EMBRYOLOGY: DISSECTION OF A CHICK EMBRYO

Introduction
Observation of early embryogenesis enhances our understanding of cell differentiation and
division, and gross anatomy. All multicellular animals have certain similarities in the pattern of
development but similarities are greatest in closely related groups. Thus, all vertebrate animals
are built about a common anatomical plan and have the same programme of development which is
driven by controlled expression of specific genes. The components of development are -

fertilization, cleavage, blastulation, gastrulation, neurulation and organogenesis.

This practical involves interpretation of the features of the embryo from fertile incubated chick
eggs. Mounts of embryos are available as slides and you will also have the chance to perform a
fresh dissection. You will make a labelled drawing of a mounted embryo and determine its stage of
development. There are also displays available for you to study. Do ask questions about these if
anything puzzles you

Background information - early development of chick embryo

Outer shell Vitelline membrane

membrane
Yolk
Inner shell
membrane

Air space
Albumin Embryonic disc

On the surface of the yolk of a freshly laid, fertile egg is a small whitish circular area, the
embryonic disc, which is about 4mm in diameter. Cleavage and blastulation occur before the
fertilized egg is laid. When the egg is laid and the temperature falls, development is arrested until
such time as the temperature is again raised by incubation. Gastrulation starts during the first day
of incubation and involves the production of the three-layered embryo by migration of cells through
the primitive streak, firstly to displace the hypoblast layer with endoderm and secondly to form
mesoderm underlying the epiblast layer, now ectoderm. The mesoderm extends to the edge of the
area pellucida and then invades the area opaqua where it contributes to extra-embryonic tissue.
This region becomes vascular (indeed is the site of red blood cell production) and is then called the
area vasculosa. At about 15 hours the notochord appears and induces formation of the neural
tube (neurulation). The segmentation of the paraxial mesoderm into somites also begins at this
time. Somitogenesis is advanced after 30 hours and the heart is formed at about 36 hours.

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Trying your own dissection and viewing a fresh dissection

1. Cut a window in the fat end of the egg shell about the size of a two pence piece. This is done by
making an initial small perforation with the point of a scissors blade and then carefully enlarging the
hole with forceps and discarding the pieces of egg shell. You should have entered the air space
under the shell and the embryo should be apparent beneath the inner shell membrane which
should still be intact.

2. Cut around the aperture to remove the inner shell membrane

3. Look at the embryo with the lens provided. Depending on the stage of development, the beating
heart and blood vessels may be visible, especially a ring of blood vessels around the perimeter of
the area vasculosa.

4. Using a pipette, apply 1% nitric acid gently around and over the exposed embryo to coagulate
the albumen and the yolk. Leave for 2 minutes

5. Cut round the embryo outside the area vasculosa using deep cutting strokes with the scissors.
The first cut can be difficult – you need to puncture the yolk. Repeat to ensure the embryo is totally
free from the membranes. This is the crucial stage: if the embryo is not freed from membranes it
will tend to pull back and tear when you try to lift it out. The whole procedure is made more difficult
by the fact that the embryo often disappears beneath the released yolk as you cut. Be patient and
don’t rush things.

6. Using the spoon spatula, lift the embryo into the bowl of water and gently wave it around to wash
off any yolk and vitelline membrane still attached.

7. Wipe the spatula and lift the free floating embryo into the jar of 5% formaldehyde. Close the lid
and leave it for 5 minutes. This fixes the tissue.

8. Quickly pour the contents of the jar into the bowl refilled with clean water. This dilutes the
formaldehyde to a safe level. Using a spatula, manipulate the embryo into the small dish. This is
made easier by immersing the dish in water.

