Chicken as a Developmental Model

Abstract

The development of a complex organism from a single cell, the fertilised egg, has fascinated people for centuries. Embryo development is highly reproducible and exquisitely regulated. How is it that all tissues and organs form in the right places and at the right time? How is the development of different organ systems coordinated, so that they all fit together correctly at the end? It is challenging to study development, because many embryos are small or inaccessible. The chick embryo is a popular model system with many experimental advantages, which include classic ‘cut and paste’ experiments and mechanistic gene function analyses. The combination of micromanipulations with gain‐ or loss‐of‐function is particularly powerful. The recent development of transgenic lines and advanced imaging techniques ensure that the chicken remains an attractive model system, which will continue to make major contributions to our understanding of molecular and cellular mechanisms controlling developmental processes.

Key Concepts

  • The chick embryo is easy to access and observe and is therefore an attractive model to study developmental processes.
  • In the early stages, chick embryo morphology is very similar to human, both are amniotes and their development is very similar.
  • The chicken and human genomes share considerable homology.
  • Classic embryological manipulations, such as tissue ablations and tissue grafts, provide information about the contribution of particular cell populations to various organs. These experiments have also told us how cells and tissues influence each other in their developmental decisions.
  • The molecular and cellular basis for many developmental processes and phenomena were first described in the chick, including limb patterning, neural crest migration, dorso‐ventral neural tube patterning, blood vessel formation, somite segmentation and left–right asymmetry.
  • Experiments in chicken, which examine the function of genes, have helped elucidate the underlying mechanisms of human genetic diseases and provide a basis for testing novel therapies.
  • Time‐lapse video microscopy can be used to image live chick embryos, either in ovo or ex ovo using embryo culture.
  • We can use chick embryos to visualise complex processes, including cell migration, cell–cell communication, cell differentiation and tissue morphogenesis, in an amniote system.

Keywords: chick; quail; vertebrate; tissue graft; microsurgery; dye injection; electroporation; embryo culture; live imaging

Figure 1. Bead implantation and detection of gene expression patterns by hybridisation or immunostaining. (a) Beads soaked in growth factors or pharmacological inhibitors or activators of signalling pathways can be implanted into developing embryos. A bead implanted into the forelimb is shown. (b) Detection of specific messenger RNA transcripts in whole‐mount embryos with anti‐sense RNA probes against Fgf‐8 (b), Myogenin = Mgn (c), or ventricular myosin heavy chain = vMHC (d). Probes incorporate a DIG‐UTP nucleotide, which is detected using an alkaline phosphatase‐coupled anti‐DIG antibody. Alkaline phosphatase enzyme converts a substrate into a coloured precipitate, which generates a localised signal that is easily detected. Different structures are indicated by arrows in the different panels, AER = apical ectodermal ridge of the limb bud, so = somite, ht = heart. Other structures also expressing these genes are not indicated. (e) Protein can be detected by antibody staining in whole mount; the fluorescent signal indicates localisation of vMHC.
Figure 2. Microinjection and electroporation of chicken embryos in EC culture or . (a) The injection set‐up consists of a stereo‐dissection microscope, micromanipulators, a pressure injector, an electroporator and a light source. (b) Close‐up of the EC‐culture dish, which contains the embryo mounted on a filter paper frame and placed on a semi‐solid medium. (c) Close‐up of the HH3 embryo in the filter paper carrier, illustrating the placement of the electrodes on either side of the primitive streak, indicated by a stippled line. (d) An embryo, which was electroporated at HH3 with a GFP‐plasmid and incubated to HH5. The GFP‐expressing cells have migrated away from the site of injection (bright green) and are now distributed in an arc shape. Many of these cells are prospective cardiac cells and will migrate to form the heart. (e) Electroporation of an embryo . Black ink injected beneath the embryo helps visualise it. (f) A chick embryo specifically expressing GFP in one half of the neural tube (nt), other structures indicated are the eye (ey) and the heart (ht), which is filled with red blood cells. (g) Section through the embryo shown in (f) shows restricted expression of the electroporated GFP plasmid on one side of the neural tube (nt).
Figure 3. Fate mapping using GFP‐transgenic chicken embryos. (a) A single somite, micro‐dissected from a GFP‐transgenic embryo, was grafted into a non‐transgenic host embryo (white arrow in b). (c) After 7 days of incubation, the somite has contributed to the vertebral column in the neck region, as indicated by the presence of GFP‐positive cells. (d) More detailed analysis shows that a single somite contributes cells (purple colour) to two different vertebrae, which are indicated by a stippled outline. The phenomenon, whereby the original segments (=somites) contribute to the posterior and anterior half of new segments (=vertebrae) is called ‘re‐segmentation’. (Courtesy of Mike McGrew, Roslin Institute, Edinburgh.)
Figure 4. Transgenic chick embryos expressing fluorescent markers, mApple (red) or eGFP (green), specifically in macrophages, under the control of the CSF1R promoter. (a) Macrophages (red) are distributed throughout the developing embryo. (b,c) Macrophages in limbs (green) at early (b) or later (c) stages of development. In (c), the limb has been co‐stained with Lysotracker, a chemical that is picked up by active macrophages that are cleaning up apoptotic cells between the forming digits. These macrophages therefore appear yellow. (d) Distribution of macrophages (red) in skeletal muscle tissue. (Courtesy of Adam Balic, David Hume and Helen Sang, Roslin Institute, Edinburgh; see also Balic ., .)
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Further Reading

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Mok, Gi F, Alrefaei, Abdulmajeed F, McColl, James, Grocott, Tim, and Münsterberg, Andrea(Jan 2015) Chicken as a Developmental Model. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0021543]