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Structural features and adaptive properties of photoreceptors in the skate retina.

In the duplex (rod/cone) retinae of most vertebrates, electrical responses associated with nocturnal vision can be recorded in relative isolation only over a limited intensity range before there is encroachment by signals from the cone system; e.g., at mesopic levels of illumination, the rods begin to saturate, and the cone mechanism is brought into play. On the other hand, the retinae of some sub-species of skate appear to contain only one class of photoreceptors, namely rods. Nevertheless, skate photoreceptors, as well as other retinal neurons, are able to respond to incremental stimuli presented on background fields so intense that 90% or more of the available rhodopsin is bleached during the exposure. These and other findings raise some doubts as to whether the skate has, in fact, an all-rod retina. In this paper, we present a body of evidence--based on the results of photochemical, anatomical, and electrophysiological studies--to support the view that only a single class of photoreceptor subserves vision in the skate retina. In addition, recent findings will be described that: a) demonstrate how the visual cells transform from sluggishly responding rods to brisk, cone-like elements, and b) may provide some insight into the functional significance of this unusual adaptive property.

Adaptation, Ocular↗

J-chain-like component in 18-S immunoglobulin of the skate Raja kenojei, a cartilaginous fish.

A J-chain was found in the IgM, an 18-S pentameric immunoglobulin, of the skate, Raja kenojei, a cartilaginous fish, by means of gel filtration-column chromatography of reduced and radioalkylated immunoglobulin, followed by alkaline-urea polyacrylamide gel electrophoresis. The J-chain showed an electrophoretic mobility equivalent to that of the light chain, and much slower than that of the J-chain in human IgM. Its electrophoretic banding patterns, however, were different from those of the light chain. Its mol. wt of 17,500 was close to that of mammalian J-chain. The J-chain was not found in another immunoglobulin of the skate, a 9-S dimer held by a noncovalent force. No immunological cross-reactivity was observed between the skate J-chain and human and chicken J-chains. In view of these findings, it was concluded that the J-chain in the skate IgM is both electrophoretically and antigenically considerably different from the J-chain in mammalian IgM.

Animals↗

Dietary intake of elite figure skating dancers.

The main objective of the present study was to examine the dietary intake and physiological status of members of the US National Figure Skating Dance Team. Study participants included seven female and seven male figure skating dance pairs. Three-day food records were collected from each dancer and analyzed using a computerized nutrient database. Height, body weight and skinfold measurements were completed on each dancer. Body fat was estimated using the Jackson et al prediction equations. Fasting blood samples were collected for biochemical analysis. Total energy intake of female dancers (1416 kcal/d, 339 kjoules/day) was considerably lower than the dietary recommendations, while the male dancers appeared to have adequate energy intake (2837 kcal/d, 679 kjoules/day). Carbohydrate intake of both male and female dancers was below the recommendations, 44% and 51%, respectively, whereas total fat and protein intake of both male and female dancers were higher than the dietary recommendations. All dancers met at least 2/3(rd) of the dietary recommendations for all micro-nutrients, except folate and zinc intake of the female dancers, which were 68% and 62% of the recommended values. Although, blood lipid concentrations and other physiological parameters of the overall group were within normal ranges, two of the dancers (14% of the dancers) had elevated blood cholesterol concentrations and low iron status. The results of the present study suggest that these elite figure skating dancers, especially females, may not be consuming adequate amounts of energy-yielding nutrients and those that influence bone health. Future studies are needed to examine the effects of nutritional status on performance of these figure skating dancers and to develop strategies to improve the nutritional status of these elite dancers.

Journal Article↗

A microtubule-binding domain in dynactin increases dynein processivity by skating along microtubules.

