Transcription profiling of acute temperature stress in the Antarctic plunderfish Harpagifer antarcticus

Harpagifer antarcticus (the Antarctic plunderfish), a shallow-water benthic fish distributed around the Antarctic Peninsula, is a member of the notothenioid family, one of whose adaptations to the cold waters of Antarctica has been the loss of the classic heat shock response. In order to gain a more comprehensive understanding of the effects of temperature stress on H. antarcticus, we constructed a liver cDNA library and a 10,371 feature microarray. This was hybridized with material from a time course series of animals held at 6°C for 48h. The resulting expression profiles show that this fish displays the classical vertebrate acute inflammatory response. There was also a pronounced signal for increased energy requirements via up-regulation of genes involved in the β oxidation of fatty acids and also a strong signature of response to oxidative stress. Genes in the latter category did not include the "classic" antioxidants such as glutathione S-transferase, but genes involved in the production of reducing potential in the form of NADPH, peroxisome proliferation via peroxisomal acyl co-enzyme A oxidase 1 and genes known to be up-regulated by hypoxia-inducible factor 1 (HIF1). These identifications provide clear support for oxygen being the whole animal limiting factor at least in acute short-term temperature challenges. The classical heat shock proteins were not up-regulated during this trial, although numerous clones for each were present on the gene chip, confirming the lack of this response in this species. These data significantly increase our knowledge of the cellular stress response from animals in this unique environment.     

Freshening effect on the osmotic response of the Antarctic spiny plunderfish Harpagifer antarcticus

Global warming is having a significant impact around the world, modifying environmental conditions in many areas, including in zones that have been thermally stable for thousands of years, such as Antarctica. Stenothermal sedentary intertidal fish species may suffer due to warming, notably if this causes water freshening from increased freshwater inputs. Acute decreases in salinity, from 33 down to 5, were used to assess osmotic responses to environmental salinity fluctuations in Antarctic spiny plunderfish Harpagifer antarcticus, in particular to evaluate if H. antarcticus is able to cope with freshening and to describe osmoregulatory responses at different levels (haematological variables, muscle water content, gene expression, NKA activity). H. antarcticus were acclimated to a range of salinities (33 as control, 20, 15, 10 and 5) for 1 week. At 5, plasma osmolality and calcium concentration were both at their lowest, while plasma cortisol and percentage muscle water content were at their highest. At the same salinity, gill and intestine Na⁺-K⁺-ATPase (NKA) activities were at their lowest and highest, respectively. In kidney, NKA activity was highest at intermediate salinities (15 and 10). The salinity-dependent NKA mRNA expression patterns differed depending on the tissue. Marked changes were also observed in the expression of genes coding membrane proteins associated with ion and water transport, such as NKCC2, CFTR and AQP8, and in the expression of mRNA for the regulatory hormone prolactin (PRL) and its receptor (PRLr). Our results demonstrate that freshening causes osmotic imbalances in H. antarcticus, apparently due to reduced capacity of both transport and regulatory mechanisms of key organs to maintain homeostasis. This has implications for fish species that have evolved in stable environmental conditions in the Antarctic, now threatened by climate change.     

Moulting reduces freeze susceptibility in the Antarctic mite Alaskozetes antarcticus (Michael)

Abstract.  The effect of moulting on the cold hardiness of the oribatid mite Alaskozetes antarcticus (Michael) is investigated. Non moulting animals show clear seasonal patterns of cold hardiness with high supercooling points (SCPs) at the peak of summer and an increasing proportion of low SCPs with declining environmental temperatures. By contrast, both field-fresh and laboratory acclimated (5 °C) mites in the moult state are consistently found to have low SCPs regardless of environmental temperature.     

