Name: morphological analysis of drosophila larval peripheral sensory neuron dendrites and axons using genetic mosaics
Uploaded: 2011-11-07T12:00:00
Description: Watch this Scientific Journal Video about Morphological Analysis of Drosophila Larval Peripheral Sensory Neuron Dendrites and Axons Using Genetic Mosaics at JoVE.com

Gli autori ringraziano RIKEN per il finanziamento. Ringraziamo anche Cagri Yalgin, Caroline Delandre, e Jay Parrish per le discussioni sui protocolli di genetica e di immunoistochimica.

1.Preparazione di reagenti <ol><li> Preparare Ca + +-libero HL3.1 saline 28. <ol><li> In mM: 70 NaCl, 5 KCl, 20 MgCl 2, 10 NaHCO 3, 5 HEPES, 115 saccarosio, trealosio e 5; pH 7,2. Filtro sterilizzare e conservare a 4 ° C. Nota: Ca + +-senza soluzione impedisce la contrazione muscolare durante la dissezione. </li></ol></li><li> Fai la poli-L-lisina (PLL) coprioggetto. <ol><li> Sciogliere 100mg PLL in acqua 4.2ml e fare aliquote 300μl in tubi Eppendorf e co...

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	<li>Grueber, W. B., Jan, L. Y., Jan, Y. 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La Drosophila larvale DA modello neurone fornisce un ottimo sistema per investigare i meccanismi genetici che controllano la morfologia dei neuroni e la formazione di circuito. MARCM è generalmente utilizzato per l&#39;etichettatura e per la generazione di mutanti DA cloni neurone. Per MARCM abbiamo utilizzare un pan-neurale (es. GAL4 C155) o DA neurone specifico driver. Utilizzando un pan-neurali del driver è possibile usare direttamente diversi stock ampiamente disponibili presso...

<table border="1"><tr><th> Nome del reagente </th><th> Azienda </th><th> Numero di catalogo </th><th> Commenti (opzionale) </th></tr><tr><td> SZX16 dissezione microscopio a fluorescenza (con GFPHQ filtro) </td><td> Olimpo </td><td> SZX16 </td><td></td></tr><tr><td> Vivere Pinza insetti </td><td> FST </td><td> 26030-10 </td><td></td></tr><tr><td> 26 millimetri x 76mm depressione vetrini </td><td> Toshinriko Co. </td><td> T8-R004 </td><td></td></tr><tr><td> Sylgard 184 (o Silpot 184) </td><td> Dow Corning </td><td> 3097358-1004 </td><td></td></tr><tr><td> Poli-L-lisina </td><td> Sigma </td><td> P-1524 </td><td> Questo prodotto si è dimostrato più efficace </td></tr><tr><td> DPX montaggio medio </td><td> Sigma </td><td> 44581 </td><td></td></tr><tr><td> Coniglio anti-GFP </td><td> Invitrogen </td><td> A-11122 </td><td> Diluizione 1:500 </td></tr><tr><td> Rat anti-CD8 </td><td> Caltag </td><td> 5H10 </td><td> Diluizione 1:200 </td></tr><tr><td> Topo anti-CD2 </td><td> AbD Serotec </td><td> MCA443R </td><td> Diluizione 1:700 </td></tr><tr><td> Topo anti-Fasciclin2 </td><td> DSHB </td><td> 1d4 </td><td> Diluizione 1:10 </td></tr></table>

morphological analysis of drosophila larval peripheral sensory neuron dendrites and axons using genetic mosaics