Microtubule-associated proteins (MAPs) use particular microtubule-binding domains that allow them to interact with microtubules in a manner specific to their individual cellular functions. Here, we have identified a highly basic microtubule-binding domain in the p150 subunit of dynactin that is only present in the dynactin members of the CAP-Gly family of proteins. Using single-particle microtubule-binding assays, we found that the basic domain of dynactin moves progressively along microtubules in the absence of molecular motors - a process we term 'skating'. In contrast, the previously described CAP-Gly domain of dynactin remains firmly attached to a single point on microtubules. Further analyses showed that microtubule skating is a form of one-dimensional diffusion along the microtubule. To determine the cellular function of the skating phenomenon, dynein and the dynactin microtubule-binding domains were examined in single-molecule motility assays. We found that the basic domain increased dynein processivity fourfold whereas the CAP-Gly domain inhibited dynein motility. Our data show that the ability of the basic domain of dynactin to skate along microtubules is used by dynein to maintain longer interactions for each encounter with microtubules.

Amino Acid Sequence↗

[Risk of injuries, symptoms of excessive strain and preventive possibilities in cross-country skiing. A comparison between classical technique and skating technique].

A standardised questionnaire was drawn up and submitted to 690 cross-country skiers active in popular (pastime) sports, competitive sports and high-performance sports, between 1990 and 1993, covering a total of 189 injuries and 1,281 cases of complaints caused by overstrain. Over and above this, muscular imbalance and postural characteristics (for example, signs of vestibular syndrome) were examined in 94 of these skiers. Three times as many skiers (69.3%) reported on complaints than skiers reporting on injuries (20.3%). Complaints concerned particularly the lower extremities (45.9%), whereas injuries were more frequent at the upper extremities (47.6%). Minor injuries predominated (77%) such as sprains and distortions (23.8%), contusions (25.9%), skin injuries (13.7%) and muscle injuries (13.7%) as well as mild complaints (45.1%) such as blisters at the hand and feet. High-performance sportsmen (14.4%) are significantly most frequently injured when employing the skating technique, whereas in competitive sports the "classical" technique results in a greater incidence of accidents (16.1%). Most accidents occur during downhill skiing, on rutty and icy tracks, collisions, due to inadequate technique, lack of balance and unskilled handling. Complaints are significantly reported by competitive (70.8%) and high-performance sports people (74.8%) for both techniques. Whereas complaints at the lower extremities occur more often during skating technique (56.8%) than during the classical technique (34.5%), complaints concerning the trunk are seen more often with the classical technique (29.7%) than with the skating technique (8.9%). Complaints are caused by icy, rutty tracks, inadequate equipment, deficient training, muscular imbalances and previous damage. A large number of pointers on how to avoid pitfalls when employing the skating and classical techniques can be obtained via analysis of the injuries and complaints.

Adolescent↗

Determinants of biliary secretion in isolated perfused skate liver.

In the isolated perfused liver of the little skate, Raja erinacea, bile flow averaged 5.07 +/- 0.58 (mean +/- SE) microliters.h-1.g liver-1 in 21 experiments at a perfusion pressure of 5.0 cm Ringer compared to 3.79 +/- 0.32 in 38 experiments at 2.5 cm (P less than 0.05). [14C]inulin readily entered skate bile. Bile-to-plasma [14C]inulin ratios corrected for delay in transit time, averaged 0.46 +/- 0.07 at 1 h and rose to 0.74 +/- 0.06 by 4 h, although bile flow remained constant. In experiments in which [14C]inulin reached equilibrium between bile and plasma, the bile-to-plasma ratio conformed to the theoretical relationship between bile flow, solvent drag, and inert solute diffusion predicted at extremely low bile flows, but demonstrated that the skate biliary tree is more permeable to inulin than that of the rat. Electron microscopic studies demonstrated that ionic lanthanum could traverse the tight junctions. However, freeze-fracture studies of junction structure did not differ qualitatively from similar studies in the rat. Partial dependence of bile flow on perfusion pressure, high bile-to-plasma inulin ratios, and permeability of the canalicular tight junctions to ionic lanthanum all suggest that the paracellular pathway may be an important component of bile formation in the skate.

2,4-Dinitrophenol↗

Cytoskeletal organization in clusters of isolated polarized skate hepatocytes: structural and functional evidence for microtubule-dependent transcytosis.

