Antarctic Notothenioids Research Articles

Synthesis and recycling of antifreeze glycoproteins in polar fishes

AbstractEvolutionary disparate Antarctic notothenioids and Arctic gadids have adapted to their freezing environments through the elaboration of essentially identical antifreeze glycoproteins (AFGPs). Here we show that this convergence of molecular identity, which evolved from unrelated parent genes, extends to convergence in physiological deployment. Both fish groups synthesize AFGPs in the exocrine pancreas from where they are discharged into the gut to inhibit the growth of ingested ice. Antifreeze glycoproteins not lost with the faeces are resorbed from the gut via the rectal epithelium, transported to the blood and ultimately secreted into the bile, from where they re-enter the gastrointestinal tract. Antifreeze glycoprotein recirculation conserves energy expenditure and explains how high levels of AFGPs reach the blood in notothenioids since, unlike Arctic gadids which also synthesize AFGP in the liver, AFGP secretion in notothenioids is directed exclusively towards the gastrointestinal lumen. Since AFGPs function by inhibiting ice crystal growth, ice must be present for them to function. The two fish groups are thus faced with an identical problem of how to deal with internal ice. Here we show that both accumulate AFGPs within ellipsoidal macrophages of the spleen, presumably adsorbed to phagocytosed ice crystals which are then held until a warming event ensues.

Ancient climate change, antifreeze, and the evolutionary diversification of Antarctic fishes

The Southern Ocean around Antarctica is among the most rapidly warming regions on Earth, but has experienced episodic climate change during the past 40 million years. It remains unclear how ancient periods of climate change have shaped Antarctic biodiversity. The origin of antifreeze glycoproteins (AFGPs) in Antarctic notothenioid fishes has become a classic example of how the evolution of a key innovation in response to climate change can drive adaptive radiation. By using a time-calibrated molecular phylogeny of notothenioids and reconstructed paleoclimate, we demonstrate that the origin of AFGP occurred between 42 and 22 Ma, which includes a period of global cooling approximately 35 Ma. However, the most species-rich lineages diversified and evolved significant ecological differences at least 10 million years after the origin of AFGPs, during a second cooling event in the Late Miocene (11.6-5.3 Ma). This pattern indicates that AFGP was not the sole trigger of the notothenioid adaptive radiation. Instead, the bulk of the species richness and ecological diversity originated during the Late Miocene and into the Early Pliocene, a time coincident with the origin of polar conditions and increased ice activity in the Southern Ocean. Our results challenge the current understanding of the evolution of Antarctic notothenioids suggesting that the ecological opportunity that underlies this adaptive radiation is not linked to a single trait, but rather to a combination of freeze avoidance offered by AFGPs and subsequent exploitation of new habitats and open niches created by increased glacial and ice sheet activity.

Metabolic and behavioural adaptations during early development of the Antarctic silverfish, Pleuragramma antarcticum

The Antarctic silverfish Pleuragramma antarcticum is a keystone species in the Ross Sea ecosystem, providing one of the major links between lower and higher trophic levels. Despite the importance of this species, surprisingly little is known of its early development and behaviour. Here, we determine the metabolic capacity of Pleuragramma embryonated eggs and larvae and make comparisons with developing stages of another notothenioid, the naked dragonfish Gymnodraco acuticeps. We also show that although large numbers of embryonated eggs of Pleuragramma are found floating among the platelet ice of Terra Nova Bay, they are able to sink prior to hatching in late spring, likely reducing the risk of exposure to the potentially lethal, ice-laden surface environment. Applying Stoke’s law, we determine the change in density required for embryonated eggs to sink at the measured rate and then consider possible mechanisms by which this might occur. Significantly, newly hatched larvae are positively gravitactic and negatively phototactic, such that their swimming behaviour also directs them away from the risk of freezing in the icy surface waters. Measurement of the acute thermal tolerance shows that Pleuragramma larvae have, on average, a sustainable swimming performance breadth of about 17°C, which is significantly greater than that of other adult notothenioids. Although it lacks significant antifreeze capacity in its early developmental stages, Pleuragramma has other attributes that may ensure survival over a wider range of environmental temperatures than other more stenothermal Antarctic notothenioids. How it might adapt to prolonged environmental change arising from phenomena such as global warming, however, requires further investigation.