Sviluppo del sistema nervoso richiede la corretta specificazione della posizione dei neuroni e l&#39;identità, seguita da accurate neurone classe specifico per lo sviluppo dendritiche e cablaggio assonale. Recentemente l&#39;arborizzazione dendritica (DA) neuroni sensoriali del sistema nervoso periferico Drosophila larvale (PNS) sono diventati potenti modelli genetici in cui per chiarire sia i meccanismi generali e specifiche della classe di differenziazione dei neuroni. Ci sono quattro principali classi di neuroni DA (I-IV) 1. Essi sono chiamati in ordine crescente di complessità dendrite pergolato, e sono specifiche della classe differenze nel controllo genetico della loro differenziazione 2-10. Il sistema DA sensoriale è un modello pratico per studiare i meccanismi molecolari alla base il controllo della morfologia dendritica 11-13 perché: 1) si può usufruire di potenti strumenti genetici disponibili nel moscerino della frutta, 2) il procuratore neurone dendrite pergolato si estende in soli 2 dimensioni sotto un ottica CLEar cuticola larvale che la rende facile la visualizzazione ad alta risoluzione in vivo, 3) la classe-specifica diversità nella morfologia dendritica facilita un&#39;analisi comparativa per trovare elementi chiave Controllando la formazione di alberi dendritici semplici vs altamente ramificata, e 4) dendritiche pergolato stereotipata forme differenti di neuroni DA facilitare morfometriche analisi statistiche. DA l&#39;attività dei neuroni modifica l&#39;uscita di un generatore di pattern locomozione larvale centrale 14-16. Le diverse classi di neuroni DA sono distinte modalità sensoriali, e la loro attivazione provoca diverse risposte comportamentali 14,16-20. Inoltre, diverse classi di inviare proiezioni stereotipo assonale nel sistema nervoso centrale Drosophila larvale nel cordone nervoso ventrale (VNC) 21. Queste proiezioni terminare con le rappresentazioni topografiche di entrambe le modalità DA neuroni sensoriali e la posizione nella parete del corpo del campo dendritiche 7,22, 23. Quindi l&#39;esame di proiezioni DA assonale può essere usata per chiarire i meccanismi alla base mappatura topografica 7,22,23, così come il cablaggio di un semplice locomozione larvale circuito modulante 14-17. Vi presentiamo qui una guida pratica per generare e analizzare i mosaici genetici 24 neuroni DA marcatura tramite MARCM (Analisi Mosaico con un marcatore cellulare reprimibile) 1,10,25 e Flp-out 22,26,27 tecniche (riassunti nella fig. 1).

Watch this Scientific Journal Video about Morphological Analysis of Drosophila Larval Peripheral Sensory Neuron Dendrites and Axons Using Genetic Mosaics at JoVE.com

Morphological Analysis of Drosophila Larval Peripheral Sensory Neuron Dendrites and Axons Using Genetic Mosaics

I neuroni sensoriali arborizzazione dendritica del<em&gt; Drosophila</em&gt; Larvale sistema nervoso periferico sono modelli utili per chiarire sia generale e classe di neuroni specifici meccanismi di differenziazione dei neuroni. Vi presentiamo una guida pratica per generare e analizzare arborizzazione dendritica dei neuroni mosaici genetici.

Analisi morfologica del Drosophila Larvale dendriti sensoriali e periferico Neuron assoni utilizzo Mosaici genetici

Nervous system development requires the correct specification of neuron position and identity, followed by accurate neuron class-specific dendritic ...

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Morphological Analysis of Drosophila Larval Peripheral Sensory Neuron Dendrites and Axons Using Genetic Mosaics

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Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

Tutta Monte immunomarcatura di recettore olfattivo neuroni nel<em&gt; Drosophila</em&gt; Antenna

Odorant molecules bind to their target receptors in a precise and coordinated manner. Each receptor recognizes a specific signal and relays this ...

Production and Isolation of Axons from Sensory Neurons for Biochemical Analysis Using Porous Filters

Produzione e isolamento degli assoni dai neuroni sensoriali per l&#39;analisi biochimica Uso dei filtri porosi

Neuronal axons use specific mechanisms to mediate extension, maintain integrity, and induce degeneration. An appropriate balance of these events is ...

Removal of Drosophila Muscle Tissue from Larval Fillets for Immunofluorescence Analysis of Sensory Neurons and Epidermal Cells

Removal of Drosophila Muscle Tissue from Larval Fillets for Immunofluorescence Analysis of Sensory Neurons and Epidermal Cells

Rimozione di<em&gt; Drosophila</em&gt; Tessuto muscolare da larvali Filetti per l&#39;analisi di immunofluorescenza dei neuroni sensoriali e cellule epidermiche

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

Using Linear Agarose Channels to Study Drosophila Larval Crawling Behavior

Using Linear Agarose Channels to Study Drosophila Larval Crawling Behavior

Utilizzando canali lineari Agarose per studiare<em&gt; Drosophila</em&gt; Comportamento larvale Crawling

Drosophila larval crawling is emerging as a powerful model to study neural control of sensorimotor behavior. However, larval crawling behavior on flat ...

A Microbiomechanical System for Studying Varicosity Formation and Recovery in Central Neuron Axons

Un sistema di Microbiomechanical per lo studio della formazione del Varicosity e del recupero in centrale del neurone assoni

Axonal varicosities are enlarged structures along the shafts of axons with a high degree of heterogeneity. They are present not only in brains with ...