Coppes and Somero CBP 2007

CBA-08049; No.
of pages: 9
ARTICLE IN PRESS
+ MODEL
Comparative Biochemistry and Physiology, Part A xx (2007) xxx xxx www.elsevier.com/locate/cbpa
Review
Biochemical adaptations of notothenioid fishes: Comparisons between cold temperate South American and New Zealand species and Antarctic species
Zulema L. Coppes Petricorena a,, George N. Somero b
b
Faculty of Chemistry Montevideo, Uruguay Hopkins Marine Station, Department of Biological Sciences, Stanford University, Pacific Grove, CA 93950-3094, USA
a
Received 17 June 2006; received in revised form 17 September 2006; accepted 29 September 2006
Abstract Fishes of the perciform suborder Notothenioidei afford an excellent opportunity for studying the evolution and functional importance of diverse types of biochemical adaptation to temperature. Antarctic notothenioids have evolved numerous biochemical adaptations to stably cold waters, including antifreeze glycoproteins, which inhibit growth of ice crystals, and enzymatic proteins with cold-adapted specific activities (kcat values) and substrate binding abilities (Km values), which support metabolism at low temperatures. Antarctic notothenioids also exhibit the loss of certain biochemical traits that are ubiquitous in other fishes, including the heat-shock response (HSR) and, in members of the family Channichthyidae, hemoglobins and myoglobins. Tolerance of warm temperatures is also truncated in stenothermal Antarctic notothenioids. In contrast to Antarctic notothenioids, notothenioid species found in South American and New Zealand waters have biochemistries more reflective of cold-temperate environments. Some of the contemporary non-Antarctic notothenioids likely derive from ancestral species that evolved in the Antarctic and later escaped to lower latitude waters when the Antarctic Polar Front temporarily shifted northward during the late Miocene. Studies of cold-temperate notothenioids may enable the timing of critical events in the evolution of Antarctic notothenioids to be determined, notably the chronology of acquisition and amplification of antifreeze glycoprotein genes and the loss of the HSR. Genomic studies may reveal how the gene regulatory networks involved in acclimation to temperature differ between stenotherms like the Antarctic notothenioids and more eurythermal species like cold-temperate notothenioids. Comparative studies of Antarctic and cold-temperate notothenioids thus have high promise for revealing the mechanisms by which temperature-adaptive biochemical traits are acquired or through which traits that cease to be of advantage under conditions of stable, near-freezing temperatures are lost during evolution. 2006 Elsevier Inc. All rights reserved.
Keywords: Antarctica; Antifreeze glycoproteins; Heat-shock response; Notothenioid; Temperature adaptation
Contents 1. 2. 3. 4. 5.
Geological and oceanographic drivers of evolution in notothenioid fishes . Characteristics of the Antarctic fish fauna: the suborder Notothenioidei . . Non-Antarctic notothenioids . . . . . . . . . . . . . . . . . . . . . . . . Antifreeze glycoproteins . . . . . . . . . . . . . . . . . . . . . . . . . . Gene loss in stably cold waters: the heat-shock response . . . . . . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
0 0 0 0 0
This paper is part of the 3rd special issue of CBP dedicated to The Face of Latin American Comparative Biochemistry and Physiology organized by Marcelo Hermes-Lima (Brazil) and co-edited by Carlos Navas (Brazil), Rene Beleboni (Brazil), Rodrigo Stabeli (Brazil), Tania Zenteno-Savn (Mexico) and the editors of CBP. This issue is dedicated to the memory of two exceptional men, Peter L. Lutz, one of the pioneers of comparative and integrative physiology, and Cicero Lima, journalist, science lover and Hermes-Lima's dad. Corresponding author. E-mail address: zucoppes@yahoo.com (Z.L. Coppes Petricorena). 1095-6433/$ - see front matter 2006 Elsevier Inc. All rights reserved. doi:10.1016/j.cbpa.2006.09.028 Please cite this article as: Coppes Petricorena, Z.L., Somero, G.N. Biochemical adaptations of notothenioid fishes: Comparisons between cold temperate South American and New Zealand species and. Comparative Biochemistry and Physiology, Part A (2007), doi:10.1016/j.cbpa.2006.09.028
ARTICLE IN PRESS
2 Z.L. Coppes Petricorena, G.N. Somero / Comparative Biochemistry and Physiology, Part A xx (2007) xxxxxx
6. Temperature adaptation of enzymatic proteins. . . . . . . . . . . . . . . . . . . . . . 7. Structural adaptations of muscle fibres: relationship between diameter and number . . 8. Genetics of notothenioids: what has been lost during evolution in stably cold waters? . Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
. . . . .
0 0 0 0 0
1. Geological and oceanographic drivers of evolution in notothenioid fishes Fishes of the perciform suborder Notothenioidei afford an excellent study system for examining how large-scale geological and oceanographic processes serve as drivers of evolution to the physical environment. The formation of the Southern Ocean, which surrounds Antarctica and includes the great embayments of the Weddell and Ross Seas, was marked by the creation of a large mass of water the planet's fourth largest ocean that is uniquely cold and thermally stable. The Southern Ocean is covered by sea ice during the winter, from the Antarctic coastline northward to approximately 60S (Gordon, 1988, 1999, 2003). The Southern Ocean is much younger than other oceans because of its origins as a result of plate tectonic activities over the past approximately 4060 million years. Two key events in the formation of the Southern Ocean were the opening of the Drake Passage between South America and the Antarctic continent, which recent analyses suggest took place approximately 41 million years ago, and the formation of the Tasmanian Gateway, which is now thought to have occurred a few millions years after the opening of the Drake Passage (Scher and Martin, 2006). The separation of these southern landmasses permitted formation of the Antarctic Circumpolar Current (ACC), the oceanographic feature of the Southern Ocean that plays a pivotal role in establishing the thermal conditions that have driven evolution of the Antarctic biota (Eastman, 1993). The ACC is the ocean's largest current. It is 21,000 km in length and transports 130 million cubic meters of water per second 100 times the flow of all the world's rivers (Gordon, 1999). The Antarctic Polar Front (APF), the northern border of the ACC between 50S and 60S, prevents mixing of the waters of the Southern Ocean with those of the Indian, Pacific and Atlantic oceans. The APF thus acts as a cold wall that inhibits mixing of the fauna of the cold temperate ocean to the north with the cold-adapted fauna of the Southern Ocean. However, this wall may not be impenetrable at all depths, for recent studies suggest that leakage of invertebrates may occur in deep water (Clarke et al., 2005). Sea temperatures of the Southern Ocean have been well below 5 C for 10 to 14 MY and they presently approach 2 C at the more southerly boundaries of the shelf (Littlepage, 1965). Annual variation in temperature of McMurdo Sound waters (78S) is between 1.9 C and 0.5 C (Hunt et al., 2003). In more northerly waters of the Antarctic Peninsula, summer temperature reach +1.5 C and winter temperatures are near 1.8 C (Dewitt, 1970). As the water column of the ACC/APF is very well mixed, temperature varies little with depth and waters are close to complete oxygen saturation.
The stably cold and oxygen-rich waters found southward of the APF would be expected to serve as important effectors of evolution in the Antarctic marine biota. One would anticipate that during the approximately 40 million years of existence of the ACC and APF, adaptations to temperature would have led to extensive differentiation of organisms endemic to waters to the north or south of the APF. Studies of the major group of Antarctic fishes, members of the perciform suborder Notothenioidei, and their cold-temperate relatives in South America and New Zealand, show this to be the case. The