The contrasting diet of Harpagifer antarcticus (Notothenioidei, Harpagiferidae) at two localities of the South Shetland Islands, Antarctica

The diet of Harpagifer antarcticus was studied at two localities of the South Shetland Islands, Antarctica. The analysis of the stomach contents of specimens collected in tide pools at Potter Cove, King George Island, indicated that gammarid amphipods (mainly Gondogeneia sp.) were the main prey of this fish, followed by polychaetes, gastropods and isopods. By contrast, the specimens from Harmony Point, Nelson Island, which were recovered from stomach contents of Antarctic shags Phalacrocorax bransfieldensis feeding at depths of 46–110 m, preyed almost exclusively on the Antarctic krill Euphausia superba. These results are discussed and compared with previous studies. Received: 27 August 1997 / Accepted: 17 November 1997     

The effect of feeding on specific soil algae on the cold-hardiness of two Antarctic micro-arthropods (Alaskozetes antarcticus and Cryptopygus antarcticus)

The effect of consuming terrestrial algae on the cold tolerance of two Antarctic micro-arthropods was examined. From the results of preferential feeding experiments, seven species of Antarctic terrestrial micro-algae were chosen and fed to two common, freeze-avoiding Antarctic micro-arthropods: the springtail Cryptopygus antarcticus (Collembola: Isotomidae), and the mite Alaskozetes antarcticus (Acari: Oribatida). Mites were very selective in their choice of food whereas the springtails were less discriminating. The ice nucleating activity of each species of alga was measured using an ice nucleator spectrometer and a differential scanning calorimeter. Pure cultures of individual species of algae had characteristic supercooling points ranging from ca. −5 to −18 °C. The effect of eating a particular alga on the supercooling point of individual micro-arthropods cultured at two different temperatures (0 and 10 °C) was examined. Neither species showed a preference for algae with low ice-nucleating activity and there was no clear correlation between the supercooling point of food material and that of the whole animal. However, feeding on certain algae such as Prasiola crispa, which contained the most active ice nucleators, decreased the cold tolerance of both species of arthropods. Accepted: 6 May 2000     

Gonopore development and sex change in the Antarctic shrimp Chorismus antarcticus (Caridea: Hippolytidae)

Chorismus antarcticus is the only protandrous hermaphrodite decapod species found on the Antarctic continental shelves. The morphological structures of the male and female gonopores were described and used to determine sex and to fix the size range at which the transition from male to female takes place. The parallel occurrence of male and female gonopores was found in all specimens. The presence of open gonopore flaps at both the third and fifth pair of pereiopods in individuals between 9 and 12 mm of carapace length (CL) is discussed as the morphological indicator of sex change. Few females occurred from 9 mm CL onwards, whereas all shrimps larger than 13 mm CL only had open female gonopores. This size corresponds exactly with the CL at which onset of oocyte development and female gonad production starts. Secondary sexual characters, the gonopore structures, seem to allow an accurate prediction of the size range at which hermaphrodite caridean decapods change sex. Received: 23 February 1996/Accepted: 15 July 1996     

A further contribution on the biology of the Antarctic flea,Glaciopsyllus antarcticus (Siphonaptera: Ceratophyllidae)

Summary Experiments showed that adult Antarctic Fleas Gladopsyllus antarcticus preferred dark over light conditions, dry over wet conditions, and fine substrate over coarse substrate. Examination of seabird chicks and collections of nest material indicated that the Southern Fulmar Fulmarus glacialoides, is the major host species of the Antarctic Flea, while Snow Petrels Pagodroma nivea, Cape Petrels Daption capense, Antarctic Petrels Thalassoica antarctica, and Wilson's Storm-Petrels Oceanites oceanicus are minor hosts. This is the first report of Antarctic Fleas occuring on either Antarctic Petrels or Wilson's Storm-Petrels. No fleas were found associated with Southern Giant Petrels Macronectes giganteus, Antarctic Skuas Catharacta maccormicki, or Adelie Penguins Pygoscelis adeliae. No live fleas were found in Southern Fulmar nest material during their period of dispersal over the winter months, supporting the hypothesis that Antarctic Fleas survive the winter period by remaining on their seabird host.     

