Isothermal gelation behavior of myofibrillar proteins from white and red chicken meat at different temperatures.

This paper provides insights in the isothermal gelation behavior of white and red chicken myofibrillar proteins (CMP) at different temperatures (20 to 80°C) and the underlying aggregation mechanism, allowing understanding of structure formation in poultry products during thermal processing. At low temperatures (20 to 60°C), an increase in aromatic surface hydrophobicity (SoANS) was found, suggesting potential formation of hydrophobic interactions between CMP. At higher temperatures (60 to 80°C), high SoANS and a significant decrease in total sulfhydryl amount (SH-amount) strongly indicate the presence of hydrophobic interactions and disulfide bonding, resulting in aggregation, as suggested by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The elastic modulus G' after 60 min isothermal heating (G'60min) significantly increased at 70 or 80°C, depending on the type of CMP. Differences in G'60min between white and red CMP were rather small at low temperatures (20 to 60°C). However, at 70°C, white CMP reached higher G'60min compared to red CMP, while the opposite was observed at 80°C. Overall, for every temperature studied, SoANS and SH-amount of red CMP were higher compared to white CMP. The differences in G'60min, SoANS, and SH-amount between white and red CMP were probably due to different isoforms. The results may help to steer quality characteristics of poultry products through intelligent choice of processing conditions.

Distribution pattern of muscle fibre types in soft palate of the dog (Canis familiaris, L.).

Distribution pattern of fibre types was studied in the muscles of the soft palate (palatinus, levator veli palatini and tensor veli palatini muscles) in the dog. The fibrillar classification was based on using histochemistry and immunohistochemistry methods: myofibrillar adenosine thriphosphatase (mATPase) to different pH of pre-incubation; nicotine adenine dinucleotide (reduced) tetrazolium reductase (NADH-TR) and finally, application of specific monoclonal antibodies against myosin heavy chain isoforms I, IIa and IIx. In the palatinus and levator veli palatini muscles, pure type I fibres and the hybrid type IIax and IIc were shown, with a checkerboard distribution in the first and a clear predominance of hybrid fibre types (about 98% of the total population) in levator veli palatini muscle. On the other hand, in the tensor veli palatini muscle, type IIx and IIm fibres were identified (fast-twitch fibres related to fast-moving muscles and the powerful jaw muscles of carnivores). The tensor veli palatini muscle had a different distribution and fibrillar composition with predominantly type IIm fibres in its central zone, whilst the peripheral zone was primarily type I and IIx fibres.

Skeletal muscle fiber type and myofibrillar proteins in relation to meat quality.

Although numerous studies have reported the relationships among muscle fiber characteristics, lean meat content and meat quality, controversial perspectives still remain. Conventional histochemical classifications may be involved in a high level of error, subjectivity and it could not clearly explain variety of myofibrillar protein isoforms. Therefore, more information is needed on how different factors, such as species, breeds, gender, nutrient conditions, physiological state of animals, and environment factors, affect ultimate meat quality in order to evaluate these uncertainness. Unfortunately, there is little information that completely covers with relationship among the muscle fiber types, myofibrillar proteins and enzymatic proteolysis. In addition to the perspective of postmortem metabolism, protein quality control in skeletal muscle and proteolytic degradation of muscle proteins during postmortem period could help to clarify this relationship. Therefore, the present review will focus on muscle fiber types, typing methods, muscle proteins and meat quality, and will summarize aspects of enzymatic view of proteasome.

Classification and development of myofiber types in the superior oblique extraocular muscle of chicken.

Precise control of contractile force of extraocular muscles is required for appropriate movements and alignment of the eyes. It is unclear how such precise regulation of contractile force is achieved during development and maturation. By using the posthatch chicken as a model, we describe and quantify critical parameters of the developing superior oblique extraocular muscle from hatching to 16 weeks of age, including contractile force, muscle mass, myofiber diameters, classification of fiber types, and distribution and quantification of mitochondria. Analysis at the light- and electron microscopic levels shows that chicken myofiber types largely correspond to their mammalian counterparts, with four fiber types in the orbital and four types in the global layer. Twitch tension muscle force and muscle mass gradually increase and stabilize at approximately 11 weeks. Tetanic tension continues to increase between 11 and 16 weeks. Myofiber diameters in both the orbital and global layer increase from hatching to six weeks, and then stabilize, whereas the myofiber number is constant after hatching. This finding suggests that muscle mass increases during late maturation due to increasing fiber length rather than fiber diameter. Quantitative ultrastructural analysis reveals continuing changes in the composition of the four muscle fiber types, suggesting ongoing fiber type conversion or differential replacement of myofiber types. Muscle fiber composition continues to change into late juvenile and adult age. Our study provides evidence for gradual, incremental, and continuing changes in avian myofiber composition and function that is similar to postnatal oculomotor maturation in visually oriented mammals such as kitten.

