bluuebunny/crossref_metadata_2025 huggingface dataset:
Hatching controlled by the circatidal clock, and the roleof the medulla terminalis in the optic peduncle of the eyestalk, in an estuarine crabSesarma haematocheir
Authors: Saigusa Masayuki
Published: April 2021

Embryos attached to the female crab Sesarma haematocheir hatch synchronously within 1 h. Hatching is also synchronized near the time of the expected nocturnal high tide. These events are governed by a single circatidal clock (or pacemaker) in the female crab. The present study examined the role of the optic peduncle of the eyestalk on hatching and hatching synchrony. Surgery was performed either from the tip of the eyestalk [to remove the region of the optic peduncle from the compound eye—retina complex to the medulla interna (MI)] or from a small triangle `window' opened on the eyestalk exoskeleton [to create lesions on the medulla terminalis (MT) of the optic peduncle]. Neither hatching nor hatching synchrony was affected by removal of the region of the optic peduncle from the compound eye—retina complex to the MI: the circatidal rhythm also remained. Removal of the MI probably caused damage to the sinus gland and the bundle of axons running from the sinus gland to the X organ. Nevertheless, maintenance of highly synchronized hatching indicates that the X organ—sinus gland system is not related to hatching. Hatching and hatching synchrony were not affected by dorsal-half cuts of the MT: the timing of hatching was not affected either. By contrast,transverse and ventral-half cuts of the MT caused severe damage to most females; hatching of many females was suppressed, while hatching of some females was either periodic, at intervals of approximately 24 h, or arrhythmic for a few days. The bundle of neuronal axons is tangled in the MT, and the axons inducing hatching pass through the ventral half of the MT. Complete incision of these axon bundles may have suppressed hatching. Incomplete incision of the axon bundle or partial damage to the neurons may have caused periodic or arrhythmic patterns of hatching. There are two possible roles for MT in hatching. One possibility is that neurons in the MT only induce hatching under the control of the circatidal pacemaker located in a site somewhere other than the optic peduncle. Another possibility is that the circatidal pacemaker is actually present in the MT. The second possibility seems more plausible. Each embryo has a special 48-49.5 h developmental program for hatching. This program could be initiated by the circatidal pacemaker in the female, and hatching synchrony may also be enhanced by the same pacemaker.
The enterins inhibit the contractile activity of the anterior aorta of Aplysia kurodai
Authors: Sasaki Kosei , Fujisawa Yuko , Morishita Fumihiro , Matsushima Osamu , Furukawa Yasuo
Published: April 2021

The anterior aorta is one of the largest blood vessels in the marine mollusc Aplysia kurodai. We examined the actions of recently identified neuropeptides, the enterins, on this blood vessel. Immunohistochemistry revealed that the enterin-immunopositive nerve fibers and varicosity-like structures are abundant in the aorta. When the enterins were applied to the aorta, the basal tonus of the arterial muscles was diminished. The enterins also decreased the contraction amplitude of the anterior aorta evoked either by the application of an Aplysia cardioactive peptide,NdWFamide, or by the stimulation of a nerve innervating the aorta (the vulvar nerve). We found that the enterins activate the 4-aminopyridine(4-AP)-sensitive K+ channels, and thereby hyperpolarize the membrane potential of the aortic muscles. In the presence of 4-AP, the enterins failed to inhibit the muscle contraction evoked by the vulvar nerve stimulation, suggesting that the inhibition is mainly due to the activation of the 4-AP-sensitive K+ channels. The inhibition of the NdWFamide-evoked contraction by the enterin was not, however, affected by 4-AP. These results suggest that the enterins are involved in inhibitory regulation of the contractile activity of the anterior aorta, and that the inhibition could be due to multiple mechanisms.
Low temperature X-ray microanalysis of calcium in a scleractinian coral:evidence of active transport mechanisms
Authors: Clode Peta L. , Marshall Alan T.
Published: April 2021

Element concentrations were measured by X-ray microanalysis in seawater(SW) compartments and mucocytes in bulk, frozen-hydrated preparations of the scleractinian coral Galaxea fascicularis. Quantitative X-ray microanalysis of polyps sampled in the daytime revealed that concentrations of the elements Na, S, K and Ca were all significantly higher in a thin (10-20μm) external SW layer adjacent to the oral ectoderm (P<0.05,<0.05, <0.0001 and <0.01, respectively) than in standard SW. In polyps sampled during night-time, concentrations of Ca and S in this external SW layer were significantly reduced (P<0.05). Ca concentration in the coelenteron and extrathecal coelenteron was significantly higher(P<0.001) than in the external SW layer, regardless of time of sampling, suggesting that Ca2+ transport across the oral epithelium occurs via an active, transcellular route. X-ray microanalyses of mucocytes revealed that the concentration of S was high and did not vary between epithelial layers, while that of Ca increased in an inward gradient toward the skeleton. We suggest that throughout the day, secreted mucus behaves as a Donnan matrix at the oral ectoderm—SW interface,facilitating intracellular Ca2+ uptake. The accumulation within internal SW compartments of high concentrations of Ca relative to standard SW levels, however, appears to be independent of mucus secretion and is likely to be a consequence of active transport processes.
Aerobic capacities in the skeletal muscles of Weddell seals: key to longer dive durations?
Authors: Kanatous S. B. , Davis R. W. , Watson R. , Polasek L. , Williams T. M. , Mathieu-Costello O.
Published: April 2021

