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

Die Autoren danken RIKEN für die Finanzierung. Wir danken auch Cagri Yalgin, Caroline Delandre und Jay Parrish für Diskussionen über genetische und Immunhistochemie-Protokollen.

1.Vorbereitung von Reagenzien <ol><li> Bereiten Ca + +-freien HL3.1 Kochsalzlösung 28. <ol><li> In mM: 70 NaCl, 5 KCl, 20 MgCl 2, 10 NaHCO 3, 5 HEPES, 115 Saccharose und 5 Trehalose; pH 7,2. Filter sterilisieren und lagern bei 4 ° C. Hinweis: Ca + +-freier Lösung verhindert Muskelkontraktion beim Präparieren. </li></ol></li><li> Machen Poly-L-Lysin (PLL) Deckgläser. <ol><li> Lösen 100mg PLL in 4.2ml Wasser und machen 300μl Aliquots in Eppendorf-Röhr...

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Die Drosophila Larve DA Neuronenmodell liefert eine hervorragende genetische System von Mechanismen zu untersuchen, dass die Kontrolle Neuron Morphologie und Schaltung Bildung. MARCM ist in der Regel für die Kennzeichnung und zur Erzeugung von Mutanten DA Neuron Klone verwendet. Für MARCM wir verwenden entweder eine pan-neuronalen (zB Gal4 C155) oder DA Neuronen-spezifische Treiber. Mit einem pan-neuronaler Fahrer ist es möglich, direkt zu verwenden mehrere Bestände weit aus öff...

<table border="1"><tr><th> Name des Reagenzes </th><th> Firma </th><th> Katalog-Nummer </th><th> Kommentare (optional) </th></tr><tr><td> SZX16 Fluoreszenz Dissektionsmikroskop (mit GFPHQ Filter) </td><td> Olymp </td><td> SZX16 </td><td></td></tr><tr><td> Live-Insect Zange </td><td> FST </td><td> 26030-10 </td><td></td></tr><tr><td> 26mm x 76mm Depression Objektträger </td><td> Toshinriko Co. </td><td> T8-R004 </td><td></td></tr><tr><td> Sylgard 184 (oder Silpot 184) </td><td> Dow Corning </td><td> 3097358-1004 </td><td></td></tr><tr><td> Poly-L-Lysin </td><td> Sigma </td><td> P-1524 </td><td> Dieses Produkt hat sich als am effektivsten </td></tr><tr><td> DPX Eindeckmedium </td><td> Sigma </td><td> 44581 </td><td></td></tr><tr><td> Rabbit anti-GFP </td><td> Invitrogen </td><td> A-11122 </td><td> Verdünnung 1:500 </td></tr><tr><td> Rat anti-CD8 </td><td> Caltag </td><td> 5h10 </td><td> Verdünnung 1:200 </td></tr><tr><td> Maus anti-CD2 </td><td> AbD serotec </td><td> MCA443R </td><td> Verdünnung 1:700 </td></tr><tr><td> Maus anti-Fasciclin2 </td><td> DSHB </td><td> 1D4 </td><td> Verdünnung 1:10 </td></tr></table>