Divergence of brain and retinal anatomy and histology in pelagic antarctic notothenioid fishes of the sister taxa Dissostichus and Pleuragramma

The neutrally buoyant Antarctic fishes of the sister taxa Dissostichus (D. eleginoides and D. mawsoni) and Pleuragramma antarcticum diverged early in the notothenioid radiation and filled different niches in the pelagic realm of the developing Southern Ocean. To assess the influence of phylogenetic and ecological factors in shaping neural morphology in these taxa, we studied the anatomy and histology of the brains and retinae, and determined the proportional weights of brain regions. With the brain of the non-Antarctic sister taxon Eleginops maclovinus as plesiomorphic, statistically significant departures in the brains of the two Antarctic taxa include reduction of the corpus cerebelli and expansion of the mesencephalon and medulla. Compared to Eleginops, both species also have a relatively smaller telencephalon, although this is significant only in Dissostichus. There are a number of apomorphic features in the brain of Pleuragramma including reduced olfactory nerves and bulbs, an extremely small corpus cerebelli and an expanded mesencephalon. Although there is not a significant difference in the relative weights of the medulla in the two taxa, the prominence of the eminentia granularis and bulging cap-like appearance of the crista cerebellaris are distinctive in Pleuragramma. Brain histology of Dissostichus and Pleuragramma reflects typical perciform patterns and the two species of Dissostichus are histologically identical. Lateral compression in Pleuragramma and notable lobation in Dissostichus also contribute to differences between the taxa. Compression in Pleuragramma is attributable to convergence on an anchovy/herring body shape and to the relatively large brain in this small fish. The less prominent pattern of lobation of the telencephalon, inferior lobes and corpus cerebelli in Pleuragramma probably reflects underlying histology, specifically a reduction in cellularity of the neuropil in the nuclei and lobes. The retinal histology of Dissostichus and Pleuragramma encompasses the extremes seen in Antarctic notothenioids. Dissostichus has a thin scotopic retina with few cones and a high degree of summation. The retina of Pleuragramma is thick and cellular with many small single cones and rods and resembles that of Eleginops. Pedomorphy has not influenced brain morphology in these species but Pleuragramma has superficial neuromasts that are pedomorphic. Although Dissostichus and Pleuragramma are sympatric in the water column, their brains and retinae are highly divergent and reflect the influences of both phylogeny and ecological partitioning of the pelagic realm. Compared to Eleginops, the relatively smaller corpus cerebelli but relatively larger medulla probably indicates, respectively, reduced activity levels of notothenioids in subzero temperatures and expansion of the mechanosensory lateral line system as a supplement to vision under conditions of reduced light. Compared to Dissostichus, Pleuragramma has reduced olfactory bulbs and corpus cerebelli and an expanded mesencephalon. The reduction of the corpus to a small round knob is consistent with physiological parameters and video observations suggesting that, although pelagic, it is relatively inactive. Because mesencephalic weights also include the valvula cerebelli, the relatively large value for Pleuragramma may be attributable to its role in integration and sensorimotor coordination of information from the highly cellular duplex retina and to integration of signals from thewell-developed octavolateralis system. The brain of Dissostichus displays considerable persistent morphology in its overall resemblance to that of Eleginops, especially the large olfactory bulbs and the relatively large caudally projecting corpus, and Dissostichus exhibits olfactory tracking ability and migratory behavior in common with Eleginops.

