Tight junctions (TJs) are an integral component of models illustrating ion transport mechanisms across fish epithelia; however, little is known about TJ proteins in fishes. Using immunohistochemical methods and Western blot analysis, we examined the localization and expression of occludin, a transmembrane TJ protein, in goldfish tissues. In goldfish gills, discontinuous occludin immunostaining was detected along the edges of secondary gill lamellae and within parts of the interlamellar region that line the lateral walls of the central venous sinus. In the goldfish intestine, occludin immunolocalized in a TJ-specific distribution pattern to apical regions of columnar epithelial cells lining the intestinal lumen. In the goldfish kidney, occludin was differentially expressed in discrete regions of the nephron. Occludin immunostaining was strongest in the distal segment of the nephron, moderate in the collecting duct and absent in the proximal segment. To investigate a potential role for occludin in the maintenance of the hydromineral balance of fishes, we subjected goldfish to 1, 2 and 4 weeks of food deprivation,...]]>
Parental care is a widespread and ecologically relevant adaptation known to enhance the developmental environment of offspring. Parental behaviors, however, may entail both costs and benefits for developing offspring. In Children's pythons (Antaresia childreni), we monitored both maternal egg-brooding behavior and intra-clutch oxygen partial pressure (PO2) in real-time to assess the effects of various brooding behaviors on PO2 in the clutch micro-environment at three stages of development. Furthermore, at the same developmental stages, we measured O2 consumption rates (VO2) of eggs at varying PO2 to determine their critical oxygen tension (i.e. the minimal PO2 that supports normal respiratory gas exchange) and to predict the impact that naturally brooded intra-clutch PO2 has on embryonic metabolism. At all three stages of development, a tightly coiled brooding posture created an intra-clutch PO2 that was significantly lower than the surrounding nest environment. Maternal postural adjustments alleviated this hypoxia, and the magnitude of such corrections increased with developmental stage. Mean intra-clutch PO2 decreased with stage of development, probably because of increasing egg VO2. Additionally, embryo critical oxygen...]]>
We employ numerical simulation to investigate the hydrodynamics of carangiform locomotion as the relative magnitude of viscous and inertial forces, i.e. the Reynolds number (Re), and the tail-beat frequency, i.e. the Strouhal number (St), are systematically varied. The model fish is a three-dimensional (3D) mackerel-like flexible body undulating with prescribed experimental kinematics of carangiform type. Simulations are carried out for three Re spanning the transitional and inertial flow regimes, Re=300 and 4000 (viscous flow), and (inviscid flow). For each Re there is a critical Strouhal number, St*, at which the net mean force becomes zero, making constant-speed self-propulsion possible. St* is a decreasing function of Re and approaches the range of St at which most carangiform swimmers swim in nature (St~0.25) only as Re approaches infinity. The propulsive efficiency at St* is an increasing function of Re while the power required for swimming is decreasing with Re. For all Re, however, the swimming power is shown to be significantly greater than that required to tow the...]]>
Jawless fishes (Agnatha; lampreys and hagfishes) most closely resemble the earliest stage in vertebrate evolution and lamprey-like animals already existed in the Lower Cambrian [about 540 million years ago (MYA)]. Agnathans are thought to have separated from the main vertebrate lineage at least 500 MYA. Hagfishes have primitive eyes, but the eyes of adult lampreys are well-developed. The southern hemisphere lamprey, Geotria australis, possesses five types of opsin genes, three of which are clearly orthologous to the opsin genes of jawed vertebrates. This suggests that the last common ancestor of all vertebrate lineages possessed a complex colour vision system. In the eyes of many bony fishes and tetrapods, well-focused colour images are created by multifocal crystalline lenses that compensate for longitudinal chromatic aberration. To trace the evolutionary origins of multifocal lenses, we studied the optical properties of the lenses in four species of lamprey (Geotria australis, Mordacia praecox, Lampetra fluviatilis and Petromyzon marinus), with representatives from all three of the extant lamprey families. Multifocal lenses...]]>
Hypoxic pulmonary vasoconstriction (HPV) is an adaptive response that diverts pulmonary blood flow from poorly ventilated and hypoxic areas of the lung to better ventilated parts, matching blood perfusion to ventilation. HPV is an ancient and highly conserved response expressed in the respiratory organs of all vertebrates. However, the underlying mechanism and the role of the endothelium remain elusive. Isolated intrapulmonary arteries (internal diameter <346 µm) from the American alligator Alligator mississippiensis were mounted in microvascular myographs for isometric tension recording. Resting vessels and vessels contracted with either serotonin (5-HT) or endothelin-1 (ET-1) were exposed to sustained (45 min) hypoxia (PO2<5 mmHg). In ET-1-contracted vessels, hypoxia induced a monophasic, sustained and fully reversible constriction, which was independent of the endothelium. In relaxed or in 5-HT-contracted vessels, hypoxia did not cause constriction. The effects of ET-1, ETA and ETB as well as the general ET-receptor antagonist were studied. ET-1 caused a contraction of the pulmonary arteries through stimulation of ETA-receptors. ETA and ETB immunoreactive staining revealed the location...]]>
The landing–take-off asymmetry of running was thought to derive from, or at least to be consistent with, the physiological property of muscle to resist stretching (after landing) with a force greater than it can develop during shortening (before take-off). In old age, muscular force is reduced, but the deficit in force is less during stretching than during shortening. The greater loss in concentric versus eccentric strength with aging led us to hypothesize that older versus younger adults would increase the landing–take-off asymmetry in running. To test this hypothesis, we measured the within-step changes in mechanical energy of the centre of mass of the body in old and young subjects. The difference between the peaks in kinetic energy attained during the fall and during the lift of the centre of mass is greater in the old subjects. The difference between the time to lift and accelerate the centre of mass (positive work) and to absorb the same amount of energy during the downward displacement...]]>
