig&eacute;nico y etiquetado ep&iacute;topos en el cerebro en desarrollo de Aedes. Entonces utilizamos los anticuerpos identificados en estudios de inmunomarcaci&oacute;n para caracterizar las caracter&iacute;sticas neuroanat&oacute;micas generales del cerebro en desarrollo y compararlos con el sistema del modelo estudiado, Drosophila melanogaster, en etapas de larvas, pupas y adultas. Adem&aacute;s, utilizamos inmunomarcaci&oacute;n para documentar el desarrollo de componentes espec&iacute;ficos del cerebro Aedes, es decir, los l&oacute;bulos de la &oacute;ptica, la subesophageal neuropil y sistema serotonin&eacute;rgico del subesophageal neuropil m&aacute;s detalladamente. Nuestro estudio revela diferencias importantes en el cerebro en desarrollo en el estado larvario en comparaci&oacute;n con la etapa pupal (y adultos) de Aedes. Los resultados tambi&eacute;n descubren interesantes similitudes y diferencias marcadas en el desarrollo del cerebro de Aedes en comparaci&oacute;n con Drosophila. Tomados en conjunto, esta investigaci&oacute;n constituye la base para futuras investigaciones celulares y moleculares del desarrollo del cerebro en este vector de la enfermedad importante.

Desarrollo en el mosquito Aedes aegypti de fiebre amarilla del cerebro: un an&aacute;lisis comparativo de immunocytochemical usando los anticuerpos Cruz-que reaccionaba de Drosophila melanogaster.

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&#x8111;&#x53D1;&#x80B2;&#x5728;&#x57C3;&#x53CA;&#x7684;&#x9EC4;&#x70ED;&#x75C5;&#x868A;&#x4F0A;&#x868A;&#xFF1A; &#x4F7F;&#x7528; cross-reacting &#x6297;&#x4F53;&#x9ED1;&#x8179;&#x679C;&#x8747;&#x6BD4;&#x8F83;&#x514D;&#x75AB;&#x7EC6;&#x80DE;&#x5316;&#x5B66;&#x5206;&#x6790;&#x3002;

Considerable effort has been directed towards understanding the organization and function of peripheral and central nervous system of disease vector mosquitoes such as Aedes aegypti. To date, all of these investigations have been carried out on adults but none of the studies addressed the development of the nervous system during the larval and pupal stages in mosquitoes. Here, we first screen a set of 30 antibodies, which have been used to study brain development in Drosophila, and identify 13 of them cross-reacting and labeling epitopes in the developing brain of Aedes. We then use the identified antibodies in immunolabeling studies to characterize general neuroanatomical features of the developing brain and compare them with the well-studied model system, Drosophila melanogaster, in larval, pupal, and adult stages. Furthermore, we use immunolabeling to document the development of specific components of the Aedes brain, namely the optic lobes, the subesophageal neuropil, and serotonergic system of the subesophageal neuropil in more detail. Our study reveals prominent differences in the developing brain in the larval stage as compared to the pupal (and adult) stage of Aedes. The results also uncover interesting similarities and marked differences in brain development of Aedes as compared to Drosophila. Taken together, this investigation forms the basis for future cellular and molecular investigations of brain development in this important disease vector.

Brain development in the yellow fever mosquito Aedes aegypti: a comparative immunocytochemical analysis using cross-reacting antibodies from Drosophila melanogaster.

