bluuebunny/crossref_metadata_2025 huggingface dataset:
Redox control in development and evolution: evidence from colonial hydroids
Authors: Blackstone Neil W.
Published: April 2021

Redox chemistry, involving the transfer of electrons and hydrogen atoms, is central to energy conversion in respiration, and the control of gene expression by redox state commonly occurs in bacteria, allowing rapid responses to environmental changes, for instance, in the food supply. Colonial metazoans often encrust surfaces over which the food supply varies in time or space; hence, in these organisms, redox control of the development of feeding structures and gastrovascular connections could be similarly adaptive, allowing colonies to adjust the timing and spacing of structures in response to a variable food supply. To investigate the possibility of redox control of colony development, the redox states of hydractiniid hydroid colonies were manipulated experimentally. As in many colonial animals, hydractiniid hydroids display a range of morphological variation from sheet-like forms (i.e. closely spaced polyps with high rates of stolon branching) to runner-like forms (i.e. widely spaced polyps with low rates of stolon branching). In the runner-like Podocoryna carnea, azide, a blocker of the electron transport chain, and dinitrophenol, an uncoupler of oxidative phosphorylation, diminished the largely polyp-driven gastrovascular flow to a similar extent. Measures of the redox state of the polyp epitheliomuscular cells using the fluorescence of NAD(P)H suggest that azide shifts the redox state in the direction of reduction, while dinitrophenol shifts the redox state in the direction of oxidation. Colony development corresponds to redox state in that azide-treated colonies were more runner-like, while dinitrophenol-treated colonies were more sheet-like. Nevertheless, the functional role of polyps in feeding and generating gastrovascular flow probably contributed to a trade-off between polyp number and size such that azide-treated colonies had few large polyps, while dinitrophenol-treated colonies had many small polyps. Regardless of the treatment, P. carnea colonies developed to maturity and produced swimming medusae in the normal fashion. In the sheet-like Hydractinia symbiolongicarpus, treatment with azide resulted in complete suppression of the development of both the stolonal mat and the blastostyles, the reproductive polyps. Azide-treated H. symbiolongicarpus colonies therefore developed in a juvenilized, runner-like manner and much resembled colonies of P. carnea. Following cessation of azide treatment in H. symbiolongicarpus, normal colony development ensued, and both a stolonal mat and blastostyles formed. In both hydroid species, relative oxidization favors sheet-like growth, while relative reduction favors runner-like growth. Since feeding triggers strong contractions of polyp epitheliomuscular cells and results in relative oxidation, this experimental evidence supports the hypothesis of adaptive redox control of colony development and evolution.
Differential activation of octopaminergic (DUM) neurones via proprioceptors responding to flight muscle contractions in the locust
Authors: Morris O. T. , Duch C. , Stevenson P. A.
Published: April 2021

The synaptic potentials generated in neuromodulatory octopaminergic dorsal unpaired median (DUM) neurones by afferents excited by twitch contractions of a dorso-ventral flight muscle were investigated in the locust. Responses to stimulation of the metathoracic wing elevator muscle 113 were obtained in locusts in which all sensory feedback from the thorax had been removed, except for feedback from the thoracic chordotonal organs, the axons of which enter via the purely sensory nerve 2. Afferents in nerve 2C, which originates from two chordotonal organs, responded reliably to twitch contractions of this flight muscle. Octopaminergic neurones innervating leg muscles (DUM5 neurones) received depolarising inputs and often spiked following stimulation of the muscle. In contrast, those innervating the wing muscles themselves (DUM3 and DUM3,4 neurones) received inhibitory inputs. The responses of DUM3,4,5 neurones, which project mainly to leg muscles, were more complex: most were excited by twitch contractions of M113 but some were inhibited. DUMDL, which innervates the dorsal longitudinal indirect flight muscles, showed no clear response. Direct stimulation of nerve 2C evoked depolarising inputs and spikes in DUM5 neurones and hyperpolarising inputs in DUM3 and DUM3,4 neurones. Our data suggest that sensory feedback from thoracic chordotonal organs, which are known to be activated rhythmically during flight, contributes to the differential activation of efferent DUM neurones observed during flight.
Isolation and characterization of a leucokinin-like peptide of Drosophila melanogaster
Authors: Terhzaz Selim , O’Connell F. Claire , Pollock Valerie P. , Kean Laura , Davies Shireen A. , Veenstra Jan A. , Dow Julian A. T.
Published: April 2021

