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First published online August 4, 2005
Journal of Experimental Biology 208, 3133-3143 (2005)
Published by The Company of Biologists 2005
doi: 10.1242/jeb.01748

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Two-dimensional gel analysis of the heat-shock response in marine snails (genus Tegula): interspecific variation in protein expression and acclimation ability

Lars Tomanek^*

Hopkins Marine Station, Stanford University, Pacific Grove, CA 93950-3094, USA

View larger version (54K): [in a new window]   Fig. 1. (A) Autoradiographs of 35S-labeled proteins following incubation to 13°C and 30°C (heat-shocked) for 2.5 h and 4 h of recovery (and labeling) at 13°C in gill tissues of 13°C-acclimated T. funebralis. Clusters of heat-induced proteins are further described in Fig. 2. (B) Heat-induced activation of synthesis of proteins between 70–90 kDa in 13°C-acclimated T. funebralis. The constitutively expressed protein spot Hsp71.8 is always found to the left of the vertical line. 14C molecular mass markers are shown on the side of the gels (A).

View larger version (46K): [in a new window]   Fig. 2. Autoradiographs of newly synthesized 35S-labelled proteins of (A) 70–88 kDa, (B) 40 kDa and (C) 24–27 kDa in T. funebralis (top), T. brunnea (middle) and T. montereyi (bottom). Proteins within a cluster are assigned according to (i) their approximate molecular mass and (ii) isoelectric point, pI (the higher the number the more basic the pI of the protein). The pattern and orientation of arrows displays matches of proteins between species. Note that not all Hsps were detected in all species, and not all Hsps were detected in all gels from a common incubation temperature within a species (for example, Hsp77.1-2 were not detected in case of T. funebralis for the particular gel shown here; additionally, not all Hsp88s were detected in T. brunnea and T. montereyi).

View larger version (27K): [in a new window]   Fig. 3. Normalized volumes (relative to total spot volume within a gel) of acute synthesis of Hsp71.1 and Hsp71.2 in response to temperature in 13°C- and 22°C-acclimated specimens of T. funebralis, T. brunnea and T. montereyi. Values are means ± 1 S.E.M. *P-value for the interaction effect of the specific acclimation and incubation temperature of either <0.05 or <0.10, located nearest the specific species symbol (see Table 1).

View larger version (30K): [in a new window]   Fig. 4. Normalized volumes (relative to total spot volume within a gel) of Hsp71.8 and Hsp74.1 in response to temperature in 13°C- and 22°C-acclimated specimens of T. funebralis, T. brunnea and T. montereyi. Values are means ± 1 S.E.M. *P-value for the interaction effect of the specific acclimation and incubation temperature of either 0.05 or 0.10, located nearest the specific species symbol (see Table 1). MEA indicates a main effect of acclimation (see Table 1 for additional P-values for the interaction effects for this species).

View larger version (26K): [in a new window]   Fig. 5. Normalized volumes (relative to total spot volume within a gel) of Hsp77.2, Hsp76.3 (T. funebralis and T. brunnea only) and Hsp72.4 (T. montereyi) in response to temperature in 13°C- and 22°C-acclimated specimens of T. funebralis, T. brunnea and T. montereyi. Values are means ± 1 S.E.M. *P-value for the interaction effect of the specific acclimation and incubation temperature of either 0.05 or 0.10, located nearest the specific species symbol (see Table 1).

View larger version (27K): [in a new window]   Fig. 6. Normalized volumes (relative to total spot volume within a gel) of Hsp88.1 and Hsp40.2 in response to temperature in 13°C- and 22°C-acclimated specimens of T. funebralis, T. brunnea and T. montereyi. *P-value for the interaction effect of the specific acclimation and incubation temperature of either 0.05 or 0.10, located nearest the specific species symbol (see Table 1).

View larger version (29K): [in a new window]   Fig. 7. Normalized volumes (relative to total spot volume within a gel) of Hsp24.1 in response to temperature in 13°C- and 22°C-acclimated specimens of T. funebralis, T. brunnea and T. montereyi. Values are means ± 1 S.E.M. *P-value for the interaction effect of the specific acclimation and incubation temperature of either 0.05 or 0.10, located nearest the specific species symbol (see Table 1). MEA indicates a main effect of acclimation (see Table 1 for additional P-values for the interaction effects for this species).

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