We investigated photoacclimation in the symbiotic Mediterranean coral Cladocora caespitosa by exposing it to three light levels (30, 80 and 250 µmol m–2 s–1), which are in the range of those recorded for this species. The coral response to a change in both light and temperature was also assessed, by subjecting coral to two treatments corresponding to winter (14°C and 30 µmol m–2 s–1) and summer (23°C and 250 µmol m–2 s–1) conditions, as measured in the Ligurian Sea. Photosynthesis, measured using both respirometry and pulse amplitude modulated (PAM) fluorometry, revealed a linear relationship only at low light levels. At higher irradiance, relative electron transport rate (rETR) approached saturation more slowly than rates of oxygen production. At constant temperature, a change in light did not induce any change in zooxanthellae (zoox) and chlorophyll (Chla+c2) concentrations (mean 3.7x106 zoox cm–2 and 14.1 µg cm–2, respectively); however, chlorophyll concentrations significantly increased under low light and temperature, probably in order to maintain a sufficient level of autotrophy. Maximal gross photosynthesis (Pgmax)...]]>
Apart from regulating insect development, juvenile hormones (JHs) play an
important role in insect reproduction, where they initiate vitellogenin
synthesis and regulate the uptake of yolk by the ovary. JH synthesis is a
tightly regulated process controlled by neurons and peptidergic neurosecretory
cells. One of the known stimulatory regulators of JH biosynthesis is
glutamate, and its N-methyl-
The nutrient amino acid transporter (NAT) subfamily is the largest subdivision of the sodium neurotransmitter symporter family (SNF; also known as SLC6; HUGO). There are seven members of the NAT population in the African malaria mosquito Anopheles gambiae, two of which, AgNAT6 and AgNAT8, preferably transport indole- and phenyl-branched substrates, respectively. The relative expression and distribution of these aromatic NATs were examined with transporter-specific antibodies in Xenopus oocytes and mosquito larval alimentary canal, representing heterologous and tissue expression systems, respectively. NAT-specific aromatic-substrate-induced currents strongly corresponded with specific accumulation of both transporters in the plasma membrane of oocytes. Immunolabeling revealed elevated expressions of both transporters in specific regions of the larval alimentary canal, including salivary glands, cardia, gastric caeca, posterior midgut and Malpighian tubules. Differences in relative expression densities and spatial distribution of the transporters were prominent in virtually all of these regions, suggesting unique profiles of the aromatic amino acid absorption. For the first time reversal of the location of a transporter between apical and basal membranes...]]>
We studied the mechanical properties of deep red aerobic muscle of yellowfin tuna (Thunnus albacares), using both in vivo and in vitro methods. In fish swimming in a water tunnel at 1–3 L s–1 (where L is fork length), muscle length changes were recorded by sonomicrometry, and activation timing was quantified by electromyography. In some fish a tendon buckle was also implanted on the caudal tendon to measure instantaneous muscle forces transmitted to the tail. Between measurement sites at 0.45 to 0.65 L, the wave of muscle shortening progressed along the body at a relatively high velocity of 1.7 L per tail beat period, and a significant phase shift (31±4°) occurred between muscle shortening and local midline curvature, both suggesting red muscle power is directed posteriorly, rather than causing local body bending, which is a hallmark of thunniform swimming. Muscle activation at 0.53 L was initiated at about 50° of the tail beat period and ceased at about 160°, where 90° is peak muscle length...]]>
Separate studies of terrestrial and aquatic locomotion are abundant, but research addressing locomotion in transitional environments (e.g. mud) is scant. The African lungfish (Protopterus annectens) moves in a gradation of water to mud conditions during seasonal droughts, and breathes air. Thus, the lungfish was an ideal organism for our study to determine the effects of a wide range of viscosities on lateral undulatory swimming and to simulate some of the muddy conditions early tetrapods may have encountered. Regardless of viscosity, several aspects of lungfish swimming were similar to those of other swimming vertebrates including: posteriorly propagated muscle activity that was unilateral and alternated between the left and right sides at each longitudinal location, and posterior increases in the amount of bending, the amplitude of muscle activity and the timing differences between muscle activity and bending. With increased viscosity (1–1000 cSt), significant increases occurred in the amount of lateral bending of the vertebral column and the amplitude of muscle activity, particularly in the most anterior sites,...]]>
Addiction is a complex psychiatric disorder considered to be a disease of the brain's natural reward reinforcement system. Repeated stimulation of the `reward' pathway leads to adaptive changes in gene expression and synaptic organization that reinforce drug taking and underlie long-term changes in behaviour. The primitive nature of reward reinforcement pathways and the near universal ability of abused drugs to target the same system allow drug-associated reward and reinforcement to be studied in non-mammalian species. Zebrafish have proved to be a valuable model system for the study of vertebrate development and disease. Here we demonstrate that adult zebrafish show a dose-dependent acute conditioned place preference (CPP) reinforcement response to ethanol or nicotine. Repeated exposure of adult zebrafish to either nicotine or ethanol leads to a robust CPP response that persists following 3 weeks of abstinence and in the face of adverse stimuli, a behavioural indicator of the establishment of dependence. Microarray analysis using whole brain samples from drug-treated and control zebrafish identified 1362 genes that...]]>
A honeybee's waggle dance is an intriguing example of multisensory convergence, central processing and symbolic information transfer. It conveys to bees and human observers the position of a relatively small area at the endpoint of an average vector in a two-dimensional system of coordinates. This vector is often computed from a collection of waggle phases from the same or different dancers. The question remains, however, of how informative a small sample of waggle phases can be to the bees, and how the spatial information encoded in the dance is actually mapped to the followers' searches in the field. Certainly, it is the variability of a dancer's performance that initially defines the level of uncertainty that followers must cope with if they were to successfully decode information in the dance. Understanding how a dancer's behaviour is mapped to that of its followers initially relies on the analysis of both the accuracy and precision with which the dancer encodes spatial information in the dance. Here...]]>
