Ole B. Nielsen

Protective effects of lactic acid on force production in rat skeletal muscle

1 During strenuous exercise lactic acid accumulates producing a reduction in muscle pH. In addition, exercise causes a loss of muscle K+ leading to an increased concentration of extracellular K+ ([K+]o). Individually, reduced pH and increased [K+]o have both been suggested to contribute to muscle fatigue. 2 To study the combined effect of these changes on muscle function, isolated rat soleus muscles were incubated at a [K+]o of 11 mm, which reduced tetanic force by 75 %. Subsequent addition of 20 mm lactic acid led, however, to an almost complete force recovery. A similar recovery was observed if pH was reduced by adding propionic acid or increasing the CO2 tension. 3 The recovery of force was associated with a recovery of muscle excitability as assessed from compound action potentials. In contrast, acidification had no effect on the membrane potential or the Ca2+ handling of the muscles. 4 It is concluded that acidification counteracts the depressing effects of elevated [K+]o on muscle excitability and force. Since intense exercise is associated with increased [K+]o, this indicates that, in contrast to the often suggested role for acidosis as a cause of muscle fatigue, acidosis may protect against fatigue. Moreover, it suggests that elevated [K+]o is of less importance for fatigue than indicated by previous studies on isolated muscles.

Increased excitability of acidified skeletal muscle : Role of chloride conductance

Generation of the action potentials (AP) necessary to activate skeletal muscle fibers requires that inward membrane currents exceed outward currents and thereby depolarize the fibers to the voltage threshold for AP generation. Excitability therefore depends on both excitatory Na+ currents and inhibitory K+ and Cl− currents. During intensive exercise, active muscle loses K+ and extracellular K+ ([K+]o) increases. Since high [K+]o leads to depolarization and ensuing inactivation of voltage-gated Na+ channels and loss of excitability in isolated muscles, exercise-induced loss of K+ is likely to reduce muscle excitability and thereby contribute to muscle fatigue in vivo. Intensive exercise, however, also leads to muscle acidification, which recently was shown to recover excitability in isolated K+-depressed muscles of the rat. Here we show that in rat soleus muscles at 11 mM K+, the almost complete recovery of compound action potentials and force with muscle acidification (CO2 changed from 5 to 24%) was associated with reduced chloride conductance (1731 ± 151 to 938 ± 64 μS/cm2, P < 0.01) but not with changes in potassium conductance (405 ± 20 to 455 ± 30 μS/cm2, P < 0.16). Furthermore, acidification reduced the rheobase current by 26% at 4 mM K+ and increased the number of excitable fibers at elevated [K+]o. At 11 mM K+ and normal pH, a recovery of excitability and force similar to the observations with muscle acidification could be induced by reducing extracellular Cl− or by blocking the major muscle Cl− channel, ClC-1, with 30 μM 9-AC. It is concluded that recovery of excitability in K+-depressed muscles induced by muscle acidification is related to reduction in the inhibitory Cl− currents, possibly through inhibition of ClC-1 channels, and acidosis thereby reduces the Na+ current needed to generate and propagate an AP. Thus short term regulation of Cl− channels is important for maintenance of excitability in working muscle.

Regulation of Na+–K+ pump activity in contracting rat muscle

1 In rat soleus muscle, high frequency electrical stimulation produced a rapid increase in intra‐cellular Na+ (Na+i) content. This was considerably larger in muscles contracting without developing tension than in muscles contracting isometrically. During subsequent rest a net extrusion of Na+ took place at rates which, depending on the frequency and duration of stimulation, approached the maximum transport capacity of the Na+–K+ pumps present in the muscle. 2 In isometrically contracting muscles, the net extrusion of Na+ continued for up to 10 min after stimulation, reducing Na+i to values 30% below the resting level (P < 0.001). This undershoot in Na+i, seen in both soleus and extensor digitorum longus muscles, could be maintained for up to 30 min and was blocked by ouabain or cooling to 0 °C. 3 The undershoot in Na+i could be elicited by direct stimulation as well as by tubocurarine‐suppressible stimulation via the motor endplate. It could not be attributed to a decrease in Na+ influx, to effects of noradrenaline or calcitonin gene‐related peptide released from nerve endings, to an increase in extracellular K+ or the formation of nitric oxide. 4 The results indicate that excitation rapidly activates the Na+–K+ pump, partly via a change in its transport characteristics and partly via an increase in intracellular Na+ concentration. This activation allows an approximately 20‐fold increase in the rate of Na+ efflux to take place within 10 s. 5 The excitation‐induced activation of the Na+–K+ pump may represent a feed‐forward mechanism that protects the Na+–K+ gradients and the membrane potential in working muscle. Contrary to previous assumptions, the Na+–K+ pump seems to play a dynamic role in maintenance of excitability during contractile activity.