The leucokinin (LK) family of neuropeptides has been found widely amongst invertebrates. A member of this family was purified from adults of the fruit fly Drosophila melanogaster. The peptide sequence for Drosophila leucokinin (DLK) was determined as Asn-Ser-Val-Val-Leu-Gly-Lys-Lys-Gln-Arg-Phe-His-Ser-Trp-Gly-amide, making it the longest member of the family characterized to date. Synthetic DLK peptide was shown to act to stimulate fluid secretion in D. melanogaster Malpighian (renal) tubules by approximately threefold, with an EC50 of approximately 10−10 mol l−1, and a secondary effect at approximately 10−7 mol l−1. DLK also acted to elevate intracellular [Ca2+ ] in the Malpighian tubules by approximately threefold, with an EC50 of 10−10 to 10−9 mol l−1. Responses were detected in stellate cells and occasionally in principal cells, although at no concentration tested did [Ca2+ ] in the principal cell increase significantly above background. In stellate cells, DLK produced a biphasic rise in intracellular [Ca2+ ] from resting levels of 80–100 nmol l−1, with a transient peak being followed by a slower rise that peaked at 200–300 nmol l−1 after 3 s, then decayed over approximately 10 s. The wide range of concentrations over which DLK acts suggests the involvement of more than one receptor. The genomic sequence encoding the DLK peptide has been identified, and the gene has been named pp. The gene resides at cytological location 70E3–70F4 of chromosome 3L. The localisation of this first Drosophila LK gene in a genetic model permits a genetic analysis of the locus.
Gas exchange and ventilation during dormancy in the tegu lizard tupinambis merianae
Authors: Andrade Denis Vieira De , Abe Augusto Shinya
Published: April 2021

The tegu lizard Tupinambis merianae exhibits an episodic ventilatory pattern when dormant at 17 °C but a uniform ventilatory pattern when dormant at 25 °C. At 17 °C, ventilatory episodes were composed of 1–22 breaths interspaced by non-ventilatory periods lasting 1.8–26 min. Dormancy at the higher body temperature was accompanied by higher rates of O2 consumption and ventilation. The increase in ventilation was due only to increases in breathing frequency with no change observed in tidal volume. The air convection requirement for O2 did not differ at the two body temperatures. The respiratory quotient was 0.8 at 17 °C and 1.0 at 25 °C. We found no consistent relationship between expired gas composition and the start/end of the ventilatory period during episodic breathing at 17 °C. However, following non-ventilatory periods of increasing duration, there was an increase in the pulmonary O2 extraction that was not coupled to an equivalent increase in elimination of CO2 from the lungs. None of the changes in the variables studied could alone explain the initiation/termination of episodic ventilation in the tegus, suggesting that breathing episodes are shaped by a complex interaction between many variables. The estimated oxidative cost of breathing in dormant tegus at 17 °C was equivalent to 52.3 % of the total metabolic rate, indicating that breathing is the most costly activity during dormancy.
Myoglobin in pelagic small cetaceans
Authors: Dolar M. L. L. , Suarez P. , Ponganis P. J. , Kooyman G. L.
Published: April 2021

Although myoglobin (Mb) is considered to contribute significantly to the oxygen and diving capacity of marine mammals, few data are available for cetaceans. Cetacean by-catch in the tuna driftnet fisheries in the Sulu Sea, Philippines, afforded the opportunity to examine Mb content and distribution, and to determine muscle mass composition, in Fraser’s (Lagenodelphis hosei) and spinner (Stenella longirostris) dolphins and a pygmy killer whale (Feresa attenuata). Age was estimated by body length determination. Stomach contents were analyzed for the presence or absence of milk and solid foods. It was hypothesized (a) that Mb concentration ([Mb]) would be higher in Fraser’s and spinner dolphins than in other small cetaceans because of the known mesopelagic distribution of their prey, (b) that [Mb] would vary among different muscles according to function during diving, and (c) that [Mb] would increase with age during development. The results were as follows. (1) Myoglobin concentrations of the longissimus muscle in adult Fraser’s (6.8–7.2 g 100 g−1 muscle) and spinner (5–6 g 100 g−1 muscle) dolphins and in an immature pygmy killer whale (5.7 g 100 g−1 muscle) were higher than those reported previously for small cetaceans. (2) [Mb] varied significantly among the different muscle types in adult dolphins but not in calves; in adults, swimming muscles had significantly higher [Mb] than did non-swimming muscles, contained 82–86 % of total Mb, and constituted 75–80 % of total muscle mass. (3) Myoglobin concentrations in Fraser’s and spinner dolphins increased with size and age and were 3–4 times greater in adults than in calves. The high Mb concentrations measured in the primary locomotory muscles of these pelagic dolphins are consistent with the known mesopelagic foraging behaviour of Fraser’s and spinner dolphins and suggest that the pygmy killer whale is also a deep-diving species. The high Mb concentrations in epaxial, hypaxial and abdominal muscle groups also support the primary locomotory functions suggested for these muscles in other anatomical studies. As in other species, the increase in [Mb] during development probably parallels the development of diving capacity.
The marble goby Oxyeleotris marmoratus activates hepatic glutamine synthetase and detoxifies ammonia to glutamine during air exposure
Authors: Jow L. Y. , Chew S. F. , Lim C. B. , Anderson P. M. , Ip Y. K.
Published: April 2021

Ammonia levels in various tissues of the marble goby Oxyeleotris marmoratus remained constant throughout a 72 h period of air exposure. The rate of ammonia excretion in these experimental fish decreased to approximately one-fifth of that of the submerged control. Ammonia was not converted to urea during air exposure because there were no significant increases in urea content in the tissues. Also, urea excretion rate was lowered to one-fiftieth that of the submerged fish. After 24 h of air exposure, there was a significant increase in muscle glutamine content, which peaked at 48 h. The increase in glutamine content could account for the decreases in the amounts of ammonia and urea excretion during air exposure. The specific activities of hepatic glutamate dehydrogenase (amination) and glutamine synthetase in these experimental fish increased threefold and thirtyfold, respectively, in comparison with the submerged controls. Thus, O. marmoratus appears to be the first known teleost that responds to air exposure by activating hepatic glutamine synthetase to detoxify internally produced ammonia.
Expansion of the tunica albuginea during penile inflation in the nine-banded armadillo (Dasypus novemcinctus)
Authors: Kelly D. A.
Published: April 2021