Insect ecdysis is a precisely coordinated series of behavioral and hormonal events that occur at the end of each molt. A great deal is known about the hormonal events that underlie this process, although less is known about the neuronal circuitry involved. In this study we identified two populations of neurons that are required for larval and adult ecdyses in the fruit fly, Drosophila melanogaster (Meigen). These neurons were identified by using the upstream region of two genes that code for atypical soluble guanylyl cyclases to drive tetanus toxin in the neurons that express these cyclases to block their synaptic activity. Expression of tetanus toxin in neurons that express Gyc-89Da blocked adult eclosion whereas expression of tetanus toxin in neurons that express Gyc-89Db prevented the initiation of the first larval ecdysis. Expression of tetanus toxin in the Gyc-89Da neurons also resulted in about 50% lethality just prior to pupariation; however, this was probably due to suffocation in the food as lethality was prevented...]]>
Brown ghost knifefish, Apteronotus leptorhynchus, are a species of weakly electric fish that produce a continuous electric organ discharge (EOD) that is used in navigation, prey capture and communication. Stereotyped modulations of EOD frequency and amplitude are common in social situations and are thought to serve as communication signals. Of these modulations, the most commonly studied is the chirp. This study presents a quantitative analysis of chirp production in pairs of free-swimming, physically interacting male and female A. leptorhynchus. Under these conditions, we found that in addition to chirps, the fish commonly produce a second signal type, a type of frequency rise called abrupt frequency rises, AFRs. By quantifying the behaviours associated with signal production, we find that Type 2 chirps tend to be produced when the fish are apart, following periods of low aggression, whereas AFRs tend to be produced when the fish are aggressively attacking one another in close proximity. This study is the first to our knowledge that quantitatively describes both...]]>
In finches husking time is non-linearly related to the ratio of seed hardness to maximal bite force. Fringillids produce larger bite force and husk relatively hard seeds faster than estrildids of similar size. This is at least partly explained by their relatively larger jaw muscle mass and a difference in husking technique. However, the effect of differences in skull geometry on bite force is unclear. In this study differences in skull morphology that may contribute to the difference in bite force between fringillids and estrildids are analyzed. The shape of the skull was described by the 3D coordinates of a set of landmarks and, after eliminating size, the effect of differences in the shape of the skull on bite force was determined using a static force model. EMG recordings of jaw muscles during seed cracking were used to validate assumptions about the muscle activation patterns used for the static bite force model. The analysis shows that most of the variation in skull...]]>
Otophysine fish possess Weberian ossicles, which connect the swimbladder to the inner ear and improve hearing ability. There is a high diversity in the morphology of the swimbladder and Weberian apparatus in catfishes, which might affect hearing. We have examined these structures in representatives of six families with large, single bladders (Ariidae, Auchenipteridae, Heptapteridae, Malapteruridae, Mochokidae, Pseudopimelodidae) and five subfamilies from two families (Callichthyidae, Loricariidae) having small, paired, encapsulated bladders. We tested their hearing abilities utilizing the non-invasive auditory evoked potential recording technique. Species with single, non-encapsulated, free airbladders possess one, three or four ossicles, whereas species with encapsulated bladders possess one or two. The relative sizes of the bladders and ossicles were significantly smaller in the latter group. All species were able to detect sound stimuli between 50 Hz and 5 kHz. Interspecific differences in hearing sensitivity varied at most by 24 dB below 1 kHz, whilst this variation increased to more than 50 dB at higher frequencies. Catfishes with free bladders had lower...]]>
The barrier to water loss from the skin of birds and mammals is localized in the stratum corneum (SC), the outer layer of the epidermis. The SC consists of corneocytes, each surrounded by a protein envelope, and a lipid compartment, formed by an extracellular matrix of lipids and by lipids covalently bound to the protein envelope. In mammals, covalently bound lipids in the SC consist of -hydroxyceramides attached to the outer surface of corneocytes. Evidence suggests that covalently bound lipids in the SC might be crucial for the establishment of a competent permeability barrier. In this study we assessed the composition of covalently bound lipids of the avian SC and their relationship to cutaneous water loss (CWL) in two populations of house sparrows, one living in the deserts of Saudi Arabia and the other in mesic Ohio. Previously, we showed that CWL of adult desert sparrows was 25% lower than that of mesic birds. In the present study we characterize covalently bound lipids...]]>
Developing animals may face a cost–benefit tradeoff during growth mediated through hormones such as glucocorticoids, as the hormone is essential for development but can have detrimental consequences. To investigate potential tradeoffs caused by brief, moderate elevations of corticosterone in avian young, we artificially elevated the hormone levels in two ways: feeding corticosterone-containing worms and applying corticosterone dermal patches. The former experiment tested the effects of an acute corticosterone elevation (25 min) on begging behavior, whereas the latter explored the effects of artificially elevated corticosterone for 24 to 48 h on growth. Corticosterone altered both begging behavior and growth of white-crowned sparrow nestlings. It increased latency to beg immediately after the treatment and suppressed growth as early as 24 h after the patch application. These experiments also showed that the effects depended on the age or types of development (e.g. gaining mass or growing feathers) that the nestlings were going through.
