Radiation Biology

Educator Guide
An Interdisciplinary Guide on Radiation Biology
for grades 9 through 12

Module 2:
Radiation Damage in Living Organisms

Revision 3
October 11, 2006
www.nasa.gov

Radiation Biology Educator Guide

Module 2: Radiation Damage in Living Organisms
Authored by:
Jon Rask1
Carol Elland2
Wenonah Vercoutere3
Edited by:
Esther Hill2
Sandy Dueck2
Graphic Design by:
Yael Kovo2
Julie Fletcher2

Give us your feedback:

To provide feedback on the
modules on-line, visit:
http://radiationbiology.arc.nasa.gov/

Module 2 Contents:
Module 2: Radiation Damage in Living Organisms ............ 1
Why is NASA Studying the Biological Effects of
Radiation? ............................................................................ 1
How Do Scientists Study Biological Change During
Spaceflight?.......................................................................... 1
Using Non-Human Organisms to Understand Radiation
Damage ................................................................................ 2
What are the Risks and Symptoms of Radiation Exposure
for Humans?......................................................................... 3
What is DNA?...................................................................... 3
What is the Structure of DNA? ............................................ 3
What is DNA's Role in Protein Production? ........................ 4
What Kinds of DNA Damage Occur Due to Radiation? ..... 5
What Kind of Damage Can High Energy Ions Cause? ........ 6
What are the Consequences of DNA Damage? ................... 7
What is the DNA Repair System?........................................ 8
How Does UV Radiation Affect Us? ................................... 8
What is Degenerative Tissue Damage? ............................... 9
Suggested Activity IIa: Biological Effects of Radiation
Damage in Plants ............................................................... 11
Suggested Activity IIb: Biological Effects of Radiation
Damage in Yeast ................................................................ 14
Suggested Activity IIc: Biological Effects of Radiation
Damage in Drosophila ....................................................... 18
Suggested Activity IId: Three-Dimensional Modeling of
DNA Damage..................................................................... 21

Appendix 1: Additional Websites................................. 23

Appendix 2: National Education Standards Module 2 .. 24
1

Enterprise Advisory Services (EASI)

NASA Ames Research Center,
Moffett Field, CA
2
Lockheed Martin, NASA Ames
Research Center, Moffett Field, CA
3
NASA Ames Research Center,
Moffett Field, CA

...............................................
Module 2: Radiation Damage in Living Organisms

...............................................

As we have discussed, space radiation can penetrate habitats, spacecraft, equipment,

spacesuits, and even astronauts themselves. The interaction of ionizing radiation with
living organisms can lead to harmful health consequences such as tissue damage, cancer,
and cataracts in space and on Earth. The underlying cause of many of these effects is
damage to Deoxyribonucleic acid (DNA). The degree of biological damage caused by
radiation depends on many factors such as radiation dose, dose rate, type of radiation, the
part of the body exposed, age, and health. In this module, we will discuss the risks and
symptoms of radiation exposure including how and why radiation causes damage, and
how the body works to repair the damage. We will also discuss how scientists study the
effects of radiation on living organisms, and why this research is important to NASA.
Why is NASA Studying the Biological Effects of Radiation?
NASA wants to keep astronauts safe and healthy during long duration space missions. To
accomplish this challenging task, NASA has identified four significant health risks due to
radiation that need to be well understood to enable the development of effective
countermeasures. The risks are described in the NASA Bioastronautics Critical Path
Roadmap, and include carcinogenesis, acute and late central nervous system risks,
chronic and degenerative tissue risks, and acute radiation risks.4 NASA scientists are
working to understand the molecular, cellular and tissue mechanisms of damage, which
include DNA damage processing, oxidative damage, cell loss through apoptosis or
necrosis, changes in the extra-cellular matrix, cytokine activation, inflammation, changes
in plasticity, and micro-lesions (clusters of damaged cells along heavy ion tracks).
Knowing that information will help researchers develop the appropriate countermeasures
(see Module 3).
How Do Scientists Study Biological Change During Spaceflight?
Because the radiation environment in
space is different than that on Earth,
the biological responses will be
different. As a result, NASA
scientists must develop space biology
experiments that are designed to
carefully study model organisms in
space. In this scenario, the organism
is sent into space and allowed to grow
and develop. This part of the
experiment is called the flight NASA Ames researchers in the Drosophila lab. Image
Credit: NASA.

http://bioastroroadmap.nasa.gov/User/risk.jsp

experiment. The same experiment is also repeated on the Earth, and this is called a
ground control. Careful analysis of both the flight experiment and ground controls are
critical to understanding the biological changes that result from spaceflight. 5
Many studies are also carried out in ground-based research. Opportunities to fly
experiments can be rare, and experiments must be well planned. Ground-based research
allows a variety of parameters to be tested so that the investigator can decide which will
be the best to focus on in a spaceflight experiment. For radiation studies, ground-based
research can also help in identifying the specific biological responses for a particular
radiation source. This is because on Earth, biological experiments can be carried out
using a source that simulates just one kind of radiation, rather than the complex mix of
radiation types that make up the space radiation environment. With a better
understanding of biological responses to space radiation, we will be able to better design
our countermeasures.
Using Non-Human Organisms to Understand Radiation Damage
To fully understand the biological response of radiation
in humans, NASA scientists begin the process by
studying model organisms. In general, biological
systems are similar across many species; studying one
animal can lead to deeper understandings of other
animals, even humans. Some animals are easier to study
than others, and those with short life cycles make it
quicker to study multigenerational genetic effects.
The fruit fly is a model organism.
Another reason these organisms are commonly used is Image Credit: NASA.
because scientists know a great deal about them. For
most model organisms, their entire genome,
physiological, and behavioral characteristics are well understood. Model organisms are
small in size, so large numbers of them can be grown and studied in a small volume,
which is very important for the confined environment aboard spacecraft. Having a large
population to study also reduces the statistical variation and makes the research more
accurate. Much of our understanding of life and human disease is because of scientists'
work with model organisms. This is also true for what is known about the biological
effects of space radiation. Examples of model organisms include bacteria, yeast, worms,
plants, fruit flies and many others. Fruit flies, like humans, have reduced ability to learn
when they are deprived of sleep. They can also sense the direction of gravity, and are
affected by radiation. Moreover, they have many things in common with humans,
including cellular processes, brain cell development, similar behaviors, and nearly
identical disease genes. In fact, there is a great deal of similarity, or homology, between
the DNA of these organisms and humans.

