The role of adrenaline as a modulator of cardiac performance in two Antarctic fishes

The present work was performed to test the hypothesis that Antarctic teleosts rely mostly on cholinergic inhibition for autonomic modulation of the heart. The effects of adrenaline on the inotropic properties on paced, isometrically contracting muscle strips were examined in two distinct Antarctic teleosts, the haemoglobinless icefish Chaenocephalus aceratus and the red-blooded Notothenia coriiceps. All tissues examined revealed a negative force-frequency relationship. Under baseline conditions C. aceratus contracted with a force twice as great as that of N. coriiceps. While the degree to which ventricular tissues responded to adrenaline varied between species, adrenergic stimulation significantly increases myocyte contraction force in this group of fishes. Contraction and relaxation times were not significantly affected by adrenaline concentration while absolute rates of contraction were. Adrenergic stimulation does not enable tissues to achieve higher contraction frequencies, but is shown to be a potent modulator of contraction force.     

Preliminary assessment of population structure in the mackerel icefish (Champsocephalus gunnari)

The mackerel icefish (Champsocephalus gunnari Lönnberg E (1905) The Fishes of the Swedish South Polar Expedition. Wiss. Ergebnisse Schwedische Südpol- Exped. 1901–1903, vol 5, p 37 is widely distributed south of the Antarctic convergence and over shelf areas surrounding sub-Antarctic Islands. In order to evaluate global population structure in this species, we examined DNA sequence variation in four mitochondrial regions and four nuclear genes in icefish from four locations in the Atlantic Ocean sector and one location in the Indian Ocean. Despite small sample sizes, mitochondrial and nuclear gene data indicated the existence of at least three genetically distinct stocks: Heard Island, South Shetland Islands, and the remaining Atlantic populations (Shag Rocks, South Georgia, and Bouvet Island). The mitochondrial and nuclear SNP markers developed here will be useful for more extensive analyses of population structure in this species.     

The complete mitochondrial genome of the mackerel icefish,Champsocephalus gunnari (Actinopterygii: Channichthyidae), with reference to the evolution of mitochondrial genomes in Antarctic notothenioids

The mackerel icefish (Champsocephalus gunnari Lönnberg, 1905) is a ray‐finned fish living in the Southern Ocean around Antarctica. We sequenced the complete mitochondrial (mt) genome of the mackerel icefish and a segment from cytochrome b to the control region (CR) in 32 individuals. The mt genome of the mackerel icefish was rearranged, containing two nicotinamide adenine dinucleotide (reduced form) dehydrogenase subunit 6 (ND6), two tRNA^Glu, and two CRs. However, variations in numbers of ND6 and tRNA^Glu were observed amongst individuals. These variations included type 1 (containing two ND6 and two tRNA^Glu), type 2 (containing one ND6, one incomplete ND6, and one tRNA^Glu), and type 3 (containing one ND6 and one tRNA^Glu). The gene orders of types 1 and 2, and variations in numbers of ND6 and tRNA^Glu were not previously found in any Antarctic notothenioids, whereas type 3 is the same as that of Racovitzia glacialis. Phylogenetic analyses of CR DNA sequences showed that duplicated CRs of the same species formed a monophyletic group, suggesting that duplication of CRs occurred in each species. The frequent duplication of mt genomes in Antarctic notothenioids is an unusual feature in vertebrates. We propose that interspecific hybridization and impairment of mismatch repair might account for the high frequency of gene duplications and rearrangement of mt genomes in Antarctic notothenioids.     

Structure and expression of genes involved in transport and storage of iron in red-blooded and hemoglobin-less antarctic notothenioids

Antarctic notothenioids are characterized by a drastic reduction of the hemoglobin content, a condition that reaches its extreme in icefish that, following a gene deletion event, are completely devoid of hemoglobin. To answer the question on what type of adaptive changes occurred in icefish to prevent accumulation of potentially dangerous ferrous iron, we investigated the genes of four proteins known to play a key role in iron metabolism. For this purpose, we cloned and sequenced the cDNAs encoding ceruloplasmin, transferrin, ferritin and divalent metal transporter 1. While the inferred amino acid sequences of transferrin from different Antarctic fish species showed a high level of similarity with the homologous proteins from other species, ceruloplasmin sequence featured amino acid substitutions affecting a copper binding site. Another peculiarity was the presence in subunit H of the icefish ferritin of the two sets of sites involved in iron oxidation and iron mineralization, which in mammals are located on two distinct ferritin subunits. Significant differences in the expression levels of the four genes were found between hemoglobinless and red-blooded notothenioids. An increased expression of ceruloplasmin mRNA in icefish was interpreted as a compensatory mechanism to prevent accumulation of ferrous iron in hemoglobinless fish. In icefish, the amounts of ferritin H-chain mRNA measured in liver, blood and head kidney were lower than in the same organs of the red-blooded fish. In the spleen of both fishes, the expression levels of ferritin H-chain were significantly lower than in the spleen of a "pink-blooded" notothenioid with an intermediate hemoglobin content. Finally, the amount of divalent metal transporter mRNA measured in the head-kidney was lower in the icefish than in the same organ of its red-blooded counterpart. These results indicate that the loss of hemoglobin in icefish is accompanied by remodulation of the iron metabolism.     

