Catalytic properties of lactate dehydrogenase in Homarus americanus.

Lobster tail and leg lactate dehydrogenases (LDH) have been characterized kinetically. The four binding sites for reduced coenzyme have been shown to be equivalent for the enzyme purified from lobster tail muscle. For the reduced form of 3-acetyl pyridineadenine dinucleotide, the K_a = 1.4 × 10⁷ M^?1 S^?1. The activity of the enzyme purified from the tail muscle is severely inhibited (90%) by high levels of pyruvate (10 mm) when assayed for pyruvate reductase activity at 11 °C; the reductase activity measured using the enzyme from the walking leg muscle was not inhibited by these high levels of pyruvate. Evidence is presented which indicates that the LDH from the tail muscle of the East Coast lobster forms an abortive ternary complex (enzyme-NAD⁺-pyruvate) which accounts for these inhibitory kinetics. The data suggest that the LDH from the tail muscles of the invertebrate lobster represents a “kinetic” heart-type l-specific LDH and that from the walking legs, a “kinetic” muscle-type l-specific LDH.     

Comparison of protein patterns in cockroach muscles innervated by fast or slow neurons

The protein composition of six of the coxal depressor muscles of the leg of the cockroach, Periplaneta americana, was analyzed by one and two dimensional SDS—polyacrylamide gel electrophoresis in conjunction with a sensitive silver stain. The proteins in each muscle were fractionated according to their solubilities in either low salt buffer, 1%NP-40, or 2%SDS. The gel patterns resulting from the electrophoresis of each of the fractions from “fast”, “slow”, and “mixed” type muscle were compared with reference to the innervation of the muscle by a particular motor neuron. More than 50 proteins were detected which were not common to all three types of muscle. These proteins were found to be distributed fairly equally amongst each of the three fractions. The large number of differentially expressed proteins observed, are not likely to be involved in specific neuromuscular recognition, but instead are probably correlated with the physiological state of the muscle.     

Coordination and neuromuscular control of rhythmic behaviors in the blue crab,Callinectes sapidus

The stereotypical courtship display (CD) behavior of the male blue crab, Callinectes sapidus, includes an unusual component: the rhythmic waving of the swimming appendages above the carapace. This behavior occurs in a unique context but it resembles two other rhythmic behaviors performed using the swimming legs: sideways swimming and backward swimming. As a first step to understanding the mechanisms that allow the expression of apparently different rhythmic motor patterns, we have examined these behaviors using slow motion video analysis and electromyography of the basal muscles of the swimming legs in freely behaving crabs. The results show that these behaviors are distinguished by four parameters: the frequency of leg waving, the phase relationship between the legs, the presence of a stationary pause in basal muscle activity combined with rotation of the distal leg during CD, and an extended range of motion of these legs during CD and backward swimming, relative to sideways swimming. EMG analysis revealed that during sideways swimming, the sequence of muscular activity between the two legs was different. In contrast, during CD and backward swimming the sequence of activity for these legs is identical.Abbreviations CD courtship display - EMGs electromyograms - CD AMP courtship display in crabs with amputated fifth legs - CD1 crabs that voluntarily used one leg to perform courtship display waving - CD 1–3 courtship waving in cycles 1–3 - CD MID courtship waving after cycles 1–3 - M-C meral-carpal joint     

Respiratory system of the primitive moth Micropterix calthella (Linnaeus) (Lepidoptera : Micropterigidae)