Artificial inflation of corpora cavernosa from the nine-banded armadillo (Dasypus novemcinctus) showed that the expansion of the tunica albuginea during erection increases both components of flexural stiffness: the second moment of area and Young’s modulus of elasticity. Folded tissue and crimped collagen fibers in the tunica albuginea permit its expansion during erection. As the tunica albuginea’s radii increase in size, its second moment of area also increases. The crimped collagen fibers permit the flaccid tunica albuginea to expand to strains of 25 % longitudinally and 15 % circumferentially, after which tissue stiffness increases by 3–4 orders of magnitude. Radial expansion of the corpus cavernosum is limited by collagenous trabeculae. The trabeculae maintain the non-circular cross section of the corpus cavernosum during erection. Restricting expansion appears to protect the corpus spongiosum and urethra from occlusion, but has the side effect of reducing the potential flexural stiffness of the corpus cavernosum by reducing the second moment of area of the tunica albuginea.
Pathways of inorganic nitrogen assimilation in chemoautotrophic bacteria–marine invertebrate symbioses: expression of host and symbiont glutamine synthetase
Authors: Lee Raymond W. , Robinson Jonathan J. , Cavanaugh Colleen M.
Published: April 2021

Symbioses between chemoautotrophic bacteria and marine invertebrates living at deep-sea hydrothermal vents and other sulfide-rich environments function autotrophically by oxidizing hydrogen sulfide as an energy source and fixing carbon dioxide into organic compounds. For chemoautotrophy to support growth, these symbioses must be capable of inorganic nitrogen assimilation, a process that is not well understood in these or other aquatic symbioses. Pathways of inorganic nitrogen assimilation were investigated in several of these symbioses: the vent tubeworms Riftia pachyptila and Tevnia jerichonana, the vent bivalves Calyptogena magnifica and Bathymodiolus thermophilus, and the coastal bivalve Solemya velum. Nitrate reductase activity was detected in R. pachyptila, T. jerichonana and B. thermophilus, but not in C. magnifica and S. velum. This is evidence for nitrate utilization, either assimilation or respiration, by some vent species and is consistent with the high levels of nitrate availability at vents. The ammonia assimilation enzymes glutamine synthetase (GS) and glutamate dehydrogenase (GDH) were detected in all symbioses tested, indicating that ammonia resulting from nitrate reduction or from environmental uptake can be incorporated into amino acids. A complicating factor is that GS and GDH are potentially of both host and symbiont origin, making it unclear which partner is involved in assimilation. GS, which is considered to be the primary ammonia-assimilating enzyme of autotrophs, was investigated further. Using a combination of molecular and biochemical approaches, host and symbiont GS were distinguished in the intact association. On the basis of Southern hybridizations, immunoreactivity, subunit size and thermal stability, symbiont GS was found to be a prokaryote GS. Host GS was distinct from prokaryote GS. The activities of host and symbiont GS were separated by anion-exchange chromatography and quantified. Virtually all activity in symbiont-containing tissue was due to symbiont GS in R. pachyptila, C. magnifica and B. thermophilus. In contrast, no symbiont GS activity was detected in the gill of S. velum, the predominant activity in this species appearing to be host GS. These findings suggest that ammonia is primarily assimilated by the symbionts in vent symbioses, whereas in S. velum ammonia is first assimilated by the host. The relationship between varying patterns of GS expression and host–symbiont nutritional exchange is discussed.
The hydrophobicity of vertebrate elastins
Authors: Chalmers G. W. G. , Gosline J. M. , Lillie M. A.
Published: April 2021

An evolutionary trend towards increasing hydrophobicity of vertebrate arterial elastins suggests that there is an adaptive advantage to higher hydrophobicity. The swelling and dynamic mechanical properties of elastins from several species were measured to test whether hydrophobicity is associated with mechanical performance. Hydrophobicity was quantified according to amino acid composition (HI), and two behaviour-based indices: the Flory–Huggins solvent interaction parameter (χ1), and a swelling index relating tissue volumes at 60 and 1 °C. Swelling index values correlated with χ1 and, for most species studied, with HI, suggesting that the different approaches used to quantify hydrophobicity are equally valid. Dynamic mechanical properties were measured both in a closed system, to control the effects of water content, and in an open system, to determine whether the increased swelling of hydrophobic materials at low temperatures offsets the direct stiffening effect of cold. There were no biologically significant differences in mechanical behaviour in either open or closed systems that could be attributed to hydrophobicity. Therefore, although the original function of hydrophobicity in an ancestral elastin may have been to produce molecular mobility, mechanical performance did not drive a subsequent increase in hydrophobicity. Higher hydrophobicities may have arisen to facilitate the manufacture of the elastic fibre.
A mechanism for branchial acid excretion in marine fish: identification of multiple Na+/H+ antiporter (NHE) isoforms in gills of two seawater teleosts
Authors: Claiborne J. B. , Blackston C. R. , Choe K. P. , Dawson D. C. , Harris S. P. , Mackenzie L. A. , Morrison-Shetlar A. I.
Published: April 2021

