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Biomedical subjects

Göran E Nilsson

Publications and source records attributed to Göran E Nilsson.

At least 19 recordsLinked to original sources

From record performance to hypoxia tolerance: respiratory transition in damselfish larvae settling on a coral reef.

The fastest swimming fishes in relation to size are found among coral reef fish larvae on their way to settle on reefs. By testing two damselfishes, Chromis atripectoralis and Pomacentrus amboinensis, we show that the high swimming speeds of the pre-settlement larvae are accompanied by the highest rates of oxygen uptake ever recorded in ectothermic vertebrates. As expected, these high rates of oxygen uptake occur at the cost of poor hypoxia tolerance. However, hypoxia tolerance is needed when coral reef fishes seek nocturnal shelter from predators within coral colonies, which can become severely hypoxic microhabitats at night. When the larvae settle on the reef, we found that they go through a striking respiratory transformation, i.e. the capacity for rapid oxygen uptake falls, while the ability for high-affinity oxygen uptake at low oxygen levels is increased. This transition to hypoxia tolerance is needed when they settle on the reef; this was strengthened by our finding that small resident larvae of Acanthochromis polyacanthus, a damselfish lacking a planktonic larval stage, do not display such a transition, being well adapted to hypoxia and showing relatively low maximum rates of oxygen uptake that change little with age.

Adaptation, Physiological↗

Evolutionary background for stress-coping styles: relationships between physiological, behavioral, and cognitive traits in non-mammalian vertebrates.

Reactions to stress vary between individuals, and physiological and behavioral responses tend to be associated in distinct suites of correlated traits, often termed stress-coping styles. In mammals, individuals exhibiting divergent stress-coping styles also appear to exhibit intrinsic differences in cognitive processing. A connection between physiology, behavior, and cognition was also recently demonstrated in strains of rainbow trout (Oncorhynchus mykiss) selected for consistently high or low cortisol responses to stress. The low-responsive (LR) strain display longer retention of a conditioned response, and tend to show proactive behaviors such as enhanced aggression, social dominance, and rapid resumption of feed intake after stress. Differences in brain monoamine neurochemistry have also been reported in these lines. In comparative studies, experiments with the lizard Anolis carolinensis reveal connections between monoaminergic activity in limbic structures, proactive behavior in novel environments, and the establishment of social status via agonistic behavior. Together these observations suggest that within-species diversity of physiological, behavioral and cognitive correlates of stress responsiveness is maintained by natural selection throughout the vertebrate sub-phylum.

Adaptation, Psychological↗

Plasticity of respiratory structures--adaptive remodeling of fish gills induced by ambient oxygen and temperature.

While a large surface area combined with short diffusion distances make fish gills well suited for gas exchange, these properties leads to costly water and ion fluxes and exposure to toxic substances and pathogens. Thus, gill morphology is likely to be a compromise between opposing demands. It has become clear that some fishes have the ability to modify gill structure in response to environmental parameters such as oxygen levels and temperature. Maybe the most dramatic example of gill plasticity is the adaptive and reversible changes in gill surface area displayed by crucian carp (Carassius carassius) and goldfish (Carassius auratus). Here, a cell mass is filling up the space between the lamellae during normoxic and cold conditions (i.e. when oxygen demands are low). Hypoxia or high temperature induce apoptosis and suppress mitosis in the interlamellar cell mass causing it to retract and the lamellae to protrude. The functional importance of oxygen and temperature induced changes in gill morphology and the underlying mechanisms are discussed.

Acclimatization↗

Behavioral indicators of stress-coping style in rainbow trout: Do males and females react differently to novelty?

