Visualization of Neural and Vascular Networks in a Chicken Embryo

Abstract

Source: Delalande, J., et.al. Dual Labeling of Neural Crest Cells and Blood Vessels Within Chicken Embryos Using Chick^GFP Neural Tube Grafting and Carbocyanine Dye DiI Injection. J. Vis. Exp. (2015)

This video demonstrates the transplantation of a GFP-labeled donor neural tube from a stage-matched transgenic chicken embryo into a recipient embryo at the level of somites one to seven, followed by vascular labeling using a lipophilic fluorescent dye. The combined approach allows for direct visualization of the embryo under a stereofluorescence microscope, enabling the study of neural graft integration and vascular network development in ovo.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Graft the Tissue

1. Using a glass micro-pipette, carefully transfer the dissected neural tube from the watch glass to the host embryo. Position the neural tube in the correct anterior-posterior orientation and gently push the explant adjacent to the excised region of the chick host using the micro-scalpel. Leave a small fragment of ectoderm attached to, or by cutting a small nick in, the dorsal surface to identify the orientation of the neural tube.
   1. If necessary, use the mi....

Results

Figure 1. Intravenous DiI injection. (A) Recommended instruments: a) CellTracker CM-DiI drop on paraf.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Fertilized chick eggs	Henry Stewart and Co, Louth, UK
Fertilized GFP chick eggs	The Transgenic Chicken Facility, The Roslin Institute, The University of Edinburgh
Egg incubator (Profi-H Hatcher)	Lyon Technologies, CA, USA	910-033
14C Incubator	Precision Cooled Incubator, Leec Ltd., Nottingham, UK	Model LT2
Stereo-microscope	LEICA	Model MZ 12.5
Digital Camera	LEICA	DC500
Image acquisition software	LEICA	IM50
Goose neck halogen cold light source	Advanced Imaging Concepts, Inc	KL 1500 LCD
Mouth tube	SIGMA - ALDRICH	A5177
Transfer pipettes, polyethylene	SIGMA - ALDRICH	Z350796
Dumont #5 forceps	Fine Science Tools	11251-30
Minutien pins	Fine Science Tools	26002-15
Dumont AA forceps, Inox Epoxy- coated	Fine Science Tools	11210-10
Sellotape (clear, 24mm width)	Any Supplier
CellTracker CM-DiI	Molecular Probes	C-7001
Borosillicate glass capillaries, thinwall without filament	Harvard apparatus	PY8 30-0035