bluuebunny/crossref_metadata_2025 huggingface dataset:
Electrospray ionisation mass spectrometric analysis of lipid restructuring in the carp (Cyprinus carpio L.) during cold acclimation
Authors: Brooks S. , Clark G. T. , Wright S. M. , Trueman R. J. , Postle A. D. , Cossins A. R. , Maclean N. M.
Published: April 2021

Cold acclimation of carp from 30°C to 10°C causes a restructuring of liver microsomal phospholipids characterised by increased proportions of monounsaturated fatty acid in phosphatidylcholine (PC) and phosphatidylethanolamine (PE). Here, we have used electrospray ionisation mass spectrometry (ESI-MS) to determine the patterns of alteration to individual molecular species compositions of PC, PE and phosphatidylinositol (PI) in response to gradually decreasing temperature. The results demonstrate that cold induces precise changes to a limited number of phospholipid species, and that these changes are distinct and different for each phospholipid class. The major change for PC was increased 16:1/22:6, but for PE the species that increased was 18:1/22:6. By contrast, the PI species that increased during cold acclimation were characterised by an sn-1 monounsaturated fatty acid in combination with arachidonoyl or eicosapentaenoyl fatty acid at the sn-2 position. Analysis of acyl distribution indicates that cold only caused the accumulation of monounsaturated fatty acids at the sn-1 and not at the sn-2 position of phospholipids. These results highlight the tight and restricted range of modifications that membranes make to their phospholipid composition in response to thermal stress.
Evidence for increased myofibrillar mobility in desmin-null mouse skeletal muscle
Authors: Shah Sameer B. , Su Fong-Chin , Jordan Kimberly , Milner Derek J. , Fridén Jan , Capetanaki Yassemi , Lieber Richard L.
Published: April 2021

Quantitative electron microscopy was used to characterize the longitudinal mobility of myofibrils during muscle extension to investigate the functional roles of skeletal muscle intermediate filaments. Extensor digitorum longus fifth toe muscles from wild-type (+/+) and desmin-null (des –/–) animals were passively stretched to varying lengths, and the horizontal displacement of adjacent Z-disks in neighboring myofibrils (Δxmyo) and average sarcomere length (SL) were calculated. At short SL (<2.20 μm), wild-type and desmin-null Δxmyo were not significantly different, although there was a trend towards greater Z-disk misalignment in muscles from knockout animals (Δxmyo 0.34±0.04 μm versus 0.22±0.09 μm; P>0.2; means ± s.e.m.). However, at higher SL (>2.90 μm), muscles from knockout animals displayed a dramatically increased Δxmyo relative to wild-type muscles (0.49±0.10 μm versus 0.25±0.07 μm; P<0.05). The results, which establish a maximum extension of the desmin network surrounding the Z-disk, provide what we believe to be the first quantitative estimation of the functional limits of the desmin intermediate filament system in the presence of an intact myofibrillar lattice. The existence of a limit on the extension of desmin suggests a mechanism for the recruitment of desmin into a network of force transmission, whether as a longitudinal load bearer or as a component in a radial force-transmission system.
Determining patterns of motor recruitment during locomotion
Authors: Wakeling James M. , Kaya Motoshi , Temple Genevieve K. , Johnston Ian A. , Herzog Walter
Published: April 2021

Motor units are the functional units of muscle contraction in vertebrates. Each motor unit comprises muscle fibres of a particular fibre type and can be considered as fast or slow depending on its fibre-type composition. Motor units are typically recruited in a set order, from slow to fast, in response to the force requirements from the muscle. The anatomical separation of fast and slow muscle in fish permits direct recordings from these two fibre types. The frequency spectra from different slow and fast myotomal muscles were measured in the rainbow trout Oncorhynchus mykiss. These two muscle fibre types generated distinct low and high myoelectric frequency bands. The cat paw-shake is an activity that recruits mainly fast muscle. This study showed that the myoelectric signal from the medial gastrocnemius of the cat was concentrated in a high frequency band during paw-shake behaviour. During slow walking, the slow motor units of the medial gastrocnemius are also recruited, and this appeared as increased muscle activity within a low frequency band. Therefore, high and low frequency bands could be distinguished in the myoelectric signals from the cat medial gastrocnemius and probably corresponded, respectively, to fast and slow motor unit recruitment. Myoelectric signals are resolved into time/frequency space using wavelets to demonstrate how patterns of motor unit recruitment can be determined for a range of locomotor activities.
Modulation of swimming in the gastropod Melibe leonina by nitric oxide
Authors: Newcomb James M. , Watson Winsor H.
Published: April 2021