Un effort consid&eacute;rable a &eacute;t&eacute; consacr&eacute; &agrave; comprendre l&#39;organisation et la fonction de p&eacute;riph&eacute;rique et le syst&egrave;me nerveux central des moustiques vecteur de maladies telles que Aedes aegypti. A ce jour, toutes ces enqu&ecirc;tes ont &eacute;t&eacute; men&eacute;es sur des adultes, mais aucune des &eacute;tudes adress&eacute;e le d&eacute;veloppement du syst&egrave;me nerveux durant les stades de larves et les nymphes de moustiques. Ici, nous l&#39;&eacute;cran tout d&#39;abord un ensemble de 30 anticorps, qui ont &eacute;t&eacute; utilis&eacute;s pour &eacute;tudier le d&eacute;veloppement du cerveau chez la drosophile et d&#39;identifier 13 d&#39;entre eux r&eacute;agissant et l&#39;&eacute;tiquetage des &eacute;pitopes sur le d&eacute;veloppement du cerveau de Aedes. Nous utilisons ensuite les anticorps identifi&eacute;s dans les &eacute;tudes d&#39;immunomarquage pour caract&eacute;riser des caract&eacute;ristiques g&eacute;n&eacute;rales neuroanatomiques du cerveau en d&eacute;veloppement et les comparer avec le syst&egrave;me de mod&egrave;le bien &eacute;tudi&eacute;s, Drosophila melanogaster, aux stades de larves, pupes et adultes. En outre, nous utilisons immunomarquage afin de documenter le d&eacute;veloppement de composants sp&eacute;cifiques du cerveau Aedes, nomm&eacute;ment les lobes optiques sous-oesophagien neuropile et le syst&egrave;me s&eacute;rotoninergique de la neuropile sous-oesophagien plus en d&eacute;tail. Notre &eacute;tude r&eacute;v&egrave;le des diff&eacute;rences importantes dans le d&eacute;veloppement du cerveau au stade larvaire par rapport &agrave; la phase nymphale (et adulte) de Aedes. Les r&eacute;sultats d&eacute;couvrent &eacute;galement int&eacute;ressants des similitudes et des diff&eacute;rences marqu&eacute;es dans le d&eacute;veloppement du cerveau de Aedes comparativement &agrave; drosophile. Globalement, cette enqu&ecirc;te constitue la base pour de futures &eacute;tudes cellulaires et mol&eacute;culaires du d&eacute;veloppement du cerveau dans ce vecteur important de maladie.

Cerveau en d&eacute;veloppement dans le moustique Aedes aegypti de fi&egrave;vre jaune : une analyse comparative immunocytochimiques r&eacute;action crois&eacute;e des anticorps de Drosophila melanogaster.

Uno sforzo considerevole &egrave; stato diretto verso la comprensione, l&#39;organizzazione e la funzione della periferica e del sistema nervoso centrale delle zanzare vettore di malattia come Aedes aegypti. Ad oggi, tutte queste indagini sono state effettuate sugli adulti, ma nessuno degli studi ha indirizzato lo sviluppo del sistema nervoso durante le fasi larvali e di pupa in zanzare. Qui, abbiamo prima schermata un set di 30 anticorpi, che sono stati usati per studiare lo sviluppo del cervello in Drosophila e identificare 13 di loro cross-reacting ed etichettatura epitopi nel cervello in via di sviluppo di Aedes. Utilizziamo quindi gli anticorpi identificati negli studi di immunomarcatura per caratterizzare la generale caratteristiche neuroanatomiche del cervello in via di sviluppo e li confronta con il sistema modello ben studiata, Drosophila melanogaster, negli stadi larvali, pupa e adulti. Inoltre, usiamo immunomarcatura per documentare lo sviluppo di componenti specifici del cervello Aedes, vale a dire i lobi ottici, il neuropilo subesophageal e serotoninergici sistema del neuropilo subesophageal pi&ugrave; in dettaglio. Il nostro studio rivela importanti differenze nel cervello in via di sviluppo nella fase larvale rispetto la fase di pupa (e adulta) di Aedes. I risultati anche scoprono interessanti analogie e marcate differenze nello sviluppo del cervello di Aedes rispetto della drosofila. Presi insieme, questa indagine costituisce la base per future indagini molecolari e cellulari dello sviluppo del cervello in questo vettore di malattie importanti.

Lo sviluppo della febbre gialla zanzara Aedes aegypti del cervello: un&#39;analisi comparativa immunocitochimica utilizzando anticorpi Traversa-reagendo da Drosophila melanogaster.