It is becoming increasingly clear that individual differences in the behavioral response to stressful situations are associated with distinct physiological profiles, and stress coping characteristics are of fundamental importance to fitness and life history. Teleost fishes display considerable variation in reproductive strategy, but sex differences in stress-coping style have not been described previously in fish. Prior to sexual maturation, the glucocorticoid response to stress is not affected by sex in salmonid fish. Nevertheless, behavior in novel and stressful situations differed between immature male and female rainbow trout (Oncorhynchus mykiss). When tested 1 week following transport to a new rearing facility, females resumed feeding after transfer to social isolation quicker than males. The locomotor response to acute confinement stress also varied between sexes, with females settling down and ceasing to move in a panic-like manner quicker than males. There was a strong correlation between behavior in the two test situations: individuals that readily resumed feeding behavior in a new environment also moved less in the acute stress test. Thus, the time to resume feeding after a stressful experience is a precise indicator of stress-coping style in salmonid fish, which is likely to reflect the dynamics of neuroendocrine stress responses. Furthermore, these observations could reflect a sex difference in the response to novel and stressful situations, which occur even in the absence of differences in glucocorticoid responsiveness.

Adaptation, Psychological↗

Temperature regulates hypoxia-inducible factor-1 (HIF-1) in a poikilothermic vertebrate, crucian carp (Carassius carassius).

Hypoxia-inducible transcription factor-1 (HIF-1) is a master regulator of hypoxia-induced gene responses. To find out whether HIF-1 function is involved in gene expression changes associated with temperature acclimation as well as in hypoxia adaptation in poikilotherms, we studied HIF-1 DNA binding activity and HIF-1alpha expression in normoxia and during hypoxia (0.7 mg l(-1) O2) in crucian carp at temperatures of 26, 18 and 8 degrees C. Temperature had a marked influence on HIF-1 in normoxia. Although HIF-1alpha mRNA levels remained unaltered, cold acclimation (8 degrees C) increased HIF-1alpha protein amounts in the liver, gills and heart and HIF-1 DNA binding activity in the heart, gills and kidney of crucian carp by two- to threefold compared to warm acclimated fish (26 degrees C). In the heart and kidney HIF-1 activity was already significantly increased in the 18 degrees C acclimated fish. Temperature also affected hypoxic regulation of HIF-1. Although hypoxia initially increased amounts of HIF-1alpha protein in all studied tissues at every temperature, except for liver at 18 degrees C, HIF-1 activity increased only in the heart of 8 degrees C acclimated and in the gills of 18 degrees C acclimated fish. At 8 degrees C HIF-1alpha mRNA levels increased transiently in the gills after 6 h of hypoxia and in the kidney after 48 h of hypoxia. In the gills at 26 degrees C HIF-1alpha mRNA levels increased after 6 h of hypoxia and remained above normoxic levels for up to 48 h of hypoxia. These results show that HIF-1 is involved in controlling gene responses to both oxygen and temperature in crucian carp. No overall transcriptional control mechanism has been described for low temperature acclimation in poikilotherms, but the present results suggest that HIF-1 could have a role in such regulation. Moreover, this study highlights interaction of the two prime factors defining metabolism, temperature and oxygen, in the transcriptional control of metabolic homeostasis in animals.

Acclimatization↗

Shrimps remove ectoparasites from fishes in temperate waters.

We have found that two very common species of North Atlantic shallow water shrimp, Palaemon adspersus and Palaemon elegans, remove and feed on ectoparasites on plaice (Pleuronectes platessa L.). The relationship could be mutualistic, as we did not observe any attempts by the fishes to feed on the shrimps. The ectoparasites removed included monogenean worms (Gyrodactylus sp.) and sea lice (Lepeophtheirus pectoralis). An experiment showed that there were 65% more Gyrodactylus parasites on the fishes that had been apart from compared with those that had been together with shrimps for 48h. Shrimps on coral reefs are known for cleaning fishes, but that shrimps in temperate waters show parasite-cleaning behaviour is, to our knowledge, a new observation.

Animals↗

HIF-1alpha and iNOS levels in crucian carp gills during hypoxia-induced transformation.