Both Na+/H+ exchange and the electrogenic extrusion of H+via an H+-ATPase have been postulated to drive acid excretion across the branchial epithelium of fishes. While the H+-ATPase/Na+ channel system appears to be the predominant mechanism in some freshwater species, it may play a reduced role in seawater and brackish-water animals, where high external Na+ concentrations may thermodynamically favor Na+/H+ exchange driven by a Na+/H+ antiporter (NHE). In this study, we used molecular and immunological methods to assess the role of NHE isoforms in the branchial epithelium of the marine long-horned sculpin (Myoxocephalus octodecimspinosus) and the euryhaline killifish (Fundulus heteroclitus).

Northern blot analysis of RNA probed with the human NHE-1 BamHI fragment suggested the presence of homologous gill NHE mRNA in sculpin. RT-PCR on gill RNA isolated from sculpin recovering from metabolic acidosis provided evidence for two distinct NHE isoforms; one with 76 % amino acid homology to mammalian NHE-2, and another 92 % homologous to trout erythrocytic β-NHE. Killifish also have transcripts with 91 % homology to β-NHE. Immunological detection using monoclonal antibodies for mammalian NHE-1 revealed a protein antigenically similar to this isoform in the gills of both species. Metabolic acidosis caused an approximately 30-fold decrease in expression of the NHE-1-like protein in sculpin. We speculate that β-NHE in the gills plays the intracellular ‘housekeeping’ roles described for mammalian NHE-1. During systemic acidosis, apical gill NHE-2 (which is sensitive to external amiloride and low [Na+]) in parallel with a dramatic suppression of basolateral NHE-1 activity enhances net ΔH+ transfers to the water.
The roles of acidosis and lactate in the behavioral hypothermia of exhausted lizards
Authors: Wagner Erica L. , Scholnick David A. , Gleeson Todd T.
Published: April 2021

We conducted this study to determine whether two of the physiological changes associated with non-sustainable exercise, elevated blood lactate levels and decreased arterial pH, contribute to the behavioral hypothermia exhibited by exhausted lizards. Dipsosaurus dorsalis were placed in a thermal gradient and their body temperatures were recorded from 08:00 to 14:00 h. At 14:00 h, animals were subjected to different experimental regimens. In the exercise (E) regimen, animals at 40 °C were forced to exercise maximally for 5 min on a treadmill. In the lactate (L) regimen, animals were infused with 11.5 ml kg−1 of 250–500 mmol l−1 sodium lactate. In the osmolarity control (O) regimen, animals were injected with 11.5 ml kg−1 of 500 mmol l−1 NaCl, and in the injection control (I) regimen, animals were injected with 11.5 ml kg−1 of 150 mmol l−1 NaCl. In the hypercapnia (H) regimen, the thermal gradient was flushed with a gas mixture containing 10 % CO2, 21 % O2 and 69 % N2, a treatment that lowers the arterial pH of D. dorsalis to a value comparable with that imposed by exhaustive exercise. A group of control (C) animals was left undisturbed in the thermal gradient for 24 h. Animals in all experimental groups were returned to the thermal gradient, and their cloacal temperatures were monitored until 08:00 h the following morning. The mean cloacal temperature of E animals underwent a significant decrease of 4–7 °C, relative to control animals, which persisted for 7 h. The mean cloacal temperatures of animals subjected to 2 h of regimen H also decreased by 3.5–9 °C and remained depressed for 12 h following the beginning of the treatment. L, O and I animals did not undergo a significant change in body temperature following treatment, and their mean body temperatures did not differ from those of C animals at any time during the experiment. The results of this study suggest that the metabolic acidosis, but not the elevated blood lactate level, that follows exhausting exercise might play a role in the behavioral hypothermia that follows exhausting exercise in D. dorsalis.
Eclosion hormone provides a link between ecdysis-triggering hormone and crustacean cardioactive peptide in the neuroendocrine cascade that controls ecdysis behavior
Authors: Gammie Stephen C. , Truman James W.
Published: April 2021

Three insect peptide hormones, eclosion hormone (EH), ecdysis-triggering hormone (ETH) and crustacean cardioactive peptide (CCAP), have been implicated in controlling ecdysis behavior in insects. This study examines the interactions between these three peptides in the regulation of the ecdysis sequence. Using intracellular recordings, we found that ETH is a potent activator of the EH neurons, causing spontaneous action potential firing, broadening of the action potential and an increase in spike peak amplitude. In turn, electrical stimulation of the EH neurons or bath application of EH to desheathed ganglia resulted in the elevation of cyclic GMP (cGMP) levels within the Cell 27/704 group (which contain CCAP). This cGMP production increases the excitability of these neurons, thereby facilitating CCAP release and the generation of the ecdysis motor program. Extracellular recordings from isolated nervous systems show that EH has no effect on nervous systems with an intact sheath. In desheathed preparations, in contrast, EH causes only the ecdysis motor output. The latency from EH application to ecdysis was longer than that after CCAP application, but shorter than that when ETH is applied to a whole central nervous system. These data, along with previously published results, support a model in which ETH causes pre-ecdysis behavior and at higher concentrations stimulates the EH neurones. EH release then facilitates the onset of ecdysis by enhancing the excitability of the CCAP neurons.
Energetics of locomotion by the australian water rat (Hydromys chrysogaster): a comparison of swimming and running in a semi-aquatic mammal
Authors: Fish F. E. , Baudinette R. V.
Published: April 2021