Images from http://quest.nasa.gov/projects/flies/expert.php

What are the Risks and Symptoms of Radiation Exposure For Humans?
It is important to note that the biological effects of acute and chronic radiation exposure
vary with the dose. An average background radiation dose received by an average person
can be approximately 3 mSv/year (including radon) without causing detectable harm
(review Module 1), while an exposure of 1
Sv/hour can result in radiation poisoning
(nausea, vomiting). A person exposed to 0.1 Sv
has roughly a 1 in 200 chance of developing
cancer later in life, while a 1 Sv dose will cause
cancer in about 1 in 20 people. Three to five Sv
received during a short period of time (minutes)
results in death in 50% of the cases. A person
that experiences a massive 10 Sv dose will risk
death in a matter of just a few days or weeks.6
Both acute and chronic exposure to such large
doses can cause bleeding and inflammation due
to lowered platelet counts. Suppressed immune Radioactive radon gas produced from the
system function and infections are possible due breakdown of uranium in the in the Earths
to lowered white blood cell counts. Reduced crust accounts for over half of the radiation
exposure to the general public. Image Credit:
fertility or permanent sterility could also result. University of Illinois Extension.
In addition to damage at the tissue, organ, and
whole organism level, radiation has the ability to
destroy molecules like DNA.
What is DNA?
DNA is the blueprint of life stored in the cells of every organism. DNA contains the code
for all the information required for the synthesis of proteins, cell reproduction, and for
organization of the tissues and organs. The information in the DNA is arranged in
sections called genes. Gene codes are read by the cells manufacturing system to make
proteins. Proteins are the building blocks for biological structures, and also the functional
machinery of the body. Therefore it is vital to our health for the structure of DNA to
remain intact.
What is the Structure of DNA?
A DNA molecule has the shape of a double helix ladder that is only ~2 nm wide. DNA is
made of individual units called nucleotides. The information in DNA is coded in paired
pyrimidine and purine nucleotides along an incredibly long molecule. A nucleotide
contains three different types of molecules: a phosphate, a ribose sugar and a base. The
backbone of the helix is made of alternating phosphate and ribose sugar molecules. The
rungs of the ladder are base pairs. Each ribose of the backbone has a base attached, which
pairs with a base that extends from the opposite backbone. There are four different types
of bases in DNA: adenine, thymine, guanine and cytosine. DNA is arranged into 23
chromosomes in human cells. If stretched out, the DNA of one chromosome, on average,
6

http://www.ccohs.ca/oshanswers/phys_agents/ionizing.html

would be about 5 cm. If all DNA in a cell were lined end to end, the molecule would
reach about 3 meters. If you took all the DNA in all the cells from one human and lined it
end to end, it would reach from the Earth to the sun 70 times!7

A drawing of DNA (left) and

RNA (right). Image Credit: The
biologycorner.com.

What is DNAs Role in Protein Production?

DNA is the storage unit for the information used to make proteins. Before any protein
manufacturing begins, the cell must transcribe DNA into another molecule. This other
messenger molecule will carry only the code for the specific gene to a ribosome, which
is the site of protein production. This messenger molecule is called Ribonucleic Acid
(RNA). The ribosome reads the gene code of a messenger RNA and manufactures
proteins by assembling long chains of amino acids together, one after another, in a
process called translation. Each amino acid is coded for by a set of three nucleotides, or a
cordon's during translation of the RNA message, the RNA molecule sequence is read
(translated) three consecutive nucleotides at a time. A protein typically consists of
hundreds of amino acids that have been joined together. For example, imagine an RNA
molecule that is 300 nucleotides long. That RNA molecule will be decoded by a
ribosome, and the ribosome will construct a protein that is a chain of exactly 100 amino
acids. A simplified chart summarizing protein production is shown below.

7
8

http://hypertextbook.com/facts/1998/StevenChen.shtml
http://www.biologycorner.com/bio1/DNA.html

What Kinds of DNA Damage Occur Due to Radiation?