]]>One of the most important decisions any animal has to make is how to respond to sensory cues that suggest an imminent attack by a predator. We measured behavioral and neural responses of juvenile crayfish to moving shadows of different velocities while the animals were searching for food. In all experiments, and independent of shadow velocity, each crayfish produced one of two discrete behavioral outputs: it either tail-flipped backwards by rapid flexion of its abdomen or it immediately stopped its forward locomotion. The probability of each behavioral response was dependent on the velocity of the shadows that were presented. While most animals responded with tail-flips to slow-moving shadows and stops were rarely observed, the number of tail-flips decreased as shadow velocity increased. Tail-flips were almost absent for very fast-moving shadows and stopping behavior became the dominating response. By using a non-invasive technique to record neural activity, we were able to identify the underlying neural circuit that controlled the observed tail-flips. All tail-flips were mediated...]]>
The evolutionary adaptations of functional genes to life at high pressure are not well understood. To elucidate the mechanisms of protein adaptation to high pressure, we cloned the myosin heavy chain (MyHC) cDNA from skeletal muscle of two deep-sea fishes, Coryphaenoides yaquinae and C. armatus, and two non-deep-sea fishes, C. acrolepis and C. cinereus. The MyHCs of deep-sea fishes have a unique structure in two loop regions, loop-1 and loop-2, in comparison with those of non-deep-sea fishes. The loop-1 region of deep-sea fishes has a Pro residue and the loop-2 region, which is an actin-binding site, is shorter than the same region in non-deep-sea fishes. The amino acid substitution in the loop-1 region is expected to be mainly involved in ATPase activity, whereas the deletion in the loop-2 region affects the association of MyHC with actin filaments at high pressure. In addition, the MyHC of deep-sea fishes has biased amino acid substitutions at core positions in the coiled-coil structure of the rod region. These...]]>
The aim of the present study was to examine the biomechanics of the lower limb during stair descent and the effects of increasing demand in two ways: by increasing step-height and by increasing body mass. Ten male subjects walked down a four-step staircase, the height of which could be altered. The step-heights were: standard (17 cm), 50% decreased, 50% increased and 75% increased. At the standard height, subjects also walked down wearing a weighted jacket carrying 20% extra body mass. Lower limb kinematics and kinetics were determined using motion capture and ground reaction forces. Also measured were gastrocnemius medialis (GM) muscle electromyography and GM muscle fascicle length using ultrasonography. GM muscle fascicles actively shortened during the touch-down phase of stair descent in all conditions, while the muscle–tendon complex (MTC), as calculated from the knee and ankle joint kinematics, lengthened. The GM muscle fascicles shortened more when step-height was increased, which corresponded to the increase in ankle joint moment. Increased body mass did not alter...]]>
A number of teleost fishes have photoreceptor mechanisms to detect linearly polarized light. We studied the neuronal mechanism underlying this ability. It was found that a polarized signal could be detected in rainbow trout (Oncorhynchus mykiss) both in the electroretinogram (ERG) and in the compound action potential (CAP) measured in the optic nerve, indicating a strong retinal contribution to the processing of polarized light. The CAP recordings showed a W-shaped sensitivity curve, with a peak at 0°, 90° and 180°, consistent with processes for both vertical and horizontal orientation. By contrast, the ERG recordings reveal a more complex pattern. In addition to the peaks at 0°, 90° and 180°, two additional peaks appeared at 45° and 135°. This result suggests a specialized contribution of the outer retina in the processing of polarized light. The spectral sensitivity of the mechanisms responsible for these intermediate peaks was studied using chromatic adaptation. Here we show that long wavelength-sensitive (LWS) cone mechanism adaptation shifted the intermediate peaks towards 90°,...]]>
Many fish change gait within their aerobically supported range of swimming speeds. The effects of acute temperature change on this type of locomotor behavior are poorly understood. Bluegill sunfish swim in the labriform mode at low speeds and switch to undulatory swimming as their swimming speed increases. Maximum aerobic swimming speed (Umax), labriform-undulatory gait transition speed (Utrans) and the relationships between fin beat frequency and speed were measured at 14, 18, 22, 26 and 30°C in bluegill acclimated to 22°C. At temperatures below the acclimation temperature (Ta), Umax, Utrans and the caudal and pectoral fin beat frequencies at these speeds were reduced relative to the acclimation level. At temperatures above Ta there was no change in these variables relative to the acclimation level. Supplementation of oxygen levels at 30°C had no effect on swimming performance. The mechanical power output of the abductor superficialis, a pectoral fin abductor muscle, was measured in vitro at the same temperatures used for the swimming experiments. At and below Ta, maximal...]]>
Previous findings in pigeons and chickens show that Ca2+ may be accumulated inside the cornified skin cells and that Ca2+ microenvironments with a lower- or higher-than-blood concentration may exist in the skin. It has been suggested that the skin may function as a secretory pathway or a reservoir for Ca2+ recycling. To test this hypothesis, we studied the dermis and epidermis of female and male chickens in vivo to find out whether cellular mechanisms exist for the accumulation, recycling or secretion of Ca2+. For calcium influx and intracellular Ca2+ release, respectively, the density of dihydropyridine receptors (DHPRs) and ryanodine receptors (RyRs) was examined, using high-affinity (–)-enantiomers of dihydropyridine and ryanodine labelled with fluorophores. To investigate Ca2+ utilization in the skin, the systemic and local activity of the enzyme alkaline phosphatase (ALP) and the concentration of ionic Ca2+ were measured in plasma and in cutaneous extracellular fluid, collected by suction blister technique. We found that both DHPRs and RyRs were present in all skin layers from...]]>