Semi-aquatic mammals occupy a precarious evolutionary position, having to function in both aquatic and terrestrial environments without specializing in locomotor performance in either environment. To examine possible energetic constraints on semi-aquatic mammals, we compared rates of oxygen consumption for the Australian water rat (Hydromys chrysogaster) using different locomotor behaviors: swimming and running. Aquatic locomotion was investigated as animals swam in a water flume at several speeds, whereas water rats were run on a treadmill to measure metabolic effort during terrestrial locomotion. Water rats swam at the surface using alternate pelvic paddling and locomoted on the treadmill using gaits that included walk, trot and half-bound. Water rats were able to run at twice their maximum swimming velocity. Swimming metabolic rate increased with velocity in a pattern similar to the ‘humps’ and ‘hollows’ for wave drag experienced by bodies moving at the water surface. Metabolic rate increased linearly during running. Over equivalent velocities, the metabolic rate for running was 13–40 % greater than for swimming. The minimum cost of transport for swimming (2.61 J N−1 m−1) was equivalent to values for other semi-aquatic mammals. The lowest cost for running (2.08 J N−1 m−1) was 20 % lower than for swimming. When compared with specialists at the extremes of the terrestrial–aquatic continuum, the energetic costs of locomoting either in water or on land were high for the semi-aquatic Hydromys chrysogaster. However, the relative costs for H. chrysogaster were lower than when an aquatic specialist attempts to move on land or a terrestrial specialist attempts to swim.
Expression of olfactory receptors during development in Xenopus laevis
Authors: Mezler Mario , Konzelmann Sidonie , Freitag Joachim , Rössler Patricia , Breer Heinz
Published: April 2021

A coordinated expression of tissue- and cell-specific genes during development is required to establish the complex functional organization of the vertebrate olfactory system. Owing to the unique features of its olfactory system and the well-characterized phases of its development, Xenopus laevis was chosen as a model organism to study the onset and the temporal and spatial patterns of expression of olfactory-specific genes. Using RT-PCR and in situ hybridization, it was found that expression of Xenopus olfactory marker protein and of class I receptors, which are thought to be responsible for the perception of water-soluble odorants, was detectable as early as stage 32, less than 2 days after fertilization. In contrast, expression of class II receptors, which are thought to recognize airborne odours, was not detected before stage 49, approximately 12 days after fertilization. The results indicate that the expression of olfactory receptors and marker protein is governed by temporally regulated cues during development.
Micromechanics of the equine hoof wall: optimizing crack control and material stiffness through modulation of the properties of keratin
Authors: Kasapi Mario A. , Gosline John M.
Published: April 2021

Small-scale components of the equine hoof wall were tested to determine their mechanical roles in the morphological hierarchy. Macroscale tensile tests conducted on samples of the inner wall tubules and intertubular material showed a sixfold difference in mean initial stiffnesses (0.47 and 0.08 GPa, respectively), indicating that the inner wall tubules stiffen the wall along its longitudinal axis. The similarity in material properties of tubule and intertubular samples from the mid-wall suggests that tubules in this region offer only minor reinforcement along the longitudinal axis.

Microscale tests conducted on rows of keratin strands from the inner wall tubules and intertubular material, and on intertubular keratin strands of the mid-wall, produced estimates of the stiffnesses of the hydrated matrix (0.03 GPa) and intermediate filament (IF; 3–4 GPa) components of the nanoscale (α-keratin) composite. The results from these tests also suggest that the properties of the keratin composite vary through the wall thickness. Birefringence measurements on inner wall and mid-wall regions agree with these observations and suggest that, although the keratin IF volume fraction is locally constant, the volume fraction changes through the thickness of the wall. These findings imply that modulation of the hoof wall properties has been achieved by varying the IF volume fraction, countering the effects of specific IF alignments which serve another function and would otherwise adversely affect the modulus of a particular region.
ATP reception and chemosensory adaptation in Tetrahymena thermophila
Authors: Kim M. Y. , Kuruvilla H. G. , Raghu S. , Hennessey T. M.
Published: April 2021

