Effect of dihydropyridines on cation transport across the skeletal muscle membrane in the frog

2,6-Dimethyl-3,5-dimethoxycarbonyl-4-(o-difluromethoxyphenyl)- 1,4-dihydropyridine (ryodipine) blocks Ca-channels in phasic muscle fibres from ileofibularis and semitendinosus muscle of Rana esculenta. Ryodipine and some other newly synthesized dihydropyridines (10(-7)-10(-4) M) exerted a slight, if any, effect on the steady-state of potassium chord conductance in isotonic K+-sulfate solution. The effluxes of potassium and rubidium from the sartorius muscle of Rana temporaria also remained unchanged after addition of 2 x 10(-4) M ryodipine. Thus, the nonspecific dihydropyridine effect on ion transport seems to be poorly expressed.     

Functional anatomy of the human saccus lacrimalis

The relations between the saccus lacrimalis and different portions of the musculus orbicularis oculi were studied in orbital regions of human fetuses sectioned into numbered series. No insertions of the pars lacrimalis or Horner's muscle on the saccus were found. These muscular fibres pass along the dorsal wall of the saccus and are separated from it by the reflex tendon of the ligamentum palpebrale mediale. The only muscular fibres that insert on the saccus are those that approach the anterior face of the saccus and the fornix. The fibres that insert on the anterior face proceed from the deep bundles of the pars preseptalis of the lower eyelids, and those that insert on the fornix derive from the deep bundles of the pars preseptalis of the upper eyelid.     

Masticatory muscles of domestic sheep and swine in ontogenesis

Age changes of morphometrical parameters of the masticatory muscles have been analyzed in domestic sheep and pigs of white large breed in the following age groups: 2-, 3-, 4-month-old fetuses, newborns, 4-month-old lambs, 10-month-old pigs, 18-month-old lambs, mature she-sheep and brood-sows. Uneven weight growth of the masticatory muscles in the sheep and pigs during the prenatal ontogenesis should be considered as a consequence of recapitulation of their phylogenesis, and in the postnatal ontogenesis it depends on changes in life conditions, type of nutrition, character of food and type of life. In newborn sheep the digastric, lateral, pterygoid and temporal muscles grow intensively, and in pigs--medial pterygoid and temporal ones. When they pass to roughage, in the former the mass of the musculus masseter major and medial pterygoid muscle increases, and in the latter--that of the musculus masseter major and temporal one. The masticatory muscles of the species studied increase in their mass especially intensively during the middle of the prenatal ontogenesis and during suckling period of their development. This should be taken into consideration in stock-breeding practice. In domestic pigs there is only one muscular belly in the digastric muscle. In sheep there are two bellies, separated one from another by means of a tendinous intersection, owing to crossing of the latter by the stylohyoid muscle.     

Effect of acetylcholine on ion transport in the frog skeletal muscle

1. The effect of acetylcholine (ACh) on the ion transport of frog (Rana esculenta) sartorius muscles was studied. ACh was applied in bathing solution, Na influx and K efflux were measured using 24Na and 42K isotopes. 2. Na influx of sartorius muscles was increased by 1 mmol/1 ACh 2-10 fold depending on the experimental arrangement. The increase was greater if Na influx was measured at the beginning of ACh depolarization. During ACh treatment the Na influx took about the same time course as the depolarization recorded extracellularly. This type of recording approximately reflects the depolarization proceeding on the sartorius muscle fibres. 3. The presence of 31 nmol/l tetrodotoxin (TTX) did not modify the degree of increase of Na influx. 4. Rate coefficients for K efflux were increased 2-5 fold by ACh. The maximum rate coefficients were obtained in the first minute of ACh treatment. 5. Increase in K loss evolves also in the presence of 31 nmol/l TTX. The increase in rate coefficients was found to be about 30% less than without TTX in the first minute of ACh action. 6. The results indicate that in the presence of ACh the observed increase in Na influx and K efflux is brought about mainly by changes in Na and K conductance induced by ACh at the end-plates rather than by the action potentials accompanying ACh depolarization.     