In contrast to terrestrial animals that function under hypoxic conditions but display the typical exercise response of increasing ventilation and cardiac output, marine mammals exercise under a different form of hypoxic stress. They function for the duration of a dive under progressive asphyxia,which is the combination of increasing hypoxia, hypercapnia and acidosis. Our previous studies on short-duration, shallow divers found marked adaptations in their skeletal muscles, which culminated in enhanced aerobic capacities that are similar to those of atheltic terrestrial mammals. The purpose of the present study was to assess the aerobic capacity of skeletal muscles from long-duration divers. Swimming and non-swimming muscles were collected from adult Weddell seals, Leptonychotes weddelli, and processed for morphometric analysis, enzymology, myoglobin concentrations and fiber-type distribution. The results showed that the skeletal muscles of Weddell seals do not have enhanced aerobic capacities compared with those of terrestrial mammals but are adapted to maintain low levels of an aerobic lipid-based metabolism, especially under the hypoxic conditions associated with diving. The lower aerobic capacity of Weddell seal muscle as compared with that of shorter-duration divers appears to reflect their energy-conserving modes of locomotion, which enable longer and deeper dives.
Electroreception in juvenile scalloped hammerhead and sandbar sharks
Authors: Kajiura Stephen M. , Holland Kim N.
Published: April 2021

The unique head morphology of sphyrnid sharks might have evolved to enhance electrosensory capabilities. The `enhanced electroreception' hypothesis was tested by comparing the behavioral responses of similarly sized carcharhinid and sphyrnid sharks to prey-simulating electric stimuli. Juvenile scalloped hammerhead sharks Sphyrna lewini and sandbar sharks Carcharhinus plumbeus oriented to dipole electric fields from the same maximum distance (approximately 30 cm) and thus demonstrated comparable behavioral-response thresholds (<1 nV cm-1). Despite the similarity of response threshold, the orientation pathways and behaviors differed for the two species. Scalloped hammerheads typically demonstrated a pivot orientation in which the edge of the cephalofoil closest to the dipole remained stationary while the shark bent its trunk to orient to the center of the dipole. By contrast, sandbars swam in a broader arc towards the center of the dipole. The different orientation patterns are attributed to the hydrodynamic properties of the cephalofoil, which enables the hammerheads to execute sharp turns at high speed. The greater trunk width of the sandbar sharks prevented them from demonstrating the same degree of flexibility. Therefore, although the sphyrnid head morphology does not appear to confer a greater sensitivity to prey-simulating dipole electric fields, it does provide(1) a greater lateral search area, which may increase the probability of prey encounter, and (2) enhanced maneuverability, which may aid in prey capture.
Elevated dietary intake of L-tryptophan counteracts the stress-induced elevation of plasma cortisol in rainbow trout (Oncorhynchus mykiss)
Authors: Lepage Olivier , Tottmar Olof , Winberg Svante
Published: April 2021

Juvenile rainbow trout were isolated in individual compartments and allowed to acclimate for 1 week, during which they were fed commercial trout pellets. The feed was then replaced by pelleted feed supplemented with L-tryptophan(TRP) at two, four or eight times the concentration in the commercial feed. Fish were fed these supplemented feeds daily to satiety for 1 week, after which half of the fish were stressed, by lowering the water level for 2 h,while the remaining fish were left undisturbed. In undisturbed fish,supplementary dietary TRP resulted in slightly elevated plasma cortisol levels. In response to the stress, fish that had been fed control feed showed elevated plasma cortisol levels, but fish fed the TRP-supplemented feed displayed a significant reduction in this stress-induced elevation of plasma cortisol levels. Plasma and brain TRP levels were elevated in fish fed TRP-supplemented feed. TRP is the precursor of the monoamine neurotransmitter serotonin. Brain serotonergic activity was elevated by stress and also tended to be increased by elevated dietary TRP intake. The central serotonergic system is involved in the control of the hypothalamic—pituitary—interrenal axis, the action of serotonin being to stimulate or inhibit this neuroendocrine axis through different projections.
Gliclazide increases insulin receptor tyrosine phosphorylation but not p38 phosphorylation in insulin-resistant skeletal muscle cells
Authors: Kumar Naresh , Dey Chinmoy S.
Published: April 2021

Sulfonylurea drugs are used in the treatment of type 2 diabetes. The mechanism of action of sulfonylureas is to release insulin from pancreatic cells and they have been proposed to act on insulin-sensitive tissues to enhance glucose uptake. The goal of the present study was to test the hypothesis that gliclazide, a second-generation sulfonylurea, could enhance insulin signaling in insulin-resistant skeletal muscle cells. We demonstrated that gliclazide enhanced insulin-stimulated insulin receptor tyrosine phosphorylation in insulin-resistant skeletal muscle cells. Although insulin receptor substrate-1 tyrosine phosphorylation was unaffected by gliclazide treatment, phosphatidylinositol 3-kinase activity was partially restored by treatment with gliclazide. No increase in 2-deoxyglucose uptake in insulin-resistant cells by treatment with gliclazide was observed. Further investigations into the mitogen-activated protein kinase (MAPK) pathway revealed that insulin-stimulated p38 phosphorylation was impaired, as compared with extracellular-signal-regulated kinase (ERK) and c-Jun N-terminal kinase(JNK), which were phosphorylated normally in insulin-resistant cells. Treatment with gliclazide could not restore p38 phosphorylation in insulin-resistant cells. We propose that gliclazide can regulate part of the insulin signaling in insulin-resistant skeletal muscle, and p38 could be a potential therapeutic target for glucose uptake to treat insulin resistance.
The notochord of hagfishMyxine glutinosa: visco-elastic properties and mechanical functions during steady swimming
Authors: Long John H. , Koob-Emunds Magdalena , Sinwell Benjamin , Koob Thomas J.
Published: April 2021