Micromolar concentrations of adenosine triphosphate (ATP) and its non-hydrolyzable analog β-γ-methylene ATP are both effective depolarizing chemorepellents in Tetrahymena thermophila. Chemorepellent behavior consists of repeated bouts of backward swimming (avoidance reactions) that can easily be quantified to provide a convenient bioassay for purinergic reception studies. Chemosensory adaptation occurs following prolonged exposure (10 min) to the repellents, and cells regain normal swimming behavior. Adaptation is specific since cells that are behaviorally adapted to either ATP or β-γ-methylene ATP still retain full responsiveness to the chemorepellents GTP and lysozyme. However, cross adaptation occurs between ATP and β-γ-methylene ATP, suggesting that they involve the same receptor. Behavioral sensitivity to both ATP and β-γ-methylene ATP is increased by the addition of Na+, but addition of either Ca2+ or Mg2+ dramatically decreases the response to ATP. These ionic effects are correlated with in vivo ATP hydrolysis, suggesting that divalent ions decrease purinergic sensitivity by activating a Ca2+- or Mg2+-dependent ecto-ATPase to hydrolyze the ATP signal. In vivo [32P]ATP binding studies and Scatchard analysis suggest that the behavioral adaptation is due to a decrease in the number of surface binding sites, as represented by decreased Bmax values. All these changes are reversible (de-adaptation) after 12 min in a repellent-free buffer. Electrophysiological analysis showed that both β-γ-methylene ATP (10 μmol l−1) and ATP (500 μmol l−1) elicited sustained, reversible depolarizations while GTP (10 μmol l−1) produced a transient depolarization, suggesting that the chemosensory response pathways for ATP and GTP reception may differ. There may be separate ATP and GTP receptors since ATP and GTP responses do not cross-adapt and ‘cold’ (unlabeled) GTP is not a good inhibitor of [32P]ATP binding. These results suggests that T. thermophila possess high-affinity surface receptors for ATP that are down-regulated during chemosensory adaptation. These ATP receptors may act as chemorepellent receptors to enable T. thermophila to recognize recently lysed cells and avoid a possibly deleterious situation. This is the simplest eukaryotic organism to show an electrophysiological response to external ATP.
A comparison of the release of a vasoactive-intestinal-peptide-like peptide and acetylcholine in the giant axon–Schwann cell preparation of the tropical squid Sepioteuthis sepioidea
Authors: Evans Peter D. , Reale Vincenzina , Merzon Rosa Maria , Villegas Jorge
Published: April 2021

A vasoactive intestinal peptide (VIP)-like peptide is released by axonal stimulation in the giant axon–Schwann cell preparation from the tropical squid Sepioteuthis sepioidea. It is also released by direct application of L-glutamate, the giant axon–Schwann cell signalling molecule in this preparation. The release of the peptide parallels the release of acetylcholine from the Schwann cells themselves in this preparation in a number of different ways. The release of both acetylcholine and the VIP-like peptide have the same threshold (between 2×10−10 and 5×10−10 mol l−1) for L-glutamate application and the same recovery time after inhibition of release by exposure of the preparation to a prolonged pulse of L-glutamate. A prolonged L-glutamate pulse of 10−8 mol l−1 releases both substances for as long as the pulse is applied to the preparation, whereas a prolonged pulse of 10−9 mol l−1 L-glutamate releases acetylcholine in the same way but releases the VIP-like peptide only transiently. The VIP-like peptide is likely to be co-released with acetylcholine from the Schwann cells.
Fuel use in flight and its dependence on nectar feeding in the hawkmoth Amphion floridensis
Authors: O’Brien Diane M.
Published: April 2021

Fuel use varies widely among insects; however, the potential determinants of variation in fuel use have not been explored experimentally. This study examines whether fuel use during tethered flight depends upon feeding status in the nectarivorous hawkmoth Amphion floridensis. Fuel use in this study is characterized by the respiratory quotient, measured at intervals during a flight using modified closed-chamber respirometry. Moths were either fed twice daily to satiation with 30 % sucrose or unfed, and their fuel use was measured during flights on the first, third and fifth day after eclosion. Flights lasted up to 30 min, with measurements taken at their onset and at 10 min intervals thereafter. Nectar feeding greatly affected fuel use in A. floridensis: fed moths relied primarily on carbohydrate, whereas unfed moths relied almost exclusively on fat reserves. Fuel use did not change during a flight, even when flights lasted 30 min or more. Males were initially more extreme than females in their response to feeding treatment: they burned more carbohydrate when fed and more fat when unfed. By the third day after eclosion, however, fuel use in males and females became identical. Rates of oxygen consumption were uncorrelated with respiratory quotient, were higher in fed moths and declined during a flight. These data indicate that fuel use in this nectarivorous hawkmoth is flexible, that carbohydrate is important as a primary flight fuel and that an understanding of ecological factors, particularly foraging habit, is critical to understanding fuel use in insects.
Plasmodium gallinaceum ookinetes adhere specifically to the midgut epithelium of Aedes aegypti by interaction with a carbohydrate ligand
Authors: Zieler Helge , Nawrocki Joseph P. , Shahabuddin Mohammed
Published: April 2021

