160 ABSTRACTS FROM THE MBL GENERAL SCIENTIFIC MEETINGS

Reference: Biol. Bull. 207: 160. (October 2004) Reference: Biol. Bull. 207: 160. (October 2004)
© 2004 Marine Biological Laboratory © 2004 Marine Biological Laboratory

Marine Plants May Polarize Remote Fucus Eggs Characterization of Anastral, Bipolar Spindle
via Luminescence Development and Atypical Cytokinesis in
Lionel F. Jaffe Ammonia-Activated Sea Urchin Eggs
Hopkins Marine Station, Pacific Grove, California John H. Henson1,2, Jessica E. Davis2,
and Marine Biological Laboratory, Woods Hole, Charles B. Shuster1,3, Christopher A. Fried1,2, and
Massachusetts Calvin R. Simerly4
1
Marine Biological Laboratory, Woods Hole,
Massachusetts
Fucus zygotes can be polarized—as shown by their later 2
Dickinson College, Carlisle, Pennsylvania
outgrowth directions— by many environmental influences. 3
New Mexico State University, Las Cruces,
These include those exerted by all tested intertidal marine New Mexico
plants (including all of the main algal phyla as well as the 4
Magee Women’s Research Institute, Pittsburgh,
flowering plant Phyllospadix) at distances of up to 5–10 mm
away from a piece of the plant in a laboratory dish. Earlier
Pennsylvania
studies inferred action via diffusing molecules. A reinves-
tigation indicates that such actions can be exerted through
The mitotic apparatus (MA) of the early sea urchin em-
glass barriers and suggests action via luminescent and,
bryo is often cited as the classic example of an aster-
perhaps, infrared signals. Since this polarizing influence is
dominated MA that is organized via the introduction of a
exerted by pieces of all tested plants, the infrared light
sperm-derived paternal centrosome. Previous studies have
signals might be emitted by luminescent bacteria growing in
indicated that artificial activation of eggs can result in
biofilms on the surfaces of all intertidal plants.
polymerization of microtubules associated with maternal
centrosomal material, with the microtubule organization
often taking the form of a monaster associated with con-
densed chromosomes. In the present study we have examined
the generation of unusual bipolar spindle-like structures in eggs
artificially activated with ammonia. Unfertilized sea urchin
(Lytechinus pictus) eggs were activated by incubation in arti-
ficial seawater containing 15 mM ammonia chloride and fixed
with cold methanol at the time appropriate for first division.
The activated eggs were stained with antibodies against tubu-
lin, actin, NuMA, centrosomal proteins, and the kinesin-like
proteins Kif2, KRP110 and KRP170, and stained cells were
viewed on a laser-scanning confocal microscope. The majority
of activated eggs demonstrated poorly organized microtubules
and/or the presence of monasters, but a subset contained atyp-
ical bipolar spindle-like structures. These spindles were small,
anastral, capable of anaphase-like chromosome segregation,
and overall very reminiscent of meiotic spindles. The spindle
matrix protein NuMA, the centrosomal protein 4D2, and the
spindle motor proteins Kif2, KRP110 and KRP170 all localized
to these structures. The ammonia-induced spindles were also
observed in living eggs using polarization optics on an LC-
PolScope (courtesy of Rudolf Oldenbourg and Grant Harris of
the Marine Biological Laboratory). The activated eggs went on
to attempt cytokinesis, which often involved unusual linear
arrays of microtubules in the region of the developing cleavage
furrow. These results suggest that microtubule motor and ma-
trix proteins have the ability to organize a meiotic-like spindle
in sea urchin eggs in the absence of the paternal centrosome.

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