To determine the possible locomotor functions of the hagfish notochord, we measured its flexural stiffness EI (N m-2) and flexural damping C (kg m3 s-1), under in vitroconditions that mimicked the body curvature and bending frequency measured during steady undulatory swimming. To assess the notochord's contribution to the mechanical behavior of the whole body, we also measured EI and C of the whole body, the body with skin removed, and the notochord with the outer fibrous sheath removed. When subjected to dynamic bending at angular frequencies from π to 6π rad s-1 and midline curvatures from 11 to 40 m-1, 1 cm in situ body segments(N=4), located at an axial position of 37% of the body length, showed significant changes in EI, C, the Young's modulus or material stiffness (E, MPa), the net work to bend the body over a cycle(W, J) and resilience (R, % energy return). When skin,muscles and the outer fibrous sheath of the notochord were removed sequentially, each structural reduction yielded significant changes in mechanical properties: C decreased when the skin was removed, E increased when the muscles were removed, and EI and R decreased when the outer fibrous sheath was removed. Although occupying only a small portion of the cross-sectional area, the notochord provides the body with 75% of its total EI and 80% of total C, by virtue of its high E, ranging from 4 to 8 MPa, which is an order of magnitude greater than that of the whole body. Thus, as the body's primary source of EI and C, the notochord determines the passive (i.e. internal, non-muscular) mechanical behavior of the swimming hagfish. EI and C covary inversely and non-linearly such that as C increases, EI decreases. However, the bending moments M (Nm) produced by each property increase proportionally, and the ratio of stiffness to damping moments, also known as the amplification ratio at resonance, is nearly invariant (approximately 7) with changes in driving frequency. If the body operates in life at or near resonance, the variables EI and C interact over a range of swimming speeds to produce passive mechanical stability.
How the viewing of familiar landscapes prior to release allows pigeons to home faster: evidence from GPS tracking
Authors: Biro Dora , Guilford Tim , Dell'Omo Giacomo , Lipp Hans-Peter
Published: April 2021

Providing homing pigeons with a 5 min preview of the landscape at familiar sites prior to release reliably improves the birds' subsequent homing speeds. This phenomenon has been taken to suggest that the visual panorama is involved in familiar-site recognition, yet the exact nature of the improvement has never been elucidated. We employed newly developed miniature Global Positioning System (GPS) tracking technology to investigate how access to visual cues prior to release affects pigeons' flight along the length of the homing route. By applying a variety of novel analytical techniques enabled by the high-resolution GPS data (track efficiency, virtual vanishing bearings,orientation threshold), we localised the preview effect to the first 1000 m of the journey. Birds denied preview of a familiar landscape for 5 min before take-off flew an initially more tortuous path, including a high incidence of circling, possibly as part of an information-gathering strategy to determine their position. Beyond the first 1000 m, no differences were found in the performance of birds with or without preview. That the effect of the visual treatment was evident only in the early part of the journey suggests that lack of access to visual cues prior to release does not result in a non-specific effect on behaviour that is maintained throughout the flight. Instead, it seems that at least some decisions regarding the direction of home can be made prior to release and that such decisions are delayed if visual access to the landscape is denied. Overall, the variety of approaches applied here clearly highlight the potential for future applications of GPS tracking technology in navigation studies.
Effects of waterborne exposure of octylphenol and oestrogen on pregnant viviparous eelpout (Zoarces viviparus) and her embryos in ovario
Authors: Rasmussen Tina H. , Andreassen Thomas K. , Pedersen Søren N. , Van der Ven Leo T. M. , Bjerregaard Poul , Korsgaard Bodil
Published: April 2021