The 1st thoracic spiracular atrium is closed by anterior and posterior muscle fibres extending between its dorsal and ventral wall. The 2nd thoracic spiracle has only a single (anterior) closing lip, movable by a muscle inserting on the wall below the spiracular aperture; this configuration may be a lepidopteran ground-plan autapomorphy. There are functional spiracles on abdominal segments I – VII, each with a closing “bow” and “lever”. There are intrinsic occlusor muscles in all abdominal spiracles and the 1st spiracle has an extrinsic (ventral) dilator. Dorsal dilator muscles or ligaments are absent. A dorsal and a ventral tracheal trunk extend from the 1st thoracic spiracle into the head; the latter supplies the mouthparts and the antenna; there is no connection between the dorsal and ventral cephalic trunk systems. There is a single series of lateral connectives between the spiracles of each side. There is a ventral tracheal commissure in both pterothoracic segments, but none in the prothorax. In each pterothoracic segment an anterior and a posterior tracheal arch give off branches to the wing and anastomose with each other on their downwards course into the leg. Wing tracheation is greatly reduced. The anterior and posterior tracheae of each wing are independent of each other. There is a dorsal commissure in abdominal segment VIII; ventral abdominal commissures are lacking in Micropterix, although present in other micropterigid genera. The terminalia are partly supplied from tracheae arising in segment VII. Air sacs occur in the tibiae only. Phylogenetic aspects of holometabolan tracheation patterns are discussed.     

The role of visual information in maintaining postural stability after the maximum exercise for the upper and lower limb muscles

The postural stability on a seesaw generating anterior–posterior instability with the eyes open (EO) and then the eyes closed (EC) in young healthy subjects (n = 28) before and 6 min after the maximum bicycle exercise (Wingate test) performed using lower limbs (“leg exercise”) or upper limbs (“hand exercise”) was investigated. It was found that “hand exercise” caused the same increase in average velocity (V, mm/s) and in the average range of sway of the centre of pressure (Qy, mm) as “leg exercise.” However, the duration of V recovery (EC: 2 min 30 s and 50 s; EO: 60 s and 40 s after “leg exercise” and “hand exercise,” respectively) and Qy (EC: 1 min 10 s and 30 s after “leg exercise” and “hand exercise,” respectively; EO: no changes from baseline) was shorter after “hand exercise.” In the presence of visual information, the increment in V decreased more than 2 times after “leg exercise” (+100.5% and + 40.5%, p < 0.01 for EC and EO, respectively) and after “hand exercise” (+73.0% and +30.3%, p < 0.01 for EC and EO, respectively). Moreover, Qy after both exercises remained at the initial level under EO conditions but significantly increased under EC conditions (+42.8%, p < 0.01 after “leg exercise” and +40.3%, p < 0.01 after “hand exercise”). Thus, the maximum exercise for the muscles of the upper limbs causes the same reduction in postural stability as analogous exercise for the muscles of the lower limbs, but the recovery period after “hand” exercise was shorter. The presence of visual information allows the baseline maintenance of postural stability and significantly reduces the strain of postural regulation while standing on a movable support after the maximum “leg exercise” and “hand exercise.”     

Trehalose metabolism in the blue crab Callinectes sapidus: Isolation of multiple structural cDNA isoforms of trehalose-6-phosphate synthase and their expression in muscles

Adult blue crab Callinectes sapidus exhibit behavioral and ecological dimorphisms: females migrating from the low salinity water to the high salinity area vs. males remaining in the same areas. The flesh basal muscle of the swimming paddle shows a dimorphic color pattern in that levator (Lev) and depressor (Dep) of females tend to be much darker than those of males, while both genders have the same light colored remoter (Rem) and promoter (Pro). The full-length cDNA sequence of four structural isoforms of trehalose-6-phosphate synthase (TPS) is isolated from chela muscles of an adult female, C. sapidus. Two isoforms of the C. sapidus TPS encode functional domains of TPS and trehalose-6-phosphorylase (TPP) in tandem as a fused gene product of Escherichia coli Ost A and Ost B. The other two isoforms contain only a single TPS domain. In both males and females, the darker (Lev + Dep) muscles exhibit greater amounts of trehalose, TPS and trehalase activities than the light colored (Rem + Pro). The fact that adult females show higher levels of trehalase activity in the basal muscles and of glucose in Lev + Dep than those of adult males suggests that there may be a metabolic dimorphism. Moreover, the involvement of trehalose in energy metabolism that was examined under the condition of strenuous swimming activity mimicked in adult females demonstrates the intrinsic trehalose metabolism in Lev + Dep, which subsequently results in hemolymphatic hyperglycemia and hyperlactemia. Our data support that trehalose serves as an additional carbohydrate source of hemolymphatic hyperglycemia in this species. Behavioral and ecological dimorphisms of C. sapidus adults may be supported by a functional dimorphism in energy metabolism.     

