Space
Biology
1
Space Biology
Research on Cells,
Animals, and Plants
in Space
Gilles Clément
International Space University
Strasbourg, France
Space
Biology
Lecture Outline
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• What is Life? Evolution of Life. Life on Mars
• The effects of gravity on cell shape and function.
Gravitational Biology
• The effects of spaceflight on development of animals.
Development Biology
• The effects of spaceflight on plants. Plant Biology
• Biotechnology in microgravity
• The biological research facilities
on board the International
Space Station
Water droplets on a plant
on board the ISS (NASA)
Space
Biology
The Evolution of Cell
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• Living cells arose on Earth by the spontaneous aggregation of
molecules about 3 ½ billion years ago
– These early (prokaryotic) cells (e.g. bacteria) are small, with
relatively simple internal structures containing DNA, proteins
and small molecules
– They replicate quickly by simply dividing
in two (a single cell can divide every
20 min. and thereby give rise to
5 billion cells in < 11 hours).
Their ability to divide quickly
(growth rate) enables these
cells to adapt rapidly to
changes in their environment
⤶⤷
– Bacteria can utilize virtually any type of organic molecule
as food (including sugars, amino acids, fats, hydrocarbons,...)
and get their energy (ATP) from chemical processes
in absence or presence of Oxygen
Space
Biology
Speaking about Contamination
• The Apollo-12 Lunar
Module landed on the Moon
156 meters away from
Surveyor-3, which landed
there 2 ½ years earlier
• The astronauts recovered
a camera of the Surveyor
spacecraft for analysis
back on Earth
• Specimen of a bacteria
(Streptococcus mitis oralis)
were still alive on the
camera
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Space
Biology
The Worldʼs Oldest Living Thing
• Spores of the Bacillus bacteria were found
during the summer of 2000 in salt crystals
buried 600 meters below ground at a cavern
in New Mexico, US
• When they were extracted from the crystals
in a laboratory and placed in a nutrient
solution, the micro-organisms revived and
began to grow
• These bacteria have survived in a
state of suspended animation
for 250 million years
• Until then, the worldʼs oldest living
survivors were thought to be
25-40 million year old bacteria spores
discovered in a bee preserved
in amber
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Space
Biology
The Evolution of Cell
• About 1 ½ billion years ago appeared larger and more complex
cells such as those found in higher animals and plants
– These eukaryotic cells (or protozoa) have a nucleus which
contains the cellʼs DNA, and a cytoplasm where most of the
cellʼs metabolic reactions occur
– They get their ATP from aerobic oxidation of food molecules
(respiration) or from sunlight (photosynthesis)
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Space
Biology
Evolution of Organisms
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(deduced from their genes sequences)
rat
toad
shrimp
maize
mushroom (multi-celled)
yeast (one-celled)
Histoplasma capsulatum
(causes lung infection)
Penicillium notatum (produces
the drug penicillin)
vertebrates
invertebrates
plants
fungi
protozoa (multi-celled)
Giardia intestinalis
(causes diarrhea)
protista
Trypanosoma brucei
(causes sleeping sickness)
Plasmodium gametocyte
(causes malaria)
extremophiles
Sulfolobus
E. coli (causes food poisoning)
Strepococcus pyrogenes
(causes strep throat)
Mycobacterium tuberculosis
(causes tuberculosis)
archaebacteria
eubacteria
Space
Biology
Life on Mars
• This 4 ½ billion-year-old rock is a
portion of a meteorite (ALH84001)
that was dislodged from Mars and
that fell to Earth in Antarctica about
16 million years ago
• It is believed to contain fossil
evidence that primitive life may
have existed on Mars more than
3 1/2 billion years ago
Possible microscopic
fossils of bacteria-like
organisms
High-resolution scanning electron
microscope images of AHL84001
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Space
Biology
Gravitational Biology
• How cells, as single unicellular organisms, or as the basic
unit of multi-cellular organisms, are sensitive to gravity?