Exposure to oestrogenic chemicals (xeno-oestrogens) may have severe effects on embryonic development. The present study investigates whether the oestrogenic endocrine disruptor 4-tert-octylphenol (4-tOP) or 17β-oestradiol (E2) is accumulated in the viviparous fish the eelpout (Zoarces viviparus) and transferred to the embryos in ovario and subsequently affects embryonic development, including gonadal differentiation. Pregnant eelpouts were exposed to nominal concentrations of 25 μgl-1 or 100 μgl-1 4-tOP (OP25 or OP100,respectively) or 0.5 μgl-1 E2 in water. During 4-tOP exposure, the compound accumulated in both plasma and ovarian fluid in a concentration-dependent manner. In the mother fish, the oestrogenic biomarkers, vitellogenin (Vtg) in plasma, Vtg mRNA in liver and oestrogen-binding activity in liver, were all induced by 4-tOP (and by E2) at an actual concentration of 14 μgl-1. E2 and 4-tOP were examined for their potency to disturb the maternal—foetal trophic relationship by disturbing the physiology of the ovary and by changing the distribution of essential nutrients normally transported to embryos during pregnancy. After exposure to E2 or 4-tOP, calcium was depleted from the ovarian fluid and the level of free amino acids available in maternal plasma was decreased. A marked overall effect on ovarian components, including the ovarian sac, ovarian fluid and embryonic mass, was evident. Embryonic growth was significantly decreased, which might in part be attributed to disturbances of the maternal—foetal trophic relationship. Marked inductions of Vtg mRNA and Vtg protein, determined by RT-PCR and immunohistochemistry, respectively, were found in embryos from the OP100 group — the only group to show considerable accumulation of an oestrogenic compound in the ovarian fluid. A different pattern of gonadal development was found in embryos from the OP100 group compared with embryos from the control, OP25 or E2 groups, in which approximately 50% had normal ovaries and 50% had normal presmptive male gonads. In the OP100 group,46% had normal ovaries but, in contrast to controls, only 22% had normal presumptive male gonads, whereas the remaining 32% had abnormal male gonads with structures resembling the endo-ovarian cavity of a female gonad. As oestrogen receptor (ER) expression was detected by in situhybridisation in early differentiating gonads, these effects could be mediated by direct interaction of the xeno-oestrogens with gonadal ER. In conclusion,this study indicates that the xeno-oestrogen 4-tOP can be transferred from the water via the mother fish to the ovarian fluid and can subsequently disturb the maternal—foetal trophic relationship and cause severe effects on embryonic development, including gonadal differentiation in ovario.
Levamisole receptor phosphorylation: effects of kinase antagonists on membrane potential responses inAscaris suumsuggest that CaM kinase and tyrosine kinase regulate sensitivity to levamisole
Authors: Trailovic Sasa M. , Robertson Alan P. , Clark Cheryl L. , Martin Richard J.
Published: April 2021

A two-micropipette current-clamp technique was used to record electrophysiological responses from the somatic muscle of Ascaris suum. Levamisole and acetylcholine were applied to the bag region of the muscle using a microperfusion system. Depolarizations produced by 10 s applications of 10 μmol l-1 levamisole or 20 s applications of 10 μmol l-1 acetylcholine were recorded. The effect on the peak membrane potential change of the kinase antagonists H-7, staurosporine, KN-93 and genistein was observed. H-7 (30 μmol l-1), a non-selective antagonist of protein kinases A, C and G but which has little effect on Ca2+/calmodulin-dependent kinase II (CaM kinase II), did not produce a significant effect on the peak response to levamisole or acetylcholine. Staurosporine (1 μmol l-1), a non-selective kinase antagonist that has effects on protein kinases A, C and G, CaM kinase and tyrosine kinase, reduced the mean peak membrane potential response to levamisole from 6.8 mV to 3.9 mV (P<0.0001) and the mean response to acetylcholine from 5.5 mV to 2.8 mV (P=0.0016). The difference between the effects of H-7 and staurosporine suggested the involvement of CaM kinase II and/or tyrosine kinase. KN-93, a selective CaM kinase II antagonist,reduced the mean peak response to levamisole from 6.2 mV to 2.7 mV(P=0.035) and the mean peak response of acetylcholine from 4.7 mV to 2.0 mV (P=0.0004). The effects indicated the involvement of CaM kinase II in the phosphorylation of levamisole and acetylcholine receptors. The effect of extracellular Ca2+ on the response to levamisole was assessed by comparing responses to levamisole in normal and in low-Ca2+ bathing solutions. The response to levamisole was greater in the presence of Ca2+, an effect that may be explained by stimulation of CaM kinase II. Genistein (90 μmol l-1), a selective tyrosine kinase antagonist, reduced peak membrane potential responses to levamisole from a mean of 6.4 mV to 3.3 mV (P=0.001). This effect indicated the involvement of tyrosine kinase in maintaining the receptor.
The influence of visual landscape on the free flight behavior of the fruit flyDrosophila melanogaster
Authors: Tammero Lance F. , Dickinson Michael H.
Published: April 2021

To study the visual cues that control steering behavior in the fruit fly Drosophila melanogaster, we reconstructed three-dimensional trajectories from images taken by stereo infrared video cameras during free flight within structured visual landscapes. Flies move through their environment using a series of straight flight segments separated by rapid turns, termed saccades, during which the fly alters course by approximately 90° in less than 100 ms. Altering the amount of background visual contrast caused significant changes in the fly’s translational velocity and saccade frequency. Between saccades, asymmetries in the estimates of optic flow induce gradual turns away from the side experiencing a greater motion stimulus, a behavior opposite to that predicted by a flight control model based upon optomotor equilibrium. To determine which features of visual motion trigger saccades, we reconstructed the visual environment from the fly’s perspective for each position in the flight trajectory. From these reconstructions, we modeled the fly’s estimation of optic flow on the basis of a two-dimensional array of Hassenstein–Reichardt elementary motion detectors and, through spatial summation, the large-field motion stimuli experienced by the fly during the course of its flight. Event-triggered averages of the large-field motion preceding each saccade suggest that image expansion is the signal that triggers each saccade. The asymmetry in output of the local motion detector array prior to each saccade influences the direction (left versus right) but not the magnitude of the rapid turn. Once initiated, visual feedback does not appear to influence saccade kinematics further. The total expansion experienced before a saccade was similar for flight within both uniform and visually textured backgrounds. In summary, our data suggest that complex behavioral patterns seen during free flight emerge from interactions between the flight control system and the visual environment.
Tissue- and development-specific induction and turnover of hsp70 transcripts from loci 87A and 87C after heat shock and during recovery inDrosophila melanogaster
Authors: Lakhotia S. C. , Prasanth K. V.
Published: April 2021