Innervation and acetylcholine sensitivity of skeletal muscle cells differentiated in vitro from chick embryo

Trypsin-dissociated myoblasts from leg muscle of 12-day chick embryos have been cultured in monolayers. After four days the muscle cultures have been confronted with fragments of the spinal cord of six-day chick embryos. Electrophysiological and morphological analysis demonstrate that characteristic neuromuscular transmission can develop in these cultures. Electrical stimulation of the cord fragment evokes contractions of innervated muscle fibers, from which end plate potentials and miniature end plate potentials with average frequency around one per second or more can be recorded. D-tubocurarine (1 μg/ml) suppresses reversibly these synaptic potentials. Non-innervated muscle fibers are sensitive to acetylcholine over all their surface, while innervated muscle fibers are sensitive at the regions where structures suggestive of motor end plate (“bulb-type”) are found. We can conclude that neuromuscular connections developed in vitro in our experiments are functional in respect of transmission of impulses but also in respect of neurotrophic influences for restriction of chemosensitivity.     

Enhanced Specificity of Minimally Modified Anti-c-myc Oligonucleotides

Abstract

The sequence-specificity of antisense oligonucleotides (ODN) against c-myc mRNA was tested by Northern blot analysis. Rat smooth muscle cells were treated with antisense or control ODN against c-myc modified by the “minimal protection strategy”. At 0.3 μM concentration the ODN show a very specific reduction in c-myc mRNA levels. Use of the “minimal protection strategy” minimizes nonspecific effects as observed for all-phosphorothioate ODN containing four consecutive guanine residues.     

Myoanatomy of the marine tardigrade Halobiotus crispae (Eutardigrada: Hypsibiidae)

The muscular architecture of Halobiotus crispae (Eutardigrada: Hypsibiidae) was examined by means of fluorescent‐coupled phalloidin in combination with confocal laser scanning microscopy and computer‐aided three‐dimensional reconstruction, in addition to light microscopy (Nomarski), scanning electron microscopy, and transmission electron microscopy (TEM). The somatic musculature of H. crispae is composed of structurally independent muscle fibers, which can be divided into a dorsal, ventral, dorsoventral, and a lateral musculature. Moreover, a distinct leg musculature is found. The number and arrangement of muscles differ in each leg. Noticeably, the fourth leg contains much fewer muscles when compared with the other legs. Buccopharyngeal musculature (myoepithelial muscles), intestinal musculature, and cloacal musculature comprise the animal's visceral musculature. TEM of stylet and leg musculature revealed ultrastructural similarities between these two muscle groups. Furthermore, microtubules are found in the epidermal cells of both leg and stylet muscle attachments. This would indicate that the stylet and stylet glands are homologues to the claw and claw glands, respectively. When comparing with previously published data on both heterotardigrade and eutardigrade species, it becomes obvious that eutardigrades possess very similar numbers and arrangement of muscles, yet differ in a number of significant details of their myoanatomy. This study establishes a morphological framework for the use of muscular architecture in elucidating tardigrade phylogeny. J. Morphol. 2009. © 2009 Wiley‐Liss, Inc.     

The eye muscles and their innervation in the gobiid fishTridentiger trigonocephalus

The morphology and innervation of the six oculomotor muscles in the gobiid fishTridentiger trigonocephalus are described. Every rectus muscle is composed of two types of muscle fibres. Muscles attach onto the cartilaginous or fibrous sclerotica. Oblique muscles attach onto the ethmoidal plate; recti muscles attach onto the parasphenoid or a thick fibrous membrane. There is no myodome. The common oculomotor nerve is composed of four bundles, the trochlear and the externus of two. The two kinds of fibres of the lateral rectus and the two distinct bundles of the nerve VI suggest a possible homology between this muscle in fishes and the lateral rectus+retractor bulbi in mammals.     

