Chlamydomonas reinhardtii is a unicellular biflagellate eucaryote. It is a theoretical account being because it contains an copiousness of conserved familial traits that extends back to pre-historic times ( Schmidt, 2006 ) . Chlamydomonas has an inducible system to use changing Ca degrees in the environment to change chemotaxis and motility. By altering Ca degrees from 10-4M to 10-7M, the end was to find cosmopolitan Ca based motion forms. Four strains of C. reinhardtii were utilised: wild type ( control ) , pf14 mt+ , p18 mt+ , and oda1 mt+ . Wild type Chlamydomonas exhibited the greatest motion forms from add-on of Ca. Pf14 mt+ , p18 mt+ , and oda1 were specifically selected due to their altered motility protein mutants. This experiment will assist to set up a specific motion phototactic form of C. reinhardtii uses to last.
C. reinhardtii is a authoritative theoretical account for cistron analysis. Despite it ‘s apparently simplistic nature, the eucaryotic algae has basal organic structures and scourge comparable to mammalian centrioles and cilia ( Leon, Rosa, et al. , 2007 ) . Cilopathies, or defects in ciliary motility and assembly in mammals, can consequence: sperm motion ( because male sperm is the closest human homolog to C. reinhardtii, sharing 75 % of the same cistrons ) , respiratory piece of land waste riddance, left-right organic structure orientation during embryosis, and coordinated motion of fluids over epithelial cells ( including respiratory piece of land, female generative system, and encephalon ventricles ) ( Sidflow and Lefebvre, 2001 and Smith, 2007 ) . Unraveling the enigma behind C. reinhardtii motor proteins would progress a assortment of subjects including: malignant neoplastic disease intervention, respiratory intervention, construct and gestation research, familial fleshiness, and kidney polycystic research ( D’Souza, J. S. , 2009 ) . Cilia cistrons have been conserved since pre-historic times, so it ‘s non a surprise that many of the unwellnesss supra would profit from bring outing cilia signaling tracts. Recognizing that a apparently simplistic eucaryote can find the destiny of so many, it makes one think that green algae is important. By finding the motion forms of C. reinhardtii, higher existences everyplace might potentially profit. Principles of scourge map in C. reinhardtii can be extended to human homolog. C. reinhardtii wild type and pf14, pf18, and Oda1 motility mutations will be compared to find the relationship between construction and motility in C. reinhardtii and possibly one twenty-four hours, worlds.
Environmental Ca concentration determines the motion of Chlamydomonas. There are five types of motion algae can exhibit: additive frontward, additive backward, coiling, clockwise handbill, and counterclockwise handbill and three possible wave form conformations: symmetric, quiescent, and asymmetric ( Bessen, M. R. , et Al. 1980. ) Research varies as to what Ca concentrations induce what specific gesture. Each cell type ( mutant or wild ) has the possible to quickly undergo one of these five types of motion once environmental Ca degrees are varied.
Testing alimentary restriction responses would uncover the mechanisms that permit C. reinhardtii, and possibly beings with similar cistrons, to digest nutritionally scarce environments. The intent of this experiment would to get down developing a incorporate scientific motion theoretical account of C. reinhardtii in response to alterations in environment, specifically calcium degrees.
To get by with low alimentary environments, beings have inducible systems that enable them to scavenge and expeditiously use restricting foods. Survival is dependent on the ability to set. Chlamydomonas reinhardtii shows extensive metabolic flexibleness that allows it to populate distinguishable environmental niches and to last fluctuations in alimentary handiness. It is a classical theoretical account due to the cell ‘s built-in copiousness of evolutionally conserved cistrons that mirrors workss and non-photosynthetic being responses. Testing alimentary restriction responses would uncover the mechanisms that permit C. reinhardtii, and beings with similar cistrons, to digest scarce environments. The intent of this experiment would to efficaciously depict the motion patterns C. reinhardtii and its mutations exhibit in harmonizing to alterations in environment, specifically calcium degrees. Variations in environmental Ca concentrations control C. reinhardtii motion, but predominating research has failed to supply agreeable informations on the motion specific Ca concentrations induces.