Haematological studies on Aethotaxis mitopteryx DeWitt,a high-Antarctic fish with a single haemoglobin

Summary The haematological parameters (haematocrit, erythrocyte number, haemoglobin concentration, MCHC, MCH, oxygen-carrying capacity, pH, p_1/2, , pCO₂, pO₂) of the Antarctic fish Aethotaxis mitopteryx DeWitt are reported. The erythrocyte number (0.39 × 10¹²/1) and the haemoglobin concentration (27.8 g/l) were found to be among the lowest values known for red-blooded Antarctic fishes. Among the species of the family Nototheniidae, this is the only one found so far to have a single haemoglobin in its blood. The results have been analysed in comparison with those of other Antarctic species, in an effort to establish correlations between the physiology of this pelagic-benthopelagic fish and its ecology. From the haematological parameters in this study and the functional properties of haemoglobin outlined in the following paper, it is suggested that A. mitopteryx has an extremely sluggish mode of life.Data presented here were collected during the European Polarstern Study (EPOS) sponsored by the European Science Foundation     

Specific immunity proteomic profile of the skin mucus of Antarctic fish Chionodraco hamatus and Notothenia coriiceps

The white-blooded Antarctic icefish is the only known vertebrate lacking oxygen-transporting haemoglobins. Fish skin mucus, as the first line of defence against pathogens, can reflect fish welfare. In this study, we analysed the skin mucus proteome profiles of the two Antarctic fish species, the white-blooded Antarctic icefish, Chionodraco hamatus, and the red-blooded Antarctic fish, Notothenia coriiceps, unfolding the different proteins by liquid chromatography coupled with tandem mass spectrometry isobaric tags for relative and absolute quantitation (iTRAQ) technology. Of the 4444 totally identified proteins, 227 differentially expressed proteins (DEPs) were found in the comparison between C. hamatus and N. coriiceps, of which 121 were upregulated and 106 were downregulated in the icefish. In the Kyoto Encyclopedia of Genes and Genomes pathway annotation, we found two pathways “Legionellosis” and “Complement and coagulation cascades” were significantly enriched, among of which innate immune candidate proteins such as C3, CASP1, ASC, F3 and C9 were significantly upregulated, suggesting their important roles in C. hamatus immune system. Additionally, the DEP protein–protein interaction network analysis and “Response to stress” GO category provided candidate biomarkers for deep understanding of the distinct immune response of the two Antarctic fish underlying the cold adaptation.     

New and rare species of snailfishes (Scorpaeniformes: Liparidae) collected during the ICEFISH cruise of 2004

During the ICEFISH cruise of 2004 13 specimens of the fish family Liparidae (Scorpaeniformes) were collected from Burdwood Bank and near South Georgia. These specimens, with the addition of three more from the King Edward Point Laboratory of the British Antarctic Survey, represent nine species. Two of these are new and are described herein: Careproctus stigmatogenus new species and C. maculosus new species; the second specimen of C. minimus Andriashev and Stein is reported and described, specimens of C. georgianus Lönnberg, C. falklandicus (Lönnberg), C. ?pallidus (Vaillant), Paraliparis copei gibbericeps Andriashev, P. gracilis Norman, and P. tetrapteryx Andriashev and Neelov are reported, and live color of C. georgianus, C. falklandicus, and P. tetrapteryx is described and shown.     

Nesting behavior of the icefish Chaenocephalus aceratus at Bouvetøya Island, Southern Ocean

We describe in situ observations on nesting by the Scotia Sea (or blackfin) icefish Chaenocephalus aceratus (Lönnberg) that constitute the first substantive evidence of egg brooding and parental care by species of the family Channichthyidae. At Boutetoya Island six fish, all apparently male, were observed guarding egg nests at depths of 141–148 m during an ROV deployment. Eggs were laid as aggregated, round masses (~20–25 cm diameter) in shallow, circular depressions (~1-m diameter, ~20-cm depth) that were probably excavated by the parent(s) to protect the nests. The fish guardians remained tenaciously in contact with the eggs despite disturbances caused by the ROV, reacting to this threat with stress and defense behaviors. Because brooding fishes are more susceptible to the population impacts from trawl fisheries, we argue that this life history should be kept in mind in designing management schemes.     

Drinking in Antarctic fishes

Drinking rates have never been measured in Antarctic fish. Drinking rates were measured in four species of notothenioid fish, including a hemoglobinless icefish, found in the near-freezing waters of the Ross Sea of the Southern Ocean. All of the fish, with the exception of the icefish, had low drinking rates and high serum osmolalities relative to temperate seawater fish. The icefish had significantly higher drinking rates and serum osmolalities relative to the Antarctic fishes containing hemoglobin, including Trematomus bernacchii. Warm acclimation of T. bernacchii, from −1.5°C to +4°C for 4 weeks, significantly increased their drinking rates 4.6-fold, significantly decreased their serum and intestinal osmolality by 11% and 12%, respectively, relative to cold-acclimated fish. These results indicate that increased drinking rates in Antarctic fish at elevated temperatures are involved in maintaining a lower serum osmolality.     

