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Tissue-specific expression of inducible and constitutive Hsp70 isoforms in the western painted turtle

Michelle Anne Scott, Marius Locke and Leslie Thomas Buck^*

Department of Zoology, University of Toronto, 25 Harbord St, Toronto, ON, Canada, M5S 3G5

View larger version (14K): [in a new window]   Fig. 4. Correlation between Hsp73 and Hsp72 expression after the 24 h dive. Mean values for basal Hsp73 expression in brain (B), heart (H), liver (L) and skeletal muscle (M) are plotted relative to the dive-induced Hsp72 expression in each tissue from Fig. 3. Linear regression of the data reveals a strong regression coefficient or r2 value of 0.82 and the equation y=-0.36x+33.8. These data are the mean ± S.E.M. of 16 basal Hsp73 and 20 dive-induced Hsp72 expression measurements; mean values are presented because it is not possible to obtain both measurements from the same animal.

View larger version (11K): [in a new window]   Fig. 1. Portions of representative two-dimensional [isoelectric focusing (IEF) and sodium dodecyl sulfate—polyacrylamide gel electrophoresis (SDS-PAGE)] blots showing the specificity of antibodies for Hsp72 and Hsp73 in turtle liver (200 µg protein). Blots were reacted with antibodies specific to Hsp72 (A), Hsp73 (B) or both Hsp73 and Hsp72 (C). Arrows indicate increasing molecular mass and pH (pH range of original blot approximately 7.4 to 4.0).

View larger version (17K): [in a new window]   Fig. 2. Expression of Hsp73 and Hsp72 following heat shock. (A) and (B) show representative one-dimensional sodium dodecyl sulfate—polyacrylamide gel electrophoresis (SDS-PAGE) western blots and the results of densitometric analyses of Hsp73 and Hsp72 expression, respectively, in turtle brain (B), heart (H), liver (L) and skeletal muscle (M) following heat shock. Values represent means ± S.E.M. of four replicate experiments and are expressed as absolute concentrations calculated from standard curves. Statistical significance (P<0.05) compared with the control is indicated by asterisks. Control western blots are shown in Fig. 3.

View larger version (31K): [in a new window]   Fig. 3. Expression of Hsp73 and Hsp72 in response to 24 h dive and 1 h recovery. (A) Representative western blots of Hsp73 and Hsp72 expression in turtle brain (B), heart (H), liver (L) and skeletal muscle (M) following dive and recovery. (B) Results of densitometric analyses of Hsp73 and Hsp72 expression. Blots from control, anoxic and recovery animals were scanned, and the net intensity of each band was determined using the Kodak 1D imaging program. Values for control, dived and recovery experiments represent means ± S.E.M. of four, five and four replicates, respectively, and are expressed as absolute concentrations calculated from standard curves. Statistical significance compared with the control is indicated by asterisks, and comparison between 24 h dived and 1 h recovery tissue is denoted by daggers (P<0.05).