Evolution of the myoglobin gene in Antarctic Icefishes (Channichthyidae)

Antarctic icefishes (Family: Channichthyidae) are the only adult vertebrate animals that do not express the circulatory oxygen-binding protein, hemoglobin. Six species of icefishes, including Dacodraco hunteri, also do not express myoglobin (Mb), an oxygen-binding protein found in ventricular tissue of Antarctic notothenioids. Sequence of the Mb gene from D. hunteri contains a duplicated TATA box in the promoter region of the Mb gene that is absent in Chionodraco rastrospinosus, an icefish species known to express Mb. The 15-base pair insertion in D. hunteri is identical to the duplicated TATA box that is thought to be directly responsible for preventing transcription of the Mb gene in C. aceratus, another icefish species that lacks Mb expression. Sequencing of the Mb promoter from the remaining channichthyid species revealed the duplicated TATA box in 14 of 16 icefish species, including 8 species known to express Mb. Based on the presence of the duplicated TATA box in promoters of both Mb-expressing and Mb-lacking species, loss of Mb expression in D. hunteri and C. aceratus is either independent of the duplicated TATA box, or the duplicated TATA box requires presence or absence of a distant regulatory element to prevent transcription.

How do Antarctic notothenioid fishes cope with internal ice? A novel function for antifreeze glycoproteins

AbstractAntarctic fishes survive freezing through the secretion of antifreeze glycoproteins (AFGPs), which bind to ice crystals to inhibit their growth. This mode of action implies that ice crystals must be present internally for AFGPs to function. The entry and internal accumulation of ice is likely to be lethal, however, so how do fishes survive in its presence? We propose a novel function for the interaction between internal ice and AFGPs, namely the promotion of ice uptake by splenic phagocytes. We show here that i) external mucus of Antarctic notothenioids contains AFGPs and thus has a potential protective role against ice entry, ii) AFGPs are distributed widely through the extracellular space ensuring that they are likely to come into immediate contact with ice that penetrates their protective barriers, and iii) using AFGP-coated nanoparticles as a proxy for AFGP adsorbed onto ice, we suggest that internal ice crystals are removed from the circulation through phagocytosis, primarily in the spleen. We argue that intracellular sequestration in the spleen minimizes the risks associated with circulating ice and enables the fish to store the ice until it can be dealt with at a later date, possibly by melting during a seasonal warming event.

Egg-guarding behaviour in the Antarctic bathydraconid dragonfish Parachaenichthys charcoti

This paper provides the first photographic evidence of nest-guarding posture in the Antarctic bathydraconid Parachaenichthys charcoti in its natural habitat. Because bathydraconids are not monophyletic and P. charcoti is included in a different clade than Gymnodraco acuticeps, there is now documentation of nest guarding in all major clades of Antarctic notothenioids except for the Artedidraconidae.

Antioxidant potential is positively correlated with mitochondrial enzyme activity in Antarctic and non-Antarctic notothenioid fishes

Antarctic notothenioid fishes possess high oxidative capacities, large amounts of intracellular lipid combined with biological membranes enriched in polyunsaturated fatty acids, all of which could make these animals susceptible to oxidative injury, particularly in the form of lipid peroxidation. The central objective in this study was to examine capacities for oxidative metabolism and total antioxidant defense in Antarctic and non-Antarctic notothenioids in order to test the hypothesis that the cold-bodied Antarctic fishes possess elevated activities of citrate synthase (CS), matched by a more robust antioxidant (AOX) defense, than non-Antarctic species. CS activities and total AOX capacities were measured in brain and heart of 4 Antarctic species and 2 non-Antarctic species collected on the 2004 ICEFISH cruise. While no statistical differences are found among Antarctic and non-Antarctic fishes in either CS or AOX capacities, AOX capacity in both tissues expands with CS activity among individuals measured when all species are combined. There is also a 4.5-fold greater AOX capacity, when normalized to CS activity, in brain than in heart indicating the requirement for extra AOX defense in a tissue well known for its particularly high levels of phospholipids more prone to lipid peroxidation.