Involvement of Delta and Nodal signals in the specification process of five types of secondary mesenchyme cells in embryo of the sea urchin, Hemicentrotus pulcherrimus

Secondary mesenchyme cells (SMCs) of the sea urchin embryo are composed of pigment cells, blastocoelar cells, spicule tip cells, coelomic pouch cells and muscle cells. To learn how and when these five types of SMCs are specified in the veg₂ descendants, Notch or Nodal signaling was blocked with γ‐secretase inhibitor or Nodal receptor inhibitor, respectively. All types of SMCs were decreased with DAPT, while sensitivity to this inhibitor varied among them. Pulse‐treatment revealed that five types of SMCs are divided into “early” (pigment cells and blastocoelar cells) and “late” (spicule tip cells, coelomic pouch cells and muscle cells) groups; the “early” group was sensitive to DAPT up to the hatching, and the “late” group was sensitive until the mesenchyme blastula stage. Judging from timing of the shift of Delta‐expressing regions, it was suggested that the “early” group and “late” groups are derived from the lower and the middle tier of veg₂ descendants, respectively. Interestingly, numbers of SMCs were also altered with SB431542; blastocoelar cells, coelomic pouch cells and circum‐esophageal muscles decreased, whereas pigment cells and spicule tip cells increased in number. Pulse‐treatment showed that the “early” group was sensitive up to the mesenchyme blastula stage, while the “late” group up to the onset of gastrulation. Thus, it became clear that precursor cells of the “early” and “late” groups, which are located in different regions in the vegetal plate, receive Delta and Nodal signals at different timings, resulting in the diversification of SMCs. Based on the obtained results, the specification processes of five types of SMCs are diagrammatically presented.     

Musculature,nervous system and glands of metasomal abdominal segments of the male of Nomia melanderi Ckll. (Hymenoptera,Apoidea)

Each muscle of the third metasomal segment of the male of Nomia melanderi Ckll. is described in detail. The points of attachment of each muscle are compared with their homologs in other pregenital segments and with their homologs in the female. The function desgnated for each muscle describes its action alone or in conjunction with other muscle(s). New names are given to genital muscles by referring in the name to their points of attachment. Each intratergal muscle has homologous points of attachment in the pregenital segments of both sexes. The median tergo-dorsoplical muscle of the seventh segment and the oblique tergo-dorsoplical muscle of the eighth segment have changed their points of attachment. The intrasternal muscles are modified to suit the needs of courtship and mating, thus they are different from their homologs in the female. The spiracular muscles are well developed in all segments except the eighth, where the sterno-spiracular muscle is absent. The extrinsic genital muscles are derived from the intrasternal muscles of the eighth and ninth segments. The parameral and volsellar muscles are reduced in number. The aedeagal muscles, except the aedeago-phallic, have retained similar points of attachment to those found in primitive Hymenoptera. The topography of the metasomal nervous system is reported in detail by following each nerve and nervule to its termination. The study shows that (at least in Nomia) the criterion of nerve-concentration should not be used alone to indicate evolutionary levels. To accommodate the morphological changes in the terminal segments the Anterior and Posterior Lateral Nerves have migrated to new locations. The pattern of nerve topography (even at the nervule level) is homologous both in the different pregenital segments and between the sexes. The fact that homology does not exist between the external genitalia of the male and the modified ovipositor of the female supports the thesis that the male genital capsule is of phallic rather than prephallic origin. A pair of intersegmental membrane glands located between the seventh and eighth sterna is described. These glands may be the source of a pheromon responsible for gregariousness among “sleeping” males.     

