Histochemical studies of muscle development in decapitated and hypophysectomized pig fetuses: blood vessel development

In Exp. 1, fetuses were decapitated in utero on d 45 of gestation and examined at 110 d of gestation; in Exp. 2, fetuses were hypophysectomized (hypox) in utero with a cauterizing needle at d 70 of gestation and examined at 110 d of gestation. Semitendinosus muscles were sampled and transverse cryostat sections were cut from the medial portion of the muscle. Quantitative analysis of lectin-stained sections showed that capillary:fiber ratios (C/F) were lower (P less than .01) for sections from decapitated and hypox fetuses than for sections from control fetuses. The superficial and deep aspects of muscle sections from control fetuses had markedly different C/F ratios (deep region, 24.1 +/- 2; superficial region, 11.2 +/- .5). Fetal decapitation and hypophysectomy abolished differences in C/F ratios between deep and superficial regions of muscle sections (hypox fetuses; deep region, 6.9 +/- 1; superficial region, 6.4 +/- .6). Capillaries in sections from control fetuses were reactive for several enzymes, whereas capillaries in sections from experimental fetuses (decapitated and hypox) were not reactive for these enzymes. The number of small arterioles was reduced in sections from experimental fetuses (decapitated and hypox) compared with sections from control fetuses. These studies demonstrate that lowering the levels of pituitary hormones in the fetus retards the development of blood vessels in muscle tissue both quantitatively and qualitatively.     

M. Articularis Coxae—a Possible Receptor Organ

The purpose of the study was to analyse the morphology of the equine m. articularis coxae with reference to its function during locomotion. The muscle was a constant finding in 50 horses of verying age nad sex. Different horses were selected for dissection of the muscle with its vessels and nerves, determining of length at different angles of the hip joint and weight. The muscle was shortest at maximal fexion, and the length incereased rapidly until a hip joint angle of 110°. Between 11° and maximal extension the increase in length was moderate. The mean weight of muscle was 2.7 g. Histological specimens were serially sectioned and the frequency of muscle spindles was recorded. The spindle density varied from 12 to 42 per gramme of muscel, with a mean of 26, and a mean length of the muscle spindles of 4.7 mm.
Myectomy was performed uni- or bilaterally on 5 horses to evaluate the influenceon the locomotion with no significant manges observed.
Considering the position, size, innervation and multitude of muscle spindles of the m. articularis coxae it is suggested that the muscle primarily has a receptor function inreporting on the torsion or "wrinkling" of the hip joint. The absence of manges in loconiotion of the myectomised horses indicates that the missing function of the muscle is taken over by other muscles.     

Effects of birth weight and postnatal nutrition on neonatal sheep: II. Skeletal muscle growth and development

This study investigated effects of birth weight and postnatal nutrition on growth and development of skeletal muscles in neonatal lambs. Low (L; mean +/- SD 2.289 +/- .341 kg, n = 28) and high (H; 4.840 +/- .446 kg, n = 20) birth weight male Suffolk x (Finnsheep x Dorset) lambs were individually reared on a liquid diet to grow rapidly (ad libitum fed, ADG 337 g, n = 20) or slowly (ADG 150 g, n = 20) from birth to live weights (LW) up to approximately 20 kg. At birth, weight of semitendinosus (ST) muscle in L lambs was 43% that in H lambs; aggregate weights of ST and seven other dissected muscles were similarly reduced. In ST muscle of L lambs, mass of DNA, RNA, and protein were also significantly reduced to levels 67, 60, and 34%, respectively, of those in H lambs. However, myofiber numbers of ST, tibialis caudalis, or soleus muscles did not differ between the L and H birth weight lambs and did not change during postnatal growth. During postnatal rearing, daily accretion rate of dissected muscle was lower in L than in H lambs. Accretion of muscle per kilogram of gain in empty body weight (EBW) was reduced in the slowly grown L lambs compared with their H counterparts, although the difference was less pronounced between the rapidly grown L and H lambs. Throughout the postnatal growth period, ST muscle of L lambs contained less DNA with a higher protein:DNA ratio at any given muscle weight than that of H lambs. Slowly grown lambs had heavier muscles at any given EBW than rapidly grown lambs. Content of DNA and protein:DNA ratio in ST muscle were unaffected by postnatal nutrition, but RNA content and RNA:DNA were greater and protein:RNA was lower at any given muscle weight in rapidly grown lambs. Results suggest that myofiber number in fetal sheep muscles is established before the presumed, negative effects of inadequate fetal nutrient supply on skeletal muscle growth and development become apparent. However, proliferation of myonuclei may be influenced by fetal nutrition in late pregnancy. Reduced myonuclei number in severely growth-retarded newborn lambs may limit the capacity for postnatal growth of skeletal muscles.     