Hypoxia inducible factor 1 alpha (HIF-1alpha) initiates expression of a wide variety of genes, some of which are involved in apoptosis and cell cycle arrest. We have previously shown that crucian carp increases its respiratory surface area 7.5-fold in response to hypoxia. This change is due to apoptosis and cell cycle arrest in specific parts of its gills. Here we have characterized crucian carp HIF-1alpha, and measured mRNA, protein and DNA binding levels during hypoxia exposure in crucian carp gills. We have also measured an HIF-1alpha-induced gene, the inducible nitric oxide synthase (iNOS), which has the ability to initiate apoptosis and cell cycle arrest. Crucian carp HIF-1alpha was found to have all critical domains known to be important for function. Comparison of the peptide sequence with other species indicated high similarity with other cyprinid fish, but a pronounced variation compared to the salmonid, rainbow trout. Further, we found HIF-1alpha protein to be stabilized during hypoxia. Further, HIF-1alpha was often present in normoxia, and showed marked individual weight-dependent variation. We found no alteration of iNOS mRNA levels during hypoxia exposure. These findings suggest HIF-1alpha involvement in hypoxia-induced change of respiratory surface area in crucian carp gills. However, its activity does not seem to be mediated through iNOS.

Amino Acid Sequence↗

Endothelin receptors in teleost fishes: cardiovascular effects and branchial distribution.

By observing gill blood flow using epi-illuminating microscopy, in parallel with cardiovascular recordings and immunohistochemistry, we have tried to identify the receptor mediating endothelin (ET) type 1 (ET1)-induced pillar cell contraction in the lamellae of the Atlantic cod (Gadus morhua). Intra-arterial injection of the specific ET(B) receptor agonist BQ-3020 induced dose-dependent increases in ventral aortic blood pressure, gill vascular resistance, and pillar cell area (indicating contraction). The specific ET(A) receptor antagonist BQ-610 did not prevent either pillar cell contraction or increased gill vascular resistance induced by ET-1 injection. The cardiovascular responses were corroborated by the detection of ET(B) receptor-like immunoreactivity (IR) associated with pillar cells in the lamellar region and in neuroendocrine cells. ET(B) receptor-like IR was also found lining the muscle layer of lamellar arterioles and filament arteries. In contrast, strong ET(A) receptor-like IR was found on branchial nerves throughout the filaments. In addition, ET-like IR was concentrated in neuroendocrine cells in the filament and lamellae. We also present data suggesting that ET-mediated pillar cell contraction is widespread among teleost fish, including Atlantic cod, rainbow trout (Oncorhynchus mykiss), sculpin (Myoxocephalus scorpius), and mackerel (Scomber scombrus). Taken together, our results suggest that an ET(B)-like receptor mediates pillar cell contraction in fishes, whereas ET(A)-like receptors may serve another function in the gill, inasmuch as ET(A) receptor-like IR is found on branchial nerves.

Animals↗

Cell proliferation and gill morphology in anoxic crucian carp.

Is DNA replication/cell proliferation in vertebrates possible during anoxia? The oxygen dependence of ribonucleotide reductase (RNR) could lead to a stop in DNA synthesis, thereby making anoxic DNA replication impossible. We have studied this question in an anoxia-tolerant vertebrate, the crucian carp (Carassius carassius), by examining 5'-bromo-2'-deoxyuridine incorporation and proliferating cell nuclear antigen levels in the gills, intestinal crypts, and liver. We exposed crucian carp to 1 and 7 days of anoxia followed by 7 days of reoxygenation. There was a reduced incidence of S-phase cells (from 12.2 to 5.0%) in gills during anoxia, which coincided with a concomitant increase of G(0) cells. Anoxia also decreased the number of S-phase cells in intestine (from 8.1 to 1.8%). No change in the fraction of S-phase cells ( approximately 1%) in liver was found. Thus new S-phase cells after 7 days of anoxia were present in all tissues, revealing a considerable rate of DNA synthesis. Subsequently, the oxygen-dependent subunit of crucian carp RNR (RNRR2) was cloned. We found no differences in amino acids involved in radical generation and availability of the iron center compared with mouse, which could have explained reduced oxygen dependence. Furthermore, the amount of RNRR2 mRNA in gills did not decrease throughout anoxia exposure. These results indicate that crucian carp is able to sustain some cell proliferation in anoxia, possibly because RNRR2 retains its tyrosyl radical in anoxia, and that the replication machinery is still maintained. Although hypoxia triggers a 7.5-fold increase of respiratory surface area in crucian carp, this response was not triggered in anoxia.