The haploid genome of Drosophila melanogaster normally carries at least five nearly identical copies of heat-shock-inducible hsp70 genes, two copies at the 87A7 and three copies at the 87C1 chromosome sites. We used in situ hybridization of the cDNA, which hybridizes with transcripts of all five hsp70 genes, and of two 3′ untranslated region (3′UTR; specific for the 87A7- and 87C1-type hsp70 transcripts) riboprobes to cellular RNA to examine whether all these copies were similarly induced by heat shock in different cell types of D. melanogaster. Our results revealed remarkable differences not only in the heat-shock-inducibility of the hsp70 genes at the 87A7 and 87C1 loci, but also in their post-transcriptional metabolism, such as the stability of the transcripts and of their 3′UTRs in different cell types in developing embryos and in larval and adult tissues. Our results also revealed the constitutive presence of the heat-shock-inducible form of Hsp70 in a subset of late spermatogonial cells from the second-instar larval stage onwards. We suggest that the multiple copies of the stress-inducible hsp70 genes do not exist in the genome of D. melanogaster only to produce large amounts of the Hsp70 rapidly and at short notice, but that they are specifically regulated in a developmental-stage-specific manner. It is likely that the cost/benefit ratio of not producing or of producing a defined amount of Hsp70 under stress conditions varies for different cell types and under different physiological conditions and, accordingly, specific regulatory mechanisms operating at the transcriptional and post-transcriptional levels have evolved.
Kinematics of diving Atlantic puffins (Fratercula arcticaL.):evidence for an active upstroke
Authors: Johansson L. Christoffer , Aldrin Björn S. Wetterholm
Published: April 2021

To examine the propulsion mechanism of diving Atlantic puffins (Fratercula arctica), their three-dimensional kinematics was investigated by digital analysis of sequential video images of dorsal and lateral views. During the dives of this wing-propelled bird, the wings are partly folded, with the handwings directed backwards. The wings go through an oscillating motion in which the joint between the radius-ulna and the hand bones leads the motion, with the wing tip following. There is a large rotary motion of the wings during the stroke, with the wings being pronated at the beginning of the downstroke and supinated at the end of the downstroke/beginning of the upstroke. Calculated instantaneous velocities and accelerations of the bodies of the birds show that, during the downstroke, the birds accelerate upwards and forwards. During the upstroke, the birds accelerate downwards and, in some sequences analysed, also forwards, but in most cases the birds decelerate. In all the upstrokes analysed, the forward/backward acceleration shows the same pattern, with a reduced deceleration or even a forward acceleration during ‘mid’ upstroke indicating the production of a forward force, thrust. Our results show that the Atlantic puffin can use an active upstroke during diving, in contradiction to previous data. Furthermore, we suggest that the partly folded wings of diving puffins might act as efficient aft-swept wingtips, reducing the induced drag and increasing the lift-to-drag ratio.

A movie is available on-line.
The pulmonary surfactant system matures upon pipping in the freshwater turtleChelydra serpentina
Authors: Johnston Sonya D. , Daniels Christopher B. , Cenzato David , Whitsett Jeffrey A. , Orgeig Sandra
Published: April 2021

Pulmonary surfactant (PS), a mixture of phospholipids (PL), neutral lipids and surfactant proteins (SP), lowers surface tension within the lung, which increases lung compliance and improves the removal of fluid at birth. Here, we have examined the expression of thyroid transcription factor-1 (TTF-1) and the surfactant protein SP-B, and also the composition of pulmonary surfactant lipids in the developing lung of the turtle Chelydra serpentina. Lavage and lung tissue were collected from late embryonic, pipped and hatchling turtles. TTF-1, a regulator of gene expression of surfactant proteins and cell differentiation in mammals, was detected using immunohistochemistry in epithelia of the gas-exchange area and conducting airways during late development. Expression declined in hatchlings. SP-B was detected in subsets of cells within the respiratory epithelium at all stages sampled. The same cell types also stained for TTF-1. Turtle surfactant lipids matured toward the end of incubation. Maximal secretion of both total phospholipids and disaturated phospholipid (DSP) occurred at the time of pipping, coincident with the onset of breathing. The DSP/PL ratio increased after pipping, whereas cholesterol levels (Chol) increased prior to pipping. This resulted in a decrease in the Chol/PL and Chol/DSP ratios after pipping. Thus, TTF-1 and SP-B appear to be highly conserved within the vertebrates. Maturation of surfactant phospholipid content occurred with the commencement of pulmonary ventilation.
Insect midgut K+ secretion: concerted run-down of apical/basolateral transporters with extra-/intracellular acidity
Authors: Zeiske Wolfgang , Meyer Heiko , Wieczorek Helmut
Published: April 2021

