Name: immunohistological labeling of microtubules in sensory neuron dendrites, tracheae, and muscles in the drosophila larva body wall
Uploaded: 2011-11-10T13:00:00
Description: Watch this Scientific Journal Video about Immunohistological Labeling of Microtubules in Sensory Neuron Dendrites, Tracheae, and Muscles in the Drosophila Larva Body Wall at JoVE.com

Wij danken RIKEN voor financiering. P10-Gal4 was een soort geschenk van Alain Vincent (Universite Paul Sabatier, Toulouse, Frankrijk).

1. Bereiding van reagentia Opmerkingen voor het begin van: Dissection en immunohistochemische kleuring worden uitgevoerd in een magnetische kamer en de larve wordt vastgepind met behulp van speciaal gevormde insect pinnen. Gedetailleerde instructies voor de bouw van een magnetische kamer, en voorbereiding van deze pinnen is te vinden in gerelateerde referenties 14,15. In het kort, is een 1x1cm vierkant gat gesneden in een magnetische plaat en een dekglaasje aangebra...

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Om te begrijpen hoe complex cel vormen zijn bereikt, is het belangrijk om te kunnen nauwkeurig test microtubuli organisatie. Hier beschrijven we een robuuste immunohistologische etikettering methode voor het testen van microtubuli organisatie van de dendritische arborization neuron zintuiglijke dendrieten. In aanvulling op vlekken sensorische neuronen, deze methode bereikt robuuste immunohistologische kleuring van luchtpijp, spieren en andere lichaamsdelen muur weefsels. Wij gebruiken dit pr...

<table border="1"><tbody><tr><th> Naam van het reagens </th><th> Vennootschap </th><th> Catalogus aantal </th><th> Reacties (Optioneel) </th></tr><tr><td> Pincet </td><td> Dumont </td><td> 11251-20 </td><td></td></tr><tr><td> Microscissors </td><td> FST </td><td> 15000-08 </td><td></td></tr><tr><td> Muis anti-α-tubuline (Clone: ​​DM1A) </td><td> Sigma </td><td> T9026 </td><td> Verdunning 1 / 1000 </td></tr><tr><td> Muis anti-Futsch (Clone: ​​22C10), supernatant </td><td> Developmental Studies Hybridoma Bank </td><td> 22C10 </td><td> Verdunning 1 / 1000 </td></tr><tr><td> Rat anti-CD8 (Clone: ​​5H10) </td><td> Caltag </td><td> MCD0800 </td><td> Verdunning 1 / 1000 </td></tr><tr><td> Alexa Fluor 488 anti-muis IgG </td><td> Invitrogen </td><td> A-11.001 </td><td> Verdunning 1 / 500 </td></tr><tr><td> Cy3 anti-Rat IgG </td><td> Jackson Immunoresearch </td><td> 712-166-150 </td><td> Verdunning 1 / 200 </td></tr></tbody></table>

immunohistological labeling of microtubules in sensory neuron dendrites, tracheae, and muscles in the drosophila larva body wall

Om te begrijpen hoe verschillen in complexe cellen vormen zijn bereikt, is het belangrijk om nauwkeurig te volgen microtubuli organisatie. De Drosophila larven lichaam muur bevat verschillende celtypen die zijn modellen om cellen en weefsels morfogenese bestuderen. Bijvoorbeeld luchtpijp worden gebruikt om de buis morfogenese 1, en ​​de dendritische arborization (DA) sensorische neuronen van de Drosophila larven hebben een primaire systeem worden voor de opheldering van algemene en neuron-class-specifieke mechanismen van dendritische differentiatie 2-5 en degeneratie zes onderzoeken . De vorm van dendriet takken kunnen sterk variëren tussen neuron klassen, en zelfs tussen verschillende takken van een enkel neuron 7,8. Genetische studies in DA neuronen suggereren dat differentieel organisatie van het cytoskelet kan morfologische verschillen in dendritische branche vorm 4,9-11 ten grondslag liggen. Wij bieden een robuuste immunologische etikettering methode om eenssay in vivo microtubuli organisatie in DA sensorische neuron dendriet prieel (figuren 1, 2, Film 1). Dit protocol geeft de dissectie en immunokleuring van de eerste instar larve, een stadium waarin actieve sensorische neuron dendriet uitgroei en vertakking organisatie is 12,13 optreedt. In aanvulling op vlekken sensorische neuronen, deze methode bereikt robuuste kenmerken van microtubuli organisatie in de spieren (Movies 2, 3), luchtpijp (Figuur 3, Movie 3), en andere lichaamsdelen muur weefsels. Het is waardevol voor onderzoekers die willen microtubuli organisatie te analyseren in situ in het lichaam muur bij het ​​onderzoek naar de mechanismen die de controle weefsels en cellen vorm.

Watch this Scientific Journal Video about Immunohistological Labeling of Microtubules in Sensory Neuron Dendrites, Tracheae, and Muscles in the Drosophila Larva Body Wall at JoVE.com

Immunohistological Labeling of Microtubules in Sensory Neuron Dendrites, Tracheae, and Muscles in the Drosophila Larva Body Wall

Om te begrijpen hoe complex cel vormen, zoals neuronale dendrieten, worden bereikt tijdens de ontwikkeling, is het belangrijk om nauwkeurig te kunnen assay microtubuli organisatie. Hier beschrijven we een robuuste immunohistologische etikettering methode om microtubule organisatie van de dendritische arborization neuron zintuiglijke dendrieten, de luchtpijp, spier-onderzoeken en andere<em&gt; Drosophila</em&gt; Larve lichaamswand weefsels.

Immunohistologische Etikettering van microtubuli in Sensory Neuron Dendrieten, luchtpijp, en de spieren in de Drosophila Larve Body Muur

To understand how differences in complex cell shapes are achieved, it is important to accurately follow microtubule organization. The Drosophila larval ...

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Neuroscience

Immunohistological Labeling of Microtubules in Sensory Neuron Dendrites, Tracheae, and Muscles in the Drosophila Larva Body Wall

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