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 wordt ondersteund door middelen van de State University van New York in Buffalo en van John R. Oishei Foundation.

1. Voorbereiding van de reagentia <ol><li> Bereid 1x Dissection buffer met 128 mM NaCl, 4mm CaCl 2, 4mm MgCl 2, 2 mM KCl, 5mM HEPES en 36mm Sucrose. PH de oplossing voor 7.2 en filteren steriliseren. </li><li> Bereid het fixatief met behulp van 01:01 verwatering van 16% formaldehyde en 1x Dissection Buffer. </li><li> Bereid 1x PBT met 1x fosfaat gebufferde zoutoplossing (PBS) bij een pH van 7,2 en Triton X-100 in een 1:100 verhouding. </li><li> Bereid 5% Bovine Serum Albumine (BSA) met 0,01% Na A...

<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>

De Drosophila larvale segmentale zenuwen zijn een krachtig systeem om mechanismen in axonaal transport te bestuderen. Als de 3 e instar larven dissectie correct wordt uitgevoerd, kan alle componenten van het CZS, PNS, en de bijbehorende weefsel, zoals de spieren en NMJs, worden onderzocht binnen een enkele larve. We hebben aangepast dit protocol bij het ene gepubliceerd door Hurd en Saxton (1996). Dit protocol kan ook worden aangepast aan andere neuronale antistoffen te testen, maar optimalisatie van...

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

Drosophila melanogaster is in opkomst als een krachtige modelsysteem voor het bestuderen van de ontwikkeling en functie van het zenuwstelsel, met name vanwege de gunstige genetica en volledig gesequenced genoom. Bovendien, het larvale zenuwstelsel is een ideaal modelsysteem om de mechanismen van axonaal transport studie als het larvale segmentale zenuwen bevatten bundels van axonen met hun cellichamen gelegen in de hersenen en hun zenuwuiteinden eindigen langs de lengte van het lichaam. Hier beschrijven we de procedure voor de visualisatie van synaptische vesicles eiwitten in larvale segmentale zenuwen. Indien correct gedaan, alle componenten van het zenuwstelsel, samen met de omliggende weefsels, zoals spieren en NMJs, blijven intact, onbeschadigd, en klaar om te worden gevisualiseerd. 3 e instar larven voeren verschillende mutaties worden ontleed, vaste, geïncubeerd met synaptische blaasje antilichamen, gevisualiseerd en vergeleken met wild type larven. Deze procedure kan worden aangepast voor verschillende synaptic of neuronale antilichamen en veranderingen in de verdeling van een verscheidenheid van eiwitten kunnen gemakkelijk worden waargenomen binnen de larvale segmentale zenuwen.

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

Drosophila melanogaster larven zorgen voor een ideaal modelsysteem om de mechanismen van het axonaal transport te onderzoeken binnen de larvale segmentale zenuwen. Met behulp van deze procedure, kan 3 e instar larven dragen verschillende mutaties worden vergeleken met wild-type larven.

Visualisatie van larvale Segmentale zenuwen in 3 Rd Instar Drosophila Larvale Voorbereidingen

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

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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

In vivo visualisatie van synaptische blaasjes Binnen Drosophila larven Segmentale Axonen

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

Mapping and Application of Enhancer-trap Flippase Expression in Larval and Adult Drosophila CNS

Mapping and Application of Enhancer-trap Flippase Expression in Larval and Adult Drosophila CNS

In kaart brengen en de toepassing van Enhancer-trap Flippase Expression in larvale en volwassen Drosophila CNS

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

Dissection and Immunohistochemistry of Larval, Pupal and Adult Drosophila Retinas

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Dissectie en Immunohistochemie van larven, popstadium en Volwassen<em&gt; Drosophila</em&gt; Netvliezen

The compound eye of Drosophila melanogaster consists of about 750 ommatidia (unit eyes). Each ommatidium is composed of about 20 cells, including ...

Forebrain Electrophysiological Recording in Larval Zebrafish

Voorhersenen Elektrofysiologische Opname in larvale zebravis

Epilepsy affects nearly 3 million people in the United States and up to 50 million people worldwide. Defined as the occurrence of spontaneous unprovoked ...

Functional Analysis of the Larval Feeding Circuit in Drosophila

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Functionele analyse van de larvenvraat Circuit in Drosophila</i

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

Live Imaging of Drosophila Larval Neuroblasts

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Stem cells divide asymmetrically to generate two progeny cells with unequal fate potential: a self-renewing stem cell and a differentiating cell. Given ...

Why Quantification Matters: Characterization of Phenotypes at the Drosophila Larval Neuromuscular Junction

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Waarom kwantificering Matters: karakterisering van fenotypes bij de<em&gt; Drosophila</em&gt; Larvale Neuromusculaire Junction

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

Using Linear Agarose Channels to Study Drosophila Larval Crawling Behavior

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Met behulp van Linear Agarose kanalen om te studeren<em&gt; Drosophila</em&gt; Larvale Crawling Behavior

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

Single-cell Resolution Fluorescence Live Imaging of Drosophila Circadian Clocks in Larval Brain Culture

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Eencellige resolutie fluorescentie leven beeldvorming van Drosophila circadiane klokken in larvale hersenen cultuur

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

Tracking Drosophila Larval Behavior in Response to Optogenetic Stimulation of Olfactory Neurons

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Bijhouden van Drosophila larvale gedrag in reactie op de stimulatie van de Optogenetic van olfactorische neuronen

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