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Noninvasive ⁶⁴Cu-ATSM and PET/CT Assessment of Hypoxia in Rat Skeletal Muscles and Tendons During Muscle Contractions

¹ Institute of Sports Medicine, Bispebjerg Hospital and Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark; ² Cluster for Molecular Imaging, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark; and ³ Department of Clinical Physiology, Nuclear Medicine and PET, Rigshospitalet, Copenhagen, Denmark

View larger version (55K): [in this window] [in a new window]   FIGURE 1.  Representative CT and PET images of hind limbs of rat immediately after unilateral electrostimulated muscle contractions. (A and C) Corresponding CT and PET images (coronal) showing clearly higher uptake of 64Cu-ATSM on stimulated left side. (C and D) Corresponding CT and PET images (sagittal) of right hind limb. Arrows mark clearly distinctive Achilles tendon.

View larger version (14K): [in this window] [in a new window]   FIGURE 2.  Changes in SUVs of 64Cu-ATSM from early to late PET scans in muscles (A and B) and tendons (C and D) in E group and EC group. Figures display changes in SUVs from early scans (0 min after contractions) to late scans (60 min after contractions). In both groups, SUVs of 64Cu-ATSM in skeletal muscles and Achilles tendons are higher in exercised hind limbs than in resting hind limbs. Furthermore, SUVs are consistently higher in tendons than in muscles in exercised and resting hind limbs. SUVs are maintained in tendons from early to late scans; however, both E group and EC group show minor (10%, P < 0.05) decrease in SUV in muscles from early to late scans. Values are means, with bars representing SEM.

View larger version (15K): [in this window] [in a new window]   FIGURE 3.  Gene expression of HIF1 in muscles (A and B) and tendons (C and D) in E group and EC group. HIF1 mRNA normalized to housekeeping genes TBP (soleus muscle) and RPL13 (Achilles tendon). Data are represented as geometric means (±SEM) of fold changes in soleus muscle and Achilles tendon subjected to isometric muscle contractions, compared with (paired) respective control samples. In skeletal muscles, HIF1 mRNA is elevated subsequent to isometric electrostimulated muscle contractions in both E group and EC group. Applied load of Achilles tendon induced by electrostimulated muscle contraction did not by itself increase HIF1 mRNA levels over resting levels. In contrast, application of cuff around contracted leg induced significant increase in HIF1 gene expression in Achilles tendon.

View larger version (12K): [in this window] [in a new window]   FIGURE 4.  Correlation between SUV of 64Cu-ATSM and gene expression of HIF1 (A) and CAIII (B) in skeletal muscles. In soleus muscle, uptake of 64Cu-ATSM expressed as SUV correlated significantly with log-transformed gene expression of both CAIII and HIF1. Correlation coefficient, P value, and 95% confidence limits are shown.