Chlamydomonas reinhardtii as a Model Organism
Over a billion old ages ago, Chlorophytes ( green algae, including Chlamydomonas ) deviated from Streptophytes ( land workss ) . C. reinhardtii is one of the lone lasting beings that still contain conserved cistrons that trace back to precursor plant-animal cells. 109 old ages of development separate C. reinhardtii and mammals, yet C. reinhardtii scourge are highly similar in construction and map to mammalian cilia and scourge. A figure of the scourge proteins in C. reinhardtii exhibit 75 % individuality and similarity to proteins with similar map in mammalian sperm ( Merchant, et al. , 2010 ) .
C. reinhardtii is a theoretical account being, despite being a unicellular algae Protozoa comprising of chondriosome, ocellus, two anterior scourge, and a chloroplast ( Harris, 2001 ) . In an optimum environment ( light, O2, CO2, and mineral salts ) , the alga behaves like a phototrophic works to bring forth its ain C. However, C. reinhardtii besides has the capacity to last in less optimum milieus ( high salt, low sulfur, no visible radiation ) . The C. reinhardtii can move as a heterotrophic, like bacteriums, and use environing C beginnings such as ethanoate for energy ( Merchant, 2010, Harris, 2001 ) . Additionally, this cell is can be cultured and maintained with easiness ( Harris, 2001 ) . With such a short life span, it is easy to coerce the being to bring forth big measure of protein of involvement or turn in a variable environment. Even though it has similarities to mammalian cells it is non about every bit susceptible to infection. The wild type cells familial stuff can be manipulated to detect alterations in normal map or construction.
C. reinhardtii uses a brace of anterior scourge, which is a flexing organelle that protrudes from the cell wall to swim and feel environmental conditions. Light perceptual experience is necessary for phototaxis, but the cell besides need to react to intense light beginning to forestall excessively much light soaking up, or photoshock. Therefore, the being ‘s ability to travel towards or off from light by whiplike system is indispensable for endurance ( Silflow and Paul Lefebvre, 2001 ) .
Scourge have the “ 9+2 ” axonemal construction, where nine outer doublet microtubules surround two interior vest microtubules. The nine doublet microtubules each consist of a complete A tubule fused to a partial B tubule in a stretched ring conformation that forms the walls of the basal organic structure ( Alberts, et al. , 2008 ) . Dyneins are located on the A tubule dwelling of an inner and outer dynein weaponries that severally control wave form and force ( Movassha, 2010 ) . Dyneins act as an interface between the outer to inner flagella constructions because are connected to radial radiuss and the A tubule.
The inner and outer dynein weaponries are forces responsible for the skiding gesture between outer-doublets that finally consequences in flagella motion. Inner dynein weaponries control the type of motion ; outer dynein weaponries control produce the round force, or how fast scourge move ( Movassha, 2010 ) . In the presence of ATP, the dyneins bind the A tubule of the next outer doublet and travel along the B tubule toward its subtractions terminal, ensuing in skiding between braces of outer doublets. The skiding gesture can non prevail due to the propinquity with other doublet microtubules and the basal organic structure. This creates possible energy buildup that is converted into scourge flexing gesture leting wave-like motion ( Alberts, et al. , 2008 ) .
The interior dynein weaponries are located along the A microtubule. They are comprised of eight heavy ironss, each heavy concatenation has the ability to bring on different verifications that control motility and moving ridge alterations. One heavy two-headed concatenation is connected to six-single headed heavy ironss to organize a web with intermediate and light ironss and alter the wave form form. One of the of import visible radiation ironss in the interior dynein arm is centrin, is a light concatenation that binds Ca. The assorted and variable interactions between the visible radiation, intermediate, and heavy ironss induce the different types of wave form the scourge exhibits ( Movassha, 2010 ) .
The outer dynein arm ‘s three heavy ironss: I± , I? and I? transfer possible energy, ATP, into kinetic energy, motion so the scourge will shift harmonizing to interior dynein wave form alterations. I± , I? and I? contains hexametrical arranged ATP-binding molecules, one time filled, they hydrolyze ATP to supply the heavy ironss with the ATP energy to bring on all in force ( Movassha, 2010 ) . Mutants in I± , I? or I? heavy ironss on the outer arm will change the speed of outer dynein weaponries along the microtubules since ATP hydrolysis will be reduced, forestalling energy to come in the dynein composite ( Heuser, 2009 ) . A specific mutant is oda1 mt+ , the ODA1 protein is absent from the outer dynein because the coding cistron encodes a stop downstream of the instigator, forestalling a docking protein to be formed. The docking protein acts as an ground tackle for the outer dynein arm onto the microtubule doublet ( Takada, 2002 ) .