Comparative morphology of the somatic musculature in species of Hexarthra and Polyarthra (Rotifera, Monogononta): Its function in appendage movement and escape behavior

Species of Hexarthra and Polyarthra are freshwater rotifers with well-known escape behaviors that result from interactions with planktonic predators. Both rotifers bear a suite of mobile appendages that function in evasive maneuvers and saltatory jumps through the water column, but the anatomical and functional bases of these actions are poorly understood. Here, we use a combination of phalloidin staining, confocal laser scanning microscopy, and video analysis to describe the morphology of the somatic muscles that supply the mobile appendages in order to understand how they function in escape behavior. Results show that species of Hexarthra, which bear six radially distributed limbs, possess a highly complex trunk musculature that supplies the inside of each limb with its own abductor and adductor muscles, i.e., a direct muscle supply. The singular dorsal and ventral limbs each receive a pair of large abductor and adductor muscles (four muscles total per limb), while the paired dorsolateral and ventrolateral limbs each receives three muscles (two abductors, one adductor per limb). Contraction of the abductor muscles creates a power stroke in the form of an anterior sweep of the limbs, which leads to a three-dimensional tumbling of the rotifer through the water column. Alternatively, species of Polyarthra possess 12 blade-like appendages that are arranged into four equal bundles; each bundle receives an indirect muscle supply that attaches to the shoulder of the paddles. A single longitudinal paddle muscle supplies each dorsolateral bundle, while a pair of longitudinal paddle muscles supplies each ventrolateral bundle. Contraction of these muscles, whether singly or in concert, functions to abduct the paddles in a power stroke, leading to rotation of the body and movement of the rotifer. The recovery stroke is hypothesized to be a multi-step process that begins with reorientation of the appendages prior to adduction, followed by contraction of various muscles to antagonize the paddle muscles. In total, these observations reveal novel complexities in the rotifer muscular system that aids our understanding of the biophysics of predator avoidance in appendage-bearing rotifers.     

Formation and function of the “Xestoleberis‐spot” in Xestoleberis hanaii (Crustacea: Ostracoda)

The crescent sculpture of the so‐called “Xestoleberis‐spot” develops inside the calcified valve of the family Xestoleberididae. Electron microscopic observations on both, intermoult and postmoult stages of Xestoleberis species reveal that the “Xestoleberis‐spot” system consists of three elements; two calcified chambers, a vesicle of electron‐dense material and an uncalcified procuticle. The formation and function of the “Xestoleberis‐spot” system are discussed. In conclusion, the “Xestoleberis‐spot” system functions as the muscle attachment site for several antennal muscles, and provides the material for chitinous fibers in the exocuticle of outer lamella. The unique cuticular structures of the family Xestoleberididae are due to the “Xestoleberis‐spot” system.     

Glycogen synthesis from lactate in skeletal muscle of the lizardDipsosaurus dorsalis

Summary The capacity of skeletal muscle to synthesize glycogen from lactate was tested in the iliofibularis muscle of the desert iguana,Dipsosaurus dorsalis. Like other reptiles,Dipsosaurus accumulates significant lactic acid concentrations following vigorous exercise. After 5 min of progressively faster treadmill running at 35°C (final speed=2.2 km/h), blood lactate concentration increased over 14 mM, which decreased 11 mM after 2 h of recovery. Blood glucose concentration remained unchanged throughout at 8.6±0.46 mM. The role that muscle gluconeogenesis might play in the removal of post-exercise lactate was evaluated. Animals were run to exhaustion at 1.5 km/h on a treadmill thermostatted at 35°C. Animals (n=43) ran 6.9±0.75 min prior to exhaustion. Animals were sacrificed and iliofibularis muscles of both hindlimbs removed and stimulated at 2 Hz for 5 min, reducing twitch tension to 6% of prestimulus tension. Fatigued muscles were then split into red and white fiber bundles and incubated 2 h or 5 h at 35°C in Ringer solution or in Ringer plus 20 mM lactate. In muscles tested in August, red fiber bundles incubated in lactate demonstrated a rate of glycogen synthesis of approximately 1 mg/(g muscle·h). In muscles tested in December, red fiber bundles synthesized glycogen at a reduced rate that was not statistically different than in fiber bundles incubated in Ringer solution without lactate. Glycogen synthesis from lactate was not evident in white fiber bundles in either August or December. The period of peak gluconeogenic capacity coincides with the field active season ofDipsosaurus. In vivo rates of lactate removal and in vitro rates of glycogen synthesis suggest that muscle gluconeogenesis may potentially account for 20% of the lactate removed during recovery from exhaustive activity.Abbreviations IF iliofibularis - FG fast twitch, glycolytic - FOG fast twitch, oxidative-glycolytic     