The hemoglobins of sub-Antarctic fishes of the suborder Notothenioidei

Fishes of the perciform suborder Notothenioidei provide an excellent opportunity for studying the evolution and functional importance of evolutionary adaptations to temperature. To understand the unique biochemical features of high-Antarctic notothenioids, it is important to improve our knowledge of these highly cold-adapted stenotherms with new information on their sub-Antarctic relatives. This paper focuses on the oxygen-transport system of two non-Antarctic species, Eleginops maclovinus and Bovichtus diacanthus. Unlike most Antarctic notothenioids, the blood of E. maclovinus and B. diacanthus displays high hemoglobin (Hb) multiplicity. E. maclovinus, the sister group of Antarctic notothenioids, has one cathodal (Hb C) and two anodal components (Hb 1, Hb 2). B. diacanthus, one of the most northern notothenioids, has three major Hbs. The multiple Hbs may have been maintained as a response to temperature differences and fluctuations of temperate waters, much larger than in the Antarctic. Although non-Antarctic notothenioids have never developed cold adaptation, the amino-acid sequence reveals high identity with the globins of Antarctic notothenioids. Hbs of sub-Antarctic notothenioids are characterised by high oxygen affinity and Root effect. Phylogenetic analyses are consistent with the hypothesis that Bovichtidae and Eleginopidae diverged after they became established in more temperate waters north of the Antarctic Polar Front.

ND6 Gene "Lost" and Found: Evolution of Mitochondrial Gene Rearrangement in Antarctic Notothenioids

Evolution of Antarctic notothenioids in the frigid and oxygen-rich Southern Ocean had led to remarkable genomic changes, most notably the gain of novel antifreeze glycoproteins and the loss of oxygen-binding hemoproteins in the icefish family. Recently, the mitochondrial (mt) NADH dehydrogenase subunit 6 (ND6) gene and the adjacent transfer RNA(Glu) (tRNA(Glu)) were also reportedly lost. ND6 protein is crucial for the assembly and function of Complex I of the mt electron transport chain that produces adenosine triphosphate (ATP) essential for life; thus, ND6 absence would be irreconcilable with Antarctic notothenioids being thriving species. Here we report our discovery that the ND6 gene and tRNA(Glu) were not lost but had been translocated to the control region (CR) from their canonical location between ND5 and cytochrome b genes. We characterized the CR and adjacent sequences of 22 notothenioid species representing all eight families of Notothenioidei to elucidate the mechanism and evolutionary history of this mtDNA rearrangement. Species of the three basal non-Antarctic families have the canonical vertebrate mt gene order, whereas species of all five Antarctic families have a rearranged CR bearing the embedded ND6 (ND6(CR)) and tRNA(Glu), with additional copies of tRNA(Thr), tRNA(Pro), and noncoding region in various lineages. We hypothesized that an initial duplication of the canonical mt region from ND6 through CR occurred in the common ancestor to the Antarctic clade, and we deduced the succession of loss or modification of the duplicated region leading to the extant patterns of mt DNA reorganization that is consistent with notothenioid evolutionary history. We verified that the ND6(CR) gene in Antarctic notothenioids is transcribed and therefore functional. However, ND6(CR)-encoded protein sequences differ substantially from basal non-Antarctic notothenioid ND6, and we detected lineage-specific positive selection on the branch leading to the Antarctic clade of ND6(CR) under the branch-site model. Collectively, the novel mt ND6(CR) genotype of the Antarctic radiation represents another major molecular change in Antarctic notothenioid evolution and may reflect an adaptive change conducive to the functioning of the protein (Complex I) machinery of mt respiration in the polar environment, driven by the advent of freezing, oxygen-rich conditions in the Southern Ocean.

Molecular pedomorphism underlies craniofacial skeletal evolution in Antarctic notothenioid fishes

BackgroundPedomorphism is the retention of ancestrally juvenile traits by adults in a descendant taxon. Despite its importance for evolutionary change, there are few examples of a molecular basis for this phenomenon. Notothenioids represent one of the best described species flocks among marine fishes, but their diversity is currently threatened by the rapidly changing Antarctic climate. Notothenioid evolutionary history is characterized by parallel radiations from a benthic ancestor to pelagic predators, which was accompanied by the appearance of several pedomorphic traits, including the reduction of skeletal mineralization that resulted in increased buoyancy.ResultsWe compared craniofacial skeletal development in two pelagic notothenioids, Chaenocephalus aceratus and Pleuragramma antarcticum, to that in a benthic species, Notothenia coriiceps, and two outgroups, the threespine stickleback and the zebrafish. Relative to these other species, pelagic notothenioids exhibited a delay in pharyngeal bone development, which was associated with discrete heterochronic shifts in skeletal gene expression that were consistent with persistence of the chondrogenic program and a delay in the osteogenic program during larval development. Morphological analysis also revealed a bias toward the development of anterior and ventral elements of the notothenioid pharyngeal skeleton relative to dorsal and posterior elements.ConclusionsOur data support the hypothesis that early shifts in the relative timing of craniofacial skeletal gene expression may have had a significant impact on the adaptive radiation of Antarctic notothenioids into pelagic habitats.