Energy liberation and chemical change in frog skeletal muscle during single isometric tetanic contractions

Recent data obtained from Rana temporaria sartorius muscles during an isometric tetanus indicate that the time-course of phosphocreatine (PC) splitting cannot account for the total energy (heat + work) liberation (Gilbert et al. 1971. J. Physiol. (Lond.) 218:)63). As this conclusion is important to an understanding of the chemical energetics of contraction, similar experments were performed on unpoisoned, oxygenated Rana pipiens sartorius muscles. The muscles were tetanized (isometrically) at 0 degrees C for 0.6, 1, or 5 s; metabolism was rapidly arrested by freezing the muscles with a specially designed hammer apparatus, and the frozen muscles were chemically analyzed. Comparable myothermal measurments were made on frogs from the same batch. Results of these experiments indicate: (a) The energy liberation parallels the PC and ATP breakdown with a proportionality constant of 10.7 kcal/mol; (b) comparably designed experiments with sartorius muscles of R. temporaria revealed that the ratio of energy liberation to PC splitting was significantly greater than that observed in R. pipiens sartorius muscles; (c) there is no systematic difference between experiments in which metabolism was arrested by the hammer apparatus and others using a conventional immersion technique.     

Effects of physostigmine on the voltage dependent ionic conductances of skeletal muscle fibres

The voltage dependent ionic conductances were studied by analysing the phase plane trajectories of action potentials evoked by electrical stimulation of the sartorius muscles of the frog (Rana esculenta). The delayed outward potassium current was measured also under voltage clamp conditions on muscle fibres of either the frog (Rana esculenta) or Xenopus laevis. On analysing the effect of physostigmine decreasing the peak amplitude, the rate of both the rising and falling phases of the action potentials, it was revealed that the alkaloid at a concentration of 1 mmol/l reduced significantly both the delayed potassium conductance and the outward ionic current values during the action potentials. The inhibition of sodium conductance and inward ionic current was less expressed. The maximum value of delayed potassium conductance measured under voltage clamp conditions was decreased by 1 mmol/l physostigmine. The time constant determined from the development of delayed potassium conductance was increased at a given membrane potential. The voltage vs. n relationship describing the membrane potential dependence of the delayed rectifier was not influenced by physostigmine. It has been concluded that physostigmine changes the time course of the action potentials by decreasing the value of both voltage dependent ionic conductances and by slowing down their kinetics. It is discussed that results obtained from the phase plane analysis of complex pharmacological effects can only be accepted with some restrictions.     

Participation of electrogenic Na+-K+-ATPase in the membrane potential of earthworm body wall muscles

The effect of Na+-K+-ATPase inhibitor ouabain on the resting membrane potential (Vm) was studied by glass microelectrodes in isolated somatic longitudinal muscles of the earthworm Lumbricus terrestris and compared with frog sartorius muscle. In earthworm muscle, Vm was -49 mV (inside negative) in a reference external solution with 4 mmol/l K+. The electrogenic participation of Na+-K+-ATPase was absent in solutions with very low concentrations of 0.01 mmol/l K+, higher in 4 and 8 mmol/l K+ (4-5 mV) and maximal (13 mV) in solutions containing 12 mmol/l K+ where Vm was -46 mV in the absence and -33 mV in the presence of 1 x 10(4) M ouabain. The electrogenic participation of Na+-K+-ATPase was much smaller in m. sartorius of the frog Rana temporaria bathed in 8 and 12 mmol/l K+. The results indicate that the Na+-K+-ATPase is an important electrogenic factor in earthworm longitudinal muscle fibres and that its contribution to Vm depends directly on the concentration of K+ in the bathing solution.     