Animals↗

Temperature alters the respiratory surface area of crucian carp Carassius carassius and goldfish Carassius auratus.

We have previously found that the gills of crucian carp Carassius carassius living in normoxic (aerated) water lack protruding lamellae, the primary site of O(2) uptake in fish, and that exposing them to hypoxia increases the respiratory surface area of the gills approximately 7.5-fold. We here examine whether this morphological change is triggered by temperature. We acclimated crucian carp to 10, 15, 20 and 25 degrees C for 1 month, and investigated gill morphology, oxygen consumption and the critical oxygen concentration at the different temperatures. As expected, oxygen consumption increased with temperature. Also at 25 degrees C an increase in the respiratory surface area, similar to that seen in hypoxia, occurred. This coincided with a reduced critical oxygen concentration. We also found that the rate of this transformation increased with rising temperature. Goldfish Carassius auratus, a close relative to crucian carp, previously kept at 25 degrees C, were exposed to 15 degrees C and 7.5 degrees C. At 7.5 degrees C the respiratory surface area of its gills was reduced by development of an interlamellar cell mass as found in normoxic crucian carp kept at 10-20 degrees C. Thus, both species alter the respiratory surface area in response to temperature. Rather than being a graded change, the results suggest that the alteration of gill morphology is triggered at a given temperature. Oxygen-binding data reveal very high oxygen affinities of crucian carp haemoglobins, particularly at high pH and low temperature, which may be prerequisites for the reduced gill respiratory surface area at low temperatures. As ambient oxygen and temperature can both induce the remodelling of the gills, the response appears primarily to be an adaptation to the oxygen demand of the fish.

Acclimatization↗

Maintained cardiac pumping in anoxic crucian carp.

Like most vertebrates, humans die within minutes when deprived of molecular oxygen (anoxia), in part because of cardiac failure. In contrast, some freshwater turtles can survive anoxia for months at low temperatures, but to do so, they drastically suppress cardiac activity and autonomic cardiovascular control. Although Carassius carassius, the crucian carp, shares this anoxia tolerance, we show that it has a unique ability among vertebrates to retain normal cardiac performance and autonomic cardiovascular regulation for at least 5 days of anoxia. These responses point to an unusual tolerance of a vertebrate heart and autonomic nervous system to prolonged anoxia.

Animals↗

Vertebrate brains at the pilot light.

While the brains of most vertebrates are unable to tolerate more than a few minutes of anoxia, some freshwater turtles (Trachemys and Chrysemys), crucian carp (Carassius carassius) and frogs (Rana pipens and Rana temporaria) can survive anoxia for hours to months. Obviously, anoxia tolerance has evolved separately several times and this is also reflected in the divergent strategies these animals utilize to survive without oxygen. The turtles and crucian carp defend their brain ATP levels and avoid a loss of ion homeostasis by reducing ATP use. In the turtles, the early release of adenosine and the activation of K(ATP) channels, a progressive release of GABA and a drastic reduction in electric activity and ion fluxes send the brain into a comatose like state. The crucian carp displays a more modest depression of ATP use, probably achieved through a moderated release of GABA and adenosine, allowing the animal to maintain physical activity in anoxia. The anoxic frog, on the other hand, seems to rely on mechanisms that greatly retard the anoxia induced fall in ATP levels and loss of ion homeostasis, so that the brain can be saved as long as the anoxia is limited to a few hours. The sequence of events characterizing the anoxic frog brain is similar to that of failing anoxic mammalian brain, although over a greatly extended time frame, allowing the frog to die slowly in anoxia, rather than survive. By contrast the only factor that limits anoxic survival in turtles and crucian carp may be the final depletion of their glycogen reserves.

Adaptation, Physiological↗

Breathing with a mouth full of eggs: respiratory consequences of mouthbrooding in cardinalfish.