In lepidopteran larvae, three transport mechanisms are involved in the active and electrogenic K+ secretion that occurs in the epithelial goblet cells of the midgut. These consist of (i) basolateral K+ channels, allowing K+ entry from the haemolymph into the cytosol, (ii) apical electrogenic K+/2H+ antiporters, which are responsible for secondary active extrusion of K+ from the cell into the gut lumen via the goblet cavity and (iii) apical V-ATPase-type proton pumps. The latter energize apical K+ exit by building up a large, cavity-positive electrical potential that drives the antiporters. Net K+ secretion (IK) can be measured as short-circuit current (Isc) across the in vitro midgut mounted in an Ussing chamber. We investigated the influence of protons on the transepithelial IK and the partial reactions of the basolateral K+ permeability (PK) and the apical, lumped ‘K+ pump’ current (IP) at various extra- and intracellular pH values. In particular, we wanted to know whether increased cellular acidity could counteract the reversible dissociation of the V-ATPase into its V1 and Vo parts, as occurs in yeast after glucose deprivation and in the midgut of Manduca sexta during starvation or moulting, thus possibly enhancing K+ transport.When intact epithelia were perfused with high-[K+] (32 mmol l–1) salines with different pH values, IK was reversibly reduced when pH values fell below 6 on either side of the epithelium. Attempts to modify the intracellular pH by pulsing with NH4+ or propionate showed that intracellular acidification caused a reduction in IK similar to that obtained in response to application of external protons. Treatment with azide, a well-known inhibitor of the mitochondrial ATP synthase, had the same effect as pulsing with ammonium or propionate with, however, much faster kinetics and higher reversibility. Breakdown of the basolateral or apical barrier using the antibiotic nystatin allowed the intracellular pH to be clamped to that of the saline facing the nystatin-treated epithelial border. Cell acidification achieved by this manipulation led to a reduction in both apical IP and basolateral PK. The transepithelial IK showed an approximately half-maximal reduction at external pH values close to 5 in intact tissues, and a similar reduction in IP and PK values was seen at an intracellular pH of 5 in nystatin-permeabilised epithelia. Thus, the hypothesized V1Vo stabilization by cell acidity is not reflected in the pH-sensitivity of IP. Moreover, all components that transport K+ are synchronously inhibited below pH 6. The significance of our findings for the midgut in vivo is discussed.
Evolution of voltage-gated Na+ channels
Authors: Goldin Alan L.
Published: April 2021

Voltage-gated Na+ channels play important functional roles in the generation of electrical excitability in most vertebrate and invertebrate species. These channels are members of a superfamily that includes voltage-gated K+, voltage-gated Ca2+ and cyclic-nucleotide-gated channels. There are nine genes encoding voltage-gated Na+ channels in mammals, with a tenth homologous gene that has not been shown to encode a functional channel. Other vertebrate and invertebrate species have a smaller number of Na+ channel genes. The mammalian genes can be classified into five branches in a phylogenetic tree, and they are localized on four chromosomes. Four of the branches representing the four chromosomal locations probably resulted from the chromosomal duplications that led to the four Hox gene clusters. These duplications occurred close to the emergence of the first vertebrates. The fifth branch probably evolved from a separate ancestral Na+ channel gene. There are two branches in the invertebrate tree, although members of only one of those branches have been demonstrated to encode functional voltage-gated Na+ channels. It is possible that the other branch may have diverged, so that its members do not represent true voltage-gated Na+ channels. Vertebrate and invertebrate Na+ channels appear to be derived from a single primordial channel that subsequently evolved independently in the two lineages.
Ticking of the clockwork cricket: the role of the escapement mechanism
Authors: Bennet-Clark H. C. , Bailey Winston J.
Published: April 2021

The ‘clockwork cricket’ model for cricket sound production suggests that the catch-and-release of the file of one forewing by the plectrum on the opposite wing act as an ‘escapement’ to provide the phasic impulses that initiate and sustain the vibration of the resonant regions of the wings from which the sounds are produced. The action of the escapement produces the familiar ticking sound of clocks.

The higher-frequency components of the songs of twelve species of cricket were analysed after removing the dominant low-frequency components and amplifying the remaining higher-frequency components. In normal song pulses of all species, the higher-frequency components showed a close phase-locking to the waveform of the dominant frequency, but the amplitude of the higher-frequency components did not correlate with that at the dominant frequency.

Anomalous pulses occurred spontaneously in the songs of several species: multimodal, interrupted or curtailed pulses are described. In all of these, the anomalous pulse envelope was associated with changes in the amplitude and/or instantaneous frequency of the higher-frequency components of the sound.

A model of the escapement suggests that the frequency of the residual components of the song depends on the symmetry of action of the plectrum on the teeth of the file.
How the efficiency of rainbow trout (Oncorhynchus mykiss) ventricular muscle changes with cycle frequency
Authors: Harwood Claire L. , Young Iain S. , Altringham John D.
Published: April 2021

Different species of animals require different cardiac performance and, in turn, their cardiac muscle exhibits different properties. A comparative approach can reveal a great deal about the mechanisms underlying myocardial contraction. Differences in myocardial Ca2+ handling between fish and mammals suggest a greater energy cost of activation in fish. Further, while there is considerable evidence that heart rate (or cycle frequency) should have a profound effect on the efficiency of teleost cardiac muscle, this effect has been largely overlooked. We set out to determine how cycle frequency affects the power output and efficiency of rainbow trout (Oncorhynchus mykiss) ventricular muscle and to relate this to the heart’s function in life. We measured power output and the rate of oxygen consumption (V̇O2) and then calculated efficiency over a physiologically realistic range of cycle frequencies.

