Problems with deflagellation of Chlamydomonas

Brian T. Greuel bgreuel at centurytel.net
Thu Oct 24 11:18:02 EST 2002


In my Molecular Cell Biology laboratory, we are studying the effects of
various reagents on the regeneration of flagella in Chlamydomonas reinhardi.
One of the persistent problems we are having, however, is that we are not
getting 100% deflagellation of Chlamydomonas cells.  In fact, less than 50%
of the cells are deflagellating, which makes the collection of data on
regeneration extremely problematic, if not impossible.  The culture medium
we are using is Medium I of Sager and Granick, which includes, among other
things, calcium chloride (40 mg/liter), that is known to be essential for
the deflagellation response.  The procedure we are using for deflagellation
is to lower the pH of the culture medium to pH 4.5 over a period of 30
seconds by the dropwise addition of 0.5 N acetic acid.  After waiting an
additional 30 seconds, the pH is restored to 6.8 by the dropwise addition of
0.5 N KOH.

Any suggestions for how I can increase my efficiency of deflagellation?

Thanks much,

Brian Greuel

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Brian T. Greuel, Ph.D.
John Brown University
Dept. of Biology, Box 3119
2000 W. University Street
Siloam Springs, AR   72761
TEL:  (479) 524-7433
FAX:  (479) 524-9548
EMAIL:  bgreuel at jbu.edu
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