Robotic lower limb exoskeletons that can alter joint mechanical power output are novel tools for studying the relationship between the mechanics and energetics of human locomotion. We built pneumatically powered ankle exoskeletons controlled by the user's own soleus electromyography (i.e. proportional myoelectric control) to determine whether mechanical assistance at the ankle joint could reduce the metabolic cost of level, steady-speed human walking. We hypothesized that subjects would reduce their net metabolic power in proportion to the average positive mechanical power delivered by the bilateral ankle exoskeletons. Nine healthy individuals completed three 30 min sessions walking at 1.25 m s–1 while wearing the exoskeletons. Over the three sessions, subjects' net metabolic energy expenditure during powered walking progressed from +7% to –10% of that during unpowered walking. With practice, subjects significantly reduced soleus muscle activity (by ~28% root mean square EMG, P<0.0001) and negative exoskeleton mechanical power (–0.09 W kg–1 at the beginning of session 1 and –0.03 W kg–1 at the end of session 3; P=0.005). Ankle...]]>
Mothers can strongly influence the development of their offspring, and if maternal resources are limited, they may influence optimal reproductive strategies. In birds, maternally deposited carotenoids are a prominent component of egg yolk and are vital for the development of the embryo. However, results of long-lasting fitness consequences of this early nutritional environment have been scarce and inconsistent. In addition, sex-biased sensitivity to different egg components is one of the mechanisms postulated to account for sex-linked environmental vulnerability during early life. However, this important aspect is usually not accounted for when investigating maternal investment in carotenoids. In this study we gave carotenoid (lutein) supplements to female Eurasian kestrels (Falco tinnunculus) before and during egg laying. The experiment increased female plasma carotenoids, but this effect was not apparent in hatchling and fledgling plasma carotenoid concentration. Also, results showed that carotenoid supplementation increased the high density lipoprotein to low density lipoprotein ratio in adult females, suggesting that dietary carotenoids may influence lipid metabolism. Furthermore, the effect of the treatment...]]>
In the last decade, there has been an increased awareness that secondary metabolites produced by marine diatoms negatively impact the reproductive success of their principal predators, the copepods. Several oxylipins, products of the enzymatic oxidation of fatty acids, are produced when these unicellular algae are damaged, as occurs during grazing. In the past, the dinoflagellate Prorocentrum minimum, which does not produce the oxylipin 2-trans,4-trans-decadienal (DD), has been used as a live carrier to calculate daily ingestion rates of this molecule by copepod crustaceans. However, since the interaction between oxylipins and live carriers is unknown, the question as to how much and for how long ingestion of these molecules affects copepod reproduction remains a critical point to understanding the functional role of such compounds at sea. In the investigation presented here we used giant liposomes (~7 µm) as a delivery system for the oxylipin DD, prepared in the same size range as copepod food and containing known amounts of DD. The aim of this work...]]>
Modulation of neural circuits in the crustacean stomatogastric nervous system (STNS) allows flexibility in the movements of the foregut musculature. The extensive repertoire of such resulting motor patterns in dietary generalists is hypothesized to permit these animals to process varied foods. The foregut and STNS of Pugettia producta are similar to those of other decapods, but its diet is more uniform, consisting primarily of kelp. We investigated the distribution of highly conserved neuromodulators in the stomatogastric ganglion (STG) and neuroendocrine organs of Pugettia, and documented their effects on its pyloric rhythm. Using immunohistochemistry, we found that the distributions of Cancer borealis tachykinin-related peptide I (CabTRP I), crustacean cardioactive peptide (CCAP), proctolin, red pigment concentrating hormone (RPCH) and tyrosine hydroxylase (dopamine) were similar to those of other decapods. For all peptides except proctolin, the isoforms responsible for the immunoreactivity were confirmed by mass spectrometry to be the authentic peptides. Only two modulators had physiological effects on the pyloric circuit similar to those seen in other species. In...]]>
In mammals, the peroxisome proliferator-activated receptor (PPAR) coactivator-1 (PGC-1) family members and their binding partners orchestrate remodelling in response to diverse challenges such as diet, temperature and exercise. In this study, we exposed goldfish to three temperatures (4, 20 and 35°C) and to three dietary regimes (food deprivation, low fat and high fat) and examined the changes in mitochondrial enzyme activities and transcript levels for metabolic enzymes and their genetic regulators in red muscle, white muscle, heart and liver. When all tissues and conditions were pooled, there were significant correlations between the mRNA for the PGC-1 coactivators (both and β) and mitochondrial transcripts (citrate synthase), metabolic gene regulators including PPAR, PPARβ and nuclear respiratory factor-1 (NRF-1). PGC-1β was the better predictor of the NRF-1 axis, whereas PGC-1 was the better predictor of the PPAR axis (PPAR, PPARβ, medium chain acyl CoA dehydrogenase). In contrast to these intertissue/developmental patterns, the response of individual tissues to physiological stressors displayed no correlations between mRNA for PGC-1 family...]]>
Here we report the partial nucleotide sequence of a reptilian uncoupling protein (repUCP) gene from the European common lizard (Lacerta vivipara). Overlapping sequence analysis reveals that the protein shows 55%, 72% and 77% sequence homology with rat UCP1, UCP2 and UCP3, respectively, and 73% with bird and fish UCPs. RepUCP gene expression was ubiquitously detected in 4°C cold-acclimated lizard tissues and upregulated in muscle tissues by a 20 h exposure to sub-zero temperatures in a supercooling state or after thawing. In parallel, we show an increase in the co-activators, peroxisome proliferator-activated receptor coactivator-1 (PGC-1) and peroxisome proliferator-activated receptors (PPAR), mRNA expression, suggesting that the mechanisms regulating UCP expression may be conserved between mammals (endotherms) and reptiles (ectotherms). Furthermore, mitochondria extracted from lizard skeletal muscle showed a guanosine diphosphate (GDP)-sensitive non phosphorylating respiration. This last result indicates an inhibition of extra proton leakage mediated by an uncoupling protein, providing arguments that repUCP is functional in lizard tissues. This result is associated with a remarkable GDP-dependent increase...]]>