Hypercapnia induced shifts in gill energy budgets of Antarctic notothenioids

Mechanisms responsive to hypercapnia (elevated CO(2) concentrations) and shaping branchial energy turnover were investigated in isolated perfused gills of two Antarctic Notothenioids (Gobionotothen gibberifrons, Notothenia coriiceps). Branchial oxygen consumption was measured under normo- versus hypercapnic conditions (10,000 ppm CO(2)) at high extracellular pH values. The fractional costs of ion regulation, protein and RNA synthesis in the energy budgets were determined using specific inhibitors. Overall gill energy turnover was maintained under pH compensated hypercapnia in both Antarctic species as well as in a temperate zoarcid (Zoarces viviparus). However, fractional energy consumption by the examined processes rose drastically in G. gibberifrons (100-180%), and to a lesser extent in N. coriiceps gills (7-56%). In conclusion, high CO(2) concentrations under conditions of compensated acidosis induce cost increments in epithelial processes, however, at maintained overall rates of branchial energy turnover.

Report on anisakid nematodes in polar regions – Preliminary results

The aim of this study is to extend our knowledge of the distribution of anisakid nematode parasites in Arctic and Antarctic polar regions. We examined vertebrate (fish) taxa characteristic of the faunas in both polar regions for the presence of parasitic nematodes. The material was collected from Svalbard (Arctic) between July and August 2008 and from King George Island (South Shetland Islands, Antarctic Peninsula) between November 2007 and January 2008. In addition, faecal, bird, and invertebrate samples were collected and examined for the presence of anisakid nematodes or eggs. Anisakis simplex s.s. was found in the body cavity of Arctic cod, and Contracaecum sp. and Pseudoterranova sp. were found in Antarctic notothenioids. Eggs of Anisakis sp. and Contracaecum sp. were recovered from the faeces of Mirounga leonina. We present the first record of the occurrence of A. simplex C in the Antarctic fishes Notothenia coriiceps and Notothenia rossii.

Some reproductive traits of the Tristan klipfish, Bovichtus diacanthus (Carmichael 1819) (Notothenioidei: Bovichtidae) from Tristan da Cunha (South Atlantic)

The reproductive biology of the Tristan klipfish, Bovichtus diacanthus, was investigated by macroscopic and histological analyses of the gonads. Fish samples were collected in tide pools at Tristan da Cunha in July 2004. Most specimens of both sexes were developing, or sexually mature, with a gonadosomatic index (GSI) of 7.0–9.2% in females and 0.2–0.6% in males. Histologically, testes showed a random distribution of spermatogonia along the lobules, a condition defined as the unrestricted spermatogonial type. Ripe males exhibited lobules with all spermatogenic stages of development from spermatogonia to spermatozoa. In mature females, the ovarian follicles consisted of three main cohorts of oocytes of different sizes; the smaller one represented by previtellogenic oocytes of 15–150 μm and the other two by yolked oocytes measuring, respectively, 300–1000 and 800–1500 μm. The overlap between the stock of advanced yolked oocytes and the early yolked oocytes was low, decreasing progressively with final maturation. As a result, B. diacanthus was considered a batch spawner, with a spawning season extending from July to August onward. Batch fecundity, based on the most advanced yolked oocytes, was 2,047–8,317 mature oocytes/female, whereas the relative fecundity was 77–141 mature oocytes/g. In the light of the phyletically basal position of bovichtids in the suborder, the reproductive traits of B. diacanthus were compared with those previously described in other Antarctic and non-Antarctic notothenioids.