Nitric oxide (NO) is a gaseous intercellular messenger produced by the enzyme nitric oxide synthase. It has been implicated as a neuromodulator in several groups of animals, including gastropods, crustaceans and mammals. In this study, we investigated the effects of NO on the swim motor program produced by isolated brains and by semi-intact preparations of the nudibranch Melibe leonina. The NO donors sodium nitroprusside (SNP, 1 mmol l–1) and S-nitroso-N-acetylpenicillamine (SNAP, 1 mmol l–1) both had a marked effect on the swim motor program expressed in isolated brains, causing an increase in the period of the swim cycle and a more erratic swim rhythm. In semi-intact preparations, the effect of NO donors was manifested as a significant decrease in the rate of actual swimming. An NO scavenger, reduced oxyhemoglobin, eliminated the effects of NO donors on isolated brains, supporting the assumption that the changes in swimming induced by donors were actually due to NO. The cGMP analogue 8-bromoguanosine 3′,5′-cyclic monophosphate (1 mmol l–1) produced effects that mimicked those of NO donors, suggesting that NO is working via a cGMP-dependent mechanism. These results, in combination with previous histological studies indicating the endogenous presence of nitric oxide synthase, suggest that NO is used in the central nervous system of Melibe leonina to modulate swimming.
Heart rate and energy expenditure of incubating wandering albatrosses: basal levels, natural variation, and the effects of human disturbance
Authors: Weimerskirch Henri , Shaffer Scott A. , Mabille Geraldine , Martin Julien , Boutard Olivier , Rouanet Jean Louis
Published: April 2021

We studied the changes in heart rate (HR) associated with metabolic rate of incubating and resting adult wandering albatrosses (Diomedea exulans) on the Crozet Islands. Metabolic rates of resting albatrosses fitted with external HR recorders were measured in a metabolic chamber to calibrate the relationship between HR and oxygen consumption (V̇O2) (V̇O2=0.074×HR+0.019, r2=0.567, P<0.001, where V̇O2 is in ml kg–1 min–1 and HR is in beats min–1). Incubating albatrosses were then fitted with HR recorders to estimate energy expenditure of albatrosses within natural field conditions. We also examined the natural variation in HR and the effects of human disturbance on nesting birds by monitoring the changes in HR. Basal HR was positively related to the mass of the individual. The HR of incubating birds corresponded to a metabolic rate that was 1.5-fold (males) and 1.8-fold (females) lower than basal metabolic rate (BMR) measured in this and a previous study. The difference was probably attributable to birds being stressed while they were held in the metabolic chamber or wearing a mask. Thus, previous measurements of metabolic rate under basal conditions or for incubating wandering albatrosses are likely to be overestimates. Combining the relationship between HR and metabolic rate for both sexes, we estimate that wandering albatrosses expend 147 kJ kg–1 day–1 to incubate their eggs. In addition, the cost of incubation was assumed to vary because (i) HR was higher during the day than at night, and (ii) there was an effect of wind chill (<0°C) on basal HR. The presence of humans in the vicinity of the nest or after a band control was shown to increase HR for extended periods (2–3 h), suggesting that energy expenditure was increased as a result of the disturbance. Lastly, males and females reacted differently to handling in terms of HR response: males reacted more strongly than females before handling, whereas females took longer to recover after being handled.
Antagonistic control of fluid secretion by the Malpighian tubules ofTenebrio molitor: effects of diuretic and antidiuretic peptides and their second messengers
Authors: Wiehart U. I. M. , Nicolson S. W. , Eigenheer R. A. , Schooley D. A.
Published: April 2021

Fluid secretion by insect Malpighian tubules is controlled by haemolymph-borne factors. The mealworm Tenebrio molitor provides the first known example of antagonistic interactions between endogenous neuropeptides acting on Malpighian tubules. The two corticotropin-releasing-factor (CRF)-related diuretic peptides previously isolated from Tenebrio molitor, Tenmo-DH37 and Tenmo-DH47, were found to stimulate Tenebrio molitor tubules in vitro in a dose-dependent manner with EC50 values of 0.12 nmol l–1 and 26 nmol l–1 respectively. However, no synergistic or additive effect was observed when these two peptides were tested simultaneously. We then investigated antagonism between second messengers: dose–response curves were constructed for stimulation of Tenebrio molitor tubules by cyclic AMP and their inhibition by cyclic GMP. When both cyclic nucleotides were included in the bathing Ringer, the stimulatory effect of cyclic AMP was neutralised by cyclic GMP. Similarly, the stimulatory effect of Tenmo-DH37 was reversed on addition of an antidiuretic peptide (Tenmo-ADF), which was recently isolated from Tenebrio molitor and acts via cyclic GMP. The cardioacceleratory peptide CAP2b, originally isolated from Manduca sexta, also increases intracellular cyclic GMP levels and inhibited fluid secretion by Tenebrio molitor tubules, with an EC50 value of 85 nmol l–1. This inhibitory effect was reversed by Tenmo-DH37. Endogenous diuretic and antidiuretic peptides, effective at low concentrations and acting via antagonistic second messengers, have the potential for fine control of secretion rates in the Malpighian tubules of Tenebrio molitor.
Ion transport across posterior gills of hyperosmoregulating shore crabs (Carcinus maenas): amiloride blocks the cuticular Na+ conductance and induces current-noise
Authors: Onken Horst , Riestenpatt Sven
Published: April 2021