Influence of maternal obesity on fetal development in pigs

Fetuses from linebred lean (L) and linebred obese (O) and reciprocal crossmatings were examined at 110 d of gestation for line, maternal and heterotic effects. There was no significant heterotic effect for any trait measured. A significant maternal effect was observed for adipose tissue lipoprotein lipase (LPL) activity and for serum triglycerides. The enzyme activity and triglycerides concentration were higher in fetuses from O dams than in fetuses from L dams. In a lipid clearance test, no maternal effect was observed for changes in serum concentrations of triglycerides and free fatty acids or in optical density (associated with the disappearance of injected Liposyn). Linebred O fetuses exhibited higher LPL activity in both the biceps femoris muscle and sc adipose tissue compared with linebred L fetuses. The LPL activity of the adipose tissue was higher than that of the skeletal muscle. The percentage of dry matter, percentage of triglycerides and protein/DNA were higher in the muscle of linebred O fetuses than in that of linebred L fetuses. Based on tissue LPL activity and on muscle compositional traits, linebred O fetuses were more mature at 110 d of gestation than were the linebred L fetuses. Maternal obesity had little detectable influence on fetal development of the pig when measured at 110 d of gestation.     

How to Open the Mouth: the Kinetic Skull, and the Jaw and Branchial Apparatus in Sharks

Aims:  Mammals cannot swallow if their necks and, hence, their mm. sternohyoidei et sternothyroidei are stretched. In order to understand the functional morphology of mastication and swallowing in mammals, it is useful to understand its evolutionary origin.
Material and Methods:  Five formalin-preserved specimens of the Spiny Dogfish (Squalus ancanthias) were used for dissection or to prepare sagittal or transverse serial sections.
Results and Conclusions:  The feeding apparatus of the shark Squalus acanthias as a representative of the cartilaginous fishes serves as an excellent model of that of an ancestral vertebrate with a kinetic skull, in which the chondrocranium is movable relative to the jaw apparatus. To open the mouth, the cranium is raised by the epibranchial musculature, the palatoquadrate cartilage (i.e., upper jaw) is entrained, and the mandibular cartilage is held in place and prevented from following the upper jaw by the coracomandibular muscle. At the same time, the branchial basket and oropharyngeal cavity are expanded and held in place by the coracohyoid and coracoarcual muscles, which prevent the hyoid and branchial arches from being entrained cranially by the raised chondrocranium. The mandible is not retracted during the opening of the mouth in sharks because the branchial region would have to be compressed instead of expanded. In the course of the evolutionary modifications leading to mammals, the heart and respiratory apparatus became decoupled from the feeding apparatus through the formation of a neck. The skull became akinetic through the integration of the upper jaw into the base of the cranium. The mouth is not opened any longer by lifting the cranium, but by depressing the mandible through a special muscle, the m. depressor mandibulae . The coracomandibular muscle lost its function as a stabilizer of the mandible during mouth opening and became modified into the lingual and geniohyoid muscles.     

Changes in muscle fiber type and expression of mRNA of myosin heavy chain isoforms in porcine muscle during pre‐ and postnatal development

The purpose of this study is to elucidate developmental changes in muscle fiber type in the pig during pre‐ and postnatal development. For this purpose, we performed a histochemical analysis for myosin adenosine triphosphatase activity to assess muscle fiber type and determined abundances of messenger RNA (mRNA) of myosin heavy chain (MHC) isoforms. Samples of Longissimus dorsi (LD) muscle were taken from fetuses on day 90 of the fetal stage. Further, samples of LD, Rhomboideus and Biceps femoris (B. femoris) muscles were taken from pigs when they were 1, 12, 26, 45 or 75 days old. Expression of MHC 2b mRNA in the LD and the B. femoris muscles rapidly and considerably increased from the late fetal stage to the early postnatal stage and this increase was associated with the development of type 2b fibers at least in the LD muscle. As shown by the rapid and considerable changes in expression of MHC 2b mRNA, it seems that a certain plasticity of muscle fiber type still remains in this developmental stage.     

