Name: functional analysis of the larval feeding circuit in drosophila
Uploaded: 2013-11-19T16:45:00
Description: Watch this Scientific Journal Video about Functional Analysis of the Larval Feeding Circuit in Drosophila at JoVE.com

Forfatterne vil gerne anerkende præsidentens Research Fund fra Saint Louis University tildelt WSN

1.. Vedligeholdelse af Befolkning Bure <ol><li> Oprethold befolkningsgrupper bure ved 25 ° C på en 12 timers lys-mørke-cyklus. Så længe kontrol-og forsøgsgrupper er udsat for de samme lysforhold, så denne teknik kan udføres i en standard laboratorium indstilling. </li><li> Tillad hunner til at lægge æg natten over på æblejuice-agarplader. </li><li> Saml nyklækkede larver ved at opretholde plader med nyligt deponerede æg ved 25 ° C i 24 timer. Placer en lille klat af gær i midten af ​​pladen for at ...

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Afvigende udvikling af det serotonerge stomatogastric kredsløb, som opstår under sen embryogenese vil påvirke dens modne funktion. Ændringer i neurit arkitektur innerverer tarmen kan korreleres med den funktionelle udgang af kredsløbet, som er fodring hastighed (målt ved munden krog sammentrækninger i en gær opløsning) (figur 1). Anvendelsen af UAS-Gal4 todelte system Drosophila gør det muligt at målrette op-eller nedreguleret ekspression af et bestemt transkript til et specifikt væ...

Drosophila er en ekstremt kraftfuld model system til at studere neurale udvikling kredsløb grund af den hurtige generationstid, lave eksperimenterende omkostninger, og evnen til at manipulere og kontrollere genetiske og miljømæssige faktorer. Neurogenese, neuronal sti fund og synaptogenesis bevares mellem mennesker og Drosophila, derfor de mekanismer i at skabe, vedligeholde og modificere neurale kredsløb bevares så godt. Klassiske neurotransmittere, såsom serotonin (5-hydroxytryptamin eller 5-HT) kan tjene som vækstfaktorer inden vedtagelsen af deres roller som signalmolekyler i den modne neurale kredsløb 1-3 Tidligere undersøgelser har vist, at urolig niveauer af 5-HT under embryogenese ændre tilslutning af modne neuroner 4. Andre har vist, at ektopisk anvendelse af 5-HT til dyrkede Helisoma neuroner undertrykke neuritudvækst samt synaptogenesen 5-7. I Drosophila er udviklingsmæssige 5-HT omvendt relateret til varicosity antal og størrelse samt graden af aborization, langs længden af neuritter, der rager til foregut fra CNS 8. Serotonerg neurotransmission har vist sig at modulere fodring adfærd i forskellige arter, herunder Drosophila 8-9. Fodring kredsløb i Drosophila er et relativt simpelt kredsløb, der kan anvendes som en model til at korrelere den funktionelle udgang (fodring) med ændringer i udviklingen af axonale projektioner fra hjernen til foregut. Schoofs et al. har vist, at Drosophila larve fodring er reguleret af centrale mønster-generatorer, der påvirker muskulaturen 10. Den specifikke muskulære anatomi ikke er helt forstået, er det blevet vist, at antennal nerve, maxillary nerve, og prothoracic tilbehør nerve er ansvarlige for de muskulære mål involveret ispiseadfærd. De fleste data, der omfatter muskulaturen og nerve anatomi hvirvelløse fodring er begrænset til Calliphora larver. Fodring på andet stadium larver kan vurderes ved tilbagetrækning af cephalopharyngeal sclerites (munden kroge), og er reproducerbar og high-throughput. De cephalopharyngeal plader innerveret af fibre fra centrale 5-HT neuroner via frontal nerve. Proventriculus eller foregut, innerveres af serotonerge fibre (recurrens nerve) at fasciculate i mellemtarmen og er ansvarlige for sammentrækninger af foregut (figur 1) 11-12. Ændringer i axonal forgrening, og antallet og størrelsen af ​​varicosities langs neurit længde, kan kvantificeres ved hjælp af immunhistokemiske teknikker. Manipulering neuronal 5-HT under udvikling, enten direkte eller indirekte, kan ændre den funktionelle produktionen af ​​denne fodring kredsløb, som kan vurderes og sammenholdes med ændringer i morphology af neurit arkitektur.

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functional analysis of the larval feeding circuit in drosophila

Serotonerge fodring kredsløb i Drosophila melanogaster-larver kan anvendes til at undersøge neurale substrater af afgørende betydning i forbindelse med udviklingen af kredsløbet. Brug den funktionelle output af kredsløbet, fodring, ændringer i neuronale arkitektur stomatogastric systemet kan visualiseres. Spiseadfærd kan optages ved at observere graden af ​​tilbagetrækning af munden kroge, som modtager innervation fra hjernen. Lokomotorisk adfærd anvendes som fysiologisk regulering for fodring, da larver bruge deres mund kroge til at krydse over en agar substrat. Ændringer i spiseadfærd kan korreleres med axonal arkitektur af neuritter innerverer tarmen. Brug immunhistokemi er det muligt at visualisere og kvantificere disse ændringer. Forkert håndtering af larver under opførsel paradigmer kan ændre data, da de er meget følsomme over for manipulationer. Korrekt billeddannelse af neurite arkitektur innerverertarmen er kritisk for nøjagtig kvantificering af antallet og størrelsen af ​​varicosities samt omfanget af branch noder. Analyse af de fleste kredsløb tillader kun til visualisering af neurite arkitektur eller adfærdsmæssige virkninger, men denne model giver en at korrelere den funktionelle output kredsløb med funktionsnedsættelser i neuronal arkitektur.

Serotonerge fodring kredsløb i Drosophila larve kan tjene som en yderst effektiv model til at observere indflydelsen af særlige faktorer på nervesystemet udvikling. Ved kvantificering fodring hastighed, er det muligt at forbinde den axonale arkitektur fodring kredsløb med dets funktionelle udgang (figur 1). Bevægelsesapparatet assay anvendes som en fysiologisk regulering for tilbagetrækninger af munden kroge, idet larver bruge deres mund kroge til at fremdrive sig over overfladen af ​?...

Watch this Scientific Journal Video about Functional Analysis of the Larval Feeding Circuit in Drosophila at JoVE.com

Functional Analysis of the Larval Feeding Circuit in Drosophila (Video) | JoVE

Fodring kredsløb i Drosophila melanogaster larver serverer en simpel endnu kraftfulde model, der tillader ændringer i fodring sats at være korreleret med ændringer i stomatogastric neurale kredsløb. Dette kredsløb består af centrale serotonerge neuroner, der sender fremspring til munden kroge samt foregut.

Funktionel analyse af larve Fodring Circuit i Drosophila</i

The serotonergic  feeding circuit in Drosophila  melanogaster larvae can be used to investigate neuronal substrates of  critical importance during the ...

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Neuroscience

Functional Analysis of the Larval Feeding Circuit in Drosophila

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Drosophila larval dendritic arborization (da) neurons are a popular model for investigating mechanisms of neuronal morphogenesis. Da neurons develop in ...

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Focal Macropatch optagelser af Synaptic strømninger fra Drosophila larve neuromuskulære Junction

Drosophila neuromuscular junction (NMJ) is an excellent model system to study glutamatergic synaptic transmission. We describe the technique of focal ...