Total Fiber Cross-sectional Area Research Articles

Semiautomatic morphometric analysis of skeletal muscle obtained by needle biopsy in older adults.

Analysis of skeletal muscle mass and composition is essential for studying the biology of age-related sarcopenia, loss of muscle mass, and function. Muscle immunohistochemistry (IHC) allows for simultaneous visualization of morphological characteristics and determination of fiber type composition. The information gleaned from myosin heavy chain (MHC) isoform, and morphological measurements offer a more complete assessment of muscle health and properties than classical techniques such as SDS-PAGE and ATPase immunostaining; however, IHC quantification is a time-consuming and tedious method. We developed a semiautomatic method to account for issues frequently encountered in aging tissue. We analyzed needle-biopsied vastus lateralis (VL) of the quadriceps from a cohort of 14 volunteers aged 74.9 ± 2.2years. We found a high correlation between manual quantification and semiautomatic analyses for the total number of fibers detected (r2 = 0.989) and total fiber cross-sectional area (r2 = 0.836). The analysis of the VL fiber subtype composition and the cross-sectional area also did not show statistically significant differences. The semiautomatic approach was completed in 10-15% of the time required for manual quantification. The results from these analyses highlight some of the specific issues which commonly occur in aged muscle. Our methods which address these issues underscore the importance of developing efficient, accurate, and reliable methods for quantitatively analyzing the skeletal muscle and the standardization of collection protocols to maximize the likelihood of preserving tissue quality in older adults. Utilizing IHC as a means of exploring the progression of disease, aging, and injury in the skeletal muscle allows for the practical study of muscle tissue down to the fiber level. By adding editing modules to our semiautomatic approach, we accurately quantified the aging muscle and addressed common technical issues.

Effect of incubation temperature on muscle growth of barramundi Lates calcarifer at hatch and post‐exogenous feeding

Muscle morphology was investigated in newly hatched barramundi Lates calcarifer larvae incubated at set temperatures (26, 29 and 31 degrees C) prior to hatching. Three days after hatching (the start of exogenous feeding), larvae from the 26 and 31 degrees C treatments were each divided into two groups and reared at that temperature or transferred over the period of several hours to 29 degrees C (control temperature). Incubation temperature significantly affected muscle cellularity in the developing embryo, with larvae incubated at 26 degrees C (mean +/-s.e. 223.3 +/- 7.9) having on average 14.4% more inner muscle fibres than those incubated at 31 degrees C (195.2 +/- 8.8) and 4.8% more than those incubated at 29 degrees C (213.5 +/- 4.7). Conversely, inner muscle fibre cross-sectional area significantly increased at the warm incubation temperature in L. calcarifer, so that the total cross-sectional muscle area was not different between treatment groups. The total cross-sectional area of superficial muscle fibres and the proportion of superficial to total fibre cross-sectional area in just hatched L. calcarifer were also affected by incubation temperature, with incubation at the cool temperature (26 degrees C) increasing both the total cross-sectional area and proportion of superficial muscle fibres. By 9 days post-hatch, the aforementioned differences were no longer significant. Similarly, there was no difference in total superficial fibre cross-sectional area between any treatment groups of L. calcarifer, whereas incubation temperature still significantly affected the proportion of superficial to total muscle fibre cross-sectional area. Larvae hatched and grown at 31 degrees C had a significantly reduced percentage of superficial muscle cross-sectional area (mean +/-s.e. 5.11 +/- 0.66%) compared with those incubated and grown at 29 degrees C (8.04 +/- 0.77%) and 26 degrees C (9.32 +/- 0.56%) and those incubated at 26 degrees C and transferred to 29 degrees C (7.52 +/- 0.53%), and incubated at 31 degrees C and transferred to 29 degrees C (6.28 +/- 0.69%). These results indicate that changes in muscle cellularity induced by raising or lowering the incubation temperature of L. calcarifer display varying degrees of persistence over developmental time. The significance of these findings to the culture of L. calcarifer is discussed.