Motile scourge contain radial radiuss, that consist of two parts: a “ chaff ” and “ spoke caput ” . The chaff part is positioned following to the interior dynein weaponries, while the chaff ‘s caput extends into the microtubule doublet. Bing near the interior dynein weaponries and stretching to the interior microtubules facilitates signal transduction ( Yang, et al. , 2006 ) . Mutants such as pf14, that lack radial radiuss exhibit unnatural motility or are paralyzed, since it relays the binding signals from the interior dynein weaponries to the microtubule doublet. Without signal transduction, dynein can no longer impel the microtubule to the axenome so any fluctuations in environmental Ca2+ concentration would non impact motion since none can be propagated with damaged/absent radial radiuss or dyneins ( Witman, et al. , 1978 ) .
Effectss of Environmental Calcium on Chlamydomonas reinhardtii
For heterotrophic photosynthetic beings, exposure to visible radiation is necessary for endurance. Plants optimize their exposure to visible radiation by turning towards it, but flagellated beings must swim toward it. C. reidhardtii cells swim to optimal light environments by modulating scourge whipping with environmental Ca. They have a Rhodesian-like photoreceptor ocellus to comprehend light strength for phototaxis ( Suzuki, et al. , 2003 ) . When visible radiation is perceived by the photoreceptor, a current propagates from the ocellus to the plasma membrane by signal transduction. The signal transduction induces alterations in the intraflagella free Ca degrees in the interior dynein arm ( Mitchell, 2000 ) . The addition in calcium concentration depolarizes the cell membrane, voltage-dependant Ca ion channels open in response, and the flow of environmental Ca rises in the cell. If the depolarisation is great plenty, the voltage-gated Ca2+ channels in the flagellar membrane unfastened and Ca2+ permeates.
Calmodulin binds the extra intraflagella Ca2+ concentration which induces a conformational alteration in the radial radius chaffs that allows signal transduction from the cardinal microtubules to the interior dynein. The interior dynein arm contains centrin, another protein that binds Ca ( Silflow and Lefebvre, 2001 ) . Bound centrin signifiers a subcomplex to an next visible radiation concatenation 4 protein which coordinates tight Ca adhering with the centrin. The visible radiation concatenation 4 interacts with the N-terminal root sphere of the I? HC, giving the heavy concatenation on the outer dynein entree to centrin, and hence Ca. The form and the size of the wave form are determined by the inner weaponries ( increasing Ca2+ is detected by centrin adhering which consequences in altering flagellar wave form ) ( Movassha, 2010 ) and can be powered by the outer dynein to increase round frequence ensuing in photoshock, swimming off from light, ( Sakato and King, 2001 ) . After a short period of clip, Ca2+ is returned to the phototaxis degree by the action of Ca2+ pumps and the cell returns to normal swimming towards visible radiation ( Mitchell, 2000 ) .
The focal point in this survey is to accurately depict alterations in Chlamydomonas whipping forms when specific sums of Ca are introduced in the planetary environment. The traveling pattern research is obscure and/or conflicting, and the purpose of this experiment is to try to sign the ambiguity. The undermentioned is comparing of theories of three Chlamydomonas experts chosen based on the sum of documents they wrote/quality of their work. Additionally, merely one paper implicitly states the quantitative ratios of motion, we want to observe the figure of cells that demonstrate different forms. It does non look logical that all the cells would be in merely one type of motion at that peculiar environment. The undermentioned ( Table 1 ) is a speedy overview of the types of Ca driven phototaxic motion forms explained in the M. Bessen ‘s, et al. , literature entitled “ Calcium Control of Waveform in Isolated Flagellar Axonemes of Chlamydomonas, ” and Sakato and King ‘s research entitled “ Calcium Regulates ATP-sensitive Microtubule Binding by Chlamydomonas Outer Arm Dynein. ”
Indicated sum of environmental Ca.
10-6-10-9 M Ca2+
10-6 M Ca2+
10-6 M Ca2+ -10-5 M
10-4 M Ca2+
10-3 M Ca2+
Sakato and King
( S and K )
Asymmetrical wave form.
Symmetrical wave form.