Mouthbrooding occurs among several groups of fishes. Although a mouth full of eggs can be expected to pose a considerable respiratory problem, to our knowledge no study has examined respiratory consequences of mouthbrooding in fishes, or how hypoxia or strenuous swimming may affect the success of this reproductive strategy. In two species of cardinalfish (Apogon fragilis and Apogon leptacanthus), from the reef at Lizard Island (Great Barrier Reef), we found that mouthbrooding significantly reduced the ability to take up oxygen at low ambient oxygen levels. While the direct energetic cost of mouthbrooding appeared insignificant at rest in well-oxygenated water, mouthbrooding significantly reduced the respiratory scope of the fishes and their capacity for sustained aerobic swimming. The males spat out their eggs in hypoxia. Interestingly, the species with the larger brood, A. fragilis, spat out the brood at a higher water [O2] than did A. leptacanthus, which had a smaller mean brood mass. Moreover, in contrast to mouthbrooding A. leptacanthus, mouthbrooding A. fragilis was unable to increase its ventilatory frequency in response to hypoxia. This suggests a trade-off situation between hypoxia tolerance and brood size. Apparently, A. fragilis has sacrificed hypoxia tolerance in favour of a large brood size to a greater extent than has A. leptacanthus.

Analysis of Variance↗

Hypoxia in paradise: widespread hypoxia tolerance in coral reef fishes.

Using respirometry, we examined the hypoxia tolerance of 31 teleost fish species (seven families) inhabiting coral reefs at a 2-5 m depth in the lagoon at Lizard Island (Great Barrier Reef, Australia). All fishes studied maintained their rate of oxygen consumption down to relatively severe hypoxia (20-30% air saturation). Indeed, most fishes appeared unaffected by hypoxia until the oxygen level fell below 10% of air saturation. This, hitherto unrecognized, hypoxia tolerance among coral reef fishes could reflect adaptations to nocturnal hypoxia in tide pools. It may also be needed to enable fishes to reside deep within branching coral at night to avoid predation. Widespread hypoxia tolerance in a habitat with such an extreme biodiversity as coral reefs indicate that there is a wealth of hypoxia related adaptations to be discovered in reef fishes.

Adaptation, Physiological↗

Anoxia tolerant brains.

While medical science has struggled to find ways to counteract anoxic brain damage with limited success, evolution has repeatedly solved this problem. The best-studied examples of anoxia-tolerant vertebrates are the crucian carp and some North American Freshwater turtles. These can survive anoxia for days to months, depending of temperature. Both animals successfully fight any major fall in brain ATP levels, but the strategies they use to accomplish this are quite divergent. The anoxic turtle suppresses brain activity to such a degree that it becomes virtually comatose. The underlying mechanisms involve closing down ion conductances and releasing GABA and adenosine. By contrast, the crucian carp remains active in anoxia, although it suppresses selected brain functions, and avoids lactate self-poisoning by producing an exotic anaerobic end-product. These animals provide unique models for studying anoxic survival mechanisms both on a molecular and physiological level.

Adaptation, Physiological↗

Coward or braveheart: extreme habitat fidelity through hypoxia tolerance in a coral-dwelling goby.

Coral reef fishes are not known for their hypoxia tolerance. The coral-dwelling goby, Gobiodon histrio, rarely leaves the shelter of its host coral colony. However, our measurements indicate that this habitat could become hypoxic on calm nights ([O(2)] minima=2-30% of air saturation) due to respiration by the coral and associated organisms. Moreover, at very low tides, the whole coral colony can be completely air exposed. Using closed respirometry in water, we found that G. histrio maintains O(2) uptake down to 18% of air saturation, and that it can tolerate at least 2 h at even lower O(2) levels. Furthermore, during air exposure, which was tolerated for more than 3 h, it upheld a rate of O(2) consumption that was 60% of that in water. The hypoxia tolerance and air breathing abilities enables this fish to stay in the safety of its coral home even when exposed to severe hypoxia or air. To our knowledge, this is the first report of hypoxia tolerance in a teleost fish intimately associated with coral reefs.