Isolated hepatocytes from the marine vertebrate Raja erinacea (the little skate) retain their structural and functional integrity as clusters of cells formed around a single tubular bile canaliculus, and therefore can be used as a model of polarized hepatocytes in situ. In this study we used confocal and conventional epifluorescence microscopy in conjunction with fluorescent markers and immunocytochemistry to examine the structure and function of the cytoskeleton in these cells. Actin filaments in the hepatocyte clusters were found cortically and also concentrated in a pericanalicular array, while microtubules appeared to radiate away from a concentration near the apical membrane of the biliary pole towards the basolateral sinusoidal surfaces. Treatment of clusters with the microtubule disrupting agent, nocodazole, resulted in the microtubules depolymerizing from the basolateral surfaces towards the apical surface, indicating that the microtubules were oriented with their plus ends at the basolateral surface and their minus ends at the apical surface. Nocodazole was also found to disrupt the ability of clusters to transcytose a fluorescent bile salt derivative into their canalicular lumens. We detected cytoplasmic dynein in skate hepatocyte homogenates by Western blotting using an anti-dynein intermediate chain antibody, and immunofluorescent staining of intact hepatocytes revealed a punctate vesicular pattern. The polarized arrangement of microtubules, the presence of cytoplasmic dynein, and the inhibition of bile salt secretion by nocodozole are consistent with the microtubule cytoskeleton playing a fundamental role in the mediation of transcytosis, endocytosis, and bile excretory function in these hepatocytes. These polarized isolated skate hepatocytes represent an excellent experimental model for the in vitro study of hepatic transport, and allow for important comparative studies aimed at elucidating the evolutionarily conserved nature of various hepatocyte structures amongst the vertebrates.

Actins↗

Morphological, physiological and biochemical observations on skate electric organ.

The electric organs of two species of skate have been examined morphologically, physiologically and biochemically. They can be easily dissociated into innervated or denervated component electrocytes by a Torpedo Ringer's solution containing 1% collagenase. Collagenase treatment did not, however, separate the Schwann cell cover capping the synaptosomes. Isolated electrocytes generate normal MEPP frequencies and show evoked responses for two days in Torpedo Ringer's. The nerve terminals retain excitability and transmitter release properties up to the time of separation. Since isolated terminals and denervated electrocytes show normal ultrastructural characteristics for up to 12 h, the skate electric organ provides several preparations which are not attainable with Torpedo tissue. Acetylcholine (ACh) content of supernatant fractions containing the synaptosomes was comparable to that found in Torpedo (sps.). Collagenase specifically eliminates the basal lamina associated with the synaptic junctional region. Neuronal cell death and synaptic terminal degeneration were also noted in the adult organs of both species. The skate electric organ is ideally suited for the study of cholinergic development and transmission.

Acetylcholine↗

Injury pattern and acceptance of passive and active injury prophylaxis for inline skating.

In a field study the injury pattern as well as the active and passive injury prophylaxis of 1036 inline skaters were evaluated. Of them, 60% had already been injured. Every 124 h an injury occurred during inline skating. While 61% of the injuries affected only soft tissue, 31% were joint distorsions, and 8% were fractures. Altogether 37% of the 626 injured skaters required medical treatment. The upper extremities are especially at risk: 78% of the fractures and 48% of the distorsions affected the fingers, wrist, forearm or elbow. It is obvious that the inline skaters' passive prophylaxis measures are still unsatisfactory. Only 17% of the questioned used complete protection, whereas 16% were wearing no protective gear at all. Of the rest, 49% were skating with wrist guards, 66% were using knee pads, and 31% elbow pads. A high percentage of injuries is due to the lack of basic knowledge and techniques. The survey revealed that only 24% can stop immediately. While 51% had only slight problems with braking, 16% admitted having great difficulties, and 7% were not able to brake at all. These numbers reveal that there are still large deficits regarding braking techniques and education on the risks of injury. Therefore, athletes who use this equipment for the first time should learn the basic techniques of skating, braking and falling with the help of a qualified instructor. Most of all those questioned wanted to have more information, and half would be interested in special educational programmes.

Adolescent↗

Response properties and biological function of the skate electrosensory system during ontogeny.