The centrally located, mitochondria-poor (CMP) and superficially located, mitochondria-rich (SMR) circular muscle fibres in the mantles of some squids provide one of the few known examples of specialization in an obliquely striated muscle. Little is known of the mechanical properties or of the mechanisms and performance consequences of specialization in these fibres. We combined morphological and physiological approaches to study specialization in the SMR and CMP fibres of the long-finned squid Doryteuthis pealeii. The mean thick filament length was 3.12±0.56 µm and 1.78±0.27 µm for the SMR and CMP fibres, respectively. The cross-sectional areas of the whole fibre and the core of mitochondria were significantly higher in the SMR fibres, but the area occupied by the myofilaments did not differ between the two fibre types. The area of sarcoplasmic reticulum visible in cross sections was significantly higher in CMP fibres than in SMR fibres. In live bundles of muscle fibres partially isolated from the mantle, mean peak isometric stress during tetanus was significantly greater...]]>
Food resources are complementary for a forager if their contribution to fitness is higher when consumed together than when consumed independently, e.g. ingesting one may reduce the toxic effects of another. The concentration of potentially toxic ethanol, [EtOH], in fleshy fruit increases during ripening and affects food choices by Egyptian fruit bats, becoming deterrent at high concentrations (>=1%). However, ethanol toxicity is apparently reduced when ingested along with some sugars; more with fructose than with sucrose or glucose. We predicted (1) that ingested ethanol is eliminated faster by bats eating fructose than by bats eating sucrose or glucose, (2) that the marginal value of fructose-containing food (food+fructose) increases with increasing [EtOH] more than the marginal value of sucrose- or glucose-containing food (food+sucrose, food+glucose), and (3) that by increasing [EtOH] the marginal value of food+sucose is incremented more than that of food+glucose. Ethanol in bat breath declined faster after they ate fructose than after eating sucrose or glucose. When food [EtOH] increased, the marginal value of...]]>
The estuarine crocodile, Crocodylus porosus, inhabits both freshwater and hypersaline waterways and maintains ionic homeostasis by excreting excess sodium and chloride ions via lingual salt glands. In the present study, we sought to investigate the phenotypic plasticity, both morphological and functional, in the lingual salt glands of the estuarine crocodile associated with chronic exposure to freshwater (FW) and saltwater (SW) environments. Examination of haematological parameters indicated that there were no long-term disruptions to ionic homeostasis with prolonged exposure to SW. Maximal secretory rates from the salt glands of SW-acclimated animals (100.8±14.7 µmol 100 g–0.7 body mass h–1) were almost three times greater than those of FW-acclimated animals (31.6±6.2 µmol 100 g–0.7 body mass h–1). There were no differences in the mass-specific metabolic rate of salt gland tissue slices from FW- and SW-acclimated animals (558.9±49.6 and 527.3±142.8 µl O2 g–1 h–1, respectively). Stimulation of the tissue slices from SW-acclimated animals by methacholine resulted in a 33% increase in oxygen consumption rate. There was no significant increase in the metabolic...]]>
Horned aphids (Cerataphidini) fight each other for access to feeding sites on leaves. An attacker attempts to force another aphid to abandon its feeding site; the victor then appears to insert its stylets into the site relinquished by the loser. This study used electrical penetration graph (EPG) recordings of Astegopteryx pallida (Van der Goot) individuals to pinpoint the benefits of fighting. We show that victors take significantly less time to commence feeding in the phloem, measured from the start of probing with their stylets, compared with aphids that initiate a new or discover an abandoned site: 9.0 versus 22.9 min, respectively. We also recorded the behaviour of aphids unencumbered with the wire necessary for EPG recordings. Those adult aphids that acquired a feeding site through fighting commenced feeding on average 20 min earlier than those that did not, taking into account the time spent searching and fighting as well as probing. This study clearly establishes that horned aphids use the exact feeding site...]]>
The correlation between ontogenetic changes in the spectral absorption characteristics of retinal photoreceptors and expression of visual pigment opsins was investigated in the black bream, Acanthopagrus butcheri. To establish whether the spectral qualities of environmental light affected the complement of visual pigments during ontogeny, comparisons were made between fishes reared in: (1) broad spectrum aquarium conditions; (2) short wavelength-reduced conditions similar to the natural environment; or (3) the natural environment (wild-caught). Microspectrophotometry was used to determine the wavelengths of spectral sensitivity of the photoreceptors at four developmental stages: larval, post-settlement, juvenile and adult. The molecular sequences of the rod (Rh1) and six cone (SWS1, SWS2A and B, Rh2A and β, and LWS) opsins were obtained and their expression levels in larval and adult stages examined using quantitative RT-PCR. The changes in spectral sensitivity of the cones were related to the differing levels of opsin expression during ontogeny. During the larval stage the predominantly expressed opsin classes were SWS1, SWS2B and Rh2A, contrasting with SWS2A, Rh2Aβ and LWS in...]]>