DNA is normally a long, continuous molecule that stores tremendous amounts of
information vital for a cell to function normally. When a DNA molecule is broken, the
long chain of information is fragmented and the original message to produce specific
proteins is lost. When DNA is broken on one strand of the double helix, it is called a
single strand break (SSB).9 If both strands of the DNA double helix are severed within 10
to 20 base pairs of each other, the break is called a double strand break (DSB). Other
forms of damage that can occur include the loss of a base, and base modification such as
oxidation. In many cases, cells are able to fix such breaks with repair systems that are
specialized for different types of damage. The damage sites that remain can cause
assembly of proteins to be stopped or started prematurely. If DNA replication occurs
before the repair system finds the damage, there is a chance that a modified nucleotide is
misread as a different nucleotide. In addition, sometimes the repair systems misread a
damaged nucleotide and replace it with the wrong nucleotide. The result in both cases is a
point mutation. A point mutation is a single change in the nucleotide sequence of a gene.
This can alter the amino acid code, so that the protein produced from the gene has a
different composition. Depending where in the protein this occurs, the altered protein
sequence can have no affect, or it can alter the protein and protein function substantially.
The result may cause cellular or tissue abnormality. In more extreme cases, the presence
of DNA damage that cannot be properly repaired can trigger apoptosis, or cell suicide
(see Module 3 for information about apoptosis and radiation countermeasures). The
individual cell is sacrificed to prevent greater damage to the whole organism.

Single Strand Break

Double Strand Break

In this drawing, the ladder-like structures represent a DNA helix of hydrogen bonded nucleotide
pairs. Image Credit: Pacific Northwest National Laboratory.

In some cases, the effects of radiation-induced DNA

damage may not be readily or immediately observable.
While some damage may not be severe enough to cause
death to a cell or organism, its effects can become
apparent several generations later. At right is a diagram
of a normal DNA molecule before and after being hit by
ionizing radiation.10
Image Credit: NASA

http://www.pnl.gov/berc/bg/fatal_lesion.html
10
http://science.nasa.gov/headlines/y2004/17feb_radiation.htm

Damage to DNA can be caused directly or indirectly. As the ion travels through material,
it will lose some of its energy to the molecules around it. Cosmic radiation contains
heavy ions, which are the nuclei of atoms with atomic weights ranging from 14 (carbon)
to 55 (Iron) or greater. This means that the atom is missing anywhere from 14 electrons
to 55 or more electrons. As this particle moves through material, it will pull electrons
from any source it can find. This ionizes the molecules along the path of the heavy ion.
Protons, alpha particles, or larger fragments can be forcibly separated from the DNA. In
addition, when the heavy ion moves through water, the hydroxide ions in water (OH-)
can be ionized, losing an electron, to give hydroxyl free radicals (OH). Such species
have a strong propensity to restore the electron pair by pulling a hydrogen atom,
complete with a single unpaired electron, from carbon-hydrogen bonds in sugars. One
excellent source for this within cells is DNA. Nucleotide modifications or removal,
Single Strand Breaks, Double Strand Breaks, or any combination of these can occur
along or near the track of a heavy ion.
What Kind of Damage Can High Energy Ions Cause?
Because of their high ionization density, heavy ions and HZE particles (high energy ions,
discussed in Module 1) can cause clusters of damage where many molecular bonds are
broken along their trajectory through the tissue. The cell's ability to repair DNA damage
becomes impaired as the severity of clustering increases. These particles can also create
damage along a long column of cells in tissue. In other words, cells will be damaged in
streaks along the path of an HZE particle. Since HZE particles are rare on Earth, the
prediction of biological risks to humans in space must rely on fundamental knowledge
gathered from biological and medical research.11
Because spaceflight radiation biology experiments are extremely expensive and
opportunities for flight are limited, NASA models spaceflight radiation exposure by
studying organisms that have been exposed to radiation produced at special facilities here
on Earth. Brookhaven National Labs
(shown at left)12 and Lawrence Livermore
National Laboratory13 are two examples of
facilities that have the capability to produce
radiation that is similar to space radiation.
This type of research greatly enhances our
understanding of the biological response to
space radiation, helps us to anticipate what
may happen during future spaceflights, and
develop countermeasures to help protect
astronauts from radiation (discussed in
Module 3). For example, scientists have
Image Credit: Brookhaven National Labs
learned that mutations, chromosomal
aberrations in plant seeds, development
11

http://haco.jsc.nasa.gov/projects/space_radiation.cfm
http://www.bnl.gov/medical/NASA/NSRL_description.asp
13
http://www.llnl.gov/
12

disturbances and malformations in small animal embryos, and even cell death in bacteria
have resulted from the traverse of a single HZE particle. 14 Examples of cellular damage
are shown in the following figure.

This diagram from Crater.bu.edu 15shows a comparison of radiation damage in three human cell nuclei
(above left). The nuclei were exposed to (A) gamma rays, (B) silicon ions, and (C) iron ions. Following
exposure, the cells were stained so the scientists could observe where DNA damage occurred. Every green
spot is a DSB. Notice that the gamma ray (electromagnetic waves) exposure in (A) produced uniform
damage, whereas cells exposed to high-energy heavy ions show DNA damage along the path traveled by
the ion. In (B) there is one track and in (C) there are three tracks. The damage tracks of ions with differing
masses are seen in (D). Note that heavier ions cause a wider path of destruction. Our understanding of
biological damage caused by heavy ions is very limited. A cell has been drawn to scale for comparison
purposes. Image Credit: crater.bu.edu.

What are the Consequences of DNA Damage?

If radiation changes the number or order of nucleotides (mutation) within a DNA
molecule, the information that is stored within the DNA is altered. This can cause
significant problems in cell structures and even their function. Even if a DNA molecule
14
15

Acta Astronaut. 1994 Nov; 32(11):749-55.