Photoreceptor ultrastructure in the Antarctic mussel shrimp Acetabulastoma (Crustacea; Ostracoda), a parasite of Glyptonotus antarcticus (Crustacea; Isopoda)

The ultrastructure of the nauplius eye of the tiny Antarctic ostracode Acetabulastoma sp. is described and conclusions about its possible function are drawn. Each of the three eye-cups measures approximately 20 μm in diameter and is optically isolated from its neighbour by screening pigments, which are contained in pigment cells behind a tapetum of concentrically arranged, ca. 1-μm-long and 0.1-μm-thick, crystals. Three and sometimes four separate rhabdoms with microvilli measuring 50–60 nm in diameter project from the concave side of the tapetum up to 5 μm deep into the eye-cup interior, which is filled by the retinula cell bodies with their spherical nuclei and various organelles. Desmosomes and microtubules are seen and light-induced cell or membrane damage was minimal. The observations suggest that the Acetabulastoma eye has photoreceptors that can tolerate an exposure to bright light and it may be used to inform its owner of the approach of danger, the depth of water, and/or the season. Accepted: 27 August 1998     

Characterisation and expression of the paired box protein 7 (Pax7) gene in polymorphic Arctic charr (Salvelinus alpinus)

Arctic charr (Salvelinus alpinus L.) from Lake Thingvallavatn, Iceland occur as four distinct morphs: large benthivorous (LB), dwarf benthivorous (DB), piscivorous (PI) and planktonivorous (PL). The morphs differ with respect to body size, head morphology, growth rate, and life history. The aim of this study was to investigate the paired box protein 7 (Pax7) gene as a candidate for such polymorphisms due to its importance in cranio-facial, skeletal muscle, and central nervous system development. No variation in coding and intronic sequences was found between morphs. We identified 10 alternate Pax7 isoforms with insertions/deletions: a four-residue (GNRT) deletion, a GEASS insertion truncated by the first serine residue (GEAS), and a thirteen-residue insertion (GQYA/TGPEYVYCGT). The latter insertion with a threonine (T) contains a putative casein kinase II (CK-2) phosphorylation site. Pax7 spatial expression patterns were identical in embryos of DB-, LB-, and PL-morphs, and were similar to those described for zebrafish Pax7c, but a difference in temporal expression for segmentation was observed between DB and LB morphs. At the end of segmentation, novel expression was observed in the mandibular region as two bilateral domains. The potential role of multiple alternative splicing of the Pax7 gene for the generation of different Arctic charr morphs is briefly discussed.     

Temporal resolution of cold acclimation and de-acclimation in the Antarctic collembolan, Cryptopygus antarcticus

Abstract.  The Antarctic collembolan, Cryptopygus antarcticus (Willem), can switch its supercooling point (SCP) between 'winter' and 'summer' modes of cold hardiness over a matter of hours. High resolution temporal scaling of the acquisition and loss of cold hardiness is undertaken by assaying changes in the proportion of animals freezing below −15 °C in response to cooling rate, acclimation temperature, and access to food and moisture. Rapid de-acclimation to the 'summer' modal state is readily achieved after 1–6 h in response to warming and access to food; however, rapid acclimation to the 'winter' modal state is only evident in response to slow cooling and narrow ranges of temperature (0–5 °C). The rapid loss of cold tolerance at higher temperatures with access to food, in particular, emphasizes this species' opportunistic responses to resource availability in the short polar summers. Cold hardiness is apparently more readily traded off against nutrient acquisition than vice versa in this maritime Antarctic species.     

The significance of the moult cycle to cold tolerance in the Antarctic collembolan Cryptopygus antarcticus

Research into the ecophysiology of arthropod cold tolerance has largely focussed on those parts of the year and/or the life cycle in which cold stress is most likely to be experienced, resulting in an emphasis on studies of the preparation for and survival in the overwintering state. However, the non-feeding stage of the moult cycle also gives rise to a period of increased cold hardiness in some microarthropods and, as a consequence, a proportion of the field population is cold tolerant even during the summer active period.In the case of the common Antarctic springtail Cryptopygus antarcticus, the proportion of time spent in this non-feeding stage is extended disproportionately relative to the feeding stage as temperature is reduced. As a result, the proportion of the population in a cold tolerant state, with low supercooling points (SCPs), increases at lower temperatures.We found that, at 5 °C, about 37% of the population are involved in ecdysis and exhibit low SCPs. At 2 °C this figure increased to 50% and, at 0 °C, we estimate that 80% of the population will have increased cold hardiness as a result of a prolonged non-feeding, premoult period. Thus, as part of the suite of life history and ecophysiological features that enable this Antarctic springtail to survive in its hostile environment, it appears that it can take advantage of and extend the use of a pre-existing characteristic inherent within the moulting cycle.     