Cross-training effect of chronic whole-body vibration exercise: a randomized controlled study

Abstract

Purpose: To determine whether unilateral leg whole-body vibration (WBV) strength training induces strength gain in the untrained contralateral leg muscle. The secondary aim was to determine the potential role of spinal neurological mechanisms regarding the effect of WBV exercise on contralateral strength training.

Materials and Methods: Forty-two young adult healthy volunteers were randomized into two groups: WBV exercise and Sham control. An isometric semi-squat exercise during WBV was applied regularly through 20 sessions. WBV training was applied to the right leg in the WBV group and the left leg was isolated from vibration. Sham WBV was applied to the right leg of participants in the Control group. Pre- and post-training isokinetic torque and reflex latency of both quadricepses were evaluated.

Results: The increase in the strength of right (vibrated) knee extensors was 9.4?±?10.7% in the WBV group (p?=?.001) and was 1.2?±?6.6% in the Control group (p?=?.724). The left (non-vibrated) extensorsvibrated) knee extensors w4?±?8.4% in the WBV group (p?=?.038), whereas it decreased by 1.4?±?7.0% in the Control (p?=?.294). The strength gains were significant between the two groups. WBV induced the reflex response of the quadriceps muscle in the vibrated ipsilateral leg and also in the non-vibrated contralateral leg, though with a definite delay. The WBV-induced muscle reflex (WBV-IMR) latency was 22.5?±?7.7?ms for the vibrated leg and 39.3?±?14.6?ms for the non-vibrated leg.

Conclusions: Chronic WBV training has an effect of the cross-transfer of strength to contralateral homologous muscles. The WBV-induced muscular reflex may have a role in the mechanism of cross-transfer strength.     

An analysis of bailer movements responsible for gill ventilation in the crab,cancer novae‐zelandiae

The movements of the bailer during normal ventilation can be resolved into two components, a cycle of pronation and supination being superimposed on a cycle of protraction and retraction. Pronation leads protraction with a phase angle of about 90° in a normal cycle. Pronation is accompanied by flexion of the bailer.

The skeletal anatomy of the bailer is such as to restrict movements of the bailer to those described above. Further the pronated and supinated positions of the limb represent the two stable positions of a skeletal click mechanism, the operation of which may help to resolve the functional duality of the promotor and remotor muscles.

This functional duality arises because the muscles are positioned so as to produce either protraction or supination of the limb. Other muscles in the limb are monofunctional. The bulk of muscle tissue responsible for protraction and supination seems to be greater than that responsible for pronation and retraction.

The sequence of muscular activity during the ventilation cycle follows that expected for a sequence, of protraction, supination, retraction and pronation. Overlap in the periods of activity of the bifunctional muscles and muscles responsible for pronation may also help to resolve the functional duality of the former.

The amplitude of bailer excursion (protraction‐retraction) is not greatly affected by changes in frequency. An advance in the onset of activity in some muscles at higher ventilation ? frequencies suggests that the system is tailored to produce a constant beat amplitude at all frequencies.

Pauses in ventilation occur with the bailer in the retracted position, and it is maintained in this position by tonic activity in the appropriate muscle. During normal ventilation the relative contraction duration of this muscle is positively correlated with cycle period, so that pauses apparently represent a prolongation of the normal retracted phase. The relative contraction durations of some other muscles are negatively correlated with cycle period. The different signs of these correlations may be related to the type of endogenous oscillator present in the central nervous system.