Lancet Oncol 2006; 7: 43135 (also see http://crater.bu.edu/Science/gcr-cancer_risk.pdf )

has had only one nucleotide deleted, that error could be perpetuated when translated into
RNA. In other words, when the RNA is produced, it will be made as if that missing
nucleotide had never existed in the first place. Interestingly, the ribosome will not know
the difference, because the cell assembles the RNA based on what it reads in the DNA.
As a result, the ribosome will assume that the information in the RNA is correct
(although we know that the nucleotide order in the gene has been shifted by one
nucleotide). Protein synthesis carries on, the triplet codons are read by the ribosome, and
amino acids are gathered and assembled into a protein structure that the DNA had not
coded for originally. In this example, a malformed protein will be constructed that could
have significant negative consequences. In summary, when the genotype (genetic
information) of a cell is changed, the phenotype (the outward observable expression of
the genetic information) may also be changed. Radiation is an environmental stimuli that
has the ability to influence whether or not a gene turns on and off, for example, some
cancer genes.
What is the DNA Repair System?
The repair system is constantly monitoring our DNA to make sure it stays intact; proteins
will congregate to sites of damage. So one way to measure DNA damage in tissues is by
staining tissue samples to look for proteins involved in the repair system. This allows
researchers to see where the damage occurred, and at how many sites. They can also
monitor how fast the repair system takes to complete its job by staining the tissues at
different times after radiation exposure. When possible, cells use the unmodified
complementary strand of the DNA as a template to recover the original information.
Without access to a template, cells use an error-prone recovery mechanism known as
translesion synthesis as a last resort. Damage to DNA alters the three-dimensional
configuration of the helix. These alterations can be detected by cellular repair
mechanisms. Once damage is localized, specific DNA repair molecules move to the site.
These molecules bind at or near the site of damage and induce other molecules to bind
and form a complex that enables the actual repair to take place. The types of molecules
involved and the mechanism of repair that assembles depend on the type of damage that
has occurred and the phase of the cell cycle that the cell is in. Some examples of specific
repair systems include: base excision repair (BER), which repairs damage due to a single
nucleotide caused by oxidation, alkylation, hydrolysis, or deamination; nucleotide
excision repair (NER), which repairs damage affecting longer strands of 2-30 bases. This
process recognizes bulky, helix-distorting changes such as thymine dimers as well as
single-strand breaks; and mismatch repair (MMR), which corrects errors of DNA
replication and recombination that result in mispaired nucleotides following DNA
replication.
How Does UV Radiation Affect Us?
There are three kinds of UV radiation. UV-A radiation (wavelengths of 320-400 nm)
plays a helpful and essential role in formation of Vitamin D by the skin. It is not absorbed
by the ozone layer, and can cause sunburn and premature aging on human skin, immune
system problems, and cataracts in our eyes. UV-B radiation (wavelengths of 290-320
nm), mostly impacts the surface of the skin. It is absorbed by DNA and the ozone layer,
and is the primary cause of sunburn, skin cancer. After exposure to UVB, the skin

increases production of the pigment melanin. This darkens the skin and protects it by
absorbing UV light. A dark tan is an indicator of extensive UV-related skin damage. The
third is UV-C, which is completely absorbed by the ozone layer and oxygen in the
atmosphere.
DNA readily absorbs UV-B radiation. In
some cases, it causes the shape of the DNA
to be changed. While cells are able to repair
this damage through the use of specialized
enzymes most of the time, sometimes
damage is permanent and the irreparable
damage has a cumulative effect that is
perpetuated from then on as previously
mentioned. UV damage can also cause a
mutation, or change in the DNA of a gene.
When this gene is transcribed and
translated into a protein, the protein may
contain an error that causes it be
misshapen, function improperly, lead to
cancer, or even cause cells to kill
themselves.16

DNA readily absorbs UV-B radiation.

Image Credit: NASA

One in five Americans will develop skin cancer and one American dies from this disease
every hour. People who have had several blistering sunburns before age 18 are at higher
risk of developing melanoma, the most serious form of skin cancer. Individuals with fair
skin and freckles have a higher risk of developing skin cancer, but dark-skinned
individuals can also get this cancer. Regardless of your skin color, exposure to UV
radiation can lead to premature aging of the skin, causing it to become thick, wrinkled
and leathery. Proteins in the lens of the eye can also be altered by radiation, leading to the
formation of cataracts that can lead to partial or complete blindness. UV radiation may
also suppress proper functioning of the bodys immune system.17
What is Degenerative Tissue Damage?
As we have discussed, ionizing radiation alters DNA such that cell repair processes, cell
cycle or cell division is affected. Low numbers of SSB or DSB may provide a trigger for
the gradual loss of cycling cells. Loss of repair mechanisms, or loss or reduction of cell
division results in tissue degeneration. This can occur in almost all tissues, including the
nervous system.
There are also radiation-induced bystander effects. These are biological responses in cells
that are not themselves directly in the path of ionizing radiation or in a field of radiation.
In fact, new studies show that an even larger portion of bystander cells, sometimes at
considerable distance from the irradiated cells, can be affected. The radiation effects can
be transmitted directly from cell to cell through channels (gap junctions) connecting cells.
16
17

http://earthobservatory.nasa.gov/Library/UVB/
http://gslc.genetics.utah.edu

Alternatively, directly hit cell can secrete factors, or signals, which travel out of the hit
cell to neighboring cells. Bystander effects have been clearly established in cell culture
systems, and a few studies are starting to provide evidence that bystander effects occur in
vivo (the natural setting). Bystander effects amplify or exaggerate the action of even low
dose radiation, so they can significantly increase radiation risk and tissue damage. This
may be particularly important when ionizing radiation hits the nervous system, where
bystander effects could lead to loss of sensory, motor, and cognitive functions.18
Degenerative tissue damage and central nervous
system damage could be particularly dangerous if
it occurs in the brain of astronauts. The damage
could cause altered behavior. Since damage to the
nervous system is not repairable, it could reduce
the ability of astronauts to work and respond to
their environment, especially in an emergency. It
could eventually lead to loss of control over their
entire body, or death.