9. Observe under the dissecting microscope

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Drawings of chick embryos

1. Make a drawing of your own embryo, identifying as many features as possible. You should be
able to see regions of the brain, the neural tube, somites, the heart, body folds, the anterior edge of
the gut portal and vitelline blood vessels

Try to determine the stage of your embryo with reference to the illustrations at the end of these
notes (Appendix 1). (You can see colour versions of the staging pictures at –

http://www.swarthmore.edu/NatSci/sgilber1/DB_lab/Chick/chick_stage.html)

Notice that you can also roughly determine the incubation age of the embryo by counting the
number of somites and adding 20 to give an answer in hours. Counting of somites is more difficult
with the later embryos because they can become obscured especially near the rostral end.

Notice also that in these embryos, there is a twisting of the head and neck to the right when viewed
dorsally. However, sometimes the embryo will have been mounted with the ventral side uppermost.
In those cases, the head will be twisted to the left and you will be looking into the gut entry. This
may be easier to understand if you look at the painted movies of chick embryos to be studied in
tomorrow's practical. In fact, looking at these movies generally while you are drawing should help
your interpretation of what you are seeing. They are available at:

https://www.eevec.vet.ed.ac.uk/vc/node.asp?ID=vcembr00

2. You also should draw a mount of an ‘early’ embryo. These are much simpler and you will be able
to draw one quickly. Depending on your choice from the slide tray, you should see features of the
primitive streak, segmented and unsegmented paraxial mesoderm, neural tube formation, the
simple linear heart, the simple 3 vesicle brain with open rostral neuropore, the anterior edge of the
gut portal. Notice if you choose one of the very early versions there may be very little to see other
than the primitive streak and the beginnings of neural tube formation

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 Your drawings

1. Labelled drawing of your ’late’ embryo:

(Give your estimate of stage, age and number of somites)

2. Labelled drawing of an ‘early’ embryo mount with an indication of the features which allow you to
define its stage

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 Appendix 1 – stages of chick embryo development

8-12 hours 13-16 hours 17-18 hours 19-20 hours

1. Fully formed 1. Hensen's node 1. Notochord Notochord
primitive streak. 2. Primitive groove 2. Hensen's node Neural groove
2. Primitive groove 3. Shield-shaped Area 3. Area pellucida Hensen's node
3. Area pellucida pellucida 4. Area vasculosa/ Area pellucida
blood islands Area vasculosa/
blood islands
paraxial mesoderm

22-26 hours 26-29 hours

21-22 hours
Notochord Notochord
(Notochord)
Neural tube Neural tube
Neural groove
Open neural groove Open neural groove
Hensen's node
Hensen's node moved Regions of brain
Area pellucida
caudally Rostral neuropore
Area vasculosa/
Area pellucida Hensen's node moved caudally
blood islands
Area vasculosa Area pellucida
Head ectoderm
Head ectoderm Area vasculosa
Region of cephalic
Four pairs of somites Subcephalic pocket
somitomeres
Anterior margin of gut Anterior margin of gut portal
portal Eight pairs of somites
5
 30-32 hours 33-35 hours 36-43 hours 44-46 hours
Notochord Neural tube Neural tube Head twisting to right
Neural tube 5 vesicle brain
Caudal open neural groove Neural tube
Open neural grove Regions of brain Optic vesicle 5 vesicle brain
3 vesicle brain Hensen's node moved Auditory vesicle Optic vesicle
Rostral neuropore caudally Area pellucida Auditory vesicle
Hensen's node moved Heart Area vasculosa Area pellucida
caudally Vitelline blood vessels Heart Heart
Area pellucida Twelve pairs of somites Nineteen pairs of Vitelline blood vessels
Area vasculosa Caudal unsegmented somites Anterior margin of gut
Vitelline blood vessels paraxial mesoderm portal
Ten pairs of somites Twenty-two pairs of
somites

47-50 hours 60 hours

Head twisted to right Head twisted to right
Amnion and amnion edge Branchial arches
lateral body folds Amnion sheath
vitelline blood vessels Amnion edge progressing caudally
heart Lateral body folds
optic vesicle Tail fold
Vitelline
6 blood vessels
Heart
Optic vesicle