Relationship between the heat resistance of different muscles and frog viability during heat selection]

An attempt was made to elucidate the relation between the survival of mature frogs Rana temporaria during sudden (unforeseen) thermal selection and the heat resistance of several muscles: m. interfalangealis, m. ileofibularis, m. sartorius, m. gastrocnemius and m. rectus abdominis. It has been found that of greater selective value are the individuals with lower heat resistance not in all muscles, but in those having lesser heat resistance, such as m. ileofibularis and m. sartorius.     

Formation of the microcirculatory bed of the muscle coat of the wall of the sigmoid colon during human fetal development]

During intrauterine life microcirculatory bed of the muscular coat is forming and developing in accordance with its differentiation, growth and functional changes at various steps of its ontogenesis. This step-like differentiation in the parts composing the microcirculatory bed and the whole microcirculatory network of longitudinal and circulatory layers of the muscular coat is genetically connected with differentiation taking place in the wall of the sigmoid colon during the developmental period studied and, in the end, reflects morphological maturation of the sigmoid colon at different stages of its individual ontogenesis.     

Light and electron microscopic study of adenosine triphosphatase activity of anuran tadpole musculature.

The histochemical activities of succinic dehydrogenase (SDH) and Ca++-activated ATPase (pHs 7.4 and 9.4) were studied in the larval tail musculature of Rana japonica, Rana catesbeiana and Rana ornativentris. The ATPase reaction product was detected by both light and electron microscopy. 'Red' and 'white' muscle fibres, as distinguished by SDH, showed high and low Ca++-ATPase reaction, respectively, at pHs 7.4, 9.4 and following preincubation in cold K2-EDTA solution. The ultrastructural investigation of Ca++-ATPase reaction at pH 7.4 by the Ca++-citrophosphate technique demonstrated electron-dense reaction product in association with A, I and 'Z' bands, intermyofibrillar (SR) compartment and the mitochondrial inner chamber. However, Pb++ precipitation technique demonstrated Mg++-activated myosin ATPase activity at pH 9.2 ultrastructurally. The present histochemical data suggest that the anuran larval tail 'red' muscle fibres are possible 'slow,' and emphasize a possible lack of correlation between the speed of contraction with their ATPase activity. Moreover, 'red' muscle fibres of the anuran tai- musculature are not equivalent to 'Type I' fibres of higher chordates.     

Stimulation of post-traumatic regeneration of x-irradiated skeletal muscles in old rats

Regeneration in murine musculus gastrocnemius was studied in animals aged 2-2.5. It was demonstrated that 20Cy irradiation caused the inhibition of necrosis resorption, suppressed the proliferative activity of muscular and connective tissues. Impulse laser therapy employed after irradiation of dissected musculus gastrocnemius potentiated fibrin resorption and markedly stimulated regeneration of muscular and connective tissues. Nonexposed disintegrated muscular tissue, when implanted to the site of injury produced a significant stimulating effect resulting in necrosis resorption, more pronounced than under laser therapy, and recovering the regenerative capacity of the exposed tissue.     

Light and electron microscopic investigation of ATPase activity in musculature during anuran tail resorption.

The histochemical activity of adenosine triphosphatase (ATPase) was studied at light and electron microscopic levels in larval tail musculature of Rana catesbeiana and Rana ornativentris during late metamorphic stages. The presence of low, moderate or dark reaction of K2-EDTA-preincubated Ca++-ATPase was correlated with the variable degree of degeneration of white fibres even at the late stage of tail resorption. The reasons for an increase in this ATPase activity in degenerating white muscle fibres are discussed. Irrespective of the degree of degeneration, all red fibres showed high ATPase reaction. During myocytolysis, it is shown that the SR vesicles accumulate electron dense amorphous material. The degree of myofibrillar disintegration correlated with decrease in ultrastructural reaction product for Mg++-ATPase. Although grouped atrophy of muscle fibres (as seen in Xenopus laevis, den Hartog Jager et al., 1973, 1975) was absent in musculature of resorptive tails, ultrastructural characteristics including proliferation of SR and dilation of its vesicles represent alteration of the normal neural influence on the skeletal muscle fibres.     