Fiber type composition and maximum shortening velocity of muscles crossing the human shoulder.

A study of the fiber type composition of fourteen muscles spanning the human glenohumeral joint was carried out with the purpose of determining the contribution of fiber types to overall muscle cross-sectional area (CSA) and to estimate the maximum shortening velocity (V(max)) of those muscles. Muscle biopsies were procured from 4 male cadavers (mean age 50) within 24 hr of death, snap frozen, mounted, and transversely sectioned (10 microm). Slides were stained for myofibrillar ATPase after alkaline preincubation. Photoimages were taken of defined areas (100 fibers) using the Bioquant system, and fiber type and CSA were measured from these images. Staining for mATPase produced three different fiber types: slow-oxidative (SO), fast-oxidative-glycolytic (FOG), and fast-glycolytic (FG). On average, the muscle fiber type composition ranged from 22 to 40% of FG, from 17 to 51% of FOG, and from 23 to 56% of SO. Twelve out of the 14 muscles had average SO proportions ranging from 35 to 50%. V(max) was calculated from the fiber type contribution relative to CSA and shortening velocity values taken from the literature. The maximum velocities of shortening presented here provide a physiological basis for the development of human shoulder musculoskeletal models suitable for predicting muscle forces for functionally relevant tasks encompassing conditions of muscle shortening and lengthening.

Testosterone mediates satellite cell activation in denervated rat levator ani muscle.

Denervation stimulates quiescent satellite cells in skeletal muscle to reenter the cell cycle. In the androgen-sensitive rat levator ani muscle (LA), this mitotic response to loss of neural input fails to occur in castrated animals. To elucidate the role of androgens in denervation-induced satellite cell proliferation, the denervated LA of castrated rats (Group A) was compared with that of animals infixed with testosterone implants after castration (Group B). Mean myofiber cross-sectional areas (Group A: 362.95 microm(2) +/- 27.74; Group B: 403.13 microm(2) +/- 53.87) and linear nuclear densities (Group A: 74.07 mm(-1) +/- 17.58; Group B: 104.13 mm(-1) +/- 4.06) were similar (P > 0.05) in both groups. The androgen-deprived myofibers of Group A, however, had a significantly lower nuclear content (271.0 +/- 74.91 vs. 1,285.80 +/- 81.74 in Group B; P < 0.05) on account of their considerably shorter mean length (3.44 mm +/- 0.29 vs. 12.31 mm +/- 0.92 in Group B; P < 0.05). The proportional representation of satellite cells in hormone-replaced, denervated muscle was more than twice that in the untreated group (Group B: 5.15 +/- 0.83% vs. Group A: 2.28 +/- 0.23%; P < 0.05). In absolute terms, the satellite cell number in Group B was approximately an order of magnitude greater than in Group A (408.4 x 10(3) vs. 38.08 x 10(3)). The results confirm the absence of testosterone as the factor responsible for the inability of satellite cells in the LA of castrated rats to respond mitotically to the withdrawal of neural input after denervation.

Satellite cell numbers in senile rat levator ani muscle.

Ageing in skeletal muscle results in motor frailty and a reduced capacity for self repair after injury. The contractile characteristics of muscle are determined principally by the myosin heavy chain (MHC) composition of its myofibers. During the restorative process, satellite cells play a central role. The present study compares the levator ani muscle of very old (32 months) and young (4 months) male WI/HicksCar rats in terms of structural integrity, MHC and satellite cell content. Myofiber typing was carried out by indirect immunohistochemistry using a panel of anti-MHC antibodies. Single myofibers for nuclear enumeration were isolated by an enzymatic technique while fiber cross-sectional areas and satellite cell frequencies were determined by computerized planimetry and electron microscopy. In both groups of rats, the myofiber population was homogeneously MHC type IIb-reactive. Cross-sectional data reflected a marked degree of atrophy in the muscle of the senile rats (710.05 +/- 63.6 microm2, compared with 1519.98 +/- 79.0 microm2 in young). The myofiber population was reduced by only about 6.7% with ageing and the representation of satellite cells, as a fraction of total sublaminal nuclei, was relatively stable (1.15 versus 1.91% in young; P > 0.05). The results indicate that ageing had a considerable atrophic effect on the levator ani muscle but induced neither MHC isoform transition nor massive depletion of the satellite cell pool. They suggest that the well-documented impairment of the restorative capacity of senile muscle could be due more to alterations in the nature of microenvironmental cues than to quantitative aspects of its cellular capacity to respond.