In contrast to mammalian cardiac muscle, in which V̇O2 increases with increasing heart rate, we found no significant change in V̇O2 in the teleost. However, power output increased by 25 % as cycle frequency was increased from 0.6 to 1.0 Hz, so net and total efficiency increased. A maximum total efficiency of 20 % was achieved at 0.8 Hz, whereas maximum power output occurred at 1.0 Hz. We propose that, since the heart operates continuously, high mechanical efficiency is a major adaptive advantage, particularly at lower heart rates corresponding to the more commonly used slower, sustainable swimming speeds. Efficiency was lower at the higher heart rates required during very fast swimming, which are used during escape or prey capture.

If a fixed amount of Ca2+ is released and then resequestered each time the muscle is activated, the activation cost should increase with frequency. We had anticipated that this would have a large effect on the total energy cost of contraction. However, since V̇O2 remains constant, less oxygen is consumed per cycle at high frequencies. We suggest that a constant V̇O2 would be observed if the amount of activator Ca2+ were to decrease with frequency. This decrease in activation energy is consistent with the decrease in the systolic intracellular Ca2+ ([Ca2+]i) transient with increasing stimulation frequency seen in earlier studies.
Oocyte apoptosis during the transition from ovary-like tissue to testes during sex differentiation of juvenile zebrafish
Authors: Uchida Daisuke , Yamashita Michiaki , Kitano Takeshi , Iguchi Taisen
Published: April 2021

Large numbers of apoptotic early diplotene oocytes were observed during the transition from ovary-like undifferentiated gonadal tissue to testes during sex differentiation in presumptive males of the zebrafish (Danio rerio). The percentage of terminal-deoxynucleotidyl-transferase-mediated dUTP nick-end labelling (TUNEL)-positive apoptotic oocytes in the gonads of presumptive males was approximately eight- to 12-fold higher than in genetic all-females. By 29 days post-hatching, all oocytes had disappeared from the gonads of presumptive males. In these males, we also observed apoptotic somatic cells in the ovarian cavity between 23 and 35 days post-hatching. Therefore, the disappearance of oocytes and the decomposition of the ovarian cavity caused by apoptosis during sex differentiation were male-specific events. In genetic all-females, apoptosis in a proportion of early diplotene oocytes was found in the undifferentiated gonads at 15–19 days post-hatching, probably as a result of programmed oocyte loss during ovarian development. These findings suggest that oocyte apoptosis is the mechanism of testicular and ovarian differentiation in zebrafish.
Peripheral oxygen transport in skeletal muscle of Antarctic and sub-Antarctic notothenioid fish
Authors: Egginton S. , Skilbeck C. , Hoofd L. , Calvo J. , Johnston I. A.
Published: April 2021

Transcellular oxygen flux was modelled mathematically in the aerobic skeletal muscles of perciform fish species living at widely different temperatures (Antarctica, sub-Antarctica and the Mediterranean Sea). Using structural data derived from stereological analysis of electron micrographs, mean fibre PO2 was calculated on the basis of temperature-corrected rates of mitochondrial respiration and oxygen diffusion. The mean muscle fibre diameter (MFD) among Antarctic notothenioids was in the range 17–61 μm and mitochondrial volume density, Vv(mit,f), was 0.27–0.53, but capillary-to-fibre ratio varied only between 1.2 and 1.5. For a mean capillary PO2 of 6 kPa, the model predicted a mean tissue PO2 in the range 0.7–5.8 kPa at the estimated maximum aerobic capacity (ṀO2max). The lowest levels of tissue oxygenation were found in the pectoral muscle fibres of the icefish Chaenocephalus aceratus, which lacks the respiratory pigments haemoglobin and myoglobin. Red-blooded notothenioids found in the sub-Antarctic had a similar muscle fine structure to those caught south of the Antarctic Convergence, with an MFD of 20–41 μm and Vv(mit,f) of 0.27–0.33, resulting in an estimated mean PO2 of 4–5 kPa at ṀO2max. Mean tissue PO2 in the sub-Antarctic icefish Champsocephalus esox, with greater MFD and Vv(mit,f), 56 μm and 0.51, respectively, was calculated to exceed 1 kPa at winter temperatures (4°C), although oxidative metabolism was predicted to be impaired at the summer maximum of 10°C. At the high end of the thermal range, related perciform species from the Mediterranean had a negligible drop in intracellular PO2 across their small-diameter fibres, to a minimum of 5.4 kPa, comparable with that predicted for Trematomus newnesi from the Antarctic (5.6 kPa) with a similar MFD. These data suggest that, within a single phylogenetic group, integrative structural adaptations potentially enable a similar degree of tissue oxygenation over a 20°C range of environmental temperature in the red-blooded notothenioids, and that this is compromised by the lack of respiratory pigments in the icefishes. The mean capillary radius was 1.5 times greater in the two icefish than in the other notothenioids, and the model simulations indicate that the evolution of wide-bore capillaries is essential to maintain tissue oxygenation in the absence of respiratory pigments.
Cn11, the first example of a scorpion toxin that is a true blocker of Na+ currents in crayfish neurons
Authors: Ramirez-Dominguez Martha E. , Olamendi-Portugal Timoteo , Garcia Ubaldo , Garcia Consuelo , Arechiga Hugo , Possani Lourival D.
Published: April 2021