Split gill lamellae and gill cuticles of shore crabs (Carcinus maenas) adapted to 10 ‰ salinity were mounted in a modified Ussing-type chamber. With NaCl saline on both sides, split gill lamellae generated a short-circuit current (Isc) of –301±16 μA cm–2 at a conductance (Gte) of 40±2 mS cm–2. The net influxes of Na+ and Cl– were 8.3±2.6 and 18.2±2.7 μmol cm–2 h–1, respectively. External amiloride (100 μmol l–1) reduced Gte to approximately 50 % of the original value at unchanged Isc; Cl– fluxes remained unaffected, whereas Na+ fluxes were markedly reduced by 70–80 %. The Isc in the presence of external amiloride was almost completely inhibited by internal ouabain. At a clamp voltage of 50 mV (outside-positive), a positive current was measured at unchanged Gte. Under these conditions, amiloride reduced the current and conductance at half-maximal concentrations of 3.6 and 2.0 μmol l–1, respectively. At outside-positive voltages, but not under short-circuit conditions, external amiloride induced Lorentzian components in the power density spectra. The amiloride-dependent changes in the corner frequency (linear) and of the low-frequency plateau (‘bell-shaped’) were as expected for channel blockade by amiloride with pseudo-first-order kinetics. With an outside-positive clamp voltage of 50 mV across isolated cuticles, a positive cuticular current (Icut) of 25 188±3791 μA cm–2 and a cuticular conductance (Gcut) of 547±76 mS cm–2 were measured. External amiloride reduced Icut and Gcut at half-maximal concentrations of 0.7 and 0.6 μmol l–1, respectively. Amiloride-induced current-noise analysis gave similar results to those observed with split gill lamellae. Ion-substitution experiments with isolated cuticles further support inhibition by external amiloride of the cuticular Na+ conductance of shore crab gills and not amiloride-sensitive transporters (Na+ channels or Na+/H+ antiports) in the apical membrane.
Shell clamping behaviour in the limpetCellana tramoserica
Authors: Ellem Gary K. , Furst John E. , Zimmerman Kenneth D.
Published: April 2021

The behaviour of clamping the shell against the substratum may play an important role in the limpet adhesion mechanism because friction generated by this behaviour resists dislodgement by shear forces. This paper describes the development of an apparatus to analyse limpet clamping activity in relation to known forces, including simulated wave activity and predator attack. The results show that Cellana tramoserica clamps its shell in a closely regulated manner consistent with an active role in the limpet adhesion mechanism. Limpets clamped sharply for several seconds in response to single disturbances such as tapping the shell. In response to more continuous disturbance simulating a concerted predator attack, limpets clamped tightly for several minutes. In response to lifting forces applied to the shell, limpets clamped at a set proportion of the lifting force, even if the lift force was a highly dynamic wave profile. This behaviour has implications for numerical models that attempt to describe limpet adhesion because it shows that limpets cannot be represented by a simple mechanical analogue and that the clamping behaviour must be accounted for if useful predictions are to be drawn.
Template-matching describes visual pattern-recognition tasks in the weakly electric fishGnathonemus petersii
Authors: Schuster Stefan , Amtsfeld Silke
Published: April 2021

Several insects use template-matching systems to recognize objects or environmental landmarks by comparing actual and stored retinal images. Such systems are not viewpoint-invariant and are useful only when the locations in which the images have been stored and where they are later retrieved coincide. Here, we describe that a vertebrate, the weakly electric fish Gnathonemus petersii, appears to use template-matching to recognize visual patterns that it had previously viewed from a fixed vantage point. This fish is nocturnal and uses its electrical sense to find its way in the dark, yet it has functional vision that appears to be well adapted to dim light conditions. We were able to train three fish in a two-alternative forced-choice procedure to discriminate a rewarded from an unrewarded visual pattern. From its daytime shelter, each fish viewed two visual patterns placed at a set distance behind a transparent Plexiglas screen that closed the shelter. When the screen was lifted, the fish swam towards one of the patterns to receive a food reward or to be directed back into its shelter. Successful pattern discrimination was limited to low ambient light intensities of approximately 10 lx and to pattern sizes subtending a visual angle greater than 3°. To analyze the characteristics used by the fish to discriminate the visual training patterns, we performed transfer tests in which the training patterns were replaced by other patterns. The results of all such transfer tests can best be explained by a template-matching mechanism in which the fish stores the view of the rewarded training pattern and chooses from two other patterns the one whose retinal appearance best matches the stored view.
Carbonic anhydrase in the midgut of larvalAedes aegypti: cloning, localization and inhibition
Authors: del Pilar Corena Maria , Seron Theresa J. , Lehman Herm K. , Ochrietor Judith D. , Kohn Andrea , Tu Chingkuang , Linser Paul J.
Published: April 2021

The larval mosquito midgut exhibits one of the highest pH values known in a biological system. While the pH inside the posterior midgut and gastric caeca ranges between 7.0 and 8.0, the pH inside the anterior midgut is close to 11.0. Alkalization is likely to involve bicarbonate/carbonate ions. These ions are produced in vivo by the enzymatic action of carbonic anhydrase. The purpose of this study was to investigate the role of this enzyme in the alkalization mechanism, to establish its presence and localization in the midgut of larval Aedes aegypti and to clone and characterize its cDNA. Here, we report the physiological demonstration of the involvement of carbonic anhydrase in midgut alkalization. Histochemistry and in situ hybridization showed that the enzyme appears to be localized throughout the midgut, although preferentially in the gastric caeca and posterior regions with specific cellular heterogeneity. Furthermore, we report the cloning and localization of the first carbonic anhydrase from mosquito larval midgut. A cDNA clone from Aedes aegypti larval midgut revealed sequence homology to α-carbonic anhydrases from vertebrates. Bioinformatics indicates the presence of at least six carbonic anhydrases or closely related genes in the genome of another dipteran, the fruit fly Drosophila melanogaster. Molecular analyses suggest that the larval mosquito may also possess multiple forms.
Evolutionary determinants of normal arterial plasma pH in ectothermic vertebrates
Authors: Burton Richard F.
Published: April 2021

