Abstract

Introduction

Materials & Methods

Results

Discussion & Conclusion

References

Discussion
Board

- The adult SERCA 1a isoform was the only SERCA 1 found in the rat diaphragm in both control and triamcinolone-treated animals

- The SERCA 1b, SERCA 2b and SERCA 3 fell below the detection limit in both control and triamcinolone-treated diaphragm

Fig.1:RT-PCR amplification of SERCA 1+2, SERCA 1 and SERCA 2 mRNA species in rat diaphragm

A. Lanes of a single amplification in a representative control and triamcinolone-treated rat

From left to right:

Lane 1: co-amplification of SERCA 1+2

Lane 2: SERCA 2 amplification product cut by Mse I into 127 bp and 67 bp fragments, SERCA 1 left intact

Lane 3: SERCA 1 amplification product cut by Nco I into 102 bp and 92 bp fragments, SERCA 2 left intact

Lane 4: Both enzymes were added. Notice complete digestion of amplification product

B. Mean and standard deviation of data from lanes 1 obtained in all animals (control, C and triamcinolone, T). Data from lanes 1 correspond to the co-amplification of SERCA 1+2 and were corrected with the data obtained after GAPDH amplification. *p < 0.05 vs control.

Notice that the levels of SERCA1+2 were significantly reduced by 70% after triamcinolone treatment (p < 0.05).

Fig. 2: RT-PCR amplification of SERCA 1 mRNA (left panel) and SERCA 2 mRNA (right panel) species in rat diaphragm (control, C and triamcinolone, T). Data are means and standard deviation obtained in all animals. These data, corrected for C/G content, are issued from values obtained in lane 4 (see example in Fig.1, left panel) for each animal. *p < 0.05 vs control.

Notice that the reduction in SERCA 1+2 levels seen in figure 1B after triamcinolone treatment resulted from a decrease in both SERCA 1 (-69%, p < 0.05) and to a lesser extent SERCA 2 (-49%, not significant) expression.

Fig. 3: Relative proportion of SERCA 1 and SERCA 2 mRNA species in rat diaphragm (control, C and triamcinolone, T). Data are means and standard deviation obtained in all animals. These data are issued from values obtained in lane 4 corresponding to the complete digest of SERCA 1+2 amplification into SERCA 1 and SERCA 2. Data are corrected for C/G content. *p < 0.05 vs control.

Notice that the relative proportion of SERCA 2 was significantly increased from 43*7% in control diaphragm to 52*4% after triamcinolone (p < 0.05).

Fig. 4:RT-PCR amplification of SERCA 2a mRNA species in rat diaphragm (control, C and triamcinolone, T). Data are means and standard deviation obtained in all animals. Data are corrected with the data obtained after GAPDH amplification. *p < 0.05 vs control.

Notice that the reduction in SERCA 2 mRNA observed in figure 2 after triamcinolone treatment was due to a decrease in SERCA 2a expression which was significantly reduced by 61% after triamcinolone treatment (*p < 0.05).

Fig. 5: RT-PCR amplification of phospholamban mRNA species in rat diaphragm (control, C and triamcinolone, T). Data are means and standard deviation obtained in all animals. Data are corrected with the data obtained after GAPDH amplification.

Notice that phospholamban expression was reduced by 31% (not significant) after triamcinolone treatment.

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