Name: revealing neural circuit topography in multi-color
Uploaded: 2011-11-14T12:00:00
Description: Watch this Scientific Journal Video about Revealing Neural Circuit Topography in Multi-Color at JoVE.com

Dette arbejde blev støttet af nye investigator startfonde fra Albert Einstein College of Medicine af Yeshiva University til RVS.

1. Levering af sporstoffer in vivo ved brug af sterile kirurgi (§ § 1,1 til 1,16) Hele proceduren anbefaler vi, at standarden sterile kirurgiske teknikker anvendes. Dette omfatter sterile handsker, kjole, og ansigtsværn. Værktøjer bør enten autoklaveres før brug eller rengøres grundigt med vand og derefter ethanol. Under operationen, og især mellem dyr, rene væv fragmenter fra de værktøjer og tørre sterilisere instrumenter med et varmt perle sterilisator (Steri 250 Cat. #...

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Vi har beskrevet de kirurgiske og tekniske oplysninger, der kræves for vellykket aksonal og dendritceller sporing ved hjælp af en roman fluorescerende-baseret tilgang til hurtigt mærkning neurale fremskrivninger i at udvikle og voksne mus. Brug af WGA-Alexa vi vise, hvordan sporstoffer og markører kan bruges til at analysere mønstrede kredsløbs topografi med høj opløsning og i tre dimensioner. Mange sporstoffer er til rådighed for sporing neuronale kredsløb, herunder Kolera toksin ...

<table border="1"><tbody><tr><td> Udstyr / Reagenser </td><td> Model / Katalognummer </td><td> Firma </td></tr><tr><td> Bead sterilisator </td><td> Model Steri 250 Kat. # 18000-45 </td><td> Fin Science Tools </td></tr><tr><td> Cauterizer </td><td> Kat. # 18000-00 </td><td> Fin Science Tools </td></tr><tr><td> Borosilikatglas kapillærer </td><td> Kat. # 300056 </td><td> Harvard Apparatur </td></tr><tr><td> To-trins Glas Mikropipette Puller </td><td> Model 001-PC-10 </td><td> Narishige </td></tr><tr><td> Mikrometer sprøjte </td><td> Kat. # GS-1100 </td><td> Gilmont Instrumenter </td></tr><tr><td> Small Animal Stereotaxic Instrument </td><td> Model 940-A </td><td> Kopf instrumentering </td></tr><tr><td> Elektrode manipulator </td><td> Model 960 </td><td> Kopf instrumentering </td></tr><tr><td> Vetcare kammer </td><td> Kat. #340508 </td><td> Harvard Apparatur </td></tr><tr><td> Varmepude </td><td> Kat. # 341241 </td><td> Harvard Apparatur </td></tr><tr><td> Leica DFC360 FX kamera </td><td> DFC360 FX </td><td> Leica </td></tr><tr><td> Leica DFC490 kamera </td><td> DFC490 </td><td> Leica </td></tr><tr><td> Leica DM5500 mikroskop </td><td> DM5500 </td><td> Leica </td></tr><tr><td> Leica DFC3000 FX kamera </td><td> DFC3000 FX </td><td> Leica </td></tr><tr><td> Leica MZ16 FA mikroskop </td><td> MZ16 FA </td><td> Leica </td></tr><tr><td> CY3 Filter </td><td> Model # 11600231 </td><td> Leica </td></tr><tr><td> FITC-filter </td><td> Model # 11513880 </td><td> Leica </td></tr><tr><td> A4 DAPI / UV-filter </td><td> Model # 11504162 </td><td> Leica </td></tr><tr><td> Hvedekim agglutinin, Alexa Fluor 488 konjugat </td><td> Kat. # W11261 </td><td> Invitrogen </td></tr><tr><td> Hvede germ agglutinin, Alexa Fluor 555 konjugat </td><td> Kat. # W32464 </td><td> Invitrogen </td></tr></tbody></table>

revealing neural circuit topography in multi-color

Neurale kredsløb er organiseret i funktionelle topografiske kort. For at visualisere komplekse kredsløb arkitektur har vi udviklet en tilgang til pålideligt mærke den globale mønster af flere topografiske fremskrivninger. Lillehjernen er en ideel model for at studere en velordnet arrangement af neurale kredsløb. For eksempel har kompartment organiseringen af ​​spinocerebellar Mossy fibre vist sig at være et uundværligt system for at studere Mossy fiber mønster. Vi har for nylig vist, at hvedekim agglutinin (WGA) konjugeret til Alexa 555 og 488 kan bruges til at spore spinocerebellar Mossy fiber fremskrivninger i at udvikle og voksne mus (Reeber et al. 2011). Vi fandt tre vigtige egenskaber, der gør WGA-Alexa sporstoffer ønskeligt redskaber til mærkning neurale fremskrivninger. Først Alexa fluorophores er intense og deres lysstyrke giver mulighed for wholemount billeddannelse direkte efter sporing. For det andet, WGA-Alexa sporstoffer mærke hele bane for at udvikle og voksne neurale projections. For det tredje, WGA-Alexa sporstoffer hurtigt transporteres i både retrograd og anterograd retninger. Her beskriver vi i detaljer, hvordan man kan forberede de sporstoffer og andre nødvendige værktøjer til, hvordan man udføre kirurgi for spinocerebellar sporing og hvordan man bedst til billede spores fremskrivninger i tre dimensioner. Sammenfattende tilbyder vi en trin-for-trin sporing protokol, som vil være nyttige for at dechifrere tilrettelæggelse og tilslutning af funktionelle kort ikke kun i lillehjernen, men også i hjernebarken, hjernestammen og rygmarven.

Watch this Scientific Journal Video about Revealing Neural Circuit Topography in Multi-Color at JoVE.com

Revealing Neural Circuit Topography in Multi-Color

Vi tilbyder en praktisk guide til at levere sporstoffer<em&gt; In vivo</em&gt; Og bruge spinocerebellar sti som en model for at demonstrere væsentlige skridt for en vellykket neurale kredsløb analyse hos mus. Vi beskriver i detaljer vores alsidige tracing-protokol, der udnytter hvedekim agglutinin (WGA) konjugeret til Alexa fluorophores.

Afslørende Neural kredsløbs topografi i Multi-Color

Neural circuits are organized into functional topographic maps. In order to visualize complex circuit architecture we developed an approach to reliably ...

Research

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Neuroscience

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