This study examined the response properties of skate electrosensory primary afferent neurons of pre-hatch embryo (8-11 weeks), post-hatch juvenile (1-8 months), and adult (> 2 year) clearnose skates (Raja eglanteria) to determine whether encoding of electrosensory information changes with age, and if the electrosense is adapted to encode natural bioelectric stimuli across life history stages. During ontogeny, electrosensory primary afferents increase resting discharge rate, spike regularity, and sensitivity at best frequency. Best frequency was at 1-2 Hz for embryos, showed an upwards shift to 5 Hz in juveniles, and a downward shift to 2-3 HZ in adults. Encapsulated embryos exhibit ventilatory movements that are interrupted by a "freeze response" when presented with weak uniform fields at 0.5 and 1 Hz. This phasic electric stimulus contains spectral information found in potentials produced by natural fish predators, and therefore indicates that the embryo electrosense can efficiently mediate predator detection and avoidance. In contrast, reproductively active adult clearnose skates discharge their electric organs at rates near the peak frequency sensitivity of the adult electrosensory system, which; facilitates electric communication during social behavior. We suggest that life-history-dependent functions such as these may shape the evolution of the low-frequency response properties for the elasmobranch electrosensory system.

Animal Communication↗

External loading does not change energy cost and mechanics of rollerski skating.

The purpose of this study was to examine the effects of external loading on the energy cost and mechanics of roller ski skating. A group of 13 highly skilled male cross-country skiers roller skied at 19.0 ( SD 0.1) km x h(-1) without additional load and with loads of 6% and 12% body mass (mb). Oxygen uptake (VO2), knee and ankle joint kinematics, roller-ski electromyogram (EMG) of the vastus lateralis and gastrocnemius lateralis muscles, and roller ski velocity were recorded during the last 40 s of each 4-min period of roller skiing. One-way repeated measures ANOVA revealed that the VO2 expressed relative to total mass (mtot), joint kinetics, eccentric-to-concentric ratio of the integrated EMG, velocity changes within a cycle, and cycle rate did not change significantly with load. The subsequent analysis of the effect of load on each resistance opposing motion suggested that the power to sustain changes in translational kinetic energy, potential energy, and overcoming rolling resistance increased proportionately with the load. The lack of a significant change in VO2/mtot with external loading was associated with a lack of marked change in external mechanical power relative to mtot. The existence of an EMG signal during the eccentric phase prior to the thrust (concentric phase), as well as the lack of significant delay between the two phases, showed that a stretch-shortening cycle (SSC) occurs in roller ski skating. Taken together, the present results would suggest that external loading up to 12% mb does not increase storage and release of elastic energy of lower limb muscles during SSC in roller ski skating.

Adult↗

GFAP-immunopositive structures in spiny dogfish, Squalus acanthias, and little skate, Raia erinacea, brains: differences have evolutionary implications.

GFAP expression patterns were compared between the brains of a spiny dogfish (Squalus acanthias) and a little skate (Raia erinacea). After anesthesia, the animals were perfused with paraformaldehyde. Serial vibratome sections were immunostained against GFAP using the avidin-biotin method. Spiny dogfish brain contained mainly uniformly-distributed, radially arranged ependymoglia. From GFAP distribution, the layered organization in both the telencephalon and the tectum were visible. In the cerebellum, the molecular and granular layers displayed conspicuously different glial structures; in the former a Bergmann glia-like population was found. No true astrocytes (i.e., stellate-shaped cells) were found. Radial glial endfeet lined all meningeal surfaces. Radial fibers also seemed to form endfeet and en passant contacts on the vessels. Plexuses of fine perivascular glial fibers also contributed to the perivascular glia. Compared with spiny dogfish brain, GFAP expression in the little skate brain was confined. Radial glia were limited to a few areas, e.g., segments of the ventricular surface of the telencephalon, and the midline of the diencephalon and mesencephalon. Scarce astrocytes occurred in every brain part, but only the optic chiasm, and the junction of the tegmentum and optic tectum contained large numbers of astrocytes. Astrocytes formed the meningeal glia limitans and the perivascular glia. No GFAP-immunopositive Bergmann glia-like structure was found. Astrocytes seen in the little skate were clearly different from the mammalian and avian ones; they had a different process system - extra large forms were frequently seen, and the meningeal and perivascular cells were spread along the surface instead of forming endfeet by processes. The differences between Squalus and Raia astroglia were much like those found between reptiles versus mammals and birds. It suggests independent and parallel glial evolutionary processes in amniotes and chondrichthyans, seemingly correlated with the thickening of the brain wall, and the growing complexity of the brain. There is no strict correlation, however, between the replacement of radial ependymoglia with astrocytes, and the local thickness of the brain wall.

