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LABORATORY EXERCISE – 7 mm FROG EMBRYO

Transverse Section
The seven-millimeter frog tadpole has well-formed external gills, a functional heart and the
oral plate is very thin and ready to break through to form the mouth. Serial transverse sections
start from the anterior end of the tadpole through the level of the liver and the following structures
should be located beginning at the anterior end.

1. The stomodeum appears as a deep invagination of the pigmented, midventral ectoderm. It is

an inverted longitudinal groove more extensive than the ultimate mouth.

2. The original nasal placodes have separated off from the ectoderm as a large pigmented nasal
pit, possessing so many cells that the cavity itself may seem to be obliterated. It may be possible
to identify some cells growing in the direction of the stomodeum later to become the tubular
connections of internal nares (choanae).

3. The epiphysis is now separated from the brain (diencephalon) as a circular knob of cells later
to become the pineal body.

4. The oral plate forms 1 to 2 cells in thickness, between the invaginating stomodeum and the
evaginating floor of the pharynx. This membrane breaks through at about the 7 mm stage to form
the mouth, the mandibular ridges.

5. Lateral to the pharynx there will appear condensations of the head mesenchyme known as the
mandibular arches.

6. Portions of the eye will come into view and it will be simple to distinguish between the outer
pigmented optic cup and the inner retina.

7. The optic stalk is now much smaller and tubular, connecting with the ventral portion of the eye
cup itself. The choroid fissure is where small nerves and blood vessels run into the eye chamber.
Note the well-formed lens.

8. The pharynx at this level is large and more rounded. Directly above the pharynx locate the
pigmented group of hyposphyseal cells which persist through a number of sections. The presence
of these cells helps in the identification of the nearby infundibulum. The thyroid gland cells will be
seen as evagination of the endodermal cells of the pharyngeal floor.

9. The second pair of visceral (hyoid) arches may be identified as those nearest the thyroid
primordium. These arches are condensations of mesenchyme cells around the pharynx. These
arches become rigid supports between the future visceral clefts, each bearing blood vessels
(aortic arches) and nerves.

10. At the level of the most posterior section of the hypophyseal cells, note that the infundibular
portion of the diencephalon has been cut off from all other parts of the brain. These brain
ectodermal cells will join those of the hypophysis to form the anterior pituitary gland. The most
dorsal cavity at this level is most likely the rhombocoel with very thin roof and nearby otic vesicles.
The mesencephalon has the dorsal thickening as its roof.

11. As the infundibulum closes off the anterior tip of the notochord will take its place
inprogressively posterior sections.

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12. Posterior to the level of the suckers locate the anterior parts of the heart. lt may be possible
to find aortic arches which carry blood from the heart through visceral arches (around the pharynx)
to the development of dorsal aortae. This anterior portion of the heart is the truncus arteriosus.
The parts of the heart can be identified as the lining endocardium, the thick-walled myocardium
and the thin outer pericardium.

13. At the level of the auditory or optic organs, there will be anlage of the 5th, 7th and 8th cranial
ganglia. The 5th ganglia are always the largest and in more posterior sections, the 7th and 8th
cranial ganglia may be identified together close to the vesicle.

14. The brain cavity above should still be the thin roofed rhombocoel, but it should be noted that
its ventro lateral walls are now very thick. This portion of the hindbrain gives rise to central ganglia
form the 5th through the 10th in the frog.

15. Lateral or ventro lateral to these head sections there will appear loose sections of tissue which
are pieces of the external gills. These gills are extremely thin and contain loops of blood vessels.
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16. Within a few sections of this level it will be noticed that the foregut cavity becomes occluded
and disappears entirely. The type of cells which normally line the gut are still obvious, but the
cavity itself has been blocked off. This blocking is called esophageal plug, which will later be
broken through to reestablish the continuous gut.

17. The paired dorsal aortae may now be seen just beneath and slightly lateral to the notochord.

18. As the gut reopens and form the ventrally directed liver diverticulum, note in the sale sections
the lateral and well-formed pronephric tubules. These come from intermediate mesoderm
between the epimere and hypomere.

19. The epimere begins to give off loose cells which encapsulate the nerve cord and notochord,
. comprising the sclerotome. The outermost loose cells of this epimere become dermatome and
will give rise to the connective tissue and blood vessels of the skin while the larger mass of
epimere becomes the segmental myotomes.

I. Illustration:
A. Label the parts of the 7mm frog embryo at the levels of the mesocoel, rhombocoel and gut

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B. Label the parts of the 7mm frog embryo at the levels of the pharynx and heart (frontal
sections)

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II. Questions for Research: (Provide references)

1. What cellular features help to identify the following: oral plate, liver, thyroid, notochord?

2. Given an isolated transverse section, how could one distinguish between the invaginations
which form the nasal pits and those which form the auditory pits?

3. The pituitary and thyroid gland are both endocrine glands. In what 3 respects do they differ
embryologically?

4. How could one identify the parts of the brain and neurocoel in isolated transverse sections?

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Name: ____________________________________ Score: _________________
Date Submitted: ____________________________

LABORATORY EXERCISE – 10 mm FROG EMBRYO

Simultaneously with the closure of the blastopore, the anus (ectodermal proctodeum) of the
frog embryo opens slightly ventral to the position of the original yolk plug. Just within the body of
such an embryo one can discern a temporary connection between the posterior end of the
neurocoel and the archenteron, the canal being called the neurenteric canal. Since this
connection is very temporary and small, you will be provided with sagittal sections to find the
evidence of this neurenteric canal which may appear closed but pigmented cells may be found
connecting the nuerocoel and archenteron.
The anterior end of the neurocoel is flexed ventrally and that it is quite bulbous, forming a
mesencephalon and the prosencephalon at the most anterior portion. This embryonic brain is
bent around the tip of the notochord. A complete neurocoel may not be evident. The notochord is
marked by vertical striations of its large, vacuolated cells and lateral to the notochord may be
seen the metameric somites.
Between the forebrain but anterior to the cavernous foregut locate a group of cells growing
inwardly between the brain cavity and the gut. These will become the hypophysis. In conjunction
with the infundibulum, will give rise to the pituitary gland. The stomodaeum is the ventro lateral
pocketing of the foregut which will give rise to the mouth. The loose mesodermal cells, which will
become organized into the heart, is located beneath the floor of the foregut and anterior to the
yolk mass.
Liver anlage – the forerunner of the liver, is postero-ventrally diverticulum of the foregut.
The bulk of the body at this stage is filled with yolk cells which continually absorbed for growth
and differentiation.

I. Illustrations:
A. Draw the whole mount of the 10 mm frog tadpole and label the parts.

B. Draw the 10 mm frog at the level of the mesencephalon, rhombencephalon, and stomach.

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II. Questions for Research (Provide references)

1. What germ layer is involved in yolk digestion? Discuss briefly.

2. What is the basis for the distinction between germ layers?

3. Since the embryo is unable to ingest food at this time, to what do you attribute the change in
size and shape from neurula to tail bud?

4. Are there any evidence of cephalization? What organ systems develop first?