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

Entwicklung des Nervensystems erfordert die korrekte Spezifikation der Neuron Position und Identität, durch genaue Neuron-Klasse-spezifische dendritische Entwicklung und axonalen Verdrahtung gefolgt. Vor kurzem hat die dendritische Verzweigung (DA) sensorischen Neuronen der Drosophila Larve peripheren Nervensystem (PNS) haben sich leistungsfähige genetische Modelle, in denen sowohl allgemeine und Klassen-spezifische Mechanismen der neuronalen Differenzierung aufzuklären. Es gibt vier Haupt-DA Neuron Klassen (I-IV) 1. Sie sind in der Reihenfolge zunehmender Dendriten Laube Komplexität, benannt und haben klassenspezifische Unterschiede in der genetischen Kontrolle ihrer Differenzierung 2-10. Die DA sensorischen Systems ist ein praktisches Modell, um die molekularen Mechanismen hinter der Kontrolle von dendritischen Morphologie 11-13 zu untersuchen, weil: 1) es Vorteile der leistungsstarken genetischen Werkzeuge in der Fruchtfliege, 2) nehmen die DA Neuron Dendriten Laube breitet sich aus in nur 2 Dimensionen unter einem optisch clear Larvencuticula macht es einfach, mit hoher Auflösung in vivo, 3 Visualisierung) der Klasse-spezifische Vielfalt in dendritische Morphologie erleichtert eine vergleichende Analyse zu den wichtigsten Elementen der Kontrolle der Bildung von einfachen vs stark verzweigte dendritische Bäume zu finden, und 4) dendritischen Dorn stereotypen Formen der verschiedenen DA Neuronen zu erleichtern morphometrische statistischen Analysen. DA Neuronen-Aktivität modifiziert die Ausgabe eines larvalen Fortbewegung central pattern generator 14-16. Die verschiedenen DA Neuron Klassen haben verschiedene sensorische Modalitäten und deren Aktivierung auslöst verschiedenen Verhaltensreaktionen 14,16-20. Außerdem verschiedene Klassen schicken axonalen Projektionen stereotyp in die Drosophila Larve des zentralen Nervensystems in der Bauchmark (VNC) 21. Diese Projektionen enden mit topographischen Darstellungen beider DA Neuron Sinnesmodalität und die Position in der Körperwand der dendritischen Bereich 7,22, 23. Daher Untersuchung von DA axonalen Projektionen können verwendet werden, um Mechanismen, die topographische Kartierung 7,22,23, sowie die Verkabelung einer einfachen Schaltung modulierenden Larven Fortbewegung 14-17 aufzuklären. Wir stellen Ihnen hier eine praktische Anleitung zur Generierung und Analyse genetischer Mosaike 24 Kennzeichnung DA Neuronen über MARCM (Mosaic Analyse mit einem reprimierbaren Zell Marker) 1,10,25 und Flp-out 22,26,27 Techniken (zusammengefasst in Abb. 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

Die dendritischen Verzweigung sensorischen Neuronen des<em&gt; Drosophila</em&gt; Larven des peripheren Nervensystems sind nützliche Modelle, um sowohl allgemeine als auch Neuron-Klasse-spezifische Mechanismen der neuronalen Differenzierung aufzuklären. Wir präsentieren eine praktische Anleitung zur Generierung und Analyse dendritischer Verzweigung Neuron genetischen Mosaiken.

Morphologische Analyse Drosophila Larven peripheren sensorischen Neuron Dendriten und Axone mit Genetic Mosaike

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

Research

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Neuroscience

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

In vivo Visualization of Synaptic Vesicles Within Drosophila Larval Segmental Axons

In vivo Visualization of Synaptic Vesicles Within Drosophila Larval Segmental Axons

In-vivo-Visualisierung von synaptischen Vesikeln Innerhalb Drosophila Larven Segmentale Axone

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

Forschung

Neurowissenschaften

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

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

Immunhistologische Markierung von Mikrotubuli in Sensory Neuron Dendriten, Tracheen und Muskeln in der Drosophila Larva Körperwand

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

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation

Visualisierung und genetische Manipulation von Dendriten und Dornen in der Maus Hirnrinde und im Hippocampus mit<em&gt; In utero</em&gt; Die Elektroporation

In utero electroporation (IUE) has become a powerful technique to study the development of different regions of the embryonic nervous system 1-5. To date ...

Voltage-sensitive Dye Recording from Axons, Dendrites and Dendritic Spines of Individual Neurons in Brain Slices

Voltage-sensitive Dye Aufnahme von Axonen, Dendriten und dendritischen Spines einzelner Neuronen in Hirnschnitten

Understanding the biophysical properties and functional organization of single neurons and how they process information is fundamental for understanding ...

Functional Analysis of the Larval Feeding Circuit in Drosophila

Functional Analysis of the Larval Feeding Circuit in Drosophila

Funktionelle Analyse der Larvenfütterung Circuit in Drosophila</i

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

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

Ganze Berge Immunmarkierung der olfaktorischen Rezeptorneuronen in der<em&gt; Drosophila</em&gt; Antennen

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

Produktion und Isolierung der Axone von Sinneszellen für Biochemische Analyse mit Porous Filter

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

Entfernung von<em&gt; Drosophila</em&gt; Muskelgewebe aus Larval Filets für Immunfluoreszenzanalyse von sensorischen Neuronen und epidermalen Zellen

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

Linear Agarose Kanäle zu studieren<em&gt; Drosophila</em&gt; Larval Krabbeln Verhalten

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

Ein Microbiomechanical-System für das Studium Varizen-Bildung und Erholung in zentralen Neuronen Axone

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