Mean values of normal arterial pH in different species of fish, amphibians and reptiles at 15 and 25°C, taken from the literature, are negatively correlated with arterial PCO2 and plasma [Na+]. At either temperature, the data accord with the hypothesis that extracellular acid–base homeostasis evolved to maintain an optimal pH at particular cell-surface sites that are similar in all species. These hypothetical sites bear fixed negative charges that attract H+, but which are partially screened by Na+; for the surface pH to be constant, the bulk interstitial pH should then vary inversely with [Na+], as is the case. At the same time, the bulk interstitial fluid must be more acid than arterial plasma by an amount that increases with decreasing arterial PCO2. With allowance made for additional screening by Ca2+ and Mg2+, the relevant cell-surface pH is probably approximately 6.2.
The use of ground-borne vibrations for prey localization in the Saharan sand vipers (Cerastes)
Authors: Young Bruce A. , Morain Malinda
Published: April 2021

Sand vipers of the genus Cerastes are specialized semi-fossorial snakes that launch predatory strikes at rodents and lizards while partially buried in the soft sand of the Saharan desert. This study attempted to document which environmental stimuli are used by these snakes as a trigger for the ambush behavior. Denervating the olfactory and vomeronasal organs produced no changes in prey capture behavior in Cerastes cerastes. Occluding the eyes of the denervated specimens resulted in significant decreases in strike distance, diversity of strike angle and strike accuracy, demonstrating the importance of visual stimuli for target acquisition in this species. Nevertheless, every olfactory-denervated, temporarily blinded specimen succeeded in capturing free-ranging mice in every trial. Presentation of chemosensory-neutral targets to the olfactory-denervated, temporarily blinded specimens resulted in similar predatory behaviors, whether the target was isothermic to the environment or heated to mammalian body temperature. Collectively, these results provide evidence for the importance of visual stimuli during foraging in C. cerastes, the first experimental evidence for foraging by vibration detection in snakes and the strongest evidence to date that snakes are capable of ‘hearing’ vibrational stimuli.
NaCl and fluid secretion by the intestine of the teleostFundulus heteroclitus: involvement of CFTR
Authors: Marshall W. S. , Howard J. A. , Cozzi R. R. F. , Lynch E. M.
Published: April 2021

Sections of posterior intestine of the euryhaline killifish Fundulus heteroclitus adapted to sea water were stimulated by the calcium ionophore ionomycin (1 μmol l–1) in combination with agents to elevate intracellular cyclic AMP levels, 0.5 mmol l–1 dibutyryl-cyclic AMP (db-cAMP) with 0.1 mmol l–1 3-isobutyl-1-methylxanthine (IBMX). Intestinal bag preparations from recently fed animals (but not from overnight unfed animals) changed from fluid absorption (+18.9±8.30 μl cm–2 h–1 , N=8) in the untreated control period to net fluid secretion after stimulation (–7.43±1.30 μl cm–2 h–1, N=8, P<0.01; means ± s.e.m.), indicative of the capacity of teleost intestine to undergo secretion. Posterior intestinal pieces mounted in vitro in Ussing-style membrane chambers showed net Cl– uptake (+2.245±0.633 μequiv cm–2 h–1, N=7) that turned to net secretion following stimulation by ionomycin + db-cAMP + IBMX (–3.809±1.22 μequiv cm–2 h–1, N=7, P<0.01). Mucosal application of the anion channel blocker 1 mmol l–1 diphenylamine-2-carboxylate (DPC) after ionomycin + db-cAMP + IBMX treatment significantly reduced serosal-to-mucosal unidirectional Cl– flux (P<0.001), net Cl– flux (P<0.05), short-circuit current (Isc, P<0.001) and tissue conductance (Gt, P<0.001), while 0.1 mmol l–1 4,4′-diisothiocyano-2,2′-stilbene-disulphonic acid (DIDS, a blocker of anion exchange) was without effect. Stimulation by db-cAMP + IBMX (no ionomycin) significantly increased unidirectional fluxes, Isc and Gt but did not produce net Cl– secretion. Ionomycin alone produced a transient increase in Isc but had no effect on Gt and caused no significant changes in unidirectional or net Cl– fluxes. Addition of db-cAMP + IBMX after ionomycin treatment produced net secretion of Cl– and large increases in unidirectional fluxes and Gt. Cystic fibrosis transmembrane conductance regulator (CFTR) was immunocytochemically localized with a monoclonal mouse antibody to the carboxy terminus and found to be present in the cytoplasm and basolateral membranes of all enterocytes and in the brush-border membrane of some cells, whereas NKCC immunofluorescence, demonstrating the presence of the Na+/K+/2Cl– cotransporter, was present in the cytoplasm and brush-border membrane. We conclude that the teleost intestine is capable of salt and fluid secretion only if intracellular Ca2+ and cyclic AMP pathways are stimulated together and that this secretion appears to involve activation of CFTR ion channels in the apical membrane of a subpopulation of enterocytes.
Metabolic rate and respiratory gas-exchange patterns in tenebrionid beetles from the Negev Highlands, Israel
Authors: Duncan Frances D. , Krasnov Boris , McMaster Megan
Published: April 2021