Animals↗

Polarity of hepatic glutathione and glutathione S-conjugate efflux, and intraorgan mercapturic acid formation in the skate.

Mechanisms of hepatic glutathione and glutathione S-conjugate efflux were investigated in isolated hepatocytes and perfused liver of the little skate (Raja erinacea). Glutathione was released by isolated skate hepatocytes at a rate of 0.12 +/- 0.03 nmol.hr-1.(mg protein)-1. In the perfused liver, glutathione concentrations in bile were high (approximately 0.7 mM) compared to hepatic tissue levels (0.61 +/- 0.11 mumol.g-1). During the first hour of perfusion, the biliary glutathione excretion rate was 3 nmol.hr-1.(g liver)-1, whereas glutathione accumulated in the recirculating perfusate at a rate of only 1.5 nmol.hr-1.(g liver)-1. Release of glutathione by isolated hepatocytes and perfused liver was not affected by the addition of acivicin, an inhibitor of gamma-glutamyltransferase (EC 2.3.2.2), to cell suspension medium or liver perfusate. 1-Chloro-2,4-dinitrobenzene (CDNB) was taken up by isolated hepatocytes, conjugated to glutathione, and released as S-(2,4-dinitrophenyl) (DNP)-glutathione. After infusion of 0.5 mumol CDNB in perfused liver, S-DNP-glutathione was concentrated in bile (0.5 mM) and was associated with choleresis. S-DNP-Conjugates of cysteinylglycine, cysteine and N-acetylcysteine, were also found in bile, suggesting intrahepatic breakdown of S-DNP-glutathione and subsequent acetylation of the resulting cysteine conjugate to form the mercapturic acid, S-DNP-N-acetylcysteine. This mercapturic acid accounted for 31% of the total S-DNP-conjugates collected in bile. In contrast, neither S-DNP-glutathione nor other S-DNP-conjugates were detected in the perfusate (less than 0.5 microM). These findings demonstrate that biliary excretion is the predominant route for efflux of glutathione and a glutathione S-conjugate from skate liver. The results also identify an intrahepatic pathway for mercapturic acid biosynthesis facilitated by biliary glutathione S-conjugate excretion.

Acetylcysteine↗

Coordination of leg muscles during speed skating.

Five speed skaters of elite performance level and six speed skaters of trained level were subjected to an inverse dynamical analysis during speed skating. Push-off forces were registered by means of special skates. Myoelectric activity (EMG) of ten leg muscles and cinematographic data were recorded. Linked segment modelling yielded net joint moments and joint powers. The speed skating technique is characterized by a typical horizontal position of the trunk and a suppression of a plantar flexion during the push-off. This technique, necessary to reduce external friction, constrains the transfer of rotation in joints to translation of the mass center of the body. In spite of constrained push-off, the EMG levels of the leg muscles show a proximo-distal temporal order which to a certain extent is comparable to that previously found in an unconstrained vertical jump. This proximo-distal sequence is also reflected by the time courses of the net moment and net power output in hip, knee and ankle joints. The temporal sequence in activation levels of activated muscles is not different between elite and trained speed skaters. The difference in performance level between these groups obviously has an origin in the ability of the elite speed skaters to realise larger net joint moments. Differences in net joint moments and in kinematics result in a higher power output and a lower air frictional force for the elite than for the trained speed skaters.

Adult↗

A power equation for the sprint in speed skating.