18

In this image, brain necrosis (unprogrammed

cell death) has occurred. Similar damage
could result from excessive radiation. Image
Credit: NASA.

Boyle, R, Radiation Biology Professional Development Course Charts, 2006.

10

...............................................
Suggested Activity IIa: Biological Effects of Radiation Damage in Plants

...............................................

Objectives:
Analyze the genetic effects of radiation in plants.
Describe changes in phenotype as a result of radiation damage.
Discuss how radiation could cause the observed effects?
Research Question:
Will irradiation affect plant growth or morphology? Dow much does the level of
irradiation affect the plants?
Discussion Questions:
1. What percentage of each irradiated group germinated for the plants you observed?
What is the average germination rate for each irradiated group for the entire class? Is
the difference in the average germination rate between each irradiated group and the
control group statistically significant?
2. What was the average height of the plants in each irradiated group when the first
flower was observed? Is the average the same for each irradiated group for the entire
class? Is the difference in the average height between each irradiated group and the
control group statistically significant?
3. What was the average number of seedpods that were present for each irradiated group
for the plants you observed? Is the average the same for each irradiated group for the
entire class? Is the difference in the number of seedpods between each irradiated
group and the control group statistically significant?
4. What was the average number of seeds in each seedpod for each irradiated group for
the plants you observed? Is the average the same for each irradiated group for the
entire class? Is the difference in the average number of seeds within the seedpods
between each irradiated group and the control group statistically significant?
5. How could you use the DNA of radiated and non-radiated plants to determine if there
were genetic effects?
6. Were all effects negative? Why? Can you observe all effects? Why or why not? What
do you think the effects on the next generation(s) will be?
Materials:
1. Irradiated Seed Set of desired type (Brassica rapa, Arabidopsis, radish,
chrysanthemums, etc. You will need to obtain them from a vendor).
2. Record book to record data.
3. Camera to record data, if desired.

11

Example: Plant Images:

The following images are the growth results of seed that were planted after having been
exposed to radiation (or no radiation, in the case of the control).

Control
16 days

50 Krad
16 days

150 Krad
16 days

500 Krad
16 days

Directions:
The exact protocol for this experiment will depend upon which vendor and specimen is
chosen. For this experiment, plant the pre-irradiated seeds following the directions
provided with the seed. Record the number of seeds planted and the date.
1.

Observe and record each day the germination rate and rate of emergence and
appearance of the seedlings of the control and irradiated types.

2.

After the recommended time for germination of the plants (on day 5 for Brassica)
record the total number of seedlings. Calculate the percentage of germination and the
percentage of emergence. Record the appearance and height of the seedlings.

3.

Each day record height, number of leaves, the general appearance of all plants, total
height when the flower first opens, and date of the first flower. As necessary, stake
up the plants. Tape 5 x 8 paper cards between the irradiated and control plants to
provide a barrier and prevent accidental pollination between groups.

4.

At the recommended time (day 14 to 18 for Brassica), use a bee-stick or a clean Qtip to place pollen from one control plant onto another control plants flowers.
Record the date of pollination.

5.

Use a bee-stick or a clean Q-tip to place pollen from one irradiated plant onto
another irradiated plants flowers. Record the date of pollination.

6.

At intervals after pollination (up to about day 40 for Brassica), make frequent
observations and record height, number of leaves, and number of seedpods.

7.

At the recommended time (between day 41 to 45 for Brassica) harvest seedpods and
count seeds found within the pods. Record number of seeds and calculate the
average number of seeds per pod.

12

Why Does NASA Study Radiation Effect in Plants?

It has been shown that plant growth is inhibited by radiation. Like mammals, the embryo
of a plant is very sensitive to radiation damage as compared to the adult.19 The rate of
seed germination is reduced, and the rate of growth is slowed.20 Excessive UV radiation
will lead to an inhibition of plant growth processes in general. Such alterations in primary
productivity (photosynthesis) can change entire ecosystems in the oceans, on land, and
even in bioregenerative life support systems
that would be aboard future spacecraft. Thus,
NASA scientists need to understand how
plants respond to radiation if future space
explorers depend upon them for nutrient
cycling and food. Experiments involving
plants in space, like the Biomass Production
System, have been a favorite of astronauts
during long-duration stays onboard the NASA scientists are looking for better ways to
grow plants both on Earth and in space.
International Space Station.21
(Note: Equipment and materials, including irradiated seeds, for this activity are
commercially available from various educational resources.)
References:
Brassica rapa: http://www.fastplants.org/
http://www.hps.org/publicinformation/ate/q1280.html

19

http://www.hps.org/publicinformation/ate/q1280.html
www.esd.ornl.gov/programs/ecorisk/documents/tm13141.pdf
21
http://liftoff.msfc.nasa.gov/news/2003/news-plants.asp
20

13

...............................................
Suggested Activity IIb: Biological Effects of Radiation Damage in Yeast

...............................................