This study correlates the pattern of external gas exchange with the diel activity of nine species of tenebrionid beetle from the Negev Desert, Israel. The study species are active throughout the summer months when daytime temperatures are high and no rain falls. There were no differences in standard metabolic rate, determined by flow-through respirometry, among the nine species. All the nocturnally active beetles exhibited a form of continuous respiration, whereas the two diurnally active and one crepuscular species exhibited a cyclic form of respiration referred to as the discontinuous gas-exchange cycle (DGC). The DGCs recorded have a long flutter period consisting of miniature ventilations, and 29–48 % of the total CO2 output occurred during this period. In this study, the flutter period played an important role in the modulation of metabolic rate, in contrast to other studies in which the burst period has been shown to be important. We suggest that the long flutter period is important in reducing respiratory water loss in arid-dwelling arthropods. This study lends support to the hypothesis that discontinuous gas exchange is important in reducing respiratory water loss from beetles that need to minimise dessication because of the high water vapour pressure gradient they experience. If the use of underground burrows were responsible for the evolution of discontinuous gas exchange, then we would expect all nine tenebrionid species to use DGCs since both the nocturnally and diurnally active species bury in the sand during periods of inactivity. We conclude that the activity patterns of the beetles are more important than their habitat associations in designating the type of respiration used.
Luminescence in ophiuroids (Echinodermata) does not share a common nervous control in all species
Authors: Dewael Y. , Mallefet J.
Published: April 2021

Study of the control mechanisms of light emission in invertebrates shows the involvement of several neurotransmitters. In ophiuroids, only one species (Amphipholis squamata) has so far been characterized for luminescence control, which seems to be cholinergic, with an influence of several excitatory and inhibitory neuromodulators (amino acids, catecholamines, neuropeptides S1 and S2, purines). The aim of this work is to investigate the nature of control mechanisms of light emission in three luminous ophiuroid species, A. filiformis, O. aranea and O. californica, in order to see whether or not they share common mechanisms. Luminescence induced by general depolarisation of tissues using KCl (200 mmol l–1) shows different patterns, according to species. Only A. filiformis emits light in response to acetylcholine. In this species, the involvement of both muscarinic and nicotinic receptors is proposed, since atropine and tubocurarine (at 10–3 mol l–1) inhibited 99 % and 71 %, respectively, of the light emitted. Study of the subtypes of cholinergic receptors involved in photogenesis revealed that several subtypes of muscarinic receptors might be involved. It was also clearly shown that ophiuroids did not share a common mechanism of nervous control of luminescence in all species.
The use of landmarks and panoramic context in the performance of local vectors by navigating honeybees
Authors: Collett Matthew , Harland Duane , Collett Thomas S.
Published: April 2021

Bees seem to use landmarks to segment familiar routes. They can associate, with a landmark, a memory that encodes the direction and distance of the path segment between that landmark and the next. The expression of the memory results in the performance of a local vector matching the distance and direction of the path segment. The memories of path segments appear to be ‘chained’ together, so that the performance of one local vector is sometimes sufficient to elicit the subsequent local vector, even in the absence of the associated landmark. We have investigated the effect of visual panoramic context on the expression of local vectors. Bees were trained to fly along a narrow channel to collect sucrose from a feeder positioned partway along it. Panoramic context was provided by various types of patterning on the walls. The channel was partitioned into different segments using landmarks of two kinds: a boundary landmark that marked a change in the pattern on one or both side-walls of the channel, and an isolated landmark, consisting of a baffle through which the bee passed, for which the wall pattern was the same before as after. In tests, we removed the feeder and analysed the search distribution of the bees for various arrangements of landmarks. Altering the spatial relationship between landmarks has different consequences for the two types of landmark. If the final boundary landmark is shifted, the centre of the search distribution shifts by approximately the same amount. Changes in the position of an isolated landmark have a weaker effect. In the absence of the final context, the search is disrupted. We suggest that for local vectors to be expressed the surrounding panoramic context needs to be appropriate. A comparison of search patterns from two different training configurations of landmarks supports the hypothesis that local vector memories merely encode route segments and that global positional coordinates are not linked to landmark memories.
Differing mechanisms of cold-induced changes in capillary supply in m. tibialis anterior of rats and hamsters
Authors: Deveci Durmus , Egginton Stuart
Published: April 2021