Effects of low and high levels of maternal nutrition consumed for the entirety of gestation on the development of muscle,adipose tissue,bone, and the organs of Wagyu cattle fetuses

This study aimed to investigate the effects of high and low levels of energy intake during the entire gestation period on the skeletal muscle development, organ development, and adipose tissue accumulation in fetuses of Wagyu (Japanese Black) cows, a breed with highly marbled beef. Cows were allocated to a high-nutrition (n = 6) group (fed 120% of the nutritional requirement) or low-nutrition (n = 6) group (fed 60% of the nutritional requirement). The cows were artificially inseminated with semen from the same sire, and the fetuses were removed by cesarean section at 260 ± 8.3 days of fetal age and slaughtered. The whole-body, total muscle, adipose, and bone masses of the fetal half-carcasses were significantly higher in the high-nutrition group than the low-nutrition group (p = 0.0018, 0.009, 0.0004, and 0.0362, respectively). Fifteen of 20 individual muscles, five of six fat depots, nine of 17 organs, and seven of 12 bones that were investigated had significantly higher masses in the high-nutrition group than the low-nutrition group. The crude components and amino acid composition of the longissimus muscle significantly differed between the low- and high-nutrition groups. These data indicate that maternal nutrition during gestation has a marked effect on the muscle, bone, and adipose tissue development of Wagyu cattle fetuses.     

Types of myofibers parasitized in experimentally induced infections with Sarcocystis cruzi and Sarcocystis capracanis

Eighteen calves were orally inoculated with either 200,000 or 225,000 sporocysts of Sarcocystis cruzi. Eight goats were orally inoculated with 20,000 sporocysts of S capracanis. Calves and goats were euthanatized at various times after inoculation, and portions of their right and left biceps femoris, right and left longissimus dorsi, myocardium, and tongue were frozen at -150 C in precooled isopentane and stored at -70 C. Frozen sections of these muscles were stained with hematoxylin and eosin, modified Gomori's trichrome, nonspecific esterase, diphosphopyridine nucleotide tetrazolium reductase, and adenosinetriphosphatase at pH 10.4 and 4.6. Muscle from the same locations was fixed in 10% neutral buffered formalin, processed for paraffin embedding, sectioned, and stained with hematoxylin and eosin. Microscopic examination of both calf and goat tissue indicated that both type I and type II muscle fibers were equally infected and that infected myofibers showed no apparent damage other than displacement by sarcocysts. Occasionally, muscle fibers within the muscle spindles contained sarcocysts.     

Relationships between tropomyosin and myosin heavy chain isoforms in bovine skeletal muscle

The composition of tropomyosin (TPM) and myosin heavy chain (MyHC) isoforms was analyzed in 10 physiologically different bovine muscles ( masseter , diaphragm, tongue, semispinalis, pectoralis profundus , biceps femoris, psoas major , semimembranosus, longissimus thoracis and semitendinosus ) to clarify the relationships between TPM and MyHC isoforms in different muscle fiber types. The content of TPM1 and TPM3 was different in muscles according to their function in muscle contraction, although the content of TPM2 was constantly about 50% of the total TPM in all muscles. The content of TPM1 was higher in semimembranosus , longissimus thoracis and semitendinosus, while that of TPM3 was higher in masseter and diaphragm. The high positive correlation between MyHC-slow content and TPM3 content ( r  = 0.92) suggested a coexpression of TPM3 and MyHC-slow isoforms in a muscle fiber. MyHC-slow and TPM3 were expressed at the same level in masseter and diaphragm, whereas there was more TPM3 than MyHC-slow in tongue and semispinalis , so it appears that the excess TPM3 in tongue and semispinalis is expressed with other MyHC isoforms. MyHC-2a was the only fast type isoform expressed in tongue and semispinalis . Therefore, the excess TPM3 was composed of myofibrils with MyHC-2a. The results suggested that a fiber expressing MyHC-2a would be regulated delicately by changing the TPM isoform types.     

Effect of nutrient restriction on calpain and calpastatin content of skeletal muscle from cows and fetuses

Calpains are crucial for the degradation of myofibrillar proteins in muscle. Calpastatin is a specific inhibitor of calpains. The objective of this study was to elucidate the effect of nutrient restriction on the activity of calpains and calpastatin in the skeletal muscle of both cows and fetuses. Beginning 30 d after conception, 20 cows were fed either a control diet consisting of native grass hay fortified with vitamins and minerals at recommendations for a mature cow to gain 0.72 kg/d or half the vitamins and minerals and millet straw at 68.1% of NEm requirements. Cows were slaughtered on d 125 of gestation, and the LM was sampled at the 12th rib for calpain and calpastatin measurement. When comparing the muscle samples from nutrient-restricted and control cows, no difference in the activity of calpain I and II was observed; however, there was a significant difference (P < 0.05) in calpastatin activity. Muscle samples from control cows had greater calpastatin content than those of nutrient-restricted cows (P < 0.05); in contrast, the calpastatin content of fetal muscle was greater in fetuses gestated by nutrient-restricted cows than those of control cows (P < 0.05). Further, there were three calpastatin isoforms of 125, 110, and 70 kD detected in fetal muscle, whereas only the110-kD isoform was detected for cow muscle. These results indicate that the activity of the calpain system in skeletal muscle is mainly controlled through the expression of calpastatin. Alternating the calpastatin content in muscle and thereby modulating calpain activity may provide a mechanism for the maintenance of fetal muscle growth during nutrient restriction, whereas skeletal muscle loss in cows is upregulated.     