In this experiment, students will explore how well sun-screening materials protect live
yeast cells from harmful UV radiation. Different sun protection factors, brands, or even
sunglasses may be used to expand the range of items tested. Note: this activity will be
referred to in Module 3 because it can also be used to demonstrate countermeasures
against radiation.
Objectives:
Discuss countermeasures for UV radiation.
Describe phenotypic changes in yeast as a result of radiation damage.
Research Question:
What is the most effective method of preventing UV damage in yeast?
Discussion Questions:
1. What are the effects of different types of sunscreen on yeast?
2. How can your health be affected by exposure to ultraviolet radiation?
3. Why use yeast to study the effects of UV radiation?
4. Do you see any differences between areas of the Petri dish? If so, describe them.
5. Did some SPFs of sunscreen protect the yeast cells better than others? Why?
6. Does yeast grow less in some areas? Does it grow more than in others? Why?
7. Does UV pass through plastic wrap? Plastic Petri dish covers?
8. Why is it important to not expose an open yeast extract dextrose agar plate for
very long? What is aseptic technique?
9. What can you conclude from the results of your experiment?
10. Describe another experiment you could carry out to obtain more information
about the effects of UV radiation on cells.
Materials:
1. Yeast-Extract Dextrose media plates (from kit, can also be made)
2. UV-sensitive yeast suspension in liquid media and wild type yeast suspension
in liquid media (this needs to be prepared from a stock sample that is
purchased from a vendor). Ensure there is enough for the number of plates
that will be plated (1 ml of cells per plate is recommended).
3. A source of UV radiation such as direct sunlight. For this or future
experiments, the radiation source could also be changed. Depending upon the
size and design of the experiment, you may want to include black lights,
halogen, or fluorescent light bulbs to determine if they also produce damaging
radiation)

14

4. Several kinds of sunscreen (each with different SPF), black paper, cloth, metal
foil, or other types of materials that can be used to experiment with UV
shielding.
5. Sterile water, sterile pipets, and sterile toothpicks
6. Plastic wrap (to cover plates)
Directions:
1. Ensure that your hands and the work area are clean. Use soap and water and wipe
your hands and your work area with alcohol and a paper towel. Good aseptic
technique will ensure that the plates do not get contaminated with other
organisms.
2. In this step you will plate the yeast suspension. You may want to do this for each
group or allow the students to perform the task. Swirl the container of UVsensitive yeast. Using aseptic technique and a sterile pipet, add 1 ml of the yeast
cell suspension uniformly on top of the agar in the Petri dish for every plate that
will be used in the experiment. Close the lid. Gently tilt and rotate the dish to
spread the liquid. If there are places the liquid does not cover, reopen the dish and
use the rounded end of a sterile toothpick to move the liquid over them. Sterile
glass beads could also be used to spread the cells across the plate. Let the liquid
soak into the agar. Place the Petri dish in a dark place for 10-20 minutes until the
liquid soaks into the media.
3. Label the dish (see the diagram at the end of this activity) by drawing lines on the
top and bottom of the dish to divide it into 4 parts (you could divide it into more
parts, depending upon the number of countermeasures you are investigating).
Label one area sun as a control, and use the other three areas to test sunscreens
or other items like cloth, foil, paper, or plastic. Ensure that one area on all plates
do not get UV exposure (cover it with black paper during UV exposure) or make
certain that at least one entire plate per group is designated as the control, which
does not get UV exposure. Label each area on both the top and the bottom of the
dish and tape the 2 halves of the Petri dish together along the side so that the lid
does not rotate. For one group (or the entire class), have the students remove the
lid and replace it with plastic wrap (tape it on tightly). This will test any possible
shielding effects of the cover.
4. Spread sunscreen on the lid of the Petri dish (or on the plastic wrap) in the places
you marked; use an equal amount in each section and spread the sunscreen
evenly. You can also use plastic, foil, etc. instead of sunscreen. If you labeled an
area no sun, tape a square of dark paper over it. Make sure you know exactly
where each sun screening material is used.
5. Expose the Petri dish to the sun or to a UV light. Vary the appropriate exposure
times for the students from 20 minutes (in midday summer sun) to as much as 4
hours (in midmorning winter sun) per dish. If you are exposing the Petri dish to
the sun, make sure that the surface of the agar is aimed directly at the sun
(perpendicular to the incoming radiation). If students are careful, the lids could be
removed and replaced with some clear plastic wrap during the exposure (to reduce
any possible shielding effects of the lid). Consider allowing one group to remove
the lid for a direct exposure.