The physiological, metabolic and anatomical adaptations of skeletal muscle to chronic cold exposure were investigated in Wistar rats (Rattus norvegicus), a species that defends core temperature, and Syrian hamsters (Mesocricetus auratus), which may adopt a lower set point under unfavourable conditions. Animals were exposed to a simulated onset of winter in an environmental chamber, progressively shortening photoperiod and reducing temperature from 12 h:12 h L:D and 22°C to 1 h:23 h L:D and 5°C over 4 weeks. The animals were left at 4°C for a further 4 weeks to complete the process of cold-acclimation. M. tibialis anterior from control (euthermic) and cold-acclimated animals of similar mass showed a significant hyperactivity-induced hypertrophy in the rat, but a small disuse atrophy in the hamster. Little evidence was found for interconversion among fibre types in skeletal muscle on cold-acclimation, and only modest differences were seen in activity of oxidative or glycolytic enzymes in either species. However, adjustments in Type II fibre size paralleled the muscle hypertrophy in rat and atrophy in hamster. Cold-induced angiogenesis was present in the rat, averaging a 28 % increase in capillary-to-fibre ratio (C:F) but, as this was balanced by fibre hypertrophy across the whole muscle, there was no change in capillary density (CD). In contrast, the C:F was similar in both groups of hamsters, whereas CD rose by 33 % in line with fibre atrophy. Within distinct regions of the m. tibialis anterior, there was a correlation between angiogenesis and fibre size in rats, in which oxygen diffusion distance increased, but not in hamsters, in which there was a reduced oxygen diffusion distance. Consequently, the change in C:F was greatest (39 %) in the glycolytic cortex region of the m. tibialis anterior in rats. We conclude that non-hibernator and hibernator rodents improve peripheral oxygen transport following cold-acclimation by different mechanisms. In rats, an increase in fibre girth was accompanied by a true angiogenesis, while the improved apparent capillary supply in hamsters was due to smaller fibre diameters. These responses are consistent with the strategies of resisting and accommodating, respectively, an annual fall in environmental temperature.
Effects of metamorphosis on the aquatic escape response of the two-lined salamander (Eurycea bislineata)
Authors: Azizi Emanuel , Landberg Tobias
Published: April 2021

Although numerous studies have described the escape kinematics of fishes, little is known about the aquatic escape responses of salamanders. We compare the escape kinematics of larval and adult Eurycea bislineata, the two-lined salamander, to examine the effects of metamorphosis on aquatic escape performance. We hypothesize that shape changes associated with resorption of the larval tail fin at metamorphosis will affect aquatic locomotor performance. Escape responses were recorded using high-speed video, and the effects of life stage and total length on escape kinematics were analyzed statistically using analysis of covariance. Our results show that both larval and adult E. bislineata use a two-stage escape response (similar to the C-starts of fishes) that consists of a preparatory (stage 1) and a propulsive (stage 2) stroke. The duration of both kinematic stages and the distance traveled during stage 2 increased with total length. Both larval and adult E. bislineata had final escape trajectories that were directed away from the stimulus. The main kinematic difference between larvae and adults is that adults exhibit significantly greater maximum curvature during stage 1. Total escape duration and the distance traveled during stage 2 did not differ significantly between larvae and adults. Despite the significantly lower tail aspect ratio of adults, we found no significant decrease in the overall escape performance of adult E. bislineata. Our results suggest that adults may compensate for the decrease in tail aspect ratio by increasing their maximum curvature. These findings do not support the hypothesis that larvae exhibit better locomotor performance than adults as a result of stronger selective pressures on early life stages.

Movie available on-line: http://www.biologists.com/JEB/movies/jeb3978.html.
Kinematics of the buccal mass during swallowing based on magnetic resonance imaging in intact, behaving Aplysia californica
Authors: Neustadter David M. , Drushel Richard F. , Chiel Hillel J.
Published: April 2021

A novel magnetic resonance imaging interface has been developed that makes it possible to image movements in intact, freely moving subjects. We have used this interface to image the internal structures of the feeding apparatus (i.e. the buccal mass) of the marine mollusc Aplysia californica. The temporal and spatial resolution of the resulting images is sufficient to describe the kinematics of specific muscles of the buccal mass and the internal movements of the main structures responsible for grasping food, the radula and the odontophore. These observations suggest that a previously undescribed feature on the anterior margin of the odontophore, a fluid-filled structure that we term the prow, may aid in opening the jaw lumen early in protraction. Radular closing during swallowing occurs near the peak of protraction as the radular stalk is pushed rapidly out of the odontophore. Retraction of the odontophore is enhanced by the closure of the lumen of the jaws on the elongated odontophore, causing the odontophore to rotate rapidly towards the esophagus. Radular opening occurs after the peak of retraction and without the active contraction of the protractor muscle 12 and is due, in part, to the movement of the radular stalk into the odontophore. The large variability between responses also suggests that the great flexibility of swallowing responses may be due to variability in neural control and in the biomechanics of the ingested food and to the inherent flexibility of the buccal mass.
Expression ofManduca sextaV-ATPase genesmvB, mvGandmvdis regulated by ecdysteroids
Authors: Reineke Stephan , Wieczorek Helmut , Merzendorfer Hans
Published: April 2021