An analysis of the start of the 500 m speed skating races during the 1988 Olympic Winter Games showed a remarkably high correlation between the acceleration of the skater in the first second of the sprint and the final time (r = -0.75). In this study a power equation is used to explain this high coefficient of correlation. The performance in speed skating is determined by the capability of external power production by the speed skater. This power is necessary to overcome the air and ice friction and to increase the kinetic energy of the skater. Numerical values of the power dissipated to air and ice friction, both dependent on speed, are obtained from ice friction and wind tunnel experiments. Using aerobic and anaerobic power production as measured during supra maximal bicycle tests of international-level speed skaters, a model of the kinetics of power production is obtained. Simulation of power production and power dissipation yields values of speed and acceleration and, finally, the performance time of the sprint during speed skating. The mean split time at 100 m and the final time at 500 m in these races, derived from simulation, were 10.57 s (+/- 0.31) and 37.82 s (+/- 0.96), respectively. The coefficient of correlation between the simulated 500 m times and the actual 500 m times was 0.90. From the results of this study it can be concluded that the distribution of the available anaerobic energy is an important factor in the short lasting events. For the same amount of anaerobic energy the better sprinters appear to be able to liberate considerably more energy at the onset of the race than skaters of lower performance level.

Biomechanical Phenomena↗

The absence of a photopic influence on the refractive development of the embryonic eye of the clearnose skate (Raja eglanteria).

The clearnose skate (Raja eglanteria) develops in an almost opaque eggcase and lays its eggs in pairs. One sibling from each of eight pairs of skates was removed from its eggcase during embryonic development, while the other sibling developed inside the eggcase. The refractive development of the eyes at hatching was examined to see if ambient light exposure during embryonic development could influence the refractive states of hatchlings. Measurements included refractive states, ocular dimensions and lens focal properties. The differences in measurements between the two groups were not significant, which would indicate that environmental light does not influence the refractive development of the embryonic skate eye.

Animals↗

Novel tachykinins from the brain of the sea lamprey, Petromyzon marinus, and the skate, Raja rhina.

Using radioimmunoassays for mammalian tachykinins, peptides with substance P-like immunoreactivity and neurokinin A-like immunoreactivity were identified in an extract of the brain of the longnose skate, Raja rhina (elasmobranch) but only a peptide with neurokinin A-like immunoreactivity was identified in the brain of the sea lamprey, Petromyzon marinus (agnathan). The primary structure of the skate peptide with substance P-like immunoreactivity (Ala-Lys-His-Asp-Lys-Phe-Tyr-Gly-Leu-Met-NH2) shows one amino acid substitution (Phe3-->His) compared with scyliorhinin I, previously isolated from dogfish brain and gut. The skate neurokinin A-related peptide (His-Lys-Leu-Gly-Ser-Phe-Val-Gly-Leu-Met-NH2) shows two substitutions (Thr3-->Leu and Asp4-->Gly) compared with mammalian neurokinin A. Although the COOH-terminus of the lamprey tachykinin (Arg-Lys-Pro-His-Pro-Lys-Glu-Phe-Val-Gly-Leu-Met-NH2) resembles neurokinin A, the presence of the strongly conserved Lys/Arg-Pro-Xaa-Pro motif at the NH2-terminus of the peptide indicates greater structural similarity with substance P. The additional arginine residue at position 1 in the peptide suggests that the lamprey is utilizing a site of posttranslational processing in the tachykinin precursor that is different from the equivalent site in mammalian and other lower vertebrate preprotachykinin(s).

Amino Acid Sequence↗

Immunoreactivity of skate electrocytes towards monoclonal antibodies against human dystrophin and dystrophin-related (DMDL) protein.

Monoclonal antibodies against human dystrophin have been used to demonstrate the existence of a dystrophin-like protein in the electrocytes of skate electric organ. This protein is also present in skate muscle and resembles that found in Torpedo electric organ. Monoclonal antibodies against a human autosomal homologue of dystrophin (DMDL protein) did not detect a similar protein in skate or Torpedo. Immunocytochemical staining of the innervated and non-innervated faces of the electrocyte membrane was obtained using the anti-dystrophin antibodies only.

Animals↗