Program of the 2001 International Worm Meeting - Sternberg Lab ...

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complexes within connective tissues and muscle
thick filaments, but also from microtubule
arrays within interphase and mitotic cells.
Effects of both molecular orientation and
symmetry upon SHG allow the signal from
species generating the second harmonic to be
decoded, by ratiometric correlation with TPEF,
to yield information on local structure below
optical resolution. Thus SHIM and DMNLM
should be valuable tools for studies of cellular
and structural biology both in C. elegans and
less GFP-friendly nematodes, as well as vital
histology and pathology in larger, more
complex phyla.
Figure: Mitiotic spindle in an one-cell embryo
expressing beta-tubuin::GFP presented in four
separate representations of DMNLM.
324. Intrinsically Thermotolerant
Substrains Of C. elegans Maintain
Their IT State, Show Increased
Longevity, And Elevated Hsp16
Levels.
Joshua P. Ellwitz, Karen A.
McFarland, Rebecca A. Buonpane,
Nicole D. Smith, Karen E. Stine,
Glenn E. White
Department of Biology/Toxicology, Academic
Honors Program, Ashland University, Ashland,
Ohio 44805
The response to external stresses has been
studied in a number of organisms including C.
elegans with the heat shock response being a
classic example. We reported at the 12th
International C. elegans Meeting (Smith et al.,
page 787) the isolation of substrains of N2
which demonstrated an intrinsic
thermotolerance (defined as a significantly
increased survival probability when taken from
either 16 or 25 o C to 37 o C for two hours as
compared to controls). These substrains were
designated N2IT# (intrinsically thermotolerant)
to distinguish these from EMS induced
mutations which also can confer intrinsic
thermotolerance. We report here that over two
years after these N2ITs were isolated, that they
are still maintaining their intrinsic
thermolerance. In our most recent experiment
(March, 2001), using N2IT6, we observed a
survival probability (SP) = 0.33 as compared to
a SP = .029 for the controls. Because of
continual concerns that perhaps it was
microenvironment that was inducing this
intrinsically thermotolerant substrain of N2s, we
tested three different "lots" of N2s obtained
from the CGC. When tested, these three
different lots of N2s also demonstrated SPs <
0.1, which implies that it is not the
microenvironment of the petri dish that is
producing these IT substrains.
We have examined one of our substrains,
designated N2IT6, for evidence of heat shock
protein induction. Western blot analysis
indicated significantly elevated hsp16 levels
when compared to unshocked controls (thanks
to Peter Candido for the anti-hsp16 antibody).
Coomassie stained SDS gels do not show any
elevated hsp70 levels. Because of the poor
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