This experiment showed that it is possible to
transplant the eye-forming region of the Axolotl embryo
without killing the host (Figure
18). Furthermore, the grafted eye precursors
continued to develop into eye-type cells. This
illustrates that the cells of a late-stage Axolotl embryo
seem to have undergone determination and are now of
particular cell lineage, reducing their potentiality. It
is therefore plausible that events of specification and
determination in the eye-forming region take place during
gastrulation.
In practice, the attrition rate of embryos was very
high. Over the period of the experiment, only about 69%
of the procedures that were attempted were successful.
Five days post-operatively, only one embryo survived with
its graft intact. This was probably due to our
inexperience. Hence, any attempts to reproduce this
experiment should allow for a high attrition rate.
This experiment could also be improved somewhat. In
order to collect more uniform data, it would be helpful
to carry out post-operative observations at standard
times in the day; however, class schedules prevented that
for this experiment. With a greater number of people
carrying out observations, taking observations at
standard times would have been possible.
In addition, any dead material should be removed from
the dishes immediately and the water changed, or if
possible, all hosts should be places in separate
containers in order to prevent contamination. It is
thought that dead and dying embryos may contaminate
other, healthy embryos present in the same dish
(Cebra-Thomas, personal communication). Again, due to
class schedules, some embryos were left in the same water
as dead or dying embryos, which may have induced their
subsequent death. This would explain why several embryos
tended to die at the same time.
For a first attempt, this experiment was quite
successful. The results and data collected were
illustrative of late-gastrulation differentiation in
Axolotl embryos, and provided worthwhile technical
experience for the students involved.
Cebra-Thomas, J. 2004. Developmental Biology Lab
Compact Disc
* Contains procedure for making eyebrow knives and
tungsten needles
*(Photographs 1-3 courtesy of J. Cebra-Thomas, 4-19 taken
using equipment from the laboratory of Dr. Cebra-Thomas)
Cebra-Thomas, J. 2004. Developmental Biology Lab
Staging Series Portfolio (personal collection)
Gilbert, S.F. 2003. Developmental Biology, 7th ed.
Sunderland, MA: Sinauer Associates; pp: 305 -395.
Gilbert, S.F. 2003.Developmental
Biology website
Hamburger, V. 1973. A Manual of Experimental
Embryology, The University of Chicago Press, Chicago,
Ill