15

6. After the exposure, wipe the sunscreen off the lid of the Petri dish. This will
reduce the mess. Remove any other materials that were tested. If the students used
the plastic wrap, just remove the wrap and replace it with the original lid. Place
the Petri dish upside down in an incubator or in a dark place and let it grow for 12 days in an incubator at 30C or 3-4 days at room temperature.
7. If desired, repeat these steps with a wild type strain as a control for comparison.
8. Compare the amount of yeast that has grown in different areas of the Petri dish
and draw conclusions.
How Do Sunscreens Work?
Sunscreens act like a very thin shield by stopping the UV radiation before it can enter the
skin and cause damage. Some sunscreens contain organic molecules (such as
oxybenzone, homosalate and PABA) that absorb UVB and/or UVA radiation. Others use
inorganic pigments (such as titanium dioxide and zinc oxide or both) that absorb, scatter,
and reflect both UVA and UVB light. Sunscreens are labeled with a Sun Protection
Factor (SPF) rating that could also be thought of as a sunburn protection factor. For
example, suppose that your skin begins to redden after 10 minutes in the sun. If you
protected it with an SPF 15 sunscreen, it would take 15 times as long, or 2.5 hours, to get
a comparable burn. Remember, SPF relates only to UVB protection; there is no standard
measurement or rating for UVA protection in the United States.
Why Does NASA Study Yeast in Space?
Like the fruit fly, ordinary baker's yeast (Saccharomyces cerevisiae) also contains genes
for DNA repair that are very similar to human genes with the same function. Therefore
we can use yeast as a model system to explore the effects of radiation on cells. Like
human cells, most yeast cells effectively repair DNA damage caused by UV radiation.
However, some yeast strains have mutations that prevent them from performing certain
types of DNA repair. Because they cannot repair all the damage to DNA, these cells
usually die after exposure to UV radiation. In addition to sensitivity to UV radiation,
yeast is also sensitive to space radiation. In a biological assessment of space radiation in
low-Earth orbit, yeast inside special experiment hardware has been shown to have a
decreased rate of survival following exposure to beta particles (electrons) and low-energy
protons.22 Other findings suggest there are highly coordinated gene expression responses
to gamma radiation.23 This knowledge is especially important when designing
countermeasures for astronauts during long-term lunar surface operations or microgravity
spacewalks.
(Note: Equipment and materials for this activity are commercially available from various
educational resources.)
References:
Another more advanced experiment example can be found at:
http://www.phys.ksu.edu/gene/d3.html
22

23

http://www.spaceflight.esa.int/users/index.cfm?act=default.page&level=11&page=2120
http://www.sanger.ac.uk/PostGenomics/S_pombe/docs/851.pdf

16

Sun
(no SPF)

SPF 15

SPF 30

SPF 45

17

...............................................
Suggested Activity IIc: Biological Effects of Radiation Damage in Drosophila

...............................................

Objectives:
Analyze the genetic effects of radiation Drosophila melanogaster
Describe phenotypic changes as a result of radiation damage.
Research Question:
What is your hypothesis for the phenotypic and genotypic results of crossing the selected
traits? What are the visible effects (physical, behavioral) of radiation on Drosophila?
Discussion Questions:
1. What is your hypothesis for the results of crossbreeding the selected traits?
2. What phenotypes and phenotypic ratio do you predict for the F1 generation?
3. What genotypes and genotypic ratio do you predict for the F1 generation?
4. What are the expected results if the F1 generation were allowed to breed?
5. Model effects of radiation with crossbreeding of known strains of Drosophila.
6. Demonstrate or predict examples of genetic/phenotypic variations as a result of
cross breeding Drosophila with different traits.
7. Compare and contrast images and video of Drosophila that have been exposed to
radiation with flies that are normal.
Materials:
1. Drosophila melanogaster of two different strains (e.g., wild-type and wingless)
2. Record book to record your findings
3. Materials to cultivate specimens (containers, food)
4. Magnifying glass or dissection microscope
5. Fly sorting materials (sorting brushes, anesthetizing tools,)
Directions:
1. Select the specific Drosophila melanogaster traits to be crossbred. This will require
some research. Mate the flies obtained for the experiment. After five days, remove the
adult flies from the container (only eggs and larva will remain in the container).
2. Observe and record the development of the larvae over the next ten days.
3. Observe the F1 generation and record the characteristics of the adults as they emerge.
When enough adults have hatched to provide a good sample, remove the adults to a
new container.

18

4. Anesthetize the adult flies (using a method that is safe for your classroom) and sort
them according the visible phenotypic traits. Record the results of your observations.
Analyze the results of your crossing the two traits.
Historical Context:
Unexpected events often determine the course of our scientific legacy. Around 1910, one
such event was the decision of Thomas Hunt Morgan, a pioneer in genetics research. He
decided to study the fruit fly, Drosophila melanogaster, instead of the costlier, preferred
rabbit specimens because there was very little funding for basic research at universities.
Fruit flies were chosen because they are small, found nearly everywhere, inexpensive to
house, and reproduced in large numbers. An additional advantage was that fruit flies
could be easily seen with only a simple hand lens. Much of Morgan's early work was
done this way. Later, microscopes were used to study Drosophila.
Morgan bred wild-type (red-eyed) fruit flies by the thousands, and his team tried to create
mutant flies with x-rays, acids, and other toxic substances. Finally, in one unaltered
lineage of flies, the researchers found a surprise. Every single fly in that line had been
born with red eyes, until one day a fly emerged from its pupa with white eyes. The image
below shows the differences in fly eye color.24 Something had spontaneously changed in
the white-eyed fly. Morgan realized that one of its genes had been altered and it had
produced a new kind of eye. 25 Morgan bred the
white-eyed fly with a red-eyed fly and got a
generation of red-eyed hybrids. And when he
bred the hybrids together, some of the
grandchildren were white-eyed. Their ratio was
three red to one white. Here was a mutation,
but one that didn't fit the current understanding.
Scientists, at that time, thought that mutations
created new species, but the fly that had
acquired the white-eyed mutation remained a
member of the same species. It could still mate
with other fruit flies, and its gene could be
Morgan's experiments involved red- and
passed
down to later generations in proper
white-eyed fruit flies like these.
Mendelian fashion.
The work of scientists such as Morgan established the science of genetics. His work
resulted in the discovery of sex-linked and autosomal genes. Autosomal genes are those
carried by any chromosome except the sex chromosome. Morgan's work with Drosophila
went on to explain linkage, two different genes being on the same chromosome and not
randomly assorted as had been understood, and the concept of crossing-over, the
transference of genes from one chromosome to another. Morgans work with Drosophila
eventually earned him the Nobel Prize in Physiology or Medicine in 1933.