&#x304B;&#x306A;&#x308A;&#x306E;&#x52AA;&#x529B;&#x306F;&#x3001;&#x75C5;&#x5A92;&#x4ECB;&#x868A;&#x30CD;&#x30C3;&#x30BF;&#x30A4;&#x30B7;&#x30DE;&#x30AB;&#x306A;&#x3069;&#x306E;&#x4E2D;&#x67A2;&#x795E;&#x7D4C;&#x7CFB;&#x306E;&#x69CB;&#x9020;&#x3068;&#x5468;&#x8FBA;&#x6A5F;&#x5668;&#x306E;&#x6A5F;&#x80FD;&#x7D44;&#x7E54;&#x306E;&#x7406;&#x89E3;&#x306E;&#x65B9;&#x306B;&#x6307;&#x793A;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x65E5;&#x4ED8;&#x3059;&#x308B;&#x306B;&#x306F;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x8ABF;&#x67FB;&#x306E;&#x3059;&#x3079;&#x3066;&#x306E;&#x5927;&#x4EBA;&#x306B;&#x884C;&#x3063;&#x305F;&#x304C;&#x3001;&#x3069;&#x308C;&#x3082;&#x7814;&#x7A76;&#x306E;&#x795E;&#x7D4C;&#x7CFB;&#x306E;&#x958B;&#x767A;&#x4E2D;&#x868A;&#x306E;&#x5E7C;&#x866B;&#x3068;&#x86F9;&#x306E;&#x6BB5;&#x968E;&#x306B;&#x5BFE;&#x51E6;&#x3057;&#x307E;&#x3059;&#x3002;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;&#x6211;&#x3005; &#x6700;&#x521D;&#x30B7;&#x30E7;&#x30A6;&#x30B8;&#x30E7;&#x30A6;&#x30D0;&#x30A8;&#x8133;&#x958B;&#x767A;&#x7814;&#x7A76;&#x3057;&#x3001;&#x305D;&#x308C;&#x3089;&#x306E; 13 &#x3092;&#x8B58;&#x5225;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x4F7F;&#x7528;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B; 30 &#x6297;&#x4F53;&#x306E;&#x30BB;&#x30C3;&#x30C8;&#x306E; cross-reacting &#x53CA;&#x3073;&#x30A8;&#x30D4;&#x30C8;&#x30FC;&#x30D7; &#x30E4;&#x30D6;&#x30AB;&#x5C5E;&#x306E;&#x4E2D;&#x306E;&#x8133;&#x3067;&#x8868;&#x793A;&#x753B;&#x9762;&#x3057;&#x307E;&#x3059;&#x3002;&#x7279;&#x5B9A;&#x306E;&#x6297;&#x4F53;&#x30A8;&#x30CA;&#x30E1;&#x30EB;&#x7814;&#x7A76;&#x958B;&#x767A;&#x9014;&#x4E0A;&#x306E;&#x8133;&#x306E;&#x4E00;&#x822C;&#x7684;&#x306A;&#x795E;&#x7D4C;&#x89E3;&#x5256;&#x5B66;&#x7684;&#x6A5F;&#x80FD;&#x3092;&#x7279;&#x5FB4;&#x3065;&#x3051;&#x308B;&#x3001;&#x3088;&#x304F;&#x7814;&#x7A76;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;&#x30E2;&#x30C7;&#x30EB;&#x306E;&#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x3067;&#x306F;&#x3001;&#x30AD;&#x30A4;&#x30ED;&#x30B7;&#x30E7;&#x30A6;&#x30