Transcript Bioluminescence

By GC
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Defined : the emission of light as a result of a chemical
reaction during which chemical energy is converted into
light energy
From a living organism that functions for its survival or
mating
Comes from Greek word “bios” for ‘living’ and Latin word
“lumen” for ‘light’, so it literally means living light
“Cold light” resulting from a specific biochemical
mechanism involving chemical processes
Specific for that organism
Found all over biosphere, phosphorescence in sea water
observed in all oceans
Bioluminescence of visible light is found in a majority of
marine organisms but is rare in terrestrial organism
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All involve an oxygen oxidation of organic molecule
(luciferin)
Catalyzed by an enzyme called luciferase
These proteins are called “photoproteins” -> oxygen is already
bound to the luciferin
 A photoprotein is a protein with a luciferin bound to it
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“Antenna proteins” -> adjust the colour of bioluminescence
likeness to proteins on similar function in photosynthesis
except they act in reverse
5 known distinct chemical classes of luciferins:
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Key organ -> photophore (light producing organ)
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Aldehydes, benzothiazoles, imidazolopyrazines, tetrapyrroles and
flavins
Seen in many luminous fish and vividly in cephalopods
Make up of complex photogenic (light emitting) cells
Bioluminescent reaction components detected in stomach,
secretory organs and liver of some organisms
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Bioluminescence: light is created through a chemical
reaction. Bioluminescence is a subset of chemiluminescence
Fluorescence: a particular atom or molecule absorbs light of
one length, but emits one of a longer length. The light going
in is usually UV photons, and comes out as visible light
Phosphorescence: requires more time to remit radiation
absorbed than fluorescence. This is because the sub atomic
reactions required to remit light occur less often than in
fluorescence.
Fluorescence
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Stories of mysterious light or fires seen over fields or mountains
were often said to be dragons or the gods
Greeks and romans first to report luminous organisms
Reports of fireflies found in early religious writing of India and
China
Earlier recordings believed to have come from these and ancient
eastern civilizations and refered to firelies and glow worms
Aristotle (384-322BC) (first to discover) decribed more than 180
marine species, first to recognize “cold light”
Later, complete and extensive descriptions of luminous organisms
published by Pliny the Elder (23-79 CE)
16th century references to bioluminescence found in literature such
as Shakespeare, specifically in hamlet who talked about “effectual
fire of the glow worm”
First book devoted to bioluminescence and chemiluminescence
published in 1555 by Conrad Gesner
1667 – Robert Boyle documented oxygen was needed for
luminescence
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Raphael Dubois performed experiment, he extracted the two
key components of bioluminescence reaction and was able
to create light and that there 2 key components= “luciferine”
and heat labile “luciferase”*
One of most eminent scientists of 20th century was
“Princeton Professor” E. Newton Harvey*
He was looking for existence of lucifernin- luciferase system
in all luminous organisms
First luciferin isolated in 1956
First photo protein isolated was the calcium activated
photoprotien aequorin in the 1960’s
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Calcium dependant photoprotein cloned in 1985
 Intensity of luminescence varies with calcium concentration
 Therefore aequorin has been used in monitoring of cell calcium
1985 firefly luciferase was cloned
Tomopteris
Firefly
Luciferin chemical structure
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All colours of the visible light spectrum; red, orange, yellow,
green, blue, indigo, violet
Different colours dependant on role the light plays and
which organism it is produced in
Many only produce one colour, however some are capable
of many colours
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Eg. Jamaican click beetle; this is due to the same luciferin substrate as the
firefly, but different luciferase structures
Bioluminescence releases a large amount of energy, not heat
Visible light radiation is equal to light wavelengths of 400700 nm
Bioluminescence max of most marine species is 460-510 nm
Terrestrial organisms mostly yellow-green BL, more yellow
Marine mostly blue-green (400-500nm) luminescence
because it travels the best through water
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Plays an important role in nature due to the darkness 200m
underwater
Location on body of bioluminescence gives clues to the
functional role of the luminescence
Attracting a mate
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Attracting prey
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eg. Fireflies flash certain patterns to either attract mates or communicate
Camouflage
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eg. Loose jaw fish, bioluminescent organs in cheeks to see in dark water
Communication
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Eg. Angler fish has a luminescent lure
Finding food
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Eg. The wave lengths attract mates, but do not draw attention to themselves