• Studies of the effects of gravity on:
– Cell Morphology
• Shape
• Structure (skeleton)
• Polarization (up-down orientation)
– Cell Function
• Secretion
• Transportation of substances
in and out the cell
• Immune response
– Cell-Cell Interaction
• Communication
• Differentiation
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Space
Biology
Results of Space Experiments
• Bacteria
– Increase in growth rate (proliferation)
– Increased resistance to antibiotics
Colonies of
bacteria (Bacillus
subtillus) cultured
on board Skylab
Bacteria grown
under the same
ambient conditions
as Skylab, but on
Earth
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Space
Biology
Results of Space Experiments
• Human Cells
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White Blood Cell
– Space anemia
– Resistance to
bacteria or virus is
altered
Red Blood Cells
Platelet
Blood draw kit
Space
Biology
Human Cells in Space
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• Reduction in the number of red blood
cells (space anemia)
– Red blood cells carry the oxygen
to the muscles
– The reduction in plasma volume
(due to fluid loss) causes an
over-abondance of oxygen-carrying
capability
– Muscles lose mass and require
less oxygen
– This over-abondance of oxygen is
detected by the kidneys
– The kidneys reduce the production of
an hormone (erythropoietin, EPO)
which, in turn, decreases
red blood cell formation
Plasma (55%)
White blood cells
Red blood cells
Space
Biology
Human Cells in Space (contʼd)
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• Resistance to bacteria or virus is altered
(immune reaction)
– Lymphocytes produce antibodies
which counteract the invading body
– Activation of lymphocytes in-vivo is
depressed after spaceflight
– Lymphocytes can be purified from
blood and activated by exposure to
various substances in culture (in vitro)
viruses
microbes
– Lymphocytes activation in-vitro is reduced by 90% in-flight
(probably due to changes in membrane structure)
– However,T-lymphocytes activation in-vitro in space is
accompanied by an increase in secretion of interferon
(a molecule which interferes with virus growth)
– Use of space for synthesis of biomolecules (biotechnology)
Space
Biology
Development Biology
• After fertilization of the amphibian eggs, two rotations occur :
– The whole egg detaches from its jelly
capsule and rotates on itself so that the
heavier vegetative pole moves downwards
– An hour or so later,
just the egg cortex
rotates by 30° relative
to the cytoplasm.
This rotation establishes
the dorso-anterior axis
and depends on a
transient array of
parallel microtubules
at the vegetal cortex
• Very roughly, the animal pole corresponds to the head,
and the vegetal pole corresponds to the dorsal side
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Space
Biology
Amphibian Development
When the cortex does not
rotate normally, the embryo
fails to develop
Cortex rotation:
The cytoplasm
rotates with respect
to the overlying
cortex by about 30
degrees.
The gray crescent is
only visible in
certain amphibian
eggs
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Space
Biology
Embryonic Development
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The early stages are
closely similar among
species (drawn to
scale)
The later stages are
more divergent (not
drawn to scale)
Fish
Salamender
Chick
Human
Space
Biology
Development in Space
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• Invertebrates
– Aquatic species less susceptible to
microgravity than terrestrial species
• Fertilization and larval development
normal in sea urchin eggs
• Formation of skeletal hard parts
(shells, spicules) which involve
calcium carbonate is altered during
development in microgravity
– Insects:
• Development abnormalities in
drosophilas bred in space
• Stick insect: reduced hatching rate of
eggs, but embryonic development
before hatching showed no major
morphological anomalies
Note: Fish is a vertebrate
Space
Biology
Invertebrates in Space
Spiders use both the wind and
gravity to determine the
required thickness of web
material
First web built in space
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Space
Biology
Development in Space (contʼd)
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• Vertebrates
– No vertebrates have been raised from
conception to sexual maturity in space
– No birds or reptiles have bred in orbit
– Fertilized chicken and quail eggs have flown: few young chick
embryos have survived; normal development of eggs launched
at later developmental stages.
– Frog eggs fertilized inflight:
• Differences in early
embryogenesis
• But tadpoles at feeding
Ground
stage are not different
from controls
Flight
Therefore, rotation of egg is not only gravity-driven.
It presumably depends on a transient array of microtubules.