24

Bhattacharya, S, presentation entitled "Basic Biology for Engineers Short Course, Unit: Nematodes and
Insects, Title of Lesson: D. melanogaster" August, 2003.
25
http://evolution.berkeley.edu/evolibrary/article/history_18

19

(Note: Equipment and materials for this activity are commercially available from various
educational resources.)
References:
Flies In Space: http://quest.nasa.gov/projects/flies/
FlyBase: http://flybase.bio.indiana.edu/
Flight Experiment: http://lis.arc.nasa.gov/lis/Hardware_App/Drosophila.html

20

................................................
Suggested Activity IId: Three-Dimensional Modeling of DNA Damage

................................................
In this activity, you will model DNA and DNA damage.
Objectives:
Visualize the three-dimensional structure of DNA
Simulate the random damage to DNA caused by radiation
Describe molecular changes as a result of radiation damage
Research Question:
What types of damage can arise from high-energy particles hitting DNA?
Discussion Questions:
What bonds are the easiest to break?
How many breaks do you get with a single hit?
How easy is it to repair the damage?
How does your choice of model affect the type of damage observed?
What type of damage is most difficult to repair?
How does this relate to the types of damage that more difficult for biological repair
systems in cells to recognize and repair?
Materials:
DNA Building kit (DNA models can be constructed using a few common kitchen items;
plastic kits can also be purchased, see references below)
Two to three partners (this activity is best done as a team)
Methods:
(1)
Construct a DNA model. Have one team member remove a bond while the others
are not looking. Ask the students to identify which bond has been broken.
Record the type of damage that occurred.
(2)

Have the students repair the damage. Repeat the exercise by removing one or
more bonds, bases or nucleotides. This represents the various types of damage
that can occur.

References:
Suggested DNA model kits
http://www.miniscience.com/projects/DNAmodel/index.html
http://www.planet-science.com/outthere/index.html?page=/outthere/diner/play/09.html
http://www.powertolearn.com/teachers/lesson_activities/science/CBV.35.E.SCI.R2.F.pdf

21

Historical Context of the Discovery of the Shape of DNA

Since DNA itself is so small, there are no methods to directly image DNA or the damage
caused by radiation. All measurements must be gathered indirectly. The first method used
to determine the structure of DNA was X-ray crystallography. This was first done in the
late 1940's, when the structure of DNA was still a mystery. At the time, X-ray
crystallography was commonly used to determine the crystalline structure for molecules
much smaller than DNA.
Just like a glass crystal will refract sunlight to produce a rainbow, X-rays directed at a
crystal formed from molecules will bounce off the repeated angles and scatter in a
specific pattern. This information can be used to calculate the repeating angles in the
crystal, which tells us about the arrangement of molecules in the crystal. Rosalind
Franklin and her colleagues were the first to learn how to prepare DNA as a crystal, and
took the first X-ray pictures of DNA using the technique of X-ray crystallography. In
1953, James Watson and Francis Crick then used these results and clues from other
measurements to painstakingly construct a three-dimensional model of the DNA
molecule. They proposed
that DNA was a double
helix, with the base pairs AT and G-C on the inside,
like rungs on a ladder.26
Today, scientists continue to
study the three-dimensional
structure of DNA, and use
computers for complex
calculations and graphical
analysis to speed up the
process of modeling and
visualizing DNA27 (right)
Image Credit: Department of Biochemistry & Molecular
and DNA damage.
Biophysics at the Washington University School of Medicine.

26
27

Watson, JD, Crick, FH, Nature vol. 171:737-738 (1953).

http://dasher.wustl.edu/ffe/images/DNA%20B-Form%20Dodecamer.png

22

Appendix 1:
Module 2: Additional Websites
NASA Quest
http://quest.nasa.gov/
The Biology Corner
http://www.biologycorner.com/bio1/DNA.html
NASA Science website
http://science.nasa.gov/
NASA Quest website for Flies In Space
http://quest.nasa.gov/projects/flies/
JSC Human Projects and Countermeasures website
http://haco.jsc.nasa.gov/projects/space_radiation.cfm
Brookhaven National Laboratory
http://www.bnl.gov/medical/NASA/NSRL_description.asp
Lawrence Livermore National Laboratory
http://www.llnl.gov/
Genetics Science Learning Center: University of Utah
http://gslc.genetics.utah.edu
Fast Plants information
http://www.fastplants.org/
Human Spaceflight Users: European Space Agency
http://www.spaceflight.esa.int/users/index.cfm?act=default.page&level=11&page=2120
Flies In Space website
http://quest.nasa.gov/projects/flies/
FlyBase
http://flybase.bio.indiana.edu/
Protein Data Bank website
http://www.rcsb.org/pdb

23

Appendix 2
National Education Standards28 by Module
Module 2: Radiation Damage in Living Organisms
Science As Inquiry
Understanding about scientific inquiry
Abilities to do scientific inquiry
Life Science
Molecular basis of heredity
Matter, energy, and organization in living systems
Physical Science
Interaction of energy and matter

28

http://lab.nap.edu/html/nses/6a.html

24

National Aeronautics and Space Administration

Ames Research Center
Moffett Field, CA 94035-1000
www.nasa.gov