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Introduction to Chlamydomonas - Brandeis University

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<strong>Introduction</strong> <strong>to</strong> <strong>Chlamydomonas</strong>:<br />

a model organism for active material<br />

Dr. Cynthia Barber<br />

Nicastro Lab, <strong>Brandeis</strong> <strong>University</strong><br />

Active Matter Workshop<br />

Feb 20th, 2013<br />

Thursday, March 7, 13


Chlamydo-what?<br />

<strong>Chlamydomonas</strong>-singled celled algae<br />

Thursday, March 7, 13


Characteristics of <strong>Chlamydomonas</strong><br />

~10um in diameter<br />

two ~12um flagella<br />

Thursday, March 7, 13


Characteristics of <strong>Chlamydomonas</strong><br />

Colored eye spots<br />

Dieckmann (2003)<br />

Thursday, March 7, 13


Characteristics of <strong>Chlamydomonas</strong><br />

cell wall- resist osmotic pressure<br />

chloroplasts- pho<strong>to</strong>synthesis<br />

pyrenoid- carbon fixation<br />

Figure 2.1 The <strong>Chlamydomonas</strong> Sourcebook, 2nd ed.<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> lifecycle<br />

The <strong>Chlamydomonas</strong> Sourcebook, 2nd ed.<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> lifecycle<br />

The <strong>Chlamydomonas</strong> Sourcebook, 2nd ed.<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> in the lab<br />

no flagella while on plates<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> in the lab<br />

grow flagella in ~1hr in liquid cultures<br />

Thursday, March 7, 13


Hundreds of species<br />

living in:<br />

soil, fresh water,<br />

salt water, snow,<br />

ice, etc.<br />

Proschold et al. (2001)<br />

Thursday, March 7, 13


Hundreds of species<br />

living in:<br />

soil, fresh water,<br />

salt water, snow,<br />

ice, etc.<br />

<strong>Chlamydomonas</strong> reinhardtii<br />

Proschold et al. (2001)<br />

Thursday, March 7, 13


Hundreds of species<br />

living in:<br />

soil, fresh water,<br />

salt water, snow,<br />

ice, etc.<br />

<strong>Chlamydomonas</strong> reinhardtii<br />

Isolated from a pota<strong>to</strong> field in<br />

Amherst, MA in 1947<br />

Proschold et al. (2001)<br />

Thursday, March 7, 13


Thursday, March 7, 13<br />

<strong>Chlamydomonas</strong> as active matter


<strong>Chlamydomonas</strong> as active matter<br />

swimming<br />

pho<strong>to</strong>taxis<br />

pho<strong>to</strong>shock<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> as active matter<br />

swimming<br />

pho<strong>to</strong>taxis<br />

pho<strong>to</strong>shock<br />

Flagella function/regulation<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> Behavior- Forward Swimming<br />

Power Stroke<br />

Recovery Stroke<br />

Weibel D B et al. PNAS 2005;102:11963-11967<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> Behavior- Forward Swimming<br />

Power Stroke<br />

Recovery Stroke<br />

~150um/s<br />

~50Hz<br />

Weibel D B et al. PNAS 2005;102:11963-11967<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> Behavior- Forward Swimming<br />

Power Stroke<br />

Recovery Stroke<br />

~150um/s<br />

~50Hz<br />

Weibel D B et al. PNAS 2005;102:11963-11967<br />

helical path rotating at a frequency of 2Hz<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> Behavior- Forward Swimming<br />

Power Stroke<br />

Recovery Stroke<br />

~150um/s<br />

~50Hz<br />

Weibel D B et al. PNAS 2005;102:11963-11967<br />

helical path rotating at a frequency of 2Hz<br />

Daniel Chen in Dogic Lab<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> Behavior- Pho<strong>to</strong>taxis<br />

Dieckmann (2003)<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> Behavior- Pho<strong>to</strong>taxis<br />

Exact behavior depends on:<br />

Wavelength<br />

Intensity<br />

State of the cell<br />

Dieckmann (2003)<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> Behavior- Pho<strong>to</strong>taxis<br />

Two types of pho<strong>to</strong>recep<strong>to</strong>rs:<br />

470nm (blue)<br />

510nm (green)<br />

Ridge (2002)<br />

Dieckmann (2003)<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> Behavior- Pho<strong>to</strong>taxis<br />

Two types of pho<strong>to</strong>recep<strong>to</strong>rs:<br />

470nm (blue)<br />

510nm (green)<br />

Ridge (2002)<br />

Cannot detect red!<br />

Dieckmann (2003)<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> Behavior- Pho<strong>to</strong>taxis<br />

The <strong>Chlamydomonas</strong> Sourcebook, 2nd ed.<br />

Thursday, March 7, 13


Thursday, March 7, 13<br />

<strong>Chlamydomonas</strong> Behavior- Pho<strong>to</strong>shock


<strong>Chlamydomonas</strong> Behavior- Pho<strong>to</strong>shock<br />

Forward<br />

Swimming<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> Behavior- Pho<strong>to</strong>shock<br />

Forward<br />

Swimming<br />

Pho<strong>to</strong>shock<br />

escape<br />

symmetrical, undulating waveform<br />

Thursday, March 7, 13<br />

Brokaw et al. 1982


Thursday, March 7, 13<br />

Creating mutants


Creating mutants<br />

UV<br />

X-rays<br />

nitrosoguanidine (MNNG)<br />

methyl methanesulfonate (MMS)<br />

ethylmethanesulfonate (EMS)<br />

5-fluorodeoxyuridine (thymidine analog)<br />

Thursday, March 7, 13


Creating mutants<br />

UV<br />

X-rays<br />

nitrosoguanidine (MNNG)<br />

methyl methanesulfonate (MMS)<br />

ethylmethanesulfonate (EMS)<br />

5-fluorodeoxyuridine (thymidine analog)<br />

Can’t specifically mutate a gene of interest<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> reinhardtii<br />

Single celled algae<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> reinhardtii<br />

Single celled algae<br />

used <strong>to</strong> study flagella dynamics<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> reinhardtii<br />

Single celled algae<br />

used <strong>to</strong> study flagella dynamics<br />

non-symmetrical forward stroke<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> reinhardtii<br />

Single celled algae<br />

used <strong>to</strong> study flagella dynamics<br />

non-symmetrical forward stroke<br />

symmetrical backward stroke<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> reinhardtii<br />

Single celled algae<br />

used <strong>to</strong> study flagella dynamics<br />

non-symmetrical forward stroke<br />

symmetrical backward stroke<br />

pho<strong>to</strong>taxis <strong>to</strong>ward medium intensity blue/green light<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> reinhardtii<br />

Single celled algae<br />

used <strong>to</strong> study flagella dynamics<br />

non-symmetrical forward stroke<br />

symmetrical backward stroke<br />

pho<strong>to</strong>taxis <strong>to</strong>ward blue/green light<br />

pho<strong>to</strong>shock/escape from high intensity blue/green light<br />

Thursday, March 7, 13


<strong>Chlamydomonas</strong> reinhardtii<br />

Single celled algae<br />

used <strong>to</strong> study flagella dynamics<br />

non-symmetrical forward stroke<br />

symmetrical backward stroke<br />

pho<strong>to</strong>taxis <strong>to</strong>ward blue/green light<br />

pho<strong>to</strong>shock/escape from high intensity blue/green light<br />

cannot detect red light<br />

Thursday, March 7, 13


Thursday, March 7, 13<br />

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