Characterization of the intestinal microbiota of two Antarctic notothenioid fish species

The fish fauna of the Southern Ocean is dominated by species of the perciform suborder Notothenioidei, which constitute 46% of fish species and 90% of biomass. Notothenioids have undergone rapid morphological and ecological diversification and developed physiological adaptations to a cold, highly oxygenated environment. Microbes inhabiting animal intestines include those that perform essential nutritional functions, but notothenioid gut microbial communities have not been investigated using cultivation-independent approaches. We analyzed bacterial 16S rRNA gene sequences obtained from the intestinal tract of Notothenia coriiceps and Chaenocephalus aceratus, which differ in their pelagic distribution and feeding strategies. Both samples showed dominance of Gammaproteobacteria (mostly Vibrionaceae), as has been reported for temperate teleost species. Both samples showed low diversity relative to that reported for other fish microbiota studies, with C. aceratus containing fewer OTUs than N. coriiceps. Despite the small sample size of this preliminary study, our findings suggest that Antarctic notothenioids carry a gut microbiota similar in composition to that of temperate fish, but exhibiting lower species-level diversity. The omnivorous N. coriiceps individual exhibited greater diversity than the exclusively carnivorous C. aceratus individual, which may indicate that increasing herbivory in fish leads to gut microbe diversification, as found in mammals. Lastly, we detected members of taxa containing known microbial pathogens, which have not been previously reported in Antarctic notothenioid fish.

Mitochondrial function in Antarctic notothenioid fishes that differ in the expression of oxygen-binding proteins

State III respiration rates were measured in mitochondria isolated from hearts of Antarctic notothenioid fishes that differ in the expression of hemoglobin (Hb) and myoglobin (Mb). Respiration rates were measured at temperatures between 2 and 40°C in Gobionotothen gibberifrons (+Hb/+Mb), Chaenocephalus aceratus (–Hb/–Mb) and Chionodraco rastrospinosus (–Hb/+Mb). Blood osmolarity was measured in all three species and physiological buffers prepared for isolating mitochondria and measuring respiration rates. Respiration rates were higher in mitochondria from G. gibberifrons compared to those from C. aceratus at 2°C, but were similar among all species at temperatures between 10 and 26°C. Respiration rates were significantly lower in icefishes at 35 and 40°C compared to G. gibberifrons. The respiratory control ratio of isolated mitochondria was lower in C. aceratus compared to G. gibberifrons at all temperatures below 35°C. At 35 and 40°C, mitochondria were uncoupled in all species. The Arrhenius break temperature of state III respiration was similar among all three species (30.5 ± 0.9°C) and higher than values previously reported for Antarctic notothenioids, likely due to the higher osmolarity of buffers used in this study. These results suggest that differences in mitochondrial structure, correlated with the expression of oxygen-binding proteins, minimally impact mitochondrial function.

Transcriptomic and genomic evolution under constant cold in Antarctic notothenioid fish

The antifreeze glycoprotein-fortified Antarctic notothenioid fishes comprise the predominant fish suborder in the isolated frigid Southern Ocean. Their ecological success undoubtedly entailed evolutionary acquisition of a full suite of cold-stable functions besides antifreeze protection. Prior studies of adaptive changes in these teleost fishes generally examined a single genotype or phenotype. We report here the genome-wide investigations of transcriptional and genomic changes associated with Antarctic notothenioid cold adaptation. We sequenced and characterized 33,560 ESTs from four tissues of the Antarctic notothenioid Dissostichus mawsoni and derived 3,114 nonredundant protein gene families and their expression profiles. Through comparative analyses of same-tissue transcriptome profiles of D. mawsoni and temperate/tropical teleost fishes, we identified 177 notothenioid protein families that were expressed many fold over the latter, indicating cold-related up-regulation. These up-regulated gene families operate in protein biosynthesis, protein folding and degradation, lipid metabolism, antioxidation, antiapoptosis, innate immunity, choriongenesis, and others, all of recognizable functional importance in mitigating stresses in freezing temperatures during notothenioid life histories. We further examined the genomic and evolutionary bases for this expressional up-regulation by comparative genomic hybridization of DNA from four pairs of Antarctic and basal non-Antarctic notothenioids to 10,700 D. mawsoni cDNA probes and discovered significant to astounding (3- to >300-fold, P < 0.05) Antarctic-specific duplications of 118 protein-coding genes, many of which correspond to the up-regulated gene families. Results of our integrative tripartite study strongly suggest that evolution under constant cold has resulted in dramatic genomic expansions of specific protein gene families, augmenting gene expression and gene functions contributing to physiological fitness of Antarctic notothenioids in freezing polar conditions.