V-ATPases are complex proteins consisting of a peripheral, ATP-hydrolysing V1 complex and a membrane-bound H+-translocating Vo complex. The plasma membrane V-ATPase from the tobacco hornworm(Manduca sexta) midgut is made up of eight different V1and four different Vo subunits. During starvation and moulting,V-ATPase activity decreases as a result of the dissociation of the V1 complex from the Vo complex. To determine whether subunit biosynthesis is reduced during periods of enzyme inactivity, we measured the transcript levels and transcriptional activities of V-ATPase genes. Northern blots revealed the downregulation of almost all V-ATPase transcripts during starvation. During moulting, transcript levels of the three V-ATPase genes examined, mvB, mvG and mvd, also decreased,and this decrease was negatively correlated with the titre of 20-hydroxyecdysone (20-HE) and positively correlated with the titre of juvenile hormone (JH). To test the biological significance of these correlations, we injected both hormones into feeding larvae and measured transcript levels several hours later. A short-term increase and a long-term decrease in levels of mRNA were observed after 20-HE injection, whereas JH injection had no significant effect. Immunohistochemical studies of the midgut epithelium revealed that 20-HE injection led to changes in goblet cell morphology and in the subcellular distribution of the V1 complex comparable with the situation during the moult and during starvation. Reporter gene assays in Sf21 cells using mvB, mvG and mvdpromoters to initiate transcription of firefly luciferase led, after incubation of the cells with 20-HE, to results comparable with those obtained in the injection experiments. These findings suggest that putative ecdysone-responsive elements are present in all three promoters. Taken together, our results suggest that the expression of V-ATPase genes is controlled in a coordinated manner by ecdysteroids.
Reflections on colourful ommatidia of butterfly eyes
Authors: Stavenga Doekele G.
Published: April 2021

The eye shine of butterflies from a large number of ommatidia was observed with a modified epi-illumination apparatus equipped with an objective lens of large numerical aperture. A few representative cases are presented: the satyrine Bicyclus anynana, the heliconian Heliconius melpomene, the small white Pieris rapae and the small copper Lycaena phlaeas. The colour of the eye shine is determined mainly by the reflectance spectrum of the tapetal mirror and the transmittance spectrum of the photoreceptor screening pigments, if present near the light-guiding rhabdom. Reflectance spectra measured from individual ommatidia show that tapetum and screening pigments are co-expressed in fixed combinations, thus determining different ommatidial classes. The classes are distributed in an irregular pattern that can be rapidly assessed with the novel epi-illumination apparatus. Many butterfly species appear to have red-reflecting ommatidia,which is interpreted to indicate the presence of red-sensitive photoreceptors.
Performance and adaptive value of tarsal morphology in rove beetles of the genusStenus(Coleoptera, Staphylinidae)
Authors: Betz Oliver
Published: April 2021

To evaluate the adaptive value of the widening of the bilobed tarsi that has paralleled the tremendous radiation of the staphylinid genus Stenus, the performance of slender versus wide tarsi has been evaluated in two different contexts: (i) locomotion on the surface of water, and (ii) climbing on vertical (plant) surfaces. Contact angle measurements at the underside of the tarsi have revealed that, irrespective of tarsus width, all the investigated species are well supported by the surface of water while walking on it. The main selective demands driving the widening of the tarsi in several lineages have instead come from their firm attachment to smooth plant surfaces. This is suggested by measurements of the maximum vertical pulling forces exerted by intact and manipulated individuals on various rough and smooth surfaces. Species with widened tarsi associated with considerably more tenet setae attain significantly higher pulling forces,particularly on smooth surfaces. The tarsal setae are of greater importance on smooth surfaces, but the claws seem to be more important on rough substrata. On substrata that combine the attributes of rough and smooth surfaces, both claws and tenent setae add significantly to the pulling forces exerted,suggesting a functional synergism. The contribution of the present study to our understanding of insect tarsal attachment to surfaces with a variety of textures is discussed.
Analysis of monoamines, adenosine and GABA in tissues of the land snailHelix lucorumand lizardAgama stellio stellioduring hibernation
Authors: Michaelidis Basile , Loumbourdis Nikolaos S. , Kapaki Elizabeth
Published: April 2021

The aim of the present study was to determine the levels of monoamines,GABA and adenosine in the brain, heart and haemolymph of the land snail Helix lucorum and in the brain, heart and blood of lizard Agama stellio stellio during long-term hibernation. We measured levels of the monoamines serotonin (5-HT) and its main metabolite 5-hydroxyindole-3-acetic acid (5-HIAA), dopamine (DA) and its metabolites dihydroxyphenylacetic acid(DOPAC) and homovanilic acid (HVA), norepinephrine (NE) and epinephrine (E). The most abundant amines detected in the brain and heart of active H. lucorum were 5-HT and DA. Of the metabolites examined only 5-HIAA was found in the brain. NE was found at very low levels but only in the brain,while E was not detected in the brain and heart. The levels of 5-HT and 5-HIAA increased in the brain and heart of H. lucorum within the first months of hibernation, showing a significant decrease thereafter. The levels of DA did not change during hibernation. The results indicated that 5-HT might be involved in preparing snails for entry into hibernation. GABA was only found in the brain of H. lucorum, and the levels were low; these levels remained during hibernation. Adenosine was present in brain and heart of H. lucorum, and during hibernation, the level of adenosine decreased significantly in the brain but remained steady in the heart. The monoamines 5-HT, DA and NE were present in the brain of active lizards A. stellio stellio, whereas E was found only at very low levels. Moreover,the metabolites 5-HIAA, DOPAC and HVA were detected in the brain of active lizards. The monoamines 5-HT, DA, NE and E were also detected in the heart and blood of active lizards. During hibernation the levels of these four monoamines were decreased significantly in the brain and heart of A. stellio stellio. In contrast, the levels of E increased in the heart and blood of hibernating lizards. Adenosine was detected in both heart and brain of active lizards, but hibernation caused a marked decrease in its levels at both tissues. GABA was found at higher levels than monoamines and adenosine in the brain of active lizards, and hibernation caused a significant increase in its levels, indicating an important role of GABA in inhibition of neuronal activity in hibernating lizards.
Trade-offs between speed and endurance in the frogXenopus laevis
Authors: Wilson Robbie S. , James Rob S. , Van Damme Raoul
Published: April 2021