Gene expression in skeletal muscle of coronary artery disease patients after concentric and eccentric endurance training

Low-intensity concentric (CET) and eccentric (EET) endurance-type training induce specific structural adaptations in skeletal muscle. We evaluated to which extent steady-state adaptations in transcript levels are involved in the compensatory alterations of muscle mitochondria and myofibrils with CET versus EET at a matched metabolic exercise intensity of medicated, stable coronary patients (CAD). Biopsies were obtained from vastus lateralis muscle before and after 8 weeks of CET (n=6) or EET (n=6). Transcript levels for factors involved in mitochondrial biogenesis (PGC-1alpha, Tfam), mitochondrial function (COX-1, COX-4), control of contractile phenotype (MyHC I, IIa, IIx) as well as mechanical stress marker (IGF-I) were quantified using an reverse-transcriptase polymerase chain reaction approach. After 8 weeks of EET, a reduction of the COX-4 mRNA level by 41% and a tendency for a drop in Tfam transcript concentration (-33%, P=0.06) was noted. This down-regulation corresponded to a drop in total mitochondrial volume density. MyHC-IIa transcript levels were specifically decreased after EET, and MyHC-I mRNA showed a trend towards a reduction (P=0.08). Total fiber cross-sectional area was not altered. After CET and EET, the IGF-I mRNA level was significantly increased. The PGC-1alpha significantly correlated with Tfam, and both PGC-1alpha and Tfam significantly correlated with COX-1 and COX-4 mRNAs. Post-hoc analysis identified significant interactions between the concurrent medication and muscular transcript levels as well as fiber size. Our findings support the concept that specific transcriptional adaptations mediate the divergent mitochondrial response of muscle cells to endurance training under different load condition and indicate a mismatch of processes related to muscle hypertrophy in medicated CAD patients.

Recovery of muscle transfers replacing the total plantar flexor muscle group in rats.

In rats, combinations of plantar flexor muscles representing approximately 20, 40, 60, and 80% of the mass of the total plantar flexor group were transferred orthotopically in the absence of synergistic muscles and allowed to recover for 120 days. We hypothesized that, compared with their individual control values for structural and functional variables, the transfers would display a hierarchical array of deficits, proportional to their initial mass and, consequently, inversely proportional to the relative load on the transfers. Surprisingly, compared with their individual control values, each muscle transfer displayed deficits of 30-40% in muscle mass, total fiber cross-sectional area, and maximum isometric force, with the exception of the smallest transfer, the plantaris (PLN) muscle, which recovered 100% of its control value for each of these variables. Therefore, except for the PLN transfer, the muscle transfers studied displayed deficits similar in magnitude to those reported for muscles transferred in the presence of synergistic muscles. The greater recovery of the PLN transfer was attributed to the relatively large requirement for force production imposed on this transfer due to the average force requirements of the total plantar flexor group.

Parallel-Fibered Muscles Transplanted with Neurovascular Repair into Bipennate Muscle Sites in Rabbits

Experiments were performed on 20 New Zealand White male rabbits. Our hypotheses were that (1) latissimus dorsi (LTD) muscles transplanted into the site of a bipennate rectus femoris (RFM) muscle with neurovascular repair would retain their parallel-fibered structure and (2) the parallel-fibered structure of latissimus dorsi grafts would reduce their total fiber cross-sectional area and adversely affect force development relative to that of bipennate rectus femoris grafts and muscles. Compared with their respective donor muscles, 120 to 150 days after grafting, latissimus dorsi and rectus femoris grafts showed no change in the number of fibers and a decrease in the mean single-fiber cross-sectional area to approximately 70 percent. The latissimus dorsi grafts, which remained parallel-fibered, developed maximum forces 34 and 23 percent of the values for fully activated rectus femoris grafts and muscles, respectively. The deficit in the maximum force of the latissimus dorsi grafts resulted primarily from the smaller total-fiber cross-sectional area as a result of the parallel-fibered structure.

The effect of hypertrophy on the properties of skeletal muscle

1. 1. The increased myofibril cross-sectional area of hypertrophied mouse soleus muscles was measured (+ 140 per cent). 2. 2. The maximum tetanic tensions of control and hypertrophied muscle were measured in vivo. 3. 3. Although there was an increase in gross tension output (+30 per cent), hypertrophy brought about a drop in tension per unit muscle mass (−24 per cent), per unit total fibre cross-sectional area (−20 per cent) and per unit total myofibril cross-sectional area (−46 per cent).