Neural induction is the first step in the formation of the brain. It is a mechanism by which Hensen’s node (organizer), instructs adjacent tissue to adopt a neural fate, i.e. to give rise to the nervous system. This video demonstrates an assay for neural induction in chick embryo.
This video demonstrates the different steps in performing an assay for neural induction; This assay is essentially used for the characterization of putative neural inducing molecules in chick, and thus can be used for a wide variety of applications, ranging from embryological micromanipulations 1-4; 6 to unraveling new signaling cascades 7,8, all aiming to the understanding of the initial step in the formation of the brain and remaining nervous system.
D.P is recipient of Ruth Kirschstein Award 1F32 DA021977-01A1 from the National Institute on Drug Abuse. This work was supported by the Margaret M. Alkek Foundation to RHF.
Material Name | Type | Company | Catalogue Number | Comment |
---|---|---|---|---|
Eggs | Animal | Charles River Laboratories | Premium Fertile | Fertilized, HH3+ (14 hr) |
Stereomicroscope | Microscope | Leica Microsystems | MZ9.5 or similar | |
Hybridization Incubator | Equipment | Robbins Scientific | M1000 | Use with inverted Pyrex dish and 500 ml ddH2O beaker |
Marsh Automatic Incubator | Equipment | Lyon | RX | |
Pyrex dish | ||||
Watchmaker’s glass 50mm | Tool | VWR | 66112-060 | |
Glass rings | Tool | Physical Plant facility | cut 4 mm thick sections of glass tubing (27 mm outer diam, 25 mm inner diam). Do not fine polish. | |
Curved Forceps (1) | Tool | Electron Microscopy Sciences | 72991-4C | |
Forceps (2) | Tool | Fine Science Tools | 11002-13 | blunt ended using sharpening Stone and 100ul mineral oil |
Fine scissors | Tool | Fine Science Tools | 14161-10 | |
Plastic dishes | Tool | Falcon | 353001 | |
Rubber Bulb | Tool | Electron Microscopy Sciences | 70980 | |
DiI | Reagent | Invitrogen | D-282 | |
Aspirator tube assembly | Tool | Sigma | A5177-5EA | |
Microelectrode puller | Equipment | Sutter Instruments | Sutter Instruments P-97 Flaming/Brown Micropipette | |
Pasteur Capillary Pipette | Tool | Electron Microscopy Sciences | 70950-12 | round edge under flame |
Culture chamber | Tool | Pioneer Plastics | 030C | |
Microcapillary tube | Tool | Sigma | P1049-1PAK | |
Microdissecting knife | Tool | Fine Science Tools | 10056-12 | Use to puncture cavities prior to in situ hybridization |
Minuten pins 0.2mm diam | Tool | Fine Science Tools | 26002-20 | Mix 1 part Curing Agent, 9 parts Base; set O/N at 37C |
Diethylpyrocarbonate (depc) | Reagent | Electron Microscopy Sciences | 15710 | |
Sylgard 184 Silicon Elastomer Curing Agent and Base | Dow Corning | 0001986475 | Mix 1 part Curing Agent, 9 parts Base; set O/N at 37C | |
Diethylpyrocarbonate (depc) | Acros Organics | 10025025 | Add 1ml depc to 1l PBS; shake; autoclave | |
16% PFA | Electron Microscopy Sciences | 15710 |