Excitability of the T‐tubular system in rat skeletal muscle: roles of K+ and Na+ gradients and Na+–K+ pump activity

Strenuous exercise causes an increase in extracellular [K+] and intracellular Na+ ([Na+]i) of working muscles, which may reduce sarcolemma excitability. The excitability of the sarcolemma is, however, to some extent protected by a concomitant increase in the activity of muscle Na+–K+ pumps. The exercise‐induced build‐up of extracellular K+ is most likely larger in the T‐tubules than in the interstitium but the significance of the cation shifts and Na+–K+ pump for the excitability of the T‐tubular membrane and the voltage sensors is largely unknown. Using mechanically skinned fibres, we here study the role of the Na+–K+ pump in maintaining T‐tubular function in fibres with reduced chemical K+ gradient. The Na+–K+ pump activity was manipulated by changing [Na+]i. The responsiveness of the T‐tubules was evaluated from the excitation‐induced force production of the fibres. Compared to control twitch force in fibres with a close to normal intracellular [K+] ([K+]i), a reduction in [K+]i to below 60 mm significantly reduced twitch force. Between 10 and 50 mm Na+, the reduction in force depended on [Na+]i, the twitch force at 40 mm K+ being 22 ± 4 and 54 ± 9% (of control force) at a [Na+]i of 10 and 20 mm, respectively (n= 4). Double pulse stimulation of fibres at low [K+]i showed that although elevated [Na+]i increased the responsiveness to single action potentials, it reduced the capacity of the T‐tubules to respond to high frequency stimulation. It is concluded that a reduction in the chemical gradient for K+, as takes place during intensive exercise, may depress T‐tubular function, but that a concomitant exercise‐induced increase in [Na+]i protects T‐tubular function by stimulating the Na+–K+ pump.

Basal ice microbiology at the margin of the Greenland ice sheet

Abstract Basal ice at the margin of the Greenland ice sheet was studied with respect to its physical characteristics and microbiological community. The basal ice contained high concentrations of dissolved ferrous Fe and must therefore be anoxic. Oxygen consumption experiments indicate that 50% of the oxidation was due to biological activity while the rest could be attributed to chemical processes, most likely weathering reactions with ferrous Fe. At least six different Fe-containing mineral sources were detected in basal ice together with potential bioavailable Fe nanoparticles. An active denitrifier population was identified due to formation of 30N-dinitrogen gas after amendment of anoxic sediment slurries with 15N-NO3 −. Sulfate reduction could not be detected. The solid ice facies contained an abundant (∼108 cells cm−3) and complex microbial community that harbored representatives of at least eight major phyla within the domain Bacteria. The clone library was dominated by members of the β-subdivision of proteobacteria of which the largest proportion was affiliated to the genus Rhodoferax that comprises facultative aerobic iron reducers. The second most abundant phylum was Bacteroidetes. The solid ice facies had many physical similarities with the overlying debris-rich banded ice facies, indicating that they formed by similar subglacial processes and harbor similar microbial communities. This study extends our knowledge of life in subglacial environments such as beneath ice sheets. GenBank accession numbers: HM439882-HM439950; HQ144215-HQ144221.

Regulation of Na+-K+ homeostasis and excitability in contracting muscles: implications for fatigue.

The performance of skeletal muscles depends on their ability to initiate and propagate action potentials along their outer membranes in response to motor signals from the central nervous system. This excitability of muscle fibres is related to the function of Na+ and K+ and Cl- channels and to steep chemical gradients for the ions across the cell membranes, i.e., the sarcolemma and T-tubular membranes. At rest, the chemical gradients for Na+ and K+ are maintained within close limits by the action of the Na+-K+ pump. During contractile activity, however, the muscles lose K+, which causes an increase in the concentration of K+ in the extracellular compartments of the body, the magnitude of which depends on the intensity of the exercise and the size of the muscle groups involved. Since the ensuing reduction in the chemical K+ gradient can have adverse effects on muscle excitability, it has repeatedly been suggested that, during intense exercise, the loss of K+ from muscle fibres can contribute to the complex set of mechanisms that leads to the development of muscle fatigue. In this review, aspects of the regulation of Na+-K+ homeostasis and excitability in contracting muscles is discussed within this context, together with the implications for the contractile function of skeletal muscles.

Geological indications for Palaeogene uplift in the eastern North Sea Basin

Abstract The timing and effect of the Cenozoic uplift of Scandinavia has been investigated using a multi-disciplinary approach involving sedimentological, seismic and biostratigraphic data from the Danish and the adjacent Norwegian parts of the North Sea Basin. It is concluded that significant uplift took place periodically throughout the Palaeogene possibly marking an earlier onset of the so-called “Neogene uplift” of Scandinavia. This conclusion is based on a number of sedimentological observations, including smectite content, grain-size variations, kaolinite thermal stabilities and T max values supported by seismic reflection geometries and biostratigraphic data. These data indicate several phases of re-working of Palaeogene and older sediments situated further to the east and northeast during the middle to late Eocene and during the middle to late Oligocene. The tectonic patterns were similar during the late Paleocene and the Oligocene with some inversion taking place, whereas no inversion has been observed during the Eocene. Main provenance areas were to the north and northeast during the Paleocene and Oligocene, whereas the Eocene sediments originate mainly from the British Isles to the west. It is proposed that Palaeogene uplift of Scandinavia was associated with regional tectonic movements along crustal zones of weakness, which were reactivated as they accommodated strain induced by the Alpine Orogeny and the opening of the North Atlantic.