Evolutionary analysis of the transferrin gene in Antarctic Notothenioidei: A history of adaptive evolution and functional divergence

Organisms living in the Southern Ocean are exposed to strong environmental constraints, especially temperature. The Perciform suborder Notothenioidei, today largely endemic to the Antarctic, has developed numerous cold-adapted characters. The most striking peculiarity of this group of fish is the drastic reduction of hemoglobin content in their blood. This condition gives rise to a low metabolic demand of iron. Recently, we have undertaken a study to add new insights on iron metabolism in hemoglobinless fish. By taking advantage to our previous studies on transferrins from Antarctic Notothenioids, in this article we compared the evolutionary properties of these sequences to those from other fish groups and mammals. Our findings based on analysis of d N/d S ratio and on the results of the relative rate ratio test suggest that the transferrin gene has undergone positive selection in fish but not in mammals. The results of type I functional divergence provide statistical evidence for shifted evolutionary rate after speciation. Finally, when superimposed onto the three-dimensional structure of transferrin, the sites identified as responsible of the divergence in Notothenioids appear to cluster preferentially at non-iron binding locations, occupying surface locations that might affect the rate of iron binding and/or release in a cold environment.

Molecular aspects of iron acquisition and storage in the cold-adapted Antarctic Notothenioids

Molecular aspects of iron acquisition and storage in the cold-adapted Antarctic Notothenioids

High mitochondrial densities in the hearts of Antarctic icefishes are maintained by an increase in mitochondrial size rather than mitochondrial biogenesis

We investigated the molecular mechanisms regulating differences in mitochondrial volume density between heart ventricles of Antarctic notothenioids that vary in the expression of hemoglobin (Hb) and myoglobin (Mb). In mammals, peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1alpha) and nuclear respiratory factor 1 (NRF-1) stimulate mitochondrial biogenesis and maintain mitochondrial density in muscle tissues. We hypothesized that these factors would also maintain mitochondrial density in the hearts of Antarctic notothenioids. The percent cell volume occupied by mitochondria is significantly lower in hearts of the red-blooded notothenioid Notothenia coriiceps (18.18+/-0.69%) in comparison with those of the icefish Chaenocephalus aceratus (36.53+/-2.07%), which lacks both Hb and cardiac Mb. Mitochondrial densities are not different between hearts of N. coriiceps and Chionodraco rastrospinosus, which lacks Hb, but whose heart expresses Mb. Despite differences in mitochondrial volume density between hearts of N. coriiceps and C. aceratus, the levels of transcripts of the genes encoding PGC-1alpha, NRF-1 and citrate synthase, and the copy number of mitochondrial DNA do not differ. Our results indicate that the high mitochondrial densities in hearts of C. aceratus may result from an increase in organelle size. The surface-to-volume ratio of mitochondria from N. coriiceps is 1.9-fold greater than that of mitochondria from C. aceratus. In addition, the levels of PGC-1alpha correlate with mitochondrial density in muscle tissues of notothenioids possessing mitochondria of similar size and morphology. Finally, the levels of PGC-1alpha are 4.6-fold higher in the aerobic pectoral adductor muscle in comparison with the glycolytic skeletal muscle of N. coriiceps. The potential physiological significance of an increase in mitochondrial size in hearts of Antarctic icefishes is discussed.