Propagation of Movement
( S and K )
1 scourge moves.
move in a forward consecutive line.
No motion cells stayed still in V-shape constellation
Move in a backwards line.
Bessen, et Al.
( B. , et al. )
Asymmetrical wave form.
Asymmetric to Symmetric
Propagation of Movement
( B. , et al. )
90 % cells swam in circle. 10 % in bottle screw form.
90 % cells switch from a handbill to consecutive way. The other 10 % swim in a stretched “ bottle screw ” constellation,
Straight, parallel line frontward.
No motion but stayed in consecutive line.
Table 1. Comparison environmental induced Ca degrees harmonizing to research.
Sakato provinces that cells in 10-4 M Ca2+ , swim with a synchronised symmetric wave form in a backward way. Therefore, the Chlamydomonas scourge must incorporate at least two Ca2+ detectors that respond to different metal concentrations and command flagellar wave form transition through changes in dynein motor map ( Sakato and King, 2003 ) . Bessen asserts at 10-4 M Ca2+ axenomes moved in consecutive parallel lines from base to tip. At 10-3 M free Ca2+ , a big per centum of the axonemes propagated seemingly symmetrical decompression sicknesss, but the all in frequence was really slow ( about 1 Hz ) and no swimming was observed ( Bessen, et al. , 1980 ) .
During normal forward swimming, the two Chlamydomonas scourge round with an asymmetric wave form. Sakato provinces, At 10-6 M Ca2+ two Chlamydomonas scourge round with an asymmetric wave form to ensue in forward swimming ( Sakato, et al. , 2006 ) . Bessen asserts that the asymmetric wave form alternatively of a swimming in a consecutive line, that cells in 10-6 M Ca2+ , 90 % of Chlamydomonas swam in a circle with a diameter of 4I?m with a velocity of 2-5 rps. When viewed from above, 90 % of the cells that touched the glass slide swam counterclockwise, and 90 % of cells in contact with the coverslip swam clockwise. The staying cells swam in a coiling “ bottle screw ” manner ( Bessen, et al. , 1980 ) .
Sakato provinces as intraflagellar Ca2+ additions from below 10-6 M Ca2+ to 10-5 M Ca2+ , cells cease to swim and scourge is quiescent in a V-shaped constellation ( Sakato and King, 2003 ) . Bessen asserts scourge ne’er pauses its whipping rythym, but over 90 % of Chlamydomonas switches from an asymmetrical to symmetrical wave form someplace between 10-5 M Ca2+ and 10-6 M Ca2+ . The other 10 % swim in a stretched “ bottle screw ” constellation, but the incline was longer than the cells than the “ bottle screw ” verification at cells in 10-6 M Ca2+ ( Bessen, et al. , 1980 ) .
At 10-6-10-9 M Ca2+ axenomes did non swim in any specific way ( Bessen, et al. , 1980 ) . flagellar motion is seemingly inhibited at such low concentrations of Ca2+ . Yet, Sakato and King, asserted that in free Ca concentrations between 10-7-10-9M Ca2+ , there is merely plenty free Ca for one scourge to be activated and swim towards visible radiation ( Sakato and King, 2003 ) .
Chlamydomonas reinhardtii shows extensive metabolic flexibleness that allows it to populate distinguishable environmental niches and to last fluctuations in alimentary handiness. The cells have developed a phototactic Ca controlled system to guarantee endurance. Waveform form of Chlamdymonoas in assorted concentrations of Calcium needs to be ascertained, there should be a method to separate if cells move helically or linearly. It does non look logical that all the cells would exhibit merely one type of motion, old surveies fail to advert the velocity the cell can travel or sum of cells in a peculiar form of motion. Video microscopy allows us to track changing calcium concentrations in wild type and mutant Chlamydomonas reinhardtii, so a gesture form should be ascertained.
In order to set up if phototactic motion has specific forms in changing sums of environmental Ca, four strains of Chlamydomonas reinhardtii ; wild-type, pf14 mt+ ( mutated radial radiuss ) , oda1 mt- ( dynein arm mutant ) , and pf 18 mt- ( cardinal microtube mutant ) will be cultured. All mutations were chosen for their motility defects, because observation of smasher traits are required to compare to wild-type consumption of environmental Ca and hence flagellar motion. All cells will be cultured at the same denseness, 1x105cells/mL, so all environmental Ca will be later eliminated by three washes in non-calcium TAP medium. Specific aliquots of Ca be reintroduced into cell civilizations and incubated for specific clip intervals. To efficaciously supervise Chlamydomonas reinhardtii motion forms, a Leica DM1000 inverted compound microscope will capture all cellular activity.