B8;&#x30E7;&#x30A6;&#x30D0;&#x30A8;&#x3001;&#x5E7C;&#x866B;&#x3001;&#x86F9;&#x3001;&#x5927;&#x4EBA;&#x306E;&#x6BB5;&#x968E;&#x3067;&#x6BD4;&#x8F03;&#x306B;&#x4F7F;&#x7528;&#x3059;&#x308C;&#x3070;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;&#x6587;&#x66F8;&#x306E;&#x30E4;&#x30D6;&#x30AB;&#x5C5E;&#x306E;&#x8133;&#x3001;&#x3059;&#x306A;&#x308F;&#x3061;&#x5149;&#x8449;&#x3001;&#x8695;&#x98DF;&#x533A;&#x753B;&#x3068;&#x30BB;&#x30ED;&#x30C8;&#x30CB;&#x30F3; &#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x8695;&#x98DF;&#x533A;&#x753B;&#x306E;&#x8A73;&#x7D30;&#x306E;&#x7279;&#x5B9A;&#x306E;&#x30B3;&#x30F3;&#x30DD;&#x30FC;&#x30CD;&#x30F3;&#x30C8;&#x306E;&#x958B;&#x767A;&#x306B;&#x30A8;&#x30CA;&#x30E1;&#x30EB;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x307E;&#x3059;&#x3002;&#x79C1;&#x305F;&#x3061;&#x306E;&#x7814;&#x7A76;&#x958B;&#x767A;&#x9014;&#x4E0A;&#x306E;&#x8133;&#x306B;&#x9855;&#x8457;&#x306A;&#x9055;&#x3044;&#x306F;&#x30E4;&#x30D6;&#x30AB;&#x5C5E;&#x306E;&#x86F9; &#xFF08;&#x3068;&#x5927;&#x4EBA;&#xFF09; &#x306E;&#x30B9;&#x30C6;&#x30FC;&#x30B8;&#x3068;&#x6BD4;&#x8F03;&#x3057;&#x3066;&#x5E7C;&#x866B;&#x306E;&#x6BB5;&#x968E;&#x3067;&#x660E;&#x3089;&#x304B;&#x306B;&#x3057;&#x307E;&#x3059;&#x3002;&#x7D50;&#x679C;&#x3082;&#x8208;&#x5473;&#x6DF1;&#x3044;&#x985E;&#x4F3C;&#x70B9;&#x3068;&#x30D2;&#x30C8;&#x30B9;&#x30B8;&#x30B7;&#x30DE;&#x30AB; &#x30B7;&#x30E7;&#x30A6;&#x30B8;&#x30E7;&#x30A6;&#x30D0;&#x30A8;&#x3068;&#x6BD4;&#x8F03;&#x3057;&#x3066;&#x306E;&#x8133;&#x306E;&#x767A;&#x9054;&#x306E;&#x5DEE;&#x7570;&#x3092;&#x660E;&#x3089;&#x304B;&#x306B;&#x3057;&#x307E;&#x3059;&#x3002;&#x4E00;&#x7DD2;&#x306B;&#x53D6;&#x3089;&#x308C;&#x3066;&#x3001;&#x3053;&#x306E;&#x8ABF;&#x67FB;&#x306F;&#x3001;&#x3053;&#x306E;&#x91CD;&#x8981;&#x306A;&#x75C5;&#x6C17;&#x306E;&#x30D9;&#x30AF;&#x30C8;&#x30EB;&#x3067;&#x5C06;&#x6765;&#x306E;&#x7D30;&#x80DE;&#x304A;&#x3088;&#x3073;&#x5206;&#x5B50;&#x306E;&#x8ABF;&#x67FB;&#x306E;&#x8133;&#x306E;&#x767A;&#x9054;&#x306E;&#x305F;&#x3081;&#x306E;&#x57FA;&#x790E;&#x3092;&#x5F62;&#x6210;&#x3057;&#x307E;&#x3059;&#x3002;