Animals↗

Hypoxic survival strategies in two fishes: extreme anoxia tolerance in the North European crucian carp and natural hypoxic preconditioning in a coral-reef shark.

Especially in aquatic habitats, hypoxia can be an important evolutionary driving force resulting in both convergent and divergent physiological strategies for hypoxic survival. Examining adaptations to anoxic/hypoxic survival in hypoxia-tolerant animals may offer fresh ideas for the treatment of hypoxia-related diseases. Here, we summarise our present knowledge of two fishes that have evolved to survive hypoxia under very different circumstances. The crucian carp (Carassius carassius) is of particular interest because of its extreme anoxia tolerance. During the long North European winter, it survives for months in completely oxygen-deprived freshwater habitats. The crucian carp also tolerates a few days of anoxia at room temperature and, unlike anoxia-tolerant freshwater turtles, it is still physically active in anoxia. Moreover, the crucian carp does not appear to reduce neuronal ion permeability during anoxia and may primarily rely on more subtle neuromodulatory mechanisms for anoxic metabolic depression. The epaulette shark (Hemiscyllium ocellatum) is a tropical marine vertebrate. It lives on shallow reef platforms that repeatedly become cut off from the ocean during periods of low tides. During nocturnal low tides, the water [O(2)] can fall by 80% due to respiration of the coral and associated organisms. Since the tides become lower and lower over a period of a few days, the hypoxic exposure during subsequent low tides will become progressively longer and more severe. Thus, this shark is under a natural hypoxic preconditioning regimen. Interestingly, hypoxic preconditioning lowers its metabolic rate and its critical P(O(2)). Moreover, repeated anoxia appears to stimulate metabolic depression in an adenosine-dependent way.

Adaptation, Physiological↗

Adenosinergic and cholinergic control mechanisms during hypoxia in the epaulette shark (Hemiscyllium ocellatum), with emphasis on branchial circulation.

Coral reef platforms may become hypoxic at night during low tide. One animal in that habitat, the epaulette shark (Hemiscyllium ocellatum), survives hours of severe hypoxia and at least one hour of anoxia. Here, we examine the branchial effects of severe hypoxia (<0.3 mg oxygen l(-1) for 20 min in anaesthetized epaulette shark), by measuring ventral and dorsal aortic blood pressure (P(VA) and P(DA)), heart rate (fh), and observing gill microcirculation using epi-illumination microscopy. Hypoxia induced a flow of blood in two parallel blood vessels, termed longitudinal vessels, in the outer borders of the free tip of the gill filament. Hypoxia also induced significant falls in fh, P(VA) and P(DA), and a biphasic change in ventilation frequency (increase followed by decrease). Adenosine injection (1 micromol kg(-1)) also initiated blood flow in the longitudinal vessels, in addition to significant drops in P(VA), P(DA) and fh, and a biphasic response in ventilation frequency (decrease followed by increase) indicating that adenosine influences ventilation. Aminophylline (10 mg kg(-1)), an A(1) and A(2) adenosine receptor antagonist, blocked the effects of adenosine injection, and also significantly reduced blood flow in the longitudinal vessels during hypoxia. In the second part of the study, we examined the cholinergic influence on the cardiovascular circulation during severe hypoxia (<0.3 mg l(-1)) using antagonists against muscarinic (atropine 2 mg kg(-1)) and nicotinic (tubocurarine 5 mg kg(-1)) receptors. Injection of acetylcholine (ACh; 1 micromol kg(-1)) into the ventral aorta caused a marked fall in fh, a large increase in P(VA), but small changes in P(DA) (suggesting increased R(gill)). Atropine was able to inhibit the branchial vascular responses to ACh but not the hypoxic bradycardia, suggesting the presence of muscarinic receptors on the heart and gill vasculature, and that the hypoxia induced bradycardia is of non-cholinergic origin. The results suggest that adenosine mediates increases in the arterio-venous circulation in the gill during hypoxia. This may serve to increase blood supply to heart and gill tissue.

Acetylcholine↗