eg. Bobtail squid blends into background
Defense against predators
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eg. Deep sea shrip vomits bioluminescent
material in direction of attacker
Deep- sea shrimp vomits bioluminescent
material
ROBERT BOYLE
 British philosopher and scientist
 In 1667 performed experiment and found that
bioluminescence didn’t occur when a known bioluminescent
type of fungus was not in the presence of air
 1672 he discovers that bioluminescence reactions require air,
when he later discovered oxygen, we realize is the oxygen
component of air that is needed
Robert Boyle
Oxygen Atom
RAPHAEL DUBOIS
 1887 he discovered luciferin and luciferase through
Raphael Dubois
experiments with clams, beetles, and other species
 He did an experiment using a clam called the common
paddock
 Ground tissue of the clam up in cold water, and light was
produced for several minutes. This showed he has extracted
the light producing chemical
 Then made a hot water extract from another clam and added
it to the cold water, which reactivated the light reaction
 A hot water extraction alone produced no light reaction
 Called the hot water extraction luciferin
 Called cold water extraction luciferase
 Noted that luciferin could only glow in the presence of
luciferase
EDMUND NEWTON HARVEY
 Greatly popularized the study of bioluminescence
 Discovered that luciferins and luciferase from different
animals are not interchangeable
 Evidence of evolution of bioluminescence to fit various
needs of different species
 Now shown that evolution of bioluminescent systems has
occurred over 30 times, which accounts for the differences in
colours and uses in different species
Humpback Angler
Fish
Jamaican Click Beetle
The discovery and study on bioluminescent organisms is the
goal of many expeditions of ocean going research vessels
and marine submersibles
COMMERCIAL APPLICATIONS
 Glowing trees to save electricity bills
 Agricultural crops that luminesce when in need of water
 Detection of bacteria in contaminated foods
 Novelty pets
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Bio-indentifies for convicts, mentally ill
 Glowing toys, glowing greeting cards
 Luminescent beer and champange
“Glofish”
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MEDICAL APPLICATIONS
 Reporter genes
 Bioluminescent Imaging*
 In vivo analysis
 Detection of bacteria
 Observations of protein to protein interacting
 Testing for genetically modified organisms*
 Water quality testing*
 Green Florescent Protein*
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Used on small animals
Real time monitoring of the progression of infections in the
same animal
Typically 2D imagery, with lower resolution
Can mark progress at different time points without
euthanizing the animal, uses animals mice than
conventional methods of progress tracking
Quicker and relatively inexpensive
Allowed specific molecular and cellular events such as cell
migration and signal transduction to be investigated in a
living, intact animal
Bioluminescence is decreased by pigmentation of organs
such and liver, spleen and the fur
 To get around this they can shave the mice
or use ones with the albino gene
 Used to study bacteria distribution,
distinguish between more and less virulent
strains and moniter antibiotic therapy
 Also used to study viruses, but because of
low resolution, it can be difficult to distinguish
between viral infections in adjacent tissues
 Used to investigate parasite infections
Mouse and BLI
 High correlation between light intensity and
amount of parasites in spleen
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For water quality/ toxicity testing
Vibrio Fischeri
Uses the bioluminescent marine bacteria ‘vibrio fischeri’
When the organism is challenged by a toxin, its breathing
pathway is disrupted, resulted in decreased
bioluminescence intensity
Now a well established and excelled genetag and protein
It can be fused to a protein of interest and fluorescence and can
be tracked within a cell to study its localization and behavior
Outstanding structural stability
Excellent for studying the cell and sub cellular processes
Cloned in 1992, expressed in various organism in 1994
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People want to know, “Does this food contain genetically
modified organisms?” and they have the right to know
Genetically Modified Organisms (GMO): genetic material (DNA
or RNA) that has been altered in ways that would not occur under
natural conditions or natural processes
Most common technique used to test for GMOS is polymerase
chain reaction (PCR)
PCR requires complex DNA extraction techniques, rapid
thermocycling, expensive equipment, and is a lengthy process
Company called Lumora came up with a breakthrough
technology to test for GMO
It’s a combination of 2 technologies; Bioluminescence and
isothermal DNA amplification
Bioluminescence used is bioluminescence real time reporter
(BART)
Organic food symbol
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Loop mediated isothermal amplification (LAMP)
BART uses luciferase and detects DNA and It lights up when it
find specific DNA and RNA sequences linked to genetic
modifications
GMO testing can be done out in the field or in a food processing
center
LAMP-BART technique requires only basic equipment for DNA
extraction, a constant temperature and simple light detection
Quicker than PCR
Good for farmers to have a cheap, reliable way to test products for
GMO’s
Lumora’s GMO detector can also be used to test for things like
salmonella in foods
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What are the biological advantage of light emission to the
animal?