Movement forms will be determined by numbering the frequences of cells that moved in a peculiar wave form and way over a peculiar clip frame. A successful experiment will find if presenting restricting supplies of the environmental factor, Ca, does hold promote traveling forms in Chlamydomonas reinhardtii phototaxis. We will besides be finding if mutants that affect normal scourge mobility have the same limited motion in the varying Ca mediums. Comparing the discrepancy samples to the control samples will qualitatively find effectivity of the experiment.
C. reinhardtii is an optimum being to analyze motility. Its genome is evocative of ascendant plant-animal predecessors every bit good as cistrons prevalent in a assortment of beings in the present twenty-four hours. C. reinhardtii can last in a assortment of environments from dirt to snow on mountaintops, and can last by phototropic or heterotrophic agencies ( Merchant, 2010, Harris, 2001 ) . Additionally, this cell is highly unsophisticated to civilization, its full life rhythm, with such a short life span, it is easy to coerce the being to bring forth big measure of protein of involvement or turn in a variable environment. Even though it has similarities to mammalian cells it is non about every bit susceptible to infection, and little sums of amphicillian can be introduced to civilizations, guaranting endurance ( Silflow and Lefebvre, 2010 ) . Wild type C. reinhardtii familial stuff can be manipulated to detect alterations in normal map or construction.
C. reinhardtii Mutants
C. reinhardtii mutations for this experiment will be: pf14 mt+ , oda1 mt- , and pf18 mt- ( cardinal microtube mutant ) each exhibit malfunctions in flagellar construction. Pf14 mt+ contains a mutant within the radial radiuss, ensuing in the deficiency of signal transduction from the dynein indicated by the lifting or falling concentrations of environmental Ca ( Smith, 2002 and Witman, et al. , 1978 ) . Since dynein is responsible for driving the movement/force of the scourge, failure of the radial radiuss to signal to microtubules would do the sliding of the microtubules to stay unchanged by any fluctuations in environmental Ca2+ concentration ( Witman, et al. , 1978 ) .
The oda1 mutation specific mutant is on the outer dynein arm, forestalling optimum motion. The ODA1 protein is absent from the outer dynein because the coding cistron encodes a stop downstream of the instigator, forestalling a docking protein to be formed. The moorage protein would usually move in complex grounding the outer dynein arm onto the microtubule doublet ( Takada, et al. , 2001 ) . The outer dynein arm generates the motion and acceleration scourge force, by hydrolysing ATP. Therefore any retardation/loss of the outer dynein arm should diminish swimming velocities even in high Ca degrees. An oda1 mutation lacks its outer dynein arm, which is responsible for how rapidly the scourge motion is produced. Theoretically, flagella force/speeds should stay similar at all Ca concentrations, since it can non alter velocities. Literature denoted that waveforms/speeds of the oda1 mutation could non mime wild-type activity in high Ca2+ photoshock forms, but it can pattern the wild-type activity in low Ca2+ ( Wargo, et al. , 2004 ) . Theoretically, this is logical, all dynein arm activity will non be ceased, but merely slowed. The outer arm dynein controls velocity and force of scourge motion, a mutant would decelerate the scourge so that any concentration of Ca would mime the slow, symmetrical motions of wild-type in low Ca environments.
Pf18 mutation lacks the cardinal brace microtubules that make up the axoneme, this defect induces flagella palsy. In theory, motility should be wholly impaired. The sliding of the microtubules in this mutation will be slower than wild type at low Ca degree. However, the skiding gesture will increase similar to wild type motion at high Ca degrees. Harmonizing to Wargo, et al. , the skiding speed of axonemes from pf18 increased as environmental Ca concentrations rose. Obviously, dynein activity can non be ceased by the deficiency of the microtubule doublet in the presence of high concentrations of Ca. However, the calcium-induced dynein activity fails if the C2 microtubule of the cardinal setup is present or if radial radius constituents are missing ( Brokwaw and Kamiyra, 1987 ) . These consequences provide grounds that dynein activity is modulated by Ca and indicate that dynein activity is regulated by the response of a peculiar enzyme to increasing concentrations of free Ca ( Wargo, et al. , 2004 ) .