Na + -K + pump stimulation restores carbacholine-induced loss of excitability and contractility in rat skeletal muscle

Intense exercise results in increases in intracellular Na+ and extracellular K+ concentrations, leading to depolarization and a loss of muscle excitability and contractility. Here, we use carbacholine to chronically activate the nicotinic acetylcholine (nACh) receptors to mimic the changes in membrane permeability, chemical Na+ and K+ gradients and membrane potential observed during intense exercise. Intact rat soleus muscles were mounted on force transducers and stimulated electrically to evoke short tetani at regular intervals. Carbacholine produced a 2.6‐fold increase in Na+ influx that was tetrodotoxin (TTX) insensitive, but abolished by tubocurarine, resulting in a significant 36% increase in intracellular Na+, and 8% decrease in intracellular K+ content. The mid region, near the motor end plate, had much larger alterations than the more distal regions of the muscle, and showed a larger membrane depolarization from −73 ± 1 to −60 ± 1 mV compared with −64 ± 1 mV. Carbacholine (10−4m) significantly reduced tetanic force to 31 ± 3% of controls, which underwent significant recovery upon application of Na+–K+ pump stimulators: salbutamol (10−5m), adrenaline (10−5m) and calcitonin gene‐related peptide (CGRP; 10−7m). The force recovery with salbutamol was accompanied by a recovery of intracellular Na+ and K+ contents, and a small but significant 4–5 mV recovery of membrane potential. Similar results were obtained using succinylcholine (10−4m), indicating that Na+–K+ pump stimulation may prevent or restore succinylcholine‐induced hyperkalaemia. The stimulation of the Na+–K+ pump allows muscle to partially recover contractility by regaining excitability through electrogenically driven repolarization of the muscle membrane.

Extracellular magnesium and calcium reduce myotonia in ClC-1 inhibited rat muscle.

Loss-of-function mutations in the ClC-1 Cl(-) channel trigger skeletal muscle hyperexcitability in myotonia congenita. For reasons that remain unclear, the severity of the myotonic symptoms can vary markedly even among patients with identical ClC-1 mutations, and may become exacerbated during pregnancy and with diuretic treatment. Since both these conditions are associated with hypomagnesemia and hypocalcemia, we explored whether extracellular Mg(2+) and Ca(2+) ([Mg(2+)]o and [Ca(2+)]o) can affect myotonia. Experimental myotonia was induced in isolated rat muscles by ClC-1 inhibition and effects of [Mg(2+)]o or [Ca(2+)]o on myotonic contractions were determined. Both cations dampened myotonia within their physiological concentration ranges. Thus, myotonic contractile activity was 6-fold larger at 0.3 than at 1.2 mM [Mg(2+)]o and 82-fold larger at 0.3 than at 1.27 mM [Ca(2+)]o. In intracellular recordings of action potentials, the threshold for action potential excitation was raised by 4-6 mV when [Mg(2+)]o was elevated from 0.6 to 3 mM, compatible with an increase in the depolarization of the membrane potential necessary to activate the Na(+) channels. Supporting this notion, mathematical simulations showed that myotonia went from appearing with normal Cl(-) channel function to disappearing in the absence of Cl(-) channel function when Na(+) channel activation was depolarized by 6 mV. In conclusion, variation in serum Mg(2+) and Ca(2+) may contribute to phenotypic variation in myotonia congenita patients.

The presence of thrust-block naled after a major surge event: Kuannersuit Glacier, West Greenland

Abstract Thrust-block naled in front of Kuannersuit Glacier, West Greenland, appears to have formed during the termination of a terrestrial surge event by a combination of enhanced winter runoff, rapid advance of the glacier terminus, and proglacial stress release by thrusting and stacking of naled blocks. This process is equivalent to the formation of thrust-block moraines. The thrust-block naled consists of at least seven thrust sheets, which are characterized by stratified ice with beds composed of a lower debris-rich lamina, an intermediate dispersed lamina and a top clean-ice lamina, and underlain by frozen outwash deposits. The thrust-block naled differs from basal stratified ice in the absence of internal deformation structures, a relatively low debris concentration, a clay-rich particle-size distribution and a preferential sorting of lighter minerals. The oxygen isotope composition of the thrust-block naled is indistinguishable from δ18O values from meteoric glacier ice and bulk meltwater, but different from basal stratified ice facies. The d–δD relationship indicates that thrust-block naled has been formed by freezing of successive thin layers of bulk waters with variable isotopic composition, whereas basal stratified ice has developed in a subglacial environment with regelation. This work shows that the association between proglacial naled and rapidly advancing glaciers may have significant consequences for the proglacial geomorphology and the interpretation of basal ice layers.