Effects of incubation temperature on muscle morphology and growth in the pacu (Piaractus mesopotamicus)

This study describes the influence of incubation temperature during initial development phase on the morphology and muscle growth characteristics in the pacu (Piaractus mesopotamicus). Pacu eggs were incubated at 25, 27, and 29 degreesC until hatching. After day 5, fish from each temperature were transferred to 5001 tanks. At hatching and after 5, 25, and 60 days, muscle samples were collected, some were frozen in liquid nitrogen and others fixed in 4% paraformaldehyde or 2.5% glutaraldehyde. These samples were used for morphological, histochemical, immunohistochemical, and morphometric analysis. At hatching, we observed a superficial monolayer of small diameter fibers, lying just beneath the skin surrounding several round cells. From day 5, we observed two distinct populations of muscle fibers distributed in two layers: (1) red-in a superficial region with aerobic activity, and following acid preincubation, high mATPase activity, and 2) white-with anaerobic activity, and following alkaline preincubation, high mATPase activity. Twenty-five days after hatching, an intermediate layer and cell proliferating zones could be seen in the dorsal fin muscle region, with intermediate characteristics. Throughout the experimental period, there was an increase in muscle mass due to new fiber recruitment in the cell proliferating zones and between the more differentiated fibers in red, intermediate, and white muscles. This was more obvious from day 25, and at 29 degreesC than at 25 and 27 degreesC. Fiber hypertrophy occurred from hatching to 60 days and was more evident from 5 to 25 days. The number of proliferating nuclei (PCNA-labelling) increased from hatching to 60 days, and was more obvious in the 29 degreesC group at 60 days. Our results show that at incubation temperatures of 25, 27 and 29 degreesC, hypertrophy was predominantly from hatching to 25 days, after that muscle growth by hyperplastic mechanism increased. The interaction of muscle hypertrophic and hyperplastic growth processes in the 29 degreesC group produced the largest fish at the end of the experiment. (C) 2004 Elsevier B.V. All rights reserved.

Histochemical and morphological characteristics of the levator ani muscle in rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ultrastructural aspects of the retractor ocular bulbi muscle in the opossum (Didelphis albiventris)

In the present study the different types of muscle fibers of the retractor ocular bulbi muscle of the South American opossum were classified according to their ultrastructural characteristics. The tridimensional characteristics of the neuromuscular junctions present: in this muscle were also demonstrated by scanning electron microscopy (SEM). Five adult opossums, three males and two females, were perfused with fixative solution through the left ventricle and their right retractor ocular bulbi muscle was prepared for the ultrastructural study of muscle fibers. The contralateral muscle was used for the study of neuromuscular junctions by SEM. Three types of fibers were detected, denoted 1r, 2r and 3r. Only simple neuromuscular junctions of the plate type were visualized by SEM.

Structural characteristics of the tendinous cord-papillary muscle junction in healthy and hypertensive rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Quantitative expression of myogenic regulatory factors MyoD and myogenin in pacu (Piaractus mesopotamicus) skeletal muscle during growth

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The use of carbon and nitrogen stable isotopes for the detection of poultry offal meal in meat-type quail feeds

The objective of the present study was to trace the inclusion of poultry offal meal (POM) in the diet of meat-type quails reared for a long period using the technique of stable isotopes. A number of 320 quails were randomly distributed into eight treatments: vegetable diet (T1), and a diet containing 8% POM were fed until the end of the experimental period (T2) or replaced by the vegetable diet on day 42 (T3), 56 (T4), 70 (T5), 84 (T6), 98 (T7), and 112 (T8). Breast muscle samples were collected from four birds randomly selected per treatment every 14 days. The obtained isotope results were submitted to multivariate analysis of variance (MANOVA) with the aid of the GLM procedure of statistical SAS program. Treatments were different from T1 when birds were sacrificed at least two weeks after the diet was changed. T2 results were different from T1 in all evaluated periods. It was concluded that it is possible to trace poultry offal meal inclusion in a strictly vegetable diet after the diet was changed for at least 14 days.