During the course of its development in the mosquito and transmission to a new vertebrate host, the malaria parasite must interact with the mosquito midgut and invade the gut epithelium. To investigate how the parasite recognizes the midgut before invasion, we have developed an in vitro adhesion assay based on combining fluorescently labelled ookinetes with isolated midgut epithelia from blood-fed mosquitoes. Using this assay, we found that Plasmodium gallinaceum ookinetes readily adhered to midguts of Aedes aegypti, mimicking the natural recognition of the epithelium by the parasite. This interaction is specific: the ookinetes preferentially adhered to the lumen (microvillar) side of the gut epithelium and did not bind to other mosquito tissues. Conversely, the binding was not due to a non-specific adhesive property of the midguts, because a variety of other cell types, including untransformed P. gallinaceum zygotes or macrogametes, did not show similar binding to the midguts. High concentrations of glycosylated (fetuin, orosomucoid, ovalbumin) or non-glycosylated (bovine serum albumin) proteins, added as non-specific competitors, failed to compete with the ookinetes in binding assays. We also found that the adhesion of ookinetes to the midgut surface is necessary for sporogonic development of the parasite in the mosquito. Antibodies and other reagents that blocked adhesion in vitro also reduced oocyst formation when these reagents were combined with mature ookinetes and fed to mosquitoes. Chemical modification of the midguts with sodium periodate at pH 5.5 destroyed adhesion, indicating that the ookinete binds to a carbohydrate ligand on the surface of the midgut. The ligand is sensitive to periodate concentrations of less than 1 mmol l−1, suggesting that it may contain sialic-acid-like sugars. Furthermore, free N-acetylneuraminic acid competed with the ookinetes in binding aasays, while other monosaccharides had no effect. However, in agreement with the current belief that adult insects do not contain sialic acids, we were unable to detect any sialic acids in mosquito midguts using the most sensitive HPLC-based fluorometric assay currently available. We postulate that a specific carbohydrate group is used by the ookinete to recognize the midgut epithelium and to attach to its surface. This is the first receptor–ligand interaction demonstrated for the ookinete stage of a malaria parasite. Further characterization of the midgut ligand and its parasite counterpart may lead to novel strategies of blocking oocyst development in the mosquito.
Ionic exchanges of turtle shell in vitro and their relevance to shell function in the anoxic turtle
Authors: Jackson Donald C. , Goldberger Zachary , Visuri Susanna , Armstrong Ryan N.
Published: April 2021

To understand more fully the role of the in vivo turtle shell in buffering lactic acid produced during prolonged anoxia, powdered turtle shell was incubated in vitro at constant pH (6.0, 6.5, 7.0, 7.5 or 8.0) in electrolyte solutions simulating extracellular fluid. Exchanges of ions and CO2 between the shell and solution were evaluated by measuring pre- and post-incubation solution concentrations of calcium, magnesium, sodium, potassium, chloride, phosphate and lactate. The production of CO2 from the shell and lactate within the shell were also measured. We observed that calcium and magnesium, but not phosphate, were released from the shell in association with CO2 and that the magnitude of release of each increased with solution acidity. The amount of acid titration required to maintain constant pH also increased as solution pH fell. The CO2 loss, in mmol, was approximately half the acid titration (in mmol), indicating that the evolved CO2 derives from carbonate. When the incubating solution contained lactate (50 mmol l−1), lactate entered the shell and again the amount entering the shell increased with solution acidity. Shell samples containing high initial lactate levels lost lactate to the solution and at high pH (7.5) acidified the solution and required NaOH titration for pH-stat control. These results are consistent with observations on anoxic turtles in vivo and confirm the important role of the shell as a source of buffer and as a storage site for lactate.
Muscle strain histories in swimming milkfish in steady and sprinting gaits
Authors: Katz Stephen L. , Shadwick Robert E. , Rapoport H. Scott
Published: April 2021

Adult milkfish (Chanos chanos) swam in a water-tunnel flume over a wide range of speeds. Fish were instrumented with sonomicrometers to measure shortening of red and white myotomal muscle. Muscle strain was also calculated from simultaneous overhead views of the swimming fish. This allowed us to test the hypothesis that the muscle shortens in phase with local body bending. The fish swam at slow speeds [U<2.6 fork lengths s−1 (=FL s−1)] where only peripheral red muscle was powering body movements, and also at higher speeds (2.6>U>4.6 FL s−1) where they adopted a sprinting gait in which the white muscle is believed to power the body movements. For all combinations of speeds and body locations where we had simultaneous measurements of muscle strain and body bending (0.5 and 0.7FL), both techniques were equivalent predictors of muscle strain histories. Cross-correlation coefficients for comparisons between these techniques exceeded 0.95 in all cases and had temporal separations of less than 7 ms on average. Muscle strain measured using sonomicrometry within the speed range 0.9–2.6 FL s−1 showed that muscle strain did not increase substantially over that speed range, while tail-beat frequency increased by 140 %. While using a sprinting gait, muscle strains became bimodal, with strains within bursts being approximately double those between bursts. Muscle strain calculated from local body bending for a range of locations on the body indicated that muscle strain increases rostrally to caudally, but only by less than 4 %. These results suggest that swimming muscle, which forms a large fraction of the body volume in a fish, undergoes a history of strain that is similar to that expected for a homogeneous, continuous beam. This has been an implicit assumption for many studies of muscle function in many fish, but has not been tested explicitly until now. This result is achieved in spite of the presence of complex and inhomogeneous geometry in the folding of myotomes, collagenous myosepta and tendon, and the anatomical distinction between red and white muscle fibers.
The importance of the feeding current for oxygen uptake in the water flea Daphnia magna
Authors: Pirow R. , Wollinger F. , Paul R. J.
Published: April 2021

