First published online February 6, 2004
Journal of Experimental Biology 207, 963-971 (2004)
Published by The Company of Biologists 2004
doi: 10.1242/jeb.00842
Desiccation and rehydration elicit distinct heat shock protein transcript responses in flesh fly pupae
Scott A. L. Hayward*,
Joseph P. Rinehart and
David L. Denlinger
Ohio State University, Department of Entomology, 318 W. 12th Ave,
Columbus, OH 43210, USA
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Fig. 1. (A) Water loss, survival and emergence time for nondiapausing pupae held at
<5% RH/25°C for up to 7 days. The puparia encasing the pupae were
either intact or opened at the anterior end. (B) Water loss and survival of
diapausing pupae that were transferred to <5% RH/25°C for different
durations starting on either day 15 or day 30 of diapause. The anterior cap of
the puparium was removed for all diapause experiments. Water loss is given as
a percentage of the initial water content (N=20 for each time point);
percent survival is based on N=30 for each time point; time to
emergence represents the number of days from pupariation to adult eclosion
(N=30 for each time point). Means ±
S.E.M. are presented.
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Fig. 2. Expression of hsp23, hsp70, hsp90 and hsc70 transcripts
in nondiapausing pupae in response to desiccation. Pupae were encased in
either (A) opened or (B) intact puparia. Lanes represent RNA samples from
pupae desiccated at <5% RH/25°C for 0–6 days or exposed to a heat
shock (HS) of 1 h at 40°C. Each sample was run in triplicate. 28s
ribosomal RNA was used as a control to confirm equal sample loading.
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Fig. 3. Expression of hsp23, hsp70, hsp90 and hsc70 transcripts
in (A) day 15 and (B) day 30 diapausing pupae. The puparium was opened in all
cases. C represents RNA from a nondesiccated, nondiapausing control, and other
lanes indicate the number of days that diapausing pupae were exposed to
desiccation (<5% RH/25°C). Each sample was run in triplicate. 28s
ribosomal RNA was used as a control to confirm equal sample loading.
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Fig. 4. Expression of hsp23, hsp70 and hsp90 transcripts during
diapause termination in day 15 diapause pupae previously desiccated at <5%
RH/25°C for 9 days. C represents RNA from a nondesiccated diapause control
sample. D represents a desiccated diapause sample (9 days at <5% RH). Other
lanes represent hours following the termination of diapause elicited by an
application of hexane. Each treatment was replicated at least twice. 28s
ribosomal RNA was used as a control to confirm equal sample loading.
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Fig. 5. Water content and survival in (A) nondiapausing and (B) diapausing pupae
desiccated at <5% RH for 4 days and then rehydrated at either constant 75%
or 100% RH for 1 or 2 days. Water content is given as a percentage of the
initial fresh mass (N=20 for each time point), and percent survival
is based on N=20 for each time point. Means ±
S.E.M. are presented. Day 0 represents the
start of each desiccation treatment for either day 4 nondiapausing pupae or
day 15 diapausing pupae.
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Fig. 6. Expression of hsp23, hsp70, hsp90 and hsc70 transcripts
in response to the rehydration of desiccated (A) nondiapausing and (B) day 15
diapausing pupae. C represents RNA from a nondesiccated control. D represents
pupae desiccated for 4 days at <5% RH. Remaining lanes represent pupae
rehydrated for 1 or 2 days at 75% or 100% RH. Each treatment was run in
triplicate. 28s ribosomal RNA was used as a control to confirm equal sample
loading.
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© The Company of Biologists Ltd 2004