Four strains of Chlamydomonas reinhardtii ; wild-type, mutant pf14 mt+ ( mutated
radial radiuss ) , mutant oda1 mt- ( dynein arm mutant ) , and pf 18 mt- ( cardinal microtube mutant ) will be cultured until they reach the same denseness of 1x105cells/mL in a standard Tris-Acetate-Phosphate ( TAP ) Medium. Cells will be centrifuged, supernatants removed, and cells resuspended three times in TAP medium without any Ca. Specific Ca aliquots runing from 10-3 to 10-7M will be reintroduced into cell civilizations, incubated for specific times, and assayed. To efficaciously assay Chlamydomonas reinhardtii motion forms, a Leica DM1000 inverted picture entering compound microscope will capture all cellular activity. The recorded picture will be reviewed in five 2nd increases utilizing MacBook iMovie to guarantee all cells will be accounted for. Movement forms will be determined by numbering the frequences of cells that moved in a peculiar wave form ( symmetrical or asymmetrical ) and way over a peculiar clip frame ( additive frontward, additive backwards, clockwise, counterclockwise, or spiral ) . The experiment needs to be performed at least twice per test for truth.
Calcium Levels Tested ( M )
Incubation Time ( proceedingss )
Control ( wt )
0, 1mM, 10 millimeter, 100mM, 1AµM, and 100AµM
20, 100 proceedingss
0, 1mM, 10 millimeter, 100mM, 1AµM, and 100AµM
20, 100 proceedingss
0, 1mM, 10 millimeter, 100mM, 1AµM, and 100AµM
20, 100 proceedingss
0, 1mM, 10 millimeter, 100mM, 1AµM, and 100AµM
20, 100 proceedingss
Table 1. Sample proving parametric quantities: clip increases for changing sums of Ca will be incubated into the each civilization.
All cells will be incubated in the same TAP medium until reach adulthood at 1x105cells/mL. All the checks will be performed with the same methods. The positive control is the wild type strain of Chlamydomonas reinhardtii exposed to every environmental Ca aliquots for each clip increases. The negative control is the pf14 mutation. It is expected these cells will exhibit small to no motility with any sum of Ca or incubation clip.
The Ca environment degrees and the clip intervals the Ca2+ is allowed to incubate will be the variable factors. Motions ( frontward, rearward, clockwise, counterclockwise, and bottle screw ) , wave form ( asymmetrical, symmetrical, and quiescent ) , and frequence will change from type to type. Mutant strains of Chlamydomonas reinhardtii with smasher motility factors will be tested to understand how environmental Ca prevents/enables flagellular mobility every bit good as justify the Chlamydomonas reinhardtii wild type consequences.
The consequences that would back up the hypothesis would be changing environmental Ca degrees shows a specific form for mobility. Since the published information was highly contradictory on what forms and wave forms would look in Chlamydomonas reinhardtii in changing sums of Ca, the consequences might be able to support/deny earlier appraisals of patterning. Wild type C. reinhardtii should exhibit the most outstanding scourge constituents and exhibit the greatest motility forms and motion rates.
Consequences that would reject our hypotheses/experimental methods include Ca degrees non impacting cell motility, no motility changes over a period of clip. Since we understand how Ca activates dyenin this would oppose our guesss. If the mutations were able to travel precisely the same as the wild type strain this would confute the scourge functionality.
In order to determine this, all free Ca ions in C. reinhardtii wild type and mutated cells will be removed so later reintroduced to specific Ca ion aliquots. The information from this survey on Chlamydomonas phototransduction should foster our apprehension of these homologous events in higher being.
pf14 has a mutant within the radial radiuss, this consequences in a deficiency of dynein-driven microtubule connexion to the axoneme. Dynein is responsible for driving the motion of the scourge and in this mutation it would be difficult to reassign the potency of dynein throughout axoneme, and hence possibly stay paralytic despite the sum of Ca degrees ( Smith, E. F. , 2002 ) . At low Ca degrees, the Oda1 mutation should exhibit similar asymmetric wave forms as the wild type, but will non be able to bring forth other types of traveling due to its losing outer dynein arm ( Wargo, M. J. , 2004 ) . The pf 18 has mutated cardinal microtubules located within the nucleus axoneme. The mutations will most likely exhibit slower sliding of microtubules than the wild type strain at lower Ca degrees, ( antonym of Oda1 ) but at a high Ca concentration ( 10-4M ) the mutant exhibit symmetric wave forms at a similar rate as the wild type ( Smith, E. F. , 2002 ) .