Altered muscle development and expression of the insulin-like growth factor system in growth retarded fetal pigs

We have used a porcine model of spontaneous differential fetal growth to investigate the effects of fetal size on muscle development. We hypothesized that altered muscle development may occur in small fetuses as a consequence of modified expression of selected genes of the insulin-like growth factor system. We examined the development of the Longissimus muscle (m. Longissimus) in small fetuses and their average sized littermates. We collected small for gestational age fetuses and their average sized sibling on days 45, 65 and 100 of gestation (term is 113-116 days). Small fetuses had significantly lower body weight at all three stages of gestation (p<0.05) and significantly reduced secondary to primary muscle fibre ratio in m. Longissimus on day 100 (p<0.05) compared to their littermates. On day 65, the expression of insulin-like growth factor receptor 1 and insulin-like growth factor binding protein 3 were significantly higher (p<0.05) in m. Longissimus of the small fetuses compared with their average sized littermates. On day 100, the expression of insulin-like growth factor receptor 1 remained significantly higher (p=0.001), in addition to significantly higher levels of insulin-like growth factor receptor 2 and insulin-like growth factor binding protein 5 in the small fetuses (p<0.05). No difference in levels of myogenin was observed between the small and average sized littermates. In conclusion, we demonstrate that reduced fetal muscle development is associated with an increased expression of several genes of the insulin-like growth factor system in small fetuses in mid to late gestation.     

Mouthpart structure and function and the feeding mechanisms of Gerres (Teleostei)

Members of the genus Gerres feed almost exclusively on benthic invertebrates and have evolved efficient protrusible jaw mechanisms and modes of feeding. The methods employed in prey capture and feeding by five species that occur in the estuaries of Natal were studied using slow-motion cine photography and results were supplemented by field observations. All species use visual cues to detect and capture prey on or just below the surface of the substrate.

The protrusible jaw mechanism is the most important feature in prey capture. Bones and muscles involved in jaw movement were studied by dissecting preserved specimens and cleaning jaws and skulls of muscle. The whole mechanism of protrusion of the mouthparts was found to be controlled by two sets of muscles acting on a modified maxilla which together with the cross-palatine ligament forms a groove along which the ascending process of the maxilla can slide when the mouthparts are protruded. The setting up of a suction pressure within the mouth further aids the uptake of items from the substrate. However, whether or not suction pressure is applied depends on the prey type.     

Prenatal Development of the Human Epicardiac Ganglia

The aim of this study was to determine the developmental anatomy of intrinsic cardiac ganglia with respect to epicardiac ganglionated nerve plexus in the human fetuses at different gestation stages. Twenty fetal hearts were investigated applying a technique of histochemistry for acetylcholinesterase to visualize the epicardiac neural ganglionated plexus with its subsequent examinations on total (non-sectioned) hearts. Most epicardiac ganglia embodied multilayered neurons and were oval in shape, but some ganglia involved neurons lying in one layer or had the irregular appearance because of their extensions along inter-ganglionic nerves. The mean ganglion area of fetuses at gestation stages of 15–40 weeks was 0.03 ± 0.008 mm². The largest epicardiac ganglia, reaching in area 0.4 mm², were concentrated on the dorsal surface of both atria. The particular fused or "dual" ganglia were identified at the gestation stages of 23–40 weeks, but they composed only 2.3 ± 0.7% of all found epicardiac ganglia. A direct positive correlation was determined between the fetal age and the ganglion area (mm²) as well as between the fetal age and the number of inter-ganglionic nerves. The revealed appearance of epicardiac ganglia in the human fetuses at 15–40 weeks of gestation confirms their prenatal development and presumable intrinsic remodelling.     