Changes in myosin expression in denervated laryngeal muscle.

The effects of chronic denervation on the myosin heavy chain (MyHC) content and muscle fiber type composition of rat laryngeal muscles are described. The posterior cricoarytenoid (PCA) and thyroarytenoid (TA) muscles were removed 3 weeks, 3 months, and 6 months after recurrent laryngeal nerve sectioning. Myofibrillar adenosine triphosphatase staining of cryostat sections was performed, and fiber type percentages were determined. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to separate MyHC isoforms, and densitometry was subsequently used for quantitative analysis. Unoperated animals served as controls. In the PCA muscle, denervation resulted in a progressive reduction in type I MyHC (the slow-contracting isoform) to an almost complete loss at 6 months, with a concomitant increase in type II MyHCs (fast-contracting isoforms, excluding type IIL). Type IIL MyHC (laryngeal-specific isoform) remained relatively constant up to 6 months after denervation. The myosin expression in the TA muscle, which contained only type II MyHCs, remained relatively constant with denervation. Changes in fiber type composition of the muscles described from tissue staining correlated with MyHC content. These findings in laryngeal muscle confirm the dependence of type I MyHC expression upon neural input, as has been found previously in limb skeletal muscles. Since the expression of all MyHCs except the IIL was modified after denervation in the PCA muscle, it is possible that the IIL isoform is maintained by factors that differ from those in the other skeletal myosins.

Motor unit recruitment and the gradation of muscle force.

The capabilities of the different types of motor units are reviewed, and their properties in a variety of muscles are discussed. Because the tension-generating capacities of motor units are so different, the order in which they are recruited will have a strong influence on the way force output of the whole muscle is graded. Activation of motor units in a random order produces a roughly linear force increase with progressive recruitment, whereas recruitment of motor units in order of increasing force produces an approximately exponential force increase as the number of active motor units increases. The latter scheme allows fine control of weak movements and rapid production of powerful movements. Motor units are shown to be well adapted to the tasks they must perform, and a "compromise" motor unit will not fulfill all the tasks demanded of it. Finally, changes in motor unit properties produced by different activity patterns and by muscle reinnervation are reviewed, and the implications for rehabilitation are discussed.

Contractile properties of skinned fibers from hypertrophied skeletal muscle.

We have previously shown deficits in specific tension (N.cm-2) in whole hypertrophied skeletal muscle (18-30%). The purpose of the present study was to determine if this intrinsic deficit exists in chemically skinned, hypertrophied single fibers, and also to measure their Ca2+ sensitivity. One group of rats (N = 4) had bilateral surgical ablation of the gastrocnemius and soleus muscles, and another group (N = 4) had the gastrocnemius and plantaris muscles ablated to induce hypertrophy in the plantaris and soleus muscles, respectively. Thirty days after surgery five fibers were dissected and analyzed from each of eight muscles per group, including control muscles. In hypertrophied plantaris and soleus muscles, fiber cross-sectional area (CSA) was 20% and 29% greater, respectively, than control values. Maximal Ca2+ activated tension (Po, pCa 4.5) was 15% greater in hypertrophied plantaris fibers and 18% greater in hypertrophied soleus fibers compared with respective control values (P < 0.05). Therefore, Po expressed per fiber CSA was slightly depressed (3-8%) in both plantaris and soleus fibers (P < 0.05). Regarding Ca2+ sensitivity, the Ca2+ concentration to elicit 50% of Po (i.e., pCa50) was significantly lower in hypertrophied soleus (58%) and plantaris (29%) fibers. This leftward shift in the force-pCa curve reflects greater Ca2+ sensitivity in hypertrophied fibers. Since the single fiber Po/CSA was only slightly decreased, these data suggest that during tetanic stimulation of whole muscle Ca2+ delivery to the contractile apparatus may be impaired.(ABSTRACT TRUNCATED AT 250 WORDS)

Morphometric analysis of the developing mouse soleus muscle.