Prince alberts, Bruce, et Al. Molecular Biology of the Cell. Vol. 5. New York: Garland Science, 2008. Print.
Bessen, M. , R. B. Fay, and G. B. Witman. “ Calcium Control of Waveform in Isolated Flagellar Axonemes of Chlamydomonas. ” The Journal of Cell Biology 86.2 ( 1980 ) : 446-55. Print.
Brokwaw, C. J. and Kamiyra, R. “ Bending forms of scourge: IV. Mutants with defects in inner and outer dynein weaponries indicate differences in dynein arm map. ” Cell Motile ( 1987 ) 8: ( 68-75 ) . Print.
D’Souza, Jacinta S. , Mohanram Gudipati, Jayashree A. Dharmadhikari, Aditya K. Dharmadhikari, Abhishek Kashyap, Manaswini Aiyer, Usha Rao, Deepak Mathur, and Basuthkar J. Rao. “ Flagella-generated Forces Reveal Gear-type Motor in Single Cells of the Green Alga, Chlamydomonas reinhardtii. ” Biochemical and Biophysical Research Communication 380 ( 2009 ) : 266-70. Science Direct. Web.
Harris, Elizabeth H. “ CHLAMYDOMONAS AS A MODEL ORGANISM. ” Annual Review of Plant Physiology and Plant Molecular Biology 52 ( 2001 ) : 363-406. Annual Reviews, 2001. Web. 6 Sept. 2010. Available from & lt ; hypertext transfer protocol: //www.annualreviews.org/doi/abs/10.1146/annurev.arplant.52.1.363? url_ver=Z39.882003 & A ; rfr_dat=cr_pub % 3Dncbi.nlm.nih.gov & A ; rfr_ids=ori % 3Arid % 3Acrossref.org & A ; journalCode=arplant.2 & gt ;
Heuser, T. , M. Raytchev, J. Krell, M. E. Porter, and D. Nicastro. “ The Dynein Regulatory Complex Is the Nexin Link and a Major Regulatory Node in Cilia and Flagella. ” The Journal of Biology 197 ( 2009 ) : 921-33. Web
Leon, Rosa, Aurora Galvan, and Emilio FernaI?ndez. “ Transgenic Microalgae as Green Cell Factories: . New York, NY: Springer Science Business Media, 2007. Print.
Merchant, Sabeeha S. , Simon E. Prochnik, and Olivier Vallon. “ The Chlamydomonas Genome Reveals the Evolution of Key Animal and Plant Functions. ” Science 318 ( 2007 ) : 245-50. PubMed Central, 24 May 2010. Web. 6 Sept. 2010. & lt ; hypertext transfer protocol: //www.ncbi.nlm.nih.gov-/pmc/articles/PMC2875087/ ? tool=pubmed & gt ;
Movassha, Tandis. “ 3-D Reconstruction of Chlamydomonas Flagellar Outer dynein Arm in the Presence of Nucleotides, Cornell Dissertation. 2010. Web.
Mitchell, Beth F. , Lotte B. Penderson, and Michael Feely, et Al. “ ATP Production in Chlamydomonas reinhardtii Flagella by Glycolytic Enzymes. ” Molecular Biology of the Cell 16 ( 2005 ) : 4509-518. PubMed Central, Oct. 2005. Web. 6 Sept. 2010. Available from & lt ; hypertext transfer protocol: //www.ncbi.nlm.nih.gov/pmc/articles/PMC1 237060/pdf/00164509.pdf & gt ; .
Pazour, G. J. , O. A. Sineshchekov, and G. B. Witman. “ Mutational Analysis of the Phototransduction Pathway of Chlamydomonas reinhardtii. ” The Journal of Cell Biology 131.2 ( 1995 ) : 427-40. Print
Smith, Elizabeth F. “ Hydin Seek: Finding a Function in Ciliary Motility – JCB. ” The Journal of Cell Biology. 22 Jan. 2007. Web. 08 Dec. 2010. & lt ; hypertext transfer protocol: //jcb.rupress.org/content/176/4/403.full & gt ; .