Muscle fibre type distribution of the thoracolumbar and hindlimb regions of horses: relating fibre type and functional role

Background

Although the majority of equine muscles have a mixed fibre type distribution indicative of diverse functional roles, the predominance of a fibre type can indicate the primary function of a muscle. The deep epaxial musculature has an important role in core spinal stability in humans, reflected as a predominantly muscle fibre type (MFT) I or postural fibre type. The fibre type of the deep epaxial musculature has not been determined in horses. The objective of the study was to determine the MFT distribution in selected muscles of thoracolumbar and hindlimb region of horses. This included deep epaxial and hypaxial muscles that were hypothesised to have a postural stabilising role. A second objective was to examine differences in MFT distribution between horses bred for endurance (Arabian) and sprinting (Quarter horse). Muscle biopsy samples were obtained from selected thoracolumbar and hind limb muscles of 5 Quarter horses, 4 Arabians, and 2 Thoroughbreds. The myosin heavy chain distribution was determined by gel electrophoresis. Mann–Whitney rank test was used to compare the proportional MFT and differences between breeds.

Results

Mm. sacrocaudalis dorsalis medialis and diaphragm had the highest proportion of MFT-I. The remaining deep epaxial muscles and the hypaxial muscle m. psoas minor had approximately equal MFT I and II proportions. Mm. psoas major, iliocostalis, longissimus dorsi and the hind limb muscles contained mostly MFT-IIX. The fibre type distribution was similar between Arabians and Quarter horses, although Quarter horses had more MFT-IIX fibres in psoas major (P = 0.02) while Arabians had more MFT-I fibres in m. longissimus dorsi (P = 0.03).

Conclusions

The fibre type distribution of the deep epaxial muscles, mm psoas minor and diaphragm varied from approximately equal MFT-I and II proportions to predominantly MFT-I suggesting a postural stabilising role possibly important in core spinal stability. In contrast the fibre type proportions of mm psoas major, iliocostalis, longissimus dorsi and the hind limb muscles were mainly MFT-II suggesting a locomotory role. Knowledge of fibre type distribution in such a clinically important area can direct diagnosis, prevention and treatment of muscular or neuromotor dysfunction.     

Nucleotide sequence of myostatin gene and its developmental expression in skeletal muscles of Japanese Black cattle

Myostatin, a member of the transforming growth factor‐β superfamily, is a well known negative regulator of skeletal muscle growth. In the present study, the 6660 bp nucleotide sequence of the myostatin gene in Japanese Black cattle (JBC), including the entire coding region of 1128 bp, was determined. The amino acid sequence deduced from the nucleotide sequence of JBC was well conserved with its sequence of other cattle, although it was found that an Α→G transition at nucleotide position 641 results in the substitution of asparagine by serine at amino acid position 214. In order to examine the expression pattern of the myostatin gene in the skeletal muscles of JBC, its expression in three skeletal muscles, Semitendinosus (ST) muscle, Biceps femoris muscle and Longissimus lumborum muscle, of fetal and calf stages was analyzed by real time polymerase chain reaction. The highest level of the myostatin expression was observed in the fetal stage. In calf stages the highest expression was observed in ST muscle compared with the other two muscles. These results suggest that a higher expression of myostatin gene, especially in the fetal stage and in ST muscle during calf stages, is involved in the arrest in skeletal muscle growth and that its functional domains and genomic structure in JBC are well conserved with those in other mammals.     

Influence of insulin and substrate concentration on protein synthetic rate in fetal tissues

The influence of fetal plasma substrate and insulin concentrations on the fractional synthesis rate or turnover of the mixed proteins in the organs of fetal lambs was calculated from measurements of the rate of uptake of L-14C lysine by the protein in the steady state in utero. The control values for protein synthesis rate in the fetal organs of Welsh mountain ewes, 130 to 140 days' pregnant, were similar to those previously observed in the fetuses of Border Leicester ewes: the mean turnover rates were 38, 72, 40 and 63 per cent per day for the brain, liver, cardiac muscle and the placenta respectively; that for skeletal muscles was 6.8 per cent per day; the corresponding half lives were 2.15, 1.27, 1.8 and 1.42 days respectively and 11.3 days for skeletal muscle. A fivefold increase in the plasma glucose levels had no influence on protein turnover rate over a six hour period. A fourfold increase in the concentration of some amino acids, following a mixed infusion, decreased turnover rate in brain, heart and liver. The half life was increased by 30 per cent, but a small reduction in half life was observed in skeletal muscle. Insulin caused a fall in intracellular lysine concentration in skeletal muscle and in plasma, which suggests that the hormone can reduce muscle protein catabolism in the fetal lamb in the last trimester of gestation. Enhancement of synthesis rate occurred in the presence of the amino acid infusions only; the half life of the proteins was reduced by a factor of three. Insulin also increased placental protein synthesis rate by 50 per cent but had no influence on the other tissues studied.