Correlation between the heat resistance of the provisional and definitive muscle tissue during the metamorphosis crisis in frogs

Using tadpoles of the lake frog Rana ridibunda Pall. during metamorphosis, a study was made of the heat resistance of the provisional muscle tissue of the tail and of that of two definitive muscles belonging to low-resistant (musculus iliofibularis) and high-resistant (musculus gastrocnemius) groups. It has been shown that during the late metamorphosis a statistically significant direct relation exists between the heat resistance of the provisional muscle tissue of the tail and definitive m. iliofibularis. A comparison with the earlier published data points to a positive correlation between each of these two characteristics and the survival of larvae at high injurious temperature.     

Phosphate metabolites in muscular contraction caused by magnetic stimulation

The metabolism of high energy phosphates during muscular contraction due to direct electrical stimulation, indirect stimulation via nerve excitation, and magnetic stimulation was studied in isolated muscles (frog sartorius muscles) by (31)P nuclear magnetic resonance ((31)P-NMR). Twitch amplitudes elicited by each stimulus were measured alternatively at 3 mm displacement loading and 5 g weight. Both the creatine/inorganic phosphate (PCr/Pi) and pH changes were more marked in direct electrical stimulation than in magnetic stimulation. The muscular contraction caused by magnetic stimulation showed less fatigue than that caused by direct electrical muscular stimulation.     

Light and electron microscopic investigation of ATPase activity in musculature during anuran tail resorption

Summary The histochemical activity of adenosine triphosphatase (ATPase) was studied at light and electron microscopic levels in larval tail musculature of Rana catesbeiana and Rana ornativentris during late metamorphic stages. The presence of low, moderate or dark reaction of K₂-EDTA-preincubated Ca⁺⁺-ATPase was correlated with the variable degree of degeneration of white fibres even at the late stage of tail resorption. The reasons for an increase in this ATPase activity in degenerating white muscle fibres are discussed. Irrespective of the degree of degeneration, all red fibres showed high ATPase reaction. During myocytolysis, it is shown that the SR vesicles accumulate electron dense amorphous material. The degree of myofibrillar disintegration correlated with decrease in ultrastructural reaction product for Mg⁺⁺-ATPase. Although grouped atrophy of muscle fibres (as seen in Xenopus laevis, den Hartog Jager et al., 1973, 1975) was absent in musculature of resorptive tails, ultrastructural characteristics including proliferation of SR and dilation of its vesicles represent alteration of the normal neural influence on the skeletal muscle fibres.     

Mathematical analysis of myelin structure of nerves of the brachial plexus in antenatal and early postnatal ontogenesis of man]

A gradual increase of the information entropy indices and a decrease of the excess percentage is observed in the course of antenatal and early postnatal ontogenesis of man. The indices of the information entropy reach their maximum and the excess percentage reaches its minimum in children younger than 3 years. At that time the myelinated fibres have the most various diameters. In the antenatal development of man the prevalence of fine (1--4 mkm) myelinated fibres makes the structure of the humeral plexus nerves more definite while in the postnatal ontogenesis their structure is less definite since the myelinated fibres of different diameter are met with almost the same probability.     

Correlation of Utrophin Levels with the Dystrophin Protein Complex and Muscle Fibre Regeneration in Duchenne and Becker Muscular Dystrophy Muscle Biopsies