&#x8133;&#x306E;&#x9EC4;&#x71B1;&#x868A;&#x30CD;&#x30C3;&#x30BF;&#x30A4;&#x30B7;&#x30DE;&#x30AB;&#x306E;&#x958B;&#x767A;: &#x30B7;&#x30E7;&#x30A6;&#x30B8;&#x30E7;&#x30A6;&#x30D0;&#x30A8; cross-reacting &#x6297;&#x4F53;&#x3092;&#x7528;&#x3044;&#x305F;&#x6BD4;&#x8F03;&#x514D;&#x75AB;&#x7D30;&#x80DE;&#x5316;&#x5B66;&#x7684;&#x5206;&#x6790;&#x3002;

&#xC0C1;&#xB2F9;&#xD55C; &#xB178;&#xB825;&#xC744; &#xC870;&#xC9C1; &#xBC0F; &#xC8FC;&#xBCC0; &#xC7A5;&#xCE58;&#xC758; &#xAE30;&#xB2A5; &#xBC0F; &#xC911;&#xC559; &#xC2E0; &#xACBD;&#xACC4; &#xC9C8;&#xBCD1; &#xBCA1;&#xD130; &#xAC19;&#xC740; Aedes aegypti &#xBAA8;&#xAE30;&#xC758; &#xC774;&#xD574;&#xB97C; &#xD5A5;&#xD55C; &#xAC10;&#xB3C5;&#xC774; &#xB418;&#xC5C8;&#xC73C;&#xBA70;. &#xC9C0;&#xAE08;&#xAE4C;&#xC9C0;, &#xBAA8;&#xB4E0; &#xC774;&#xB7EC;&#xD55C; &#xC870;&#xC0AC;&#xB294; &#xC2E4;&#xC2DC; &#xC131;&#xC778;&#xC5D0; &#xD558;&#xC9C0;&#xB9CC; &#xC5F0;&#xAD6C; &#xC911; &#xB204;&#xAD6C;&#xB3C4; &#xD574;&#xACB0; &#xBAA8;&#xAE30;&#xC758; &#xC560;&#xBC8C;&#xB808;&#xC640; &#xBC88;&#xB370;&#xAE30; &#xB2E8;&#xACC4; &#xB3D9;&#xC548; &#xC2E0;&#xACBD; &#xC2DC;&#xC2A4;&#xD15C;&#xC758; &#xAC1C;&#xBC1C;. &#xC5EC;&#xAE30;, &#xC6B0;&#xB9AC;&#xB294; &#xBA3C;&#xC800; cross-reacting &#xACE0; Aedes &#xAC1C;&#xBC1C; &#xB450;&#xB1CC;&#xC5D0; &#xC788;&#xB294; epitopes &#xB77C;&#xBCA8; 30 &#xD56D; &#xCCB4;, &#xCD08;&#xD30C;&#xB9AC;, &#xB450;&#xB1CC; &#xAC1C;&#xBC1C;&#xC744; &#xC5F0;&#xAD6C; &#xD558; &#xACE0; 13 &#xADF8;&#xB4E4;&#xC744; &#xC2DD;&#xBCC4; &#xD558;&#xB294; &#xC9D1;&#xD569; &#xD654;&#xBA74;. &#xC6B0;&#xB9AC;&#xB294; &#xB2E4;&#xC74C; &#xC77C;&#xBC18; &#xD574;&#xBD80;&#xD559; &#xAE30;&#xB2A5; &#xAC1C;&#xBC1C; &#xB450;&#xB1CC;&#xC758; &#xD2B9;&#xC131;&#xD654; &#xACE0;, &#xBC88;&#xB370;&#xAE30;, &#xC560;&#xBC8C;&#xB808; &#xBC0F; &#xC131;&#xC778; &#xB2E8;&#xACC4;&#xC5D0;&#xC11C; Drosophila melanogaster &#xC774;&#xC57C;&#xAE30;&#xB3C4; &#xBAA8;&#xB378; &#xC2DC;&#xC2A4;&#xD15C;&#xACFC; &#xBE44;&#xAD50; immunolabeling &#xC5F0;&#xAD6C;&#xC5D0;&#xC11C; &#xD655;&#xC778; &#xB41C; &#xD56D; &#xCCB4;&#xB97C; &#xC0AC;&#xC6A9; &#xD569;&#xB2C8;&#xB2E4;. &#xB610;&#xD55C;, immunolabeling &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xBB38;&#xC11C; Aedes &#xB1CC;, &#xC989; &#xC2DC; &#xB3CC;&#xCD9C;&#xBD80;, subesophageal neuropil &#xBC0F; &#xC790;&#xC138;&#xD788; subesophageal neuropil serotonergic &#xC2DC;&#xC2A4;&#xD15C;&#xC758; &#xD2B9;&#xC815; &#xAD6C;&#xC131; &#xC694;&#xC18C; &#xAC1C;&#xBC1C;. &#xC6B0;&#xB9AC;&#xC758; &#xC5F0;&#xAD6C;&#xB294; Aedes &#xBC88;&#xB370;&#xAE30; (&#xADF8;&#xB9AC;&#xACE0; &#xC131;&#xC778;) &#xB2E8;&#xACC4;&#xC5D0; &#xBE44;&#xD574; &#xC560;&#xBC8C;&#xB808; &#xB2E8;&#xACC4;&#xC5D0;&#xC11C; &#xAC1C;&#xBC1C; &#xB450;&#xB1CC;&#xC5D0;&#xC11C; &#xB208;&#xC5D0; &#xB744;&#xB294; &#xCC28;&#xC774; &#xBCF4;&#xC5EC;&#xC900;&#xB2E4;. &#xACB0;&#xACFC; &#xB610;&#xD55C; &#xD765;&#xBBF8;&#xB85C;&#xC6B4; &#xC720;&#xC0AC;&#xC810;&#xACFC; &#xCD08;&#xD30C;&#xB9AC;&#xC5D0; &#xBE44;&#xD574; Aedes&#xC758; &#xB450;&#xB1CC; &#xAC1C;&#xBC1C;&#xC5D0; &#xD45C;&#xC2DC; &#xB41C; &#xCC28;&#xC774; &#xBC1D;&#xD788;&#xAE30;. &#xD568;&#xAED8; &#xCD2C;&#xC601;,&#xC774; &#xC870;&#xC0AC;&#xAC00; &#xC911;&#xC694; &#xD55C; &#xC9C8;&#xBCD1; &#xBCA1;&#xD130;&#xC5D0; &#xB450;&#xB1CC; &#xAC1C;&#xBC1C;&#xC758; &#xBBF8;&#xB798; &#xC138;&#xD3EC;&#xC9C8;&#xC774; &#xACE0; &#xBD84;&#xC790; &#xC870;&#xC0AC;&#xC5D0; &#xB300; &#xD55C; &#xAE30;&#xCD08;&#xB97C; &#xD615;&#xC131;.