One of the most interesting trade-offs within the vertebrate locomotor system is that between speed and endurance capacity. However, few studies have demonstrated a conflict between whole-animal speed and endurance within a vertebrate species. We investigated the existence of trade-offs between speed and endurance capacity at both the whole-muscle and whole-animal levels in post-metamorphs of the frog Xenopus laevis. The burst-swimming performance of 55 frogs was assessed using a high-speed digital camera, and their endurance capacity was measured in a constant-velocity swimming flume.

The work-loop technique was used to assess maximum power production of whole peroneus muscles at a cycle frequency of 6 Hz, while fatigue-resistance was determined by recording the decrease in force and net power production during a set of continuous cycles at 2 Hz. We found no significant correlations between measures of burst swimming performance and endurance capacity, suggesting that there is no trade-off between these two measures of whole-animal performance. In contrast, there was a significant negative correlation between peak instantaneous power output of the muscles at 6 Hz and the fatigue-resistance of force production at 2 Hz (other correlations between power and fatigue were negative but non-significant). Thus, our data support the suggestion that a physiological conflict between maximum power output and fatigue resistance exists at the level of vertebrate muscles. The apparent incongruence between whole-muscle and whole-animal performance warrants further detailed investigation and may be related to factors influencing both whole-muscle and whole-animal performance measures.
Copper uptake across rainbow trout gills
Authors: Grosell Martin , Wood Chris M.
Published: April 2021

Several components of branchial copper uptake were identified in juvenile freshwater rainbow trout (Oncorhynchus mykiss) using 64Cu. On the basis of competitive interactions between sodium and copper uptake,inhibition of copper uptake by a proton pump inhibitor (bafilomycin A1, 2μmol l-1) and a Na+ channel blocker (phenamil, 100μmol l-1), it appears that a proportion of the branchial copper uptake occurs via an apical Na+ channel. This sodium-sensitive copper uptake demonstrates saturation kinetics, with a Km of 7.1 nmol l-1 and a Jmax of 21.2 pmol g-1 h-1, and is characterized by an IC50 of 104 μmol l-1 sodium. On the basis of residual copper uptake in the presence of high sodium concentrations (20 mmol l-1) and differential inhibition of sodium and copper uptake by phenamil (100 μmol l-1), a sodium-insensitive component of copper uptake is also present in trout gills. It demonstrates saturation kinetics with a comparably low Km (9.6 nmol l-1) but a lower maximum transport capacity (Jmax=3.5 pmol g-1 h-1)than the sodium-insensitive system. Sodium uptake exhibits saturation kinetics with a Km of 69 μmol l-1. Copper reduced branchial sodium transport affinity but increased the maximal sodium transport capacity.
Buoyancy and maximal diving depth in penguins
Authors: Sato Katsufumi , Naito Y. , Kato A. , Niizuma Y. , Watanuki Y. , Charrassin J. B. , Bost C.-A. , Handrich Y. , Le Maho Y.
Published: April 2021

Using a newly developed data logger to measure acceleration, we demonstrate that free-ranging king and Adélie penguins only beat their flippers substantially during the first part of descent or when they were presumed to be chasing prey at the bottom of dives. Flipper beating stopped during the latter part of ascent: at 29±9 % (mean ± S.D.) of dive depth(mean dive depth=136.8±145.1 m, N=425 dives) in king penguins,and at 52±20 % of dive depth (mean dive depth=72.9±70.5 m, N=664 dives) in Adélie penguins. Propulsive swim speeds of both species were approximately 2 m s-1 during dives; however, a marked increase in speed, up to approximately 2.9 m s-1, sometimes occurred in king penguins during the passive ascending periods. During the prolonged ascending, oblique ascent angle and slowdown near the surface may represent one way to avoid the potential risk of decompression sickness. Biomechanical calculations for data from free-ranging king and Adélie penguins indicate that the air volume of the birds (respiratory system and plumage) can provide enough buoyancy for the passive ascent. When comparing the passive ascents for shallow and deep dives, there is a positive correlation between air volume and the depth of the dive. This suggests that penguins regulate their air volume to optimize the costs and benefits of buoyancy.
Drosophilaas a new model organism for the neurobiology of aggression?
Authors: Baier Andrea , Wittek Britta , Brembs Björn
Published: April 2021

We report here the effects of several neurobiological determinants on aggressive behaviour in the fruitfly Drosophila melanogaster. This study combines behavioural, transgenic, genetic and pharmacological techniques that are well established in the fruitfly, in the novel context of the neurobiology of aggression. We find that octopamine, dopamine and a region in the Drosophila brain called the mushroom bodies, all profoundly influence the expression of aggressive behaviour. Serotonin had no effect. We conclude that Drosophila, with its advanced set of molecular tools and its behavioural richness, has the potential to develop into a new model organism for the study of the neurobiology of aggression.