Expression of warm temperature acclimation-related protein 65-kDa (Wap65) mRNA, and physiological changes with increasing water temperature in black porgy, Acanthopagrus schlegeli

We isolated the warm temperature acclimation-related protein 65-kDa (Wap65) cDNA from the liver of black porgy and investigated the expression by increasing water temperature in black porgy, Acanthopagrus schlegeli. Black porgy Wap65 full-length cDNA consists of 1,338 nucleotides, including an open reading frame, predicted to encode a protein of 425 amino acids and showed high homology to pufferfish (79%), Medaka (73%), carp (70%), and goldfish (68%) Wap65. Increase in water temperature (20 degrees C --> 30 degrees C; 1 degrees C/day) induced the rise of Wap65 mRNA expression in liver of black porgy. Also, the levels of cortisol and glucose in plasma were significantly higher at 30 degrees C than at 20 degrees C. To determine the high water temperature stressor specificity of the induction of Wap65, black porgy were transferred from seawater (SW) to freshwater (FW) for 24 hr. Wap65 expression was not detected when the fish were transferred from SW to FW (in fish transferred from SW to FW), although the levels of cortisol and glucose in plasma were increased. These results suggest that increase in Wap65 gene is related to high water temperature stress and play important roles in high water temperature environment of black porgy.     

Characterisation of the S-adenosylmethionine decarboxylase (SAMDC) gene of potato

S-adenosylmethionine decarboxylase (SAMDC) is involved in the biosynthesis of the polyamines, spermidine and spermine. Recently, we reported the isolation of a putative cDNA clone of the SAMDC clone of potato (Plant Mol Biol 20; 641–651). In order to confirm that the potato genes does encode SAMDC, a complementation experiment with a yeast strain that possesses a null mutation in the SAMDC gene was performed. The yeast strain contains a deletion-insertion mutation in the SAMDC gene and has an absolute requirement for the addition of exogenous spermidine for growth. When the full-length potato cDNA was expressed in the mutant yeast strain there was no longer a requirement for exogenous spermidine. Immunoblotting experiments suggest that the potato SAMDC gene product has an apparent molecular mass of 39 kDa. Expression of the SAMDC gene was high in the young and actively dividing tissues and low in the mature and non-dividing tissues of both vegetative and reproductive organs. Additionally, isolation and characterisation of the corresponding genomic clone is reported. The gene has one intron in its 5-untranslated sequence but otherwise the transcribed portion is identical to the cDNA clone.     

Marsupium morphology and brooding biology of the Antarctic giant isopod,Glyptonotus antarcticus Eights 1853 (Crustacea,Isopoda, Chaetiliidae)

The marsupium of the Antarctic giant isopod, Glyptonotus antarcticus Eights 1853, has unique characteristics. It deviates in its proportions and construction from the so called Idotea type of brood pouch found in other marine isopod species; some features even remind you of the so called Porcellio type of marsupium which is typical of terrestrial isopods. Details are described by scanning electron microscopy. From our observations and from reviewed data, we suggest that, during the prolonged incubation of brood, adelphophagy contributes to the nourishment of the developing young, and assume that additional nutrition is provided by maternal secretion. These suggestions would partly explain marsupial conditions in G. antarcticus as an adaptation to slow development in an adverse environment. Two species of epizoa, a turbellarian and a fish leech, were found to occur on G. anarcticus in the vicinity of the marsupium.Contribution No. 561 of the Alfred-Wegener-Institute of Polar and Marine Research (AWI Bremerhaven)