Morphological, histochemical and morphometric study of the myotomal muscle tissue of the pacu (Piaractus mesopotamicus Holmberg 1887: Serrasalminae, Characidae, Teleostei)

Histochemical, ultrastructural and morphometric methods were used to study growth patterns of red, pink and white muscle fibres and their relation to body weight and total length in the fast-growing freshwater fish Piaractus mesopotamicus Holmberg. The correlations amongst body weight, body length and diameter of red, pink and white fibres were low. From 10-15 to 40 50 cm, body weight increased 102.7 times, while the diameter of each type of fibro increased by factors of 0.94, 0.74 and 0.70, respectively. Muscle fibres revealed different morphological and histochemical stages of maturation. The frequencies of < 20 mu-m fibres of red, pink and white muscle tissue in the youngest and oldest classes were 64.5 and 11.0, 38.2 and 7.7 and 24.0 and 1.4%, respectively. In 30-40 cm fish, the frequency of < 20 mu-m fibres in the red and pink tissue was 24.5 and 25.5%, while in the white tissue it was 11.5%. During sexual maturity (40-50 cm), the recruitment of < 20 pm fibres in white muscle was 1.4%. Muscle fibres of this species showed continuous growth by both hyperplastic and hypertrophic mechanisms, and hyperplasia was particularly active in the juvenile phase.

Morphological aspects of neuromuscular junctions and gene expression of nicotinic acetylcholine receptors (nAChRs) in skeletal muscle of rats with heart failure

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Differential flight muscle development in workers, queens and males of the eusocial bees, Apis mellifera and Scaptotrigona postica

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Muscle damage slows oxygen uptake kinetics during moderate-intensity exercise performed at high pedal rate

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Anatomy of smooth muscle cells in nonmalignant and malignant human prostate tissue

Differently graded areas of human prostate adenocarcinoma were examined after Masson's trichrome staining or immunohistochemistry for smooth muscle alpha-actin, type IV collagen and laminin. In addition, the ultrastructure of the prostatic smooth muscle cells (SMC) during glandular proliferation and epithelial invasion in selected tumors was studied. The SMC formed a thick layer below the epithelial structures in unaffected areas and were closely associated with each other in homotypic interactions. As the tumor grade increased, the SMC gradually lost interactions with each other and became atrophic. With the growth of the epithelial compartment, the SMC initially segregated to the tumor periphery and the intercellular spaces increased. In high grade tumors, the epithelial cancer cells invaded the spaces between the SMC. Immunohistochemical analysis of the basal membrane revealed increased disruption of the usually thick basal membrane, which became thinner and faintly stained with each of the antibodies used. We conclude that most SMC become atrophic following epithelial invasion in human tumors and that degradation of the basal membrane is an important factor in this process. At the ultrastructural level, different SMC phenotypes occur in prostatic tissues during epithelial invasion. Interconversion between these phenotypes is suggested and a probable relationship among them is proposed.

DISTRIBUTION OF DIFFERENT FIBER TYPES OF M-EXTENSOR CARPI RADIALIS LONGUS OF THE RAT

As revealed by the NADH-diaphorase and myosine ATPase, the M. extensor carpi radialis longus of the rat possesses at least 3 main kinds of fibres, with different distribution on the superficial and deep portions of the muscle. The superficial portion revealed that 67.68 % are FG (fast-twitch-glycolytic) fibres, 14.72 % are FOG (fast-twitch-oxidative) fibres and 17.60 % are SO (slow-twitch-glycolytic) fibres. Already the deep portion revealed that 71.29 % are SO (slow-twitch-glycolytic) fibres, 17.46 % are FOG (fast-twitch-oxidative-glycolytic) fibres and 11.25 % are FG (fast-twitch-glycolytic) fibres. The miosine ATPase reaction was used to demonstrate contracting characteristics. These findings suggest that the movements of fast contraction of the M. extensor carpi radialis longus are predominant.