Sciaenid fishes are important models of fish sound production, but investigations into their auditory abilities are limited to acoustic pressure measurements on five species. In this study, we used auditory brainstem response (ABR) to assess the pressure and particle acceleration thresholds of six sciaenid fishes commonly found in Chesapeake Bay, eastern USA: weakfish (Cynoscion regalis), spotted seatrout (Cynoscion nebulosus), Atlantic croaker (Micropogonias undulatus), red drum (Sciaenops ocellatus), spot (Leiostomus xanthurus) and northern kingfish (Menticirrhus saxatilis). Experimental subjects were presented with pure 10 ms tone bursts in 100 Hz steps from 100 Hz to 1.2 kHz using an airborne speaker. Sound stimuli, monitored with a hydrophone and geophone, contained both pressure and particle motion components. Sound pressure and particle acceleration thresholds varied significantly among species and between frequencies; audiograms were notably flatter for acceleration than pressure at low frequencies. Thresholds of species with diverticulae projecting anteriorly from their swim bladders (weakfish, spotted seatrout, and Atlantic croaker) were typically but not significantly lower than those of species lacking such...]]>
The transition from juvenile to adult in the Australian western rock lobster, Panulirus cygnus (George), is preceded by a mass migration from inshore nursery reefs to offshore breeding grounds. Associated with this migration is a moult which results in the animals that are due to migrate undergoing characteristic colour change from deep red to pale pink, known as the `white' phase, which is believed to be triggered by environmental factors. To investigate this phenomenon, the colour change of wild-caught animals was measured over two separate years in response to two important modifiers of crustacean shell colour, dietary carotenoid and background substrate colour. Changes in shell colour during this colour transition period were influenced more greatly by other factors independent of diet or background substrate and no mass colour change was induced during this time. Shell colour measurement and carotenoid quantification confirmed the presence of animals similar to wild-caught `whites', regardless of the treatment. From these experimental observations we infer that the `white' phase of...]]>
To investigate the diving heart rate (fH) response of the emperor penguin (Aptenodytes forsteri), the consummate avian diver, birds diving at an isolated dive hole in McMurdo Sound, Antarctica were outfitted with digital electrocardiogram recorders, two-axis accelerometers and time depth recorders (TDRs). In contrast to any other freely diving bird, a true bradycardia (fH significantly <fH at rest) occurred during diving [dive fH (total beats/duration)=57±2 beats min–1, fH at rest=73±2 beats min–1 (mean ± s.e.m.)]. For dives less than the aerobic dive limit (ADL; duration beyond which [blood lactate] increases above resting levels), dive fH=85±3 beats min–1, whereas fH in dives greater than the ADL was significantly lower (41±1 beats min–1). In dives greater than the ADL, fH reached extremely low values: fH during the last 5 mins of an 18 min dive was 6 beats min–1. Dive fH and minimum instantaneous fH during dives declined significantly with increasing dive duration. Dive fH was independent of swim stroke frequency. This suggests that progressive bradycardia and peripheral vasoconstriction (including isolation of...]]>
The ability to navigate long distances to find rewarding flowers and return home is a key factor in the survival of honeybees (Apis mellifera). To reliably perform this task, bees combine both odometric and landmark cues, which potentially creates a dilemma since environments rich in odometric cues might be poor in salient landmark cues, and vice versa. In the present study, honeybees were provided with differential conditioning to images of complex natural scenes, in order to determine if they could reliably learn to discriminate between very similar scenes, and to recognise a learnt scene from a novel distractor scene. Choices made by individual bees were modelled with signal detection theory, and bees demonstrated an ability to discriminate between perceptually similar target and distractor views despite similar spatiotemporal content of the images. In a non-rewarded transfer test bees were also able to recognise target stimuli from novel distractors. These findings indicate that visual processing in bees is sufficiently accurate for recognising views of complex scenery...]]>
Studies of limb bone loading during terrestrial locomotion have focused primarily on birds and mammals. However, data from a broader functional and phylogenetic range of species are critical for understanding the evolution of limb bone function and design. Turtles are an interesting lineage in this context. Although their slow walking speeds and robust limb bones might lead to low locomotor forces and limb bone stresses similar to other non-avian reptiles, their highly sprawled posture could produce high bending loads, leading to high limb bone stresses similar to those of avian and mammalian species, as well as high torsion. To test between these possibilities, we evaluated stresses experienced by the femur of river cooter turtles (Pseudemys concinna) during terrestrial walking by synchronizing measurements of three-dimensional joint kinematics and ground reaction forces (GRFs) during isolated hindlimb footfalls. Further, we evaluated femoral safety factors for this species by comparing our locomotor stress calculations with the results of mechanical property tests. The net GRF magnitude at peak tensile bone...]]>
The vocal sac is a visually conspicuous attribute of most male frogs, but its role in visual communication has only been demonstrated recently in diurnally displaying frogs. Here we characterized the spectral properties of the inflated vocal sac of male túngara frogs (Physalaemus pustulosus), a nocturnal species, and túngara visual sensitivity to this cue across reproductive state and sex. We measured the spectral and total reflectance of different male body regions, including inflated and non-inflated vocal sacs, along with samples of the visual background against which males are perceived. Inflated vocal sacs were the most reflective of all body parts, being one log unit more reflective than background materials. We utilized an optomotor drum with black stripes and stripes that mimicked the spectral reflectance of the inflated vocal sacs with various nocturnal light intensities to measure the visual sensitivity thresholds of males, non-reproductive females and reproductive females. All three groups exhibited visual sensitivities corresponding to intensities below moonless conditions in open habitats or at the...]]>