A novel crustacean toxin (Cn11) was isolated and characterized from the venom of the Mexican scorpion Centruroides noxius Hoffmann. It contains 63 amino acid residues and is stabilized by four disulphide bridges. It is lethal to crustaceans (Cambarellus montezumae), less toxic to insects (crickets) and non-toxic to mammals (mice) at the doses assayed. In neurons isolated from the X organ–sinus gland system of the crayfish Procambarus clarkii, it blocks the Na+ currents with an estimated Km of 320 nmol l–1, without affecting the Ca2+ and K+ currents. The voltage-gated tetrodotoxin-sensitive Na+ current was recorded from X organ neurons in culture 24 h after plating using the whole-cell clamp configuration. The Na+ current was isolated by blocking Ca2+ currents with Cd2+ and Cs+ and K+ currents with tetraethylammonium and 4-aminopyridine. Under control conditions, the Na+ currents were activated at –40 mV with a maximum amplitude at 0 mV. In the presence of 1 μmol l–1 Cn11, the Na+ current amplitude was reduced by 75 % without apparent modifications to the gating mechanism. These findings suggest that Cn11 selectively blocks a Na+ channel. It is the first representative of a new group of scorpion toxins specific for this molecular target.
Visual pigments and oil droplets in diurnal lizards
Authors: Loew Ellis R. , Fleishman Leo J. , Foster Russell G. , Provencio Ignacio
Published: April 2021

We report microspectrophotometric (MSP) data for the visual pigments and oil droplets of 17 species of Caribbean anoline lizard known to live in differing photic habitats and having distinctly different dewlap colors. The outgroup Polychrus marmoratus was also examined to gain insight into the ancestral condition. Except for Anolis carolinensis, which is known to use vitamin A2 as its visual pigment chromophore, all anoline species examined possessed at least four vitamin-A1-based visual pigments with maximum absorbance (λmax) at 564, 495,455 and 365 nm. To the previously reported visual pigments for A. carolinensis we add an ultraviolet-sensitive one withλ max at 365 nm. Five common classes of oil droplet were measured, named according to apparent color and associated with specific cone classes — yellow and green in long-wavelength-sensitive (LWS) cones,green only in medium-wavelength-sensitive (MWS) cones and colorless in short-wavelength-sensitive (SWS) and ultraviolet-sensitive (UVS) cones. MSP data showed that the colorless droplet in the SWS cone had significant absorption between 350 and 400 nm, while the colorless droplet in the UVS cone did not. The pattern for Polychrus marmoratus was identical to that for the anoles except for the presence of a previously undescribed visual cell with a rod-like outer segment, a visual pigment with a λmaxof 497 nm and a colorless oil droplet like that in the UVS cones. These findings suggest that anoline visual pigments, as far as they determine visual system spectral sensitivity, are not necessarily adapted to the photic environment or to the color of significant visual targets (e.g. dewlaps).
Effects of temperature on sustained swimming performance and swimming kinematics of the chub mackerelScomber japonicus
Authors: Dickson Kathryn A. , Donley Jeanine M. , Sepulveda Chugey , Bhoopat Lisa
Published: April 2021

The effects of a 6°C difference in water temperature on maximum sustained swimming speed, swimming energetics and swimming kinematics were measured in the chub mackerel Scomber japonicus (Teleostei:Scombridae), a primarily coastal, pelagic predator that inhabits subtropical and temperate transition waters of the Atlantic, Pacific and Indian Oceans. New data for chub mackerel acclimated to 18°C are compared with published data from our laboratory at 24°C. Twelve individuals acclimated to each of two temperatures (15.6-26.3 cm fork length, FL, and 34-179g at 18°C; 14.0-24.7 cm FL and 26-156g at 24°C) swam at a range of speeds in a temperature-controlled Brett-type respirometer, at the respective acclimation temperature. At a given fish size, the maximum speed that S. japonicus was able to maintain for a 30-min period, while swimming steadily using slow, oxidative locomotor muscle (Umax,c),was significantly greater at 24 than at 18°C (52.5-97.5 cm s-1at 18°C and 70-120 cm s-1 at 24°C). At a given speed and fish size, the rate of oxygen consumption(V̇O2) was significantly higher at 24 than at 18°C because of a higher net cost of transport (1073-4617 J km-1 kg-1 at 18°C and 2708-14895 J km-1 kg-1 at 24°C). Standard metabolic rate, calculated by extrapolating the logV̇O2versus swimming speed relationship to zero speed, did not vary significantly with temperature or fish mass (126.4±67.2 mg O2 h-1 kg-1 at 18°C and 143.2±80.3 mg O2 h-1 kg-1 at 24°C; means ±S.D., N=12). Swimming kinematics was quantified from high-speed (120 Hz) video recordings analyzed with a computerized, two-dimensional motion-analysis system. At a given speed and fish size, there were no significant effects of temperature on tail-beat frequency, tail-beat amplitude or stride length, but propulsive wavelength increased significantly with temperature as a result of an increase in propulsive wave velocity. Thus, the main effects of temperature on chub mackerel swimming were increases in both Umax,c and the net cost of swimming at 24°C. Like other fishes, S. japonicus apparently must recruit more slow,oxidative muscle fibers to swim at a given sustainable speed at the lower temperature because of the reduced power output. Thus, the 24°C mackerel reach a higher speed before they must recruit the fast, glycolytic fibers,thereby increasing Umax,c at 24°C. By quantifying in vivo the effects of temperature on the swimming performance of an ectothermic species that is closely related to the endothermic tunas, this study also provides evidence that maintaining the temperature of the slow,oxidative locomotor muscle at 6°C or more above ambient water temperature in tunas should significantly increase sustainable swimming speeds, but also increase the energetic cost of swimming, unless cardiac output limits muscle performance.