Smith, Elizabeth F. “ Regulation of Flagellar Dynein by Calcium and a Role for an Axonemal Calmodulin and Calmodulin- Dependent Kinase. ” Molecular Biology of the Cell 13 ( 2002 ) : 3303-313. Print.
Sakato, M. , and S. M. King. “ Calcium Regulates ATP-sensitive Microtubule Binding by Chlamydomonas Outer Arm Dynein. ” Journal of Biological Chemistry 278.44 ( 2003 ) : 43571-3579. Print.
Schmidt, Melanie, Gebner Gunther, and Luff Matthias, et Al. “ Proteomic Analysis of the Eyespot of Chlamydomonas reinhardtii Provides Novel Insights into Its Components and Tactic Motions. ” Plant Cell 18.8 ( 2006 ) : 1908-930. PubMed Central, Aug. 2006. Web. 6 Sept. 2010. Available from & lt ; hypertext transfer protocol: //www.ncbi.nlm.nih.gov/pmc/articles/PMC1533972/ ? mtool=pubmed & gt ; .
Silflow, Carolyn D. , and Paul A. Lefebvre. “ Assembly and Motility of Eukaryotic Cilia and Flagella. Lessons from Chlamydomonas reinhardtii. ” Plant Physiol 122 ( 2001 ) : 1500-507. American Society of Plant Biologists, Dec. 2001. Web. 6 Sept. 2010. Available from & lt ; hypertext transfer protocol: //www. plantphysiol.org/cgi/content/full/127/4/1500 & gt ;
Stauber, Einar J. , and Michael Hippler. “ Chlamydomonas reinhardtii Proteomics. ” Plant Physiology and Biochemistry 42 ( 2004 ) : 989-1001. Science Direct. Web.
Suzuki, Takeshi, Kenta Yamasaki, Satoshi Fujita, et Al. “ Archaeal-type Rhodopsin in Chlamydomonas: Model Structure and Intracellular Localization. ” Biochemical and Biophysical Research Communication 301 ( 2003 ) : 711-17. ScienceDirect. Web.
Takada, Saeko, Curtis G. Wilkerson, Kenichi Wakabayashi, Ritsu Kamiya, and George B. Whitman. “ The Outer Dynein Arm-Docking Complex: Composition and Characterization of a Subunit ( Oda1 ) Necessary for Outer Arm Assembly. ” Microbiology of the Cell 13.3 ( 2002 ) : 1015-029. Mar. 2002. Web. 15 Sept. 2010.
Wagner, V. , G. Gessner, I. Heiland, M. Kaminski, S. Hawat, K. Scheffler, and M. Mittag. “ Analysis of the Phosphoproteome of Chlamydomonas reinhardtii Provides New Insights into Various Cellular Pathways. ” Eukaryotic Cell 5.3 ( 2006 ) : 457-68. Print.
Wargo, M. J. , et Al. “ Analysis of Microtubule Sliding Patterns in Chlamydomonas Flagellar Axonemes Reveals Dynein Activity on Specific Doublet Microtubules. ” Journal of Cell Science 117 ( 2004 ) : 2533-544. Print.
Witman, G.B. , Plummer, J. , and Sander, J. “ Chlamydomonas flagellar mutations missing radial radiuss and cardinal tubules. ” 1978, THE JOURNAL OF CELL BIOLOGY 76 ( 1978 ) :729-747.
Wilson, Nedra F. , Janaki K. Iyer, Julie A. Buchheim, and William Meek. “ Regulation of Flagellar Length in Chlamydomonas. ” Seminars in Cell & A ; Development Biology 19 ( 2008 ) : 494-501. Science Direct. Web.
Yang, Pinfen, Dennis R. Diene, Chun Yang, Takahiro Kohno, Gregory J. Pazouur, Jennifer M. Dienes, Stephen M. King, Winfield S. Sale, Ritsu Kamiya, Joel L. Roesnbaulm Rosenbaum, and George B. Witman. “ Radial Spoke Proteins of Chlamydomonas Flagella. ” Journal of Cell Science. The Company of Biologists, 28 Feb. 2006. Web. 5 Oct. 2010. & lt ; hypertext transfer protocol: //www.ncbi.nlm.nih.gov/pmc/articles/P M C1973137/ & gt ; .