At most joints, there is a redundancy of muscle function. For any given movement, there are a wide range of possible solutions to the problem of how force is shared among muscle synergists. A better understanding of how force is shared among muscle synergists can provide insight into the mechanics and control of movement. We examined force sharing in the gastrocnemius of wild turkeys (Meleagris gallopavo), using strain gauges bonded to bony tendons. Force was measured separately in the lateral (LG) and medial (MG) heads of the gastrocnemius, to evaluate force sharing. We also used inverse dynamics to determine the total force required during swing phase. To determine whether the pattern of force sharing varied depending on the mechanical task, we used running speed (1 to 3.5 m s–1) and limb loading (30 and 60 g added tarsometatarsal mass) to vary the force required at the intertarsal joint. We found that the distribution of force between these two heads varied depending on the...]]>
Following the identification and confirmation of the substructures of the leading-edge vortex (LEV) system on flapping wings, it is apparent that the actual LEV structures could be more complex than had been estimated in previous investigations. In this experimental study, we reveal for the first time the detailed three-dimensional (3-D) flow structures and evolution of the LEVs on a flapping wing in the hovering condition at high Reynolds number (Re=1624). This was accomplished by utilizing an electromechanical model dragonfly wing flapping in a water tank (mid-stroke angle of attack=60°) and applying phase-lock based multi-slice digital stereoscopic particle image velocimetry (DSPIV) to measure the target flow fields at three typical stroke phases: at 0.125T (T=stroke period), when the wing was accelerating; at 0.25T, when the wing had maximum speed; and at 0.375T, when the wing was decelerating. The result shows that the LEV system is a collection of four vortical elements: one primary vortex and three minor vortices, instead of a single conical or tube-like...]]>
We investigated the distribution of serotonin (5-HT)-containing neuroepithelial cells (NECs), the putative O2 sensing cells, in the gills of four species of fish: trout (Oncorhynchus mykiss), goldfish (Carassius auratus), trairão (Hoplias lacerdae) and traira (Hoplias malabaricus) using immunohistochemical markers for 5-HT, synaptic vesicles and neural innervation. We found that all fish had a cluster of innervated, serotonergic NECs at the filament tips, but there were species-specific distributions of serotonin-containing NECs within the primary gill filaments. Trout gill filaments had a greater number of serotonin-containing NECs than both trairão and traira, whereas goldfish primary filaments had none. Serotonin-containing NECs in the secondary lamellae were most numerous in goldfish, present in trairão and traira, but absent in trout. Those found in the primary filament were generally associated with the efferent filamental artery. Innervated, serotonin-containing cells (NECs or Merkel-like cells) were also found in the gill rakers of trout and goldfish although vesicular serotonin was only found in the gill rakers of goldfish. These differences in serotonergic NEC distribution appear...]]>
The nature of the courtship signalling used by a species is shaped by many factors, one of which is its habitat. Male blue crabs, Callinectes sapidus, have a courtship display in which they elevate their body by standing high on their legs, open their chelae and paddle their swimming legs. This courtship display is not reported in other swimming (portunid) crabs and is rarely expressed in laboratory experiments on male blue crabs. In this study, we characterised this display, which we call `courtship stationary paddling', and distinguished it from other types of paddling. To explain the species specificity of courtship stationary paddling, we hypothesised that this behaviour is an adaptation to low visibility and abundant refuges in the habitat of blue crabs, and that this behaviour enhances chemical signalling when females are relatively inaccessible to males. We used particle imaging velocimetry to visualise water currents generated during courtship stationary paddling, showing that it created water currents directed away from the male and towards the...]]>
In social insects, individuals typically recognize and behave aggressively towards alien conspecifics, thereby maintaining colony integrity. This is presumably achieved via a nestmate recognition system in which cuticular compounds, usually cuticular hydrocarbons (CHC), of genetic and/or environmental origin serve as recognition cues. Most invasive populations of the Argentine ant, Linepithema humile (Mayr), display minimal nestmate–non-nestmate discrimination, resulting in low levels of intraspecific aggression allowing free movement of workers and queens among nests. However, invasive L. humile in the southeastern United States show relatively high levels of intraspecific aggression, and selectively adopt non-nestmate queens. Using behavioral assays and gas chromatography, we found an association between non-nestmate queen adoption and similarity of the CHC profiles of adopted and host colony queens. Also, nestmate and non-nestmate queen CHC profiles became more similar after adoption by queenless colonies. Furthermore, queens treated with non-nestmate queen CHC had distinct CHC profiles and were generally attacked by nestmate workers. We suggest that in L. humile, CHC are used as queen recognition cues, and...]]>
We performed experiments on male Australian painted dragon lizards (Ctenophorus pictus) to test the hypothesis that carotenoids can scavenge reactive oxygen species (ROS), protecting the organism from oxidative stress, and that this capacity is reflected in skin colours involved in signalling. Subsequent to 4 weeks of carotenoid treatment we used flow cytometry to analyse unspecified ROS (H2O2, singlet oxygen, superoxide and peroxynitrite level), hereafter termed ROS, and baseline superoxide specifically (bSO in peripheral blood cells). Mean background levels of ROS and bSO did not differ between carotenoid-treated and control males. bSO, which represents the superoxide level in un-manipulated blood, was negatively correlated with colour development in all males, regardless of carotenoid treatment. Thus, carotenoid intake does not reduce circulating levels of ROS or bSO, suggesting that carotenoids are inefficient antioxidants in vivo and, therefore, are unlikely to provide a direct link between oxidative stress and colouration.
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