Duchenne muscular dystrophy is a severe and currently incurable progressive neuromuscular condition, caused by mutations in the DMD gene that result in the inability to produce dystrophin. Lack of dystrophin leads to loss of muscle fibres and a reduction in muscle mass and function. There is evidence from dystrophin-deficient mouse models that increasing levels of utrophin at the muscle fibre sarcolemma by genetic or pharmacological means significantly reduces the muscular dystrophy pathology. In order to determine the efficacy of utrophin modulators in clinical trials, it is necessary to accurately measure utrophin levels and other biomarkers on a fibre by fibre basis within a biopsy section. Our aim was to develop robust and reproducible staining and imaging protocols to quantify sarcolemmal utrophin levels, sarcolemmal dystrophin complex members and numbers of regenerating fibres within a biopsy section. We quantified sarcolemmal utrophin in mature and regenerating fibres and the percentage of regenerating muscle fibres, in muscle biopsies from Duchenne, the milder Becker muscular dystrophy and controls. Fluorescent immunostaining followed by image analysis was performed to quantify utrophin intensity and β-dystrogylcan and ɣ –sarcoglycan intensity at the sarcolemma. Antibodies to fetal and developmental myosins were used to identify regenerating muscle fibres allowing the accurate calculation of percentage regeneration fibres in the biopsy. Our results indicate that muscle biopsies from Becker muscular dystrophy patients have fewer numbers of regenerating fibres and reduced utrophin intensity compared to muscle biopsies from Duchenne muscular dystrophy patients. Of particular interest, we show for the first time that the percentage of regenerating muscle fibres within the muscle biopsy correlate with the clinical severity of Becker and Duchenne muscular dystrophy patients from whom the biopsy was taken. The ongoing development of these tools to quantify sarcolemmal utrophin and muscle regeneration in muscle biopsies will be invaluable for assessing utrophin modulator activity in future clinical trials.     

Effect of hypokinesia and muscle training on catecholamine levels of the myocardium of rats in postnatal ontogeny

Following muscular training and hypokinesia in postnatal ontogenesis (10 weeks) rats were examined by fluorometry for the content of catecholamines at rest and after extreme exertion (swimming). Regular muscular training led to an increase in catecholamine concentration in the myocardium. Hypokinesia decelerated catecholamine accumulation by the myocardium. A single extreme swimming exercise brought about a decrease in catecholamine concentration in the myocardium. The minimal adrenaline concentration was the same, being equal to 0.04 microgram/g crude tissue whatever the age and locomotion pattern.     

Structural and histochemical characteristics of different variants of the perimuscularization of the femoral vein in an experiment

Structural and histochemical changes in the region of perivenous muscular constrictors created by means of experimental perimuscularization of the femoral vein were studied and compared. Thirty-two mongrel dogs were operated on. In 18 dogs, perimuscularization was made by placing the femoral vein beneath a flap from the medial head of the musculus quadriceps femoris. In 14 dogs, the proximal segment of the dissected femoral vein was introduced through a tunnel formed in the medial head of the musculus quadriceps femoris followed by restoration of the vein continuity by an angiography apparatus. It has been shown that the method of the femoral vein introduction through the muscular tunnel is very promising, since it results in minimal histostructural changes in the vessel and the adjacent muscle.     

Mechanism of the reduction of Meckel's cartilage in man

The mechanism of reduction of the anterior end of Meckel's cartilage was studied in human embryos, with the following findings: 1. Meckel's cartilage is surrounded, from the outside and from below, by newly formed mandibular bone over the extent of the insertion of the musculus mylohyoideus. 2. Blood vessels from the newly formed bone penetrate Meckel's cartilage and break it down in the same way as in enchondral ossification of cartilaginous models of other bones. 3. The anlagen of the musculus mylohyoideus and musculus genioglossus are at first inserted on Meckel's cartilage; further muscle fibres, formed on the under surface of the two muscles, are inserted on the newly formed bone of the rudimentary mandible. Parallel to this process, the fibres on the upper surface of the muscles, which were originally inserted on Meckel's cartilage, disappear. The two processes combined lead to transposition of the insertions of the two muscles from Meckel's cartilage to the mandible. 4. In the area of the resorbed Meckel's cartilage, a minimum number of bone trabeculae are formed at the time of its resorption. The space left by Meckel's cartilage is taken over chiefly by the primitive medullary cavity of the rudimentary mandible, medially to the canal for the nerve and blood vessels.