Evolution history?
Metabolic/ dietary source of the luciferins?
What are the control mechanisms for light flashing?
Discovering- luminous mollusc, a roman delicacy, the
bioluminescence mechanism of it is still not completely
solved
•Deep-sea Shrimp
http://www.nytimes.com/imagepages/2011/12/20/science/20JPBIO1_SPAN.html
•Chemical equations and fluorescence animations:
http://www.lifesci.ucsb.edu/~biolum/chem/
•Tomopteris
http://discovermagazine.com/photos/1-8-marine-creatures-that-light-up-the-sea
•Chemical Structure drawing of Luciferin
http://www.biotium.com/product/applications/Enzyme_substrates/price_and_info.asp?item=10100&layer1=D;&layer2=D01;
•Robert Boyle
http://www.friedpost.com/sciencetech/greatest-science-discoveris-boyle%E2%80%99s-law-1977.html
•Raphael Dubois
http://en.wikipedia.org/wiki/File:Raphael_Dubois_(1849-1929).jpg
•Oxygen Atom
http://www.historyforkids.org/scienceforkids/chemistry/atoms/oxygen.htm
•Bioluminescent Imaging mouse
http://www.caliperls.com/products/preclinical-imaging/?gclid=CMiigb2T_q8CFWwDQAodmVZCzg
•Glofish
http://fish-blog.co.uk/News/manufacturer_supports_glofish_industry/
•Vibrio Fischeri
http://microbewiki.kenyon.edu/index.php/Vibrio_fischeri
•Bioluminescent Imaging mouse
http://www.caliperls.com/products/preclinical-imaging/?gclid=CMiigb2T_q8CFWwDQAodmVZCzg
•Organic food stamp
http://www.google.ca/imgres?um=1&hl=en&sa=N&biw=1366&bih=569&tbm=isch&tbnid=pcYC93zfdGquhM:&imgrefurl=http://www.omafra.gov.on.ca/e
nglish/crops/organic/certification.htm&docid=MuinqDkP2Ld1M&imgurl=http://www.omafra.gov.on.ca/english/crops/organic/certificationf10.jpg&w=600&h=616&ei=JSGwT6XnFarE6QG1rqCvCQ&zoom=1
•Humpback Angler Fish
http://written-4u.blogspot.ca/2007/01/allah-creator-sustainer-surely-it-is.html
•Jamaican Click Beetle
http://www.asknature.org/media/image/6970
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Beecher, C. (2012, May 7). Breakthrough Offers Promise of Improved GMO Testing. In Food Safety News.
Retrieved May 9, 2012, from http://www.foodsafetynews.com/2012/05/breakthrough-offers-promise-of-improvedgmo-testing/
Binger, J. M. (2007, March 14). Bioluminescence. In Center for Biophotonics Science and Technology. Retrieved
May 6, 2012, from cbst.ucdavis.edu/education/courses/spring-2007./bingerfinaldraft.doc
Hutchens, M., & Luker, G. D. (2007, June 24). Applications of bioluminescence imaging to the study of
infectious diseases. Retrieved May 7, 2012, from Wiley Online Library (10.1111/j.1462-5822.2007.00995.x).
John, L. (08). Basic Bioluminescence. In Photobiological Sciences Online. Retrieved May 7, 2012, from
http://www.photobiology.info/LeeBasicBiolum.html
Lee, J. (08). A History of Bioluminescence. In Photobiological Sciences Online. Retrieved May 6, 2012, from
http://www.photobiology.info/HistBiolum.html
Lumora welcomes expressions of interest for GMO test commercialisation after BART technology demonstrates
ability to detect genetically modified contamination of crops rapidly and at very low levels o. (2012, April 30). In
Lumora . Retrieved May 9, 2012, from
http://79.170.44.87/lumora.co.uk/index.php?option=com_content&view=article&id=100:lumora-welcomesexpressions-of-interest-for-gmo-test-commercialisation-after-bart-technology-demonstrates-ability-to-d