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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

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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

View larger version (25K): [in a new window]   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.

View larger version (69K): [in a new window]   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.

View larger version (64K): [in a new window]   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.

View larger version (73K): [in a new window]   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.

View larger version (17K): [in a new window]   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.

View larger version (66K): [in a new window]   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.