Dan Matthew Burias BIO 120 Developmental Biology

Exercise 1
Fertilization and Early Development of Sea Urchin

Question:
1. Describe the action of 0.5M potassium chloride (KCl) solution in sea urchin spawning.

The K+ ions depolarize a muscle on the outer surface of the alveolar ovary and testis (Vacquier,
2011). It stimulates the gonad wall to contract; ripe gametes emerge from the gonopores
surrounding the anus on the aboral (up) side of the animal (Center for Cell Dynamics, 2004).

References:

Center for Cell Dynamics (2004). Sea Urchin Embryology Lab. Available at
http://rusty.fhl.washington.edu/celldynamics/downloads/methods/urchinlab.html

Vacquier, V. D. (2011). Laboratory on Sea Urchin Fertilization. Molecular Reproduction &

Development. 78:553–564. DOI 10.1002/mrd.21360

2. What conditions might affect cleavage from taking place?

Sea urchin development can be altered by a series of environmental changes. One of these
changes is temperature. The sea urchin embryo development is temperature-dependent, with the
process occurring at a faster rate and demonstrating abnormal developments at warmer than ideal
temperatures, and a slower rate, and even cell death, taking place at cooler than optimal
temperatures (Carballo & Graye, 2001).

Pattern of embryonic cleavage is also determined both by the position of the mitotic spindles
and by the amount and distribution of yolk. Yolk tends to inhibit cleavage. It slows it down or
actually prevents complete cleavage. Yolk is an adaptation of those animals that go through more
or less of embryogenesis isolated from any food supply. Some animals, like the sea urchin, have
relatively little yolk because they rapidly develop into a free swimming larval form that acquires
nutrients from their environment. Other animals such as marsupials are born prematurely, but are
provided nourishment in a parental pouch. Placental mammals develop a specialized organ through
which the embryo is nourished throughout development and so also have little yolk
(courses.biology.utah.edu).

Sources:

Carballo, Laura and Bryan Graye. (2001). The effect of temperature on the embryonic development
of sea urchins (Lytechinus variegatus). Retrieved from
http://www.swarthmore.edu/NatSci/sgilber1/DB_lab/Urchin/Temperaature/discussion.html
Developmental Biology 3230 (2015). Retrieved from
http://courses.biology.utah.edu/bastiani/3230/DB%20Lecture/Lectures/a6Cleav.html

3. What conditions may affect the rate at which cleavage and embryonic development takes
place.

Effect of Gravity (Marthy, 1997):

 In sea urchin eggs, fertilisation under real µg conditions occurs in a normal monospermic way and with
a high (normal) fertilisation rate.
 Eggs fertilised in µg develop normally on the ground to an advanced pluteus stage.
  The unspectacular results have to be interpreted in a positive way. It is reassuring to know that a classic
animal egg model clearly shows that the initial crucial processes for creating new life are not disturbed
in µg.
 Despite the brief duration of µg, sounding rocket flights are well suited to developmental biology
studies, with the proviso that clearly defined processes are studied (e.g. fertilisation sensu stricto, a
specific developmental step such as egg cleaving, and onset of gastrulation).
 It is an open question whether the well known and considerable regulation capacities of the sea urchin
egg mask or compensate for some temporary µg effects at fertilisation during subsequent embryonic
and larval development.
 It is also an open question whether eggs fertilised and kept under continued µg conditions would be
able to develop normally. New space flight opportunities are necessary and therefore solicited.

Effect of Temperature:

In a study on temperature effects on sea urchin development, elevated temperature conditions, +48 0C
reduced cleavage by 40 per cent and +60C by a further 20 per cent. Normal gastrulation fell below 4 per cent at
+60C. At 260C, development was impaired (Byrne et. al., 2009)

Effect of Heavy Metals (Copper):

In study conducted by (Huska and Myers, N.D.) The development of sea urchin embryos is negatively
affected in an environment containing excess copper. The rate of cleavage appeared to slow in the embryos
exposed to 10 mM and 20 mM copper chloride. At time point 55 minutes, >60% of the embryos in the 0 and 5
mM copper solutions progressed to the 2-cell stage as expected. At the same time point, only 34% of the
embryos developing in the 10 mM and 20 mM copper chloride solutions reached this stage. Normal embryonic
development to the 4-cell stage was observed after 80 minutes in both the 0 mM and 5 mM copper chloride
solutions. However, contradictory to our hypothesis, the embryos in the 5 mM solution appeared to divide
faster, indicated by the presence of 8-cell embryos. The 8-cell embryo was also observed in the 10 mM copper
chloride environment; however, the cells appeared distorted and out of place. We also noted similar
abnormalities in the only 8-cell embryo in the 20 mM solution. Development also appeared to halt in the 20 mM
solution; only 16% of the total cells progressed beyond first cleavage. The abnormalities and slow growth were
only observed in higher concentrations of copper chloride, suggests that the heavy metal has adverse effects on
early cleavage events within the embryo.

In a similar study performed by Cebra-Thomas (2004), temperature had an effect on the cleavage rates
of developing sea urchin embryos. Generally, the higher the environmental temperature, the faster the embryo
divided. At 37°C, the sea urchin eggs reached the first cell cleavage at 40 minutes after fertilization, and cleaved
the second time at 65 minutes after fertilization. Abnormalities in the gastrulating embryo were observed at
37°C. Under ideal conditions, 22°C, the embryos first cleavage cycle occurred at 60 minutes after fertilization
and the second cleavage cycle at 100 minutes. At 14 °C, the cleavage rate was dramatically slower, with the first
division taking place at 95 minutes after fertilization. Only 50% of the embryos continued to divide, in which
they reached the second cleavage at 205 minutes after fertilization.

REFERENCES:

Byrne, M., Ho, M., Selvakumaraswamy, P., Nguyen, H. D., Dworjanyn, S. A. and Davis, A. R. (2009).
Temperature, but not pH, compromises sea urchin fertilization and early development under near-
future climate change scenarios. Proceedings of the Royal Society B. 276, 1883–1888.
doi:10.1098/rspb.2008.1935

Marthy, H. J. (1997). Sea Urchin Eggs under Microgravity Conditions. European Space Agency.
http://www.esa.int/esapub/sp/sp1206/marth.htm
Huska, J. and Myers, B. (N.D.). Sea Urchin Embryonic Cleavage Rates are Directly Affected by Environmental
Copper Concentrations.
http://sites.millersville.edu/jcebrathomas/cebra_thomas/DB_lab/Urchin/Copper_Urchin.htm

Cebra-Thomas (2004). Effects of temperature on Sea Urchin Cleavage.

http://www.swarthmore.edu/NatSci/sgilber1/DB_lab/Urchin/Temperaature/results.html