&#xD669;&#xC5F4;&#xBCD1; &#xBAA8;&#xAE30; Aedes aegypti &#xAC1C;&#xBC1C; &#xBE0C;&#xB808;&#xC778;: Drosophila melanogaster&#xC5D0;&#xC11C; cross-reacting &#xD56D; &#xCCB4;&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xBE44;&#xAD50; immunocytochemical &#xBD84;&#xC11D;.

Despite the devastating impact of mosquito-borne illnesses on human health, surprisingly little is known about mosquito developmental biology, including development of the olfactory system, a tissue of vector importance. Analysis of mosquito olfactory developmental genetics has been hindered by a lack of means to target specific genes during the development of this sensory system. In this investigation, chitosan/siRNA nanoparticles were used to target semaphorin-1a (sema1a) during olfactory system development in the dengue and yellow fever vector mosquito Aedes aegypti. Immunohistochemical analyses and anterograde tracing of antennal sensory neurons, which were used to track the progression of olfactory development in this species, revealed antennal lobe defects in sema1a knockdown fourth instar larvae. These findings, which correlated with a larval odorant tracking behavioral phenotype, identified previously unreported roles for Sema1a in the developing insect larval olfactory system. Analysis of sema1a knockdown pupae also revealed a number of olfactory phenotypes, including olfactory receptor neuron targeting and projection neuron defects coincident with a collapse in the structure and shape of the antennal lobe and individual glomeruli. This study, which is to our knowledge the first functional genetic analysis of insect olfactory development outside of D. melanogaster, identified critical roles for Sema1a during Ae. aegypti larval and pupal olfactory development and advocates the use of chitosan/siRNA nanoparticles as an effective means of targeting genes during post-embryonic Ae. aegypti development. Use of siRNA nanoparticle methodology to understand sensory developmental genetics in mosquitoes will provide insight into the evolutionary conservation and divergence of key developmental genes which could be exploited in the development of both common and species-specific means for intervention.

Disruption of Aedes aegypti olfactory system development through chitosan/siRNA nanoparticle targeting of semaphorin-1a.

Essentially nothing is known about the genetic regulation of olfactory system development in vector mosquitoes, which use olfactory cues to detect blood meal hosts. Studies in Drosophila melanogaster have identified a regulatory matrix of transcription factors that controls pupal/adult odorant receptor (OR) gene expression in olfactory receptor neurons (ORNs). However, it is unclear if transcription factors that function in the D. melanogaster regulatory matrix are required for OR expression in mosquitoes. Furthermore, the regulation of OR expression during development of the larval olfactory system, which is far less complex than that of pupae/adults, is not well understood in any insect, including D. melanogaster. Here, we examine the regulation of OR expression in the developing larval olfactory system of Aedes aegypti, the dengue vector mosquito.

Chitosan/siRNA nanoparticle targeting demonstrates a requirement for single-minded during larval and pupal olfactory system development of the vector mosquito Aedes aegypti.

Despite the devastating impact of mosquito-borne illnesses on human health, very little is known about mosquito developmental biology, including development of the mosquito visual system. Mosquitoes possess functional adult compound eyes as larvae, a trait that makes them an interesting model in which to study comparative developmental genetics. Here, we functionally characterize visual system development in the dengue and yellow fever vector mosquito Aedes aegypti, in which we use chitosan/siRNA nanoparticles to target the axon guidance gene semaphorin-1a (sema1a).

Role of semaphorin-1a in the developing visual system of the disease vector mosquito Aedes aegypti.

Most animal species exhibit sexually dimorphic behaviors, many of which are linked to reproduction. A number of these behaviors, including blood feeding in female mosquitoes, contribute to the global spread of vector-borne illnesses. However, knowledge concerning the genetic basis of sexually dimorphic traits is limited in any organism, including mosquitoes, especially with respect to differences in the developing nervous system.

Keshava Mysore

Department of Medical and Molecular Genetics

Indiana University School of Medicine

Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae

Keshava Mysore

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Medical and Molecular Genetics

Indiana University School of Medicine

Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae

Department of Medical and Molecular Genetics