The major embryonic axis forms independently of gravity
Space
Biology
Development in Space (contʼd)
• Vertebrates (contʼd)
– Tadpoles born in space have difficulties
to inflate their lungs in microgravity
– Tadpoles born in space exhibit larger
visual-oriented responses during 9 days postflight
– Fish (Medaka) larvae raised in space swim normally when
tested on Earth (Ijiri,1997)
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Space
Biology
Development in Space (contʼd)
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• Vertebrates (contʼd)
– The Dorsal Light Response (DLR) of fish is a simple model
of visual-vestibular control of posture
– On Earth, the DLR is amplified in fish with no otoliths
– In microgravity, the DLR is dominant in most animals
Arrow show the direction of coming light
Normal carp on Earth
Carp with otoliths of the vestibular system removed on Earth
Space
Biology
Development in Space (contʼd)
• Mammals
– Flown pregnant rats gave birth to normal neonates
after flight
– During postflight delivery, flight dams have twice as many
abdominal contractions as the ground controls
– Flown neonate rats show persistent slower weight-bearing
behaviors (walking, surface righting) postflight. Therefore,
gravity is required for critical developmental periods
The Neurolab mission was
16-day long
Walking/righting of neonates rats in 0-G
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Space
Biology
Behavioral Studies in Space
• Little neurobehavioral research has been done in
microgravity with vertebrates, juvenile or adult
• On Earth, jellyfish normally float upright, and pulse their
•
•
•
mantles upward. In 0-g they were either quiet or pulsed in
large loops
On board Skylab, killifish first swam in somersaults' loops
and spirals. 18 days later, they had ceased to swim
erratically, and relied on light as a indication of up and
down
In 0-g during parabolic flight, pigeons flap their wings and
rotate in place without any forward motion. They do not
seem to have any ability to control their flying behavior
Quail chicks were hatched on Mir. If held in a cosmonaut's
hand, they would not be bothered by 0-g and peck food.
When turned loose, they rapidly rotated and made
somersaults, loops, and spirals
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Space
Biology
Space Research in Plant Biology
• Mechanism of gravity perception
(gravitropism)
• Development of closed ecological
life support systems
• Plants respond to environmental
stimuli such as light (phototropism),
water (hydrotropism), and magnetic
or electric fields. These responses
are masked on Earth by the
overriding response of plants to
gravity
• Role of the absence of 24 hour cycles in light
and temperature on circadian rhythms in plants.
The generation of the circadian rhythms is likely to involve
the membrane transport systems, and these systems are
affected by microgravity
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Space
Biology
Gravity Perception in Plants
Plants have gravity-sensing organs in their roots, which
involve the sedimentation of particles (statoliths)
Zea maize root cap
On Earth, in a
root placed
vertically, the
statoliths (black
particles) are
sedimented at
the bottom end
of the cell
When the root is placed
horizontally for 3 hours, the
statoliths are now sedimented
onto the lateral walls of the cell
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Space
Biology
Gravity Perception in Plants
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Removal of the root abolishes the capacity to detect gravity
Zea maize
• Is it the movement of the statoliths
through the cytoplasm, or the
pressure they exert on other (lower)
cellular components, that is involved in
graviception?
• What is the threshold for gravity
perception?
Space
Biology
Plant Development in 0-g
1-G
0-G
apical stem
leaf
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0-G Effects
Changes in orientation
of stem and leaves
axillary bud
stem
adventilious roots
More adventilious roots
secondary root
Faster growth of
secondary roots
primary root
Changes in orientation
of secondary roots
Reduction of the primary
root growth
root cap
From Perbal (2001)
Loss of apical
dominance
Space
Biology
Roots grow randomly in 0-g,
but can be reoriented uniformly on
exposure to 1-g for as little as 3 hours
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0-g
g
1g
On-board centrifuge
Adapted from G. Perbal (1992)
0-g
1-g for 3 hrs
Space
Biology
Plant Biology in Space — Results
• On-board centrifuge experiments
have demonstrated that the
minimum force that is sensed by
plant organs is in the range of
1/1000th of g
• The root is able to perceive its
orientation with respect to a linear
acceleration vector and to generate
a signal of curvature in less
than 30 seconds
• The reproductive phase is completed in microgravity when the
culture conditions (gas and liquid exchanges) are adequate
• Whether or not a seedling growing from the beginning in
microgravity can flower and produce normal seeds remains a
matter of debate
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Space
Biology
Plant Reproduction