In the planktonic crustacean Daphnia magna (Branchiopoda, Cladocera), different views exist on the mechanism of respiratory gas exchange, ranging from gill breathing to general integumentary respiration. The presumed structures for specialized gas exchange are located ventrally within the filter chamber, which is continuously perfused with the ambient medium for food gathering. To localize respiratory gas exchange in D. magna, we determined the contribution of the feeding current to total oxygen transport. Combining microscopy with special optical techniques, we used a phosphorescent oxygen-sensitive dye for oxygen partial pressure measurements and applied fluorescent microspheres for flow-rate analysis. Appendage beat rate was determined by digital image-processing. All experiments were carried out on hypoxia-adapted animals with a body length ranging from 2.3 to 2.7 mm at 20 °C. External measurement revealed oxygen depletion in the ventral body region but essentially no change at posterior, lateral and dorsal positions. The difference between the inflow and outflow of the feeding current was 13.0 mmHg (1.73 kPa). The flow rate of the feeding current ranged from 1.2 to 5.2 ml h−1 and showed a close correlation with appendage beat rate, which varied from 310.4 to 460.7 beats min−1. Model calculations based on the Fick principle suggest that oxygen extraction from the feeding current satisfies most of the total of oxygen requirement of D. magna.
The force–velocity properties of a crustacean muscle during lengthening
Authors: Josephson Robert K. , Stokes Darrell R.
Published: April 2021

Muscle force during active lengthening was characterized for scaphognathite levator muscle L2B from the crab Carcinus maenas. The muscle was tetanically stimulated and, during the peak of the contraction, stretched at constant velocity. The total strain was approximately 4 %, the strain rates ranged from 0.03 to 1.6 muscle lengths s−1 (L s−1), and the temperature was 15 °C.

Force increased throughout stretch. During low-velocity stretch, up to approximately 0.3 L s−1, force rose during isovelocity stretch along an approximately exponential trajectory. The asymptotic force approached during the stretch increased and the time constant of the response decreased with increasing strain rate. With stretch at 0.6 L s−1 and greater, the force increased to a distinct yield point, reached after a strain of approximately 1 %, after which force continued to increase but with a slope approximately one-quarter as great as that before yield. Because force changes continuously during constant-velocity lengthening, the adequate descriptor for the force–velocity relationship in a lengthening crab muscle is not a two-dimensional force–velocity curve, but rather a three-dimensional force–velocity–time or force–velocity–strain surface.

Stimulating muscle L2B at 20 Hz or 50 Hz gives a smoothly fused tetanic contraction in which muscle activation is only partial and the plateau force reached is less than that at the optimum stimulus frequency of approximately 100 Hz. The force–velocity relationships of a partially activated muscle are not simply those of a fully activated one scaled down in proportion to the reduction in the maximum isometric force. At low stretch velocities, the asymptotic force approached is larger in proportion to the pre-stretch isometric tension, and the time constant of the force increase is greater, in partially activated than in fully activated muscles. At high stretch velocities, the force at yield relative to the pre-stretch force, and the relative values of the slopes of the force increase before and after yield, are all greater in partially activated than in fully activated muscles, while the strain at yield is smaller.
Does behavioural hypothermia promote post-exercise recovery in cold-submerged frogs?
Authors: Tattersall Glenn J. , Boutilier Robert G.
Published: April 2021

At the low temperatures of the overwintering environment of the frog Rana temporaria, small changes in ambient temperature have large effects on metabolism and behaviour, especially since Q10 values are often greatly elevated in the cold. How the overwintering aquatic frog copes with variable thermal environments in terms of its overall activity metabolism and recovery from pursuit by predators is poorly understood, as is the role of behavioural thermoregulation in furthering recovery from intense activity. Exhaustive exercise was chosen as the method of evaluating activity capacity (defined by time to exhaustion, total distance swum and number of leg contractions before exhaustion) and was determined at 1.5 and 7 °C. Other cohorts of frogs were examined at both temperatures to determine the metabolic (acid–base, lactate, glucose, ATP and creatine phosphate) and respiratory responses to exercise in cold-submerged frogs. Finally, temperature preference before and after exercise was determined in a thermal gradient to define the importance of behavioural thermoregulation on the recovery rates of relevant metabolic and respiratory processes. Activity capacity was significantly reduced in frogs exercised at 1.5 versus 7 °C, although similar levels of tissue acid–base metabolites and lactate were reached. Blood pH, plasma and lactate levels recovered more rapidly at 1.5 °C than at 7 °C; however, intracellular pH and the recovery of tissue metabolite levels were independent of temperature. Resting aerobic metabolic rates were strongly affected by temperature (Q10=3.82); however, rates determined immediately after exercise showed a reduced temperature sensitivity (Q10=1.67) and, therefore, a reduced factorial aerobic scope. Excess oxygen consumption recovered to resting values after 5–6.25 h, and 67 % recovery times tended to be slightly faster at the lower temperatures. Exercise in the cold, therefore, provided an immediately higher factorial scope, which could be involved in the faster rate of recovery of blood lactate levels in the colder frogs. In addition, exercise significantly lowered the preferred temperature of the frogs from 6.7 to 3.6 °C for nearly 7 h, after which they returned to their normal, unstressed preferred temperatures. Thus, a transient behavioural hypothermia in the skin-breathing, overwintering frog may be an important strategy for minimising post-exercise stress and maintaining aerobic metabolism during recovery from intense activity.