Effect of mechanical cycling on the push-out bond strength of fiber posts adhesively bonded to human root dentin

This study evaluated the effect of mechanical cycling on the bond strength of fiber posts bonded to root dentin. The hypotheses examined were that bond strength is not changed after fatigue testing and bond strength does not present vast variations according to the type of fiber post. Sixty crownless, single-rooted human teeth were endodontically treated, with the space prepared at 12 mm. Thirty specimens received a quartz fiber post (Q-FRC (DT Light-Post), and the remaining 30 specimens received a glass fiber post (G-FRC) (FRC Postec Plus). All the posts were resin luted (All Bond+Duolink), and each specimen was embedded in a cylinder with epoxy resin. The specimens were divided into six groups: G1-Q-FRC+no cycling, G2- Q-FRC+20,000 cycles (load: 50N, angle of 45 degrees; frequency: 8Hz); G3- Q-FRC+2,000,000 cycles; G4- G-FRC+no cycling; G5- G-FRC+20,000 cycles; G6- GFRC+2,000,000 cycles. The specimens were cut perpendicular to their long axis, forming 2-mm thick disc-samples, which were submitted to the push-out test. ANOVA (alpha=.05) revealed that: (a) QFRC (7.1 +/- 2.2MPa) and G-FRC (6.9 +/- 2.1MPa) were statistically similar (p=0.665); (b) the no cycling groups (7.0 +/- 2.4MPa), 20,000 cycles groups (7.0 +/- 2.1MPa) and 2,000,000 cycles groups (7.0 +/- 2.0MPa) were statistically similar (p=0.996). It concluded that mechanical cycling did not affect the bond strength of two fiber posts bonded to dentin.

CONCERTED ACTION OF THE HIGH-AFFINITY CALCIUM-BINDING SITES IN SKELETAL-MUSCLE TROPONIN-C

Mutants of each of the four divalent cation binding sites of chicken skeletal muscle troponin C (TnC) were constructed using site directed mutagenesis to convert Asp to Ala at the first coordinating position in each site. With a view to evaluating the importance of site-site interactions both within and between the N- and C-terminal domains, in this study the mutants are examined for their ability to associate with other components of the troponin-tropomyosin regulatory complex and to regulate thin filaments. The functional effects of each mutation in reconstitution assays are largely confined to the domain in which it occurs, where the unmutated site is unable to compensate for the defect, Thus the mutants of sites I and II bind to the regulatory complex but are impaired in ability to regulate tension and actomyosin ATPase activity, whereas the mutants of sites III and IV regulate activity but are unable to remain bound to thin filaments unless Ca2+ is present. When all four sites are intact, free Mg2+ causes a 50-60-fold increase in TnC's affinity for the other components of the regulatory complex, allowing it to attach firmly to thin filaments. Calcium can replace Mg2+ at a concentration ratio of 1:5000, and at this ratio the Ca2 . TnC complex is more tightly bound to the filaments than the Mg2 . TnC form, In the C-terminal mutants, higher concentrations of Ca2+ (above tension threshold) are required to effect this transformation than in the recombinant wild-type protein, suggesting that the mutants reveal an attachment mediated by Ca2+ in the N-domain sites.

Complete staining of nerve fiber and myoneural junctions with acetylthiocholine and silver

We describe a combined stain for simultaneous demonstration of the preterminal axons and cholinesterase activity at myoneural junctions of mammalian muscles. This technique employs acetylthiocholine iodide as the substrate for cholinesterase activity and silver nitrate impregnation of preterminal axons. The procedure is rapid, simple and uses fresh muscles. Intramuscular nerves, preterminal axons and myoneural junctions are stained simultaneously brown or black with minimal background staining of connective tissue and muscle fibers.