• LDEF: Long-Duration Exposure Facility
– 12 million tomato seeds in space for 6 years
– Post-flight measurement of germination
Germination %
Flight
73.8
Ground Control
70.3
– Conclusion: Seeds remain viable in space
• Cell Division and Chromosomal Damage in
embryos of cultured Hemerocallis (daylily)
Cells in division (%)
Cells in metaphase (%)
Chromosome damage (%)
Double nuclei (%)
Ground
2.6
31.3
0
0
Flight
0.4
11.3
1.7
3.1
– Conclusion: The space environment
(microgravity and/or radiation) results
in reduced cell division and increased
chromosomal damage
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Space
Biology
Plant Reproduction
Seed pod
(silique)
formed in
space
Pre-flight
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Pistil of flower
grown in space
Flowers formed in space
were normal in appearance
Ovules from siliques
formed in space
Post-flight
(11 days)
Pollen germinated and pollen
tubes grew towards style
• Recent experiments showed that reproduction proceeded
normally through the stage of an immature seed
• Past failures were probably due to less than adequate
horticultural conditions
Space
Biology
Bioprocessing
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• Biotechnology is an applied biological science
that involves the research, manipulation, and
manufactoring of biological molecules,
tissues, and living organisms
• Biotechnology has a critical role in health,
agriculture, and environmental protection
Flowering on board the ISS (NASA)
• Space biotechnology is focused on protein crystal growth, cell and
tissue culture, and the fundamental mechanisms of cell growth
and secretion
• Microgravity offers a unique environment that re-orders the forces
exerted on cells. The response of cells to this re-ordering provides
novel insights into fundamental cellular mechanisms. Cells
unloaded from gravity may perform to our advantage in tissue
formation
• The response of cells to microgravity, combined with advances in
molecular biology and genetics, offers the opportunity to explore
new strategies in applied science and contribute to public health
Space
Biology
Biotechnology in Microgravity
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• Research Areas
– Protein crystal growth
– Cell and tissue culture
Lysozyme x150
– Manufacture of biological materials
• Research Themes
– Molecular structure of proteins
and viruses
– Structure-based drug design
– In-vitro drug testing
– New technologies for
bioprocessing (in particular
isolation and purification)
Satellite Tobacco Mosaic Virus
(STMV) crystals grown in 0 g
Courtesy of ESA
Space
Biology
ESA Biolab onboard ISS
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Space
Biology
ISS Centrifuge Accommodation Module
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Space
Biology
Biology Research onboard ISS
• Cell Culture Unit*
• Aquatic Habitat*
• Advanced
•
•
•
Animal Habitat*
Plant Research
Unit*
Insect Habitat**
Egg Incubator**
Research in cell and tissue biology
Capability to maintain and monitor animal and
plant cell and tissue culture for up to 30 days
Egg to egg generation studies for examination
of life stages. Can accommodate small fresh
water organisms for up to 90 days
Housing for up to 6 rats or 12 pregnant mice
for studies of mammals development
Studies of growth and development
for plant specimens up to 30 cm (root+shoot)
Multigenerational and radiation biology
Incubation and development of small reptilian
and avian eggs prior to hatching
* can be used on the 2.5-m diameter centrifuge (0.01-2.0 g)
** equipped with internal 0.01-1.5 g centrifuges
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Space
Biology
Research Opportunities
• International Life Science Research Announcement
(ILSRA) coordinated between the ISS partners
• This ILSRA is released annually and includes several
research opportunities
– Life Sciences themes using the ISS
– Research in biology using sounding rockets
– Research in biology using biosatellites
– Utilization of the bed rest facilities to prepare for
human physiology projects on the ISS
– Utilization of the centrifuge facilities to prepare for
artificial gravity projects
– Utilization of the specific environment and living
conditions of polar stations
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Space
Biology
Reading Material
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• Clément G (2005) Fundamentals of Space Medicine.
Dordrecht: Springer
• Clément G, K Slenzka (2006) Fundamentals of Space Biology.
El Segundo: Microcosm Press, New York: Springer
• Dutemple L (2000) The Complete Idiotʼs Guide to Life Sciences.
Indianapolis, IN: Alpha Books
• Oser H, Battrick B (eds) (1989) Life Sciences Research in Space.
Noordwijk: ESA Publication Divisions, ESA SP-1105
• Future Biotechnology Research on the International Space Station
(2000). Commission on Physical Sciences, Mathematics, and
Applications, Space Studies Board. Washington DC: The National
Academies Press.
Available online at URL:
http://www.nap.edu/books/03090697/html
• http://astrobiology.arc.nasa.gov/genomics
/technologies/available_hardware.html