Nitric oxide synthase type I (nNOS), vascular endothelial growth factor (VEGF) and myoglobin-like expression in skeletal muscle of Antarctic icefishes (Notothenioidei: Channichthyidae)

The Antarctic icefishes Channichthyidae lack haemoglobin and are thought to lack myoglobin (Mb) in their skeletal muscle as well. Due to the absence of both respiratory pigments, icefishes may present a variety of physiological adaptations in their skeletal muscles. In mammals, molecular responses to limiting oxygen availability in the skeletal muscle include, among others, the over expression of nitric oxide synthases (NOS), such as type I (neuronal nNOS) and type III (endothelial eNOS), as well as the vascular endothelial growth factor (VEGF). In this paper, we evaluated by western blot analysis whether the skeletal muscle of haemoglobin-less icefishes expresses in a constitutive manner higher levels of the type I and type III NOS isoforms and VEGF. Our results demonstrate that haemoglobin-less icefish of the family Channichthyidae do indeed present higher expression of the type I NOS isoform compared with red-blooded Antarctic fish species of other families of the same suborder Notothenioidei. In contrast, VEGF was not over-expressed. Moreover, we show that some icefish species, thought previously to lack Mb in oxidative muscles, actually present Mb-like immunoreactivity in their skeletal muscle.     

A new species of eel cods Muraenolepis trunovi sp. nova (Muraenolepididae) from the Lazarev Sea with redescription of lectotypes Muraenolepis marmorata Günther, 1880 and M. microps (Lönnberg, 1905)

A new species of eel cods Muraenolepis trunovi sp. nova (Muraenolepididae) from the Lazarev Sea is described. The type specimen is caught at the depth 730–860 m. The new species differs from all known species in very elongated first dorsal and pectoral fins. With respect to the body proportions and the number of rays in the second dorsal fin and the anal fin, it is related to M. marmorata but clearly differs from the latter in a greater number of vertebrae (73 vs. 67–69), in the reduced upper lateral line (to one sensory tube), and in complete absence of the lower lateral line on the body. The lectotypes of Muraenolepis marmorata Günther, 1880 (British Natural History Museum, BNHM) and M. marmoratus microps (Lönnberg, 1905) (Naturhistoriska Riksmuseum, NRM) are redescribed.     

Heart structure and ventricular ultrastructure of hemoglobin- and myoglobin-free icefish Channichthys rhinoceratus

Summary The structural and ultrastructural characteristics of the heart of Channichthys rhinoceratus, an antarctic teleost devoid of respiratory pigments, are described and compared with those obtained from the red-blooded related species Notothenia rossii.The heart of the icefish is characterized by a spongy myocardium supplied with a highly developed arterial coronary system. This vasculature includes a subepicardial system and an extensive intratrabecular capillary network. Arterial hilar network and Thebesian vessels may also be present. The bulbus arteriosus shows unusually large spheroid structures located in the middle layer of the wall.Both white- and red-blooded species display comparable myocardial cell morphology and organelle distribution. However, the mitochondrial cristae of the former are more densely packed and the sarcolemma possesses numerous caveolae. A large proportion of non-contractile cells is also found in the icefish ventricular wall.     

Is cold the new hot? Elevated ubiquitin-conjugated protein levels in tissues of Antarctic fish as evidence for cold-denaturation of proteins in vivo

Levels of ubiquitin (Ub)-conjugated proteins, as an index of misfolded or damaged proteins, were measured in notothenioid fishes, with both Antarctic (Trematomus bernacchii, T. pennellii, Pagothenia borchgrevinki) and non-Antarctic (Notothenia angustata, Bovichtus variegatus) distributions, as well as non-notothenioid fish from the Antarctic (Lycodichthys dearborni, Family Zoarcidae) and New Zealand (Bellapiscis medius, Family Tripterygiidae), in an effort to better understand the effect that inhabiting a sub-zero environment has on maintaining the integrity of the cellular protein pool. Overall, levels of Ub-conjugated proteins in cold-adapted Antarctic fishes were significantly higher than New Zealand fishes in gill, liver, heart and spleen tissues suggesting that life at sub-zero temperatures impacts protein homeostasis. The highest tissue levels of ubiquitinated proteins were found in the spleen of all fish. Ub conjugate levels in the New Zealand N. angustata, more closely resembled levels measured in other Antarctic fishes than levels measured in other New Zealand species, likely reflecting their recent shared ancestry with Antarctic notothenioids.     

The heart of the Antarctic icefish as paradigm of cold adaptation

The stenothermal Antarctic fishes, particularly the hemoglobinless icefish, have developed biochemical, metabolic and morpho-functional features of cardiac performance that can help to decipher some mechanisms underlying cardiac cold adaptation. Examples taken from different levels of cardiac biology in Antarctic fish as a paradigm of cold adaptation include: the function of myoglobin in the icefish species that either express or do not express this pigment; the metabolic and ultrastructural reshaping of the myocardiocytes; and the intrinsic mechanical characteristics of the icefish heart ventricle as a low rate, low pressure and high volume pump.