The pattern of organogenesis of the soleus muscle of the 129 ReJ mouse was evaluated quantitatively using spaced, serial, ultrathin sections and computer-assisted morphometric analysis. Muscles from 14-, 16-, and 18-day in utero mice and muscles of 1- and 5-day-old mice were analyzed to determine age-related alterations in the maximal girth and length of the muscle, number of myotubes, cluster frequency, and the lengths and diameters of myotubes. Primary myotubes are found in the muscle at 14 days in utero. There is little de novo myotube formation between 14 and 16 days in utero, this interval being principally one of primary myotube growth and maturation. The interval between 16 and 18 days in utero is marked by extensive secondary myotube formation, with more myotubes being formed during this period than in any period studied. Morphometric data support the hypothesis that secondary generation myotubes use primary myotubes as a scaffold on which they are formed. Morphometric data also confirm the hypothesis that cluster formation and cluster dispersal occur concurrently during the prenatal period. Secondary myotubes continue to form until birth. At birth, the soleus muscle contains the adult number of myofibers. The first 5 days postnatally are marked by myofiber growth and maturation.

A new method for myofibrillar Ca++-ATPase reaction based on the use of metachromatic dyes: its advantages in muscle fibre typing.

A new method for Ca++-ATPase reaction in human muscle fibres is presented as an alternative to previous ATPase stains. The method is based on the use of metachromatic dyes, namely Azure A and Toluidine Blue, and has the advantages of speed, ease of performance and production of an elegant and clearcut fibre typing. The method distinguishes fibre types because of their metachromatic or orthochromatic staining, due to their different content of phosphate after incubation in the reaction medium. The comparison of serial sections stained by cationic dyes and by ammonium sulphide revealed close correspondence of fibre typing. Fibre type differentiation was also obtained with Acridine Orange; however this method was less reproducible.

A histochemical study of myofiber types in muscle of the growing pig.

The conversion of myofiber types was studied in the masseter, trapezius, longissimus, rectus femoris and vastus intermedius muscles of pigs at birth and 2, 4, 8 and 16 weeks of age. The histochemical reaction for ATPase was used to classify myofibers as type I (acid-stable, alkali-labile) or type II (alkali-stable, acid labile). Type II myofibers were divided further into categories IIA and IIB. Subtypes SM, SS and MS were intermediate to types I and II and were classified on the basis of pH sensitivity of the ATPase reaction. In the longissimus, rectus femoris and masseter muscles, the proportion of type II myofibers decreased and the proportion of type I myofibers increased from birth to 8 weeks of age, and little change occurred thereafter. These three muscles had more type II than type I myofibers, regardless of age. At birth the trapezius and vastus intermedius muscles both and more type II than type I myofibers, but during development, the proportion of type I myofibers increased greatly while that of type II decreased markedly. At 16 weeks, the trapezius had about equal proportions of type II and type l myofibers, but the vastus intermedius had a much higher proportion of type I than type II myofibers. A flow diagram was developed to illustrate the proposed conversion of type II to type I myofibers via the intermediate stages identified as subtypes MS, SS and SM. Development of the type I grouped pattern and the association of myofiber proportions with functions in the masseter muscle are discussed.

Myofibrillar differences among mammalian skeletal muscle fibres at the ultrastructural level. A comparison of immunocytochemical and morphometrical parameters.

In the light microscope two types (I, II) of skeletal muscle fibres can be distinguished with antibodies against myosin isozymes. At the ultrastructural level a difference in Z-line width has led to muscle fibre classification. In this study we distinguish at the ultrastructural level between type I and type II fibres of the M. soleus and M. plantaris of adult mice using ultracryosections with immuno-ferritin and antisera against myosin isozymes. Muscle fibres of the M. plantaris are identified as type II fibres and the fibres of the M. soleus are divided in type I and type II fibres. In the immunologically identified fibres the filament overlap in the Z-line was measured. The type II fibres of the M. plantaris have narrow Z-lines, whereas type I and type II fibres of the M. soleus have wide Z-lines. We conclude that a classification of fibres based on Z-line width differs from the type I/type II classification. The antimyosin antibodies react exclusively with the A-band. In serial sections the myosin isozymes can be identified unambiguously. This is a prerequisite for further studies of myosin isozyme distribution in "mixed" muscle fibres.