Name: visualization of larval segmental nerves in 3rd instar drosophila larval preparations
Uploaded: 2010-09-29T17:00:00
Description: Watch this Scientific Journal Video about Visualization of Larval Segmental Nerves in 3rd Instar Drosophila Larval Preparations at JoVE.com

SG è sostenuto da fondi del State University di New York a Buffalo e da John R. Oishei Foundation.

1. Preparazione dei reagenti <ol><li> Preparare 1x buffer Dissection a 128 mM NaCl, CaCl 2 4mm, 4 mm MgCl 2, 2 mM KCl, HEPES 5 mm e 36 mm saccarosio. Il pH della soluzione a 7,2 e filtro sterilizzare. </li><li> Preparare il fissativo con diluizione 1:1 del 16% di formaldeide e Buffer Dissection 1x. </li><li> Preparare 1x PBT utilizzando 1x soluzione tampone fosfato (PBS) a pH di 7,2 e Triton X-100 in un rapporto 1:100. </li><li> Preparare il 5% di albumina sierica bovina (BSA) con 0,01% Na azide ...

<ol>
	<li>Gunawardena, S., Goldstein, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disruption+of+Axonal+Transport+and+Neuronal+Viability+by+Amyoid+Precursor+Protein+Mutations+in+Drosophila.">Disruption of Axonal Transport and Neuronal Viability by Amyoid Precursor Protein Mutations in Drosophila.</a> Neuron. 32 (2), 389-401 (2001).</li><li>Gunawardena, S., Her, L. S., Brusch, R. G., Laymon, R. A., Niesman, I. R., Gordesky-Gold, B., Sintasath, L., Bonini, N. M., Goldstein, L. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disruption+of+axonal+transport+by+loss+of+huntingtin+or+expression+of+pathogenic+polyQ+proteins+in+Drosophila.">Disruption of axonal transport by loss of huntingtin or expression of pathogenic polyQ proteins in Drosophila.</a> Neuron. 40 (1), 1-2 (2003).</li><li>Hurd, D. D., Saxton, W. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinesin+mutations+cause+motor+neuron+disease+phenotypes+by+disrupting+fast+axonal+transport+in+Drosophila.">Kinesin mutations cause motor neuron disease phenotypes by disrupting fast axonal transport in Drosophila.</a> Genetics. 144 (3), 1075-1085 (1996).</li><li>Zinsmaier, K. E., Eberle, K. K., Buchner, E., Walter, N., Benzer, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Paralysis+and+early+death+in+cysteine+string+protein+mutants+of+Drosophila.">Paralysis and early death in cysteine string protein mutants of Drosophila.</a> Science. 263, 977-980 (1994).</li></ol>

I nervi larvale Drosophila segmentale sono un potente sistema per lo studio dei meccanismi di trasporto assonale. Se il 3 ° instar dissezione larvale viene eseguita correttamente, tutte le componenti del sistema nervoso centrale, PNS, e tessuti associati, come i muscoli e NMJs, possono essere esaminate all&#39;interno di una singola larva. Abbiamo adattato questo protocollo da quella pubblicata da Hurd e Saxton (1996). Questo protocollo può essere adattato anche a test altri anticorpi neuronali, ma...

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visualization of larval segmental nerves in 3rd instar drosophila larval preparations

Drosophila melanogaster sta emergendo come un potente sistema modello per studiare lo sviluppo e la funzione del sistema nervoso, in particolare a causa della sua genetica e conveniente genoma completamente sequenziato. Inoltre, il sistema nervoso larvale è un sistema modello ideale per studiare i meccanismi del trasporto assonale, come i nervi larvale segmentale contenere fasci di assoni con i loro corpi cellulari situato all&#39;interno del cervello e le loro terminazioni nervose finale lungo la lunghezza del corpo. Qui si descrive la procedura per la visualizzazione di proteine ​​delle vescicole sinaptiche all&#39;interno larvale nervi segmentale. Se fatto correttamente, tutte le componenti del sistema nervoso, con relativi tessuti come i muscoli e NMJs, rimane intatto, integro e pronto per essere visualizzato. 3 ° larve instar portatori di mutazioni diverse sono sezionati, fisso, incubate con anticorpi delle vescicole sinaptiche, visualizzati e confrontati con le larve wild-type. Questa procedura può essere adattata per diversi anticorpi diversi sinaptica o neuronale e cambiamenti nella distribuzione di una varietà di proteine ​​può essere facilmente osservata entro larvale nervi segmentale.

Watch this Scientific Journal Video about Visualization of Larval Segmental Nerves in 3rd Instar Drosophila Larval Preparations at JoVE.com

Visualization of Larval Segmental Nerves in 3rd Instar Drosophila Larval Preparations

Larve di Drosophila melanogaster fornire un sistema modello ideale per studiare i meccanismi di trasporto assonale in nervi larvale segmentale. Utilizzando questa procedura, 3 ° larve instar portatori di mutazioni varie può essere paragonato a larve wild-type.

Visualizzazione dei nervi larvali segmentale in 3 Rd Instar Drosophila I preparativi larvali

Drosophila melanogaster is emerging as a powerful model system for studying the development and function of the nervous system, particularly because of ...

Research

JoVE Journal

Neuroscience

Visualization of Larval Segmental Nerves in 3rd Instar Drosophila Larval Preparations

In vivo Visualization of Synaptic Vesicles Within Drosophila Larval Segmental Axons

In vivo Visualization of Synaptic Vesicles Within Drosophila Larval Segmental Axons

Nella visualizzazione in vivo delle vescicole sinaptiche In Drosophila assoni larvali segmentale

Elucidating the mechanisms of axonal transport has shown to be very important in determining how defects in long distance transport affect different ...

Ricerca

Neuroscienze

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The Gal4/ UAS binary method is powerful for gene and neural circuitry manipulation in Drosophila. For most neurobiological studies, however, Gal4 ...

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Most studies on morphogenesis rely on qualitative descriptions of how anatomical traits are affected by the disruption of specific genes and genetic ...

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

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Cella singola risoluzione fluorescenza Live Imaging di Drosophila orologi circadiani nel cervello larvale cultura

The circadian pacemaker circuit orchestrates rhythmic behavioral and physiological outputs coordinated with environmental cues, such as day/night cycles. ...

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Il comportamento larvale della drosofila in risposta alla stimolazione optogenetica dei neuroni olfattivi di rilevamento

The ability of insects to navigate toward odor sources is based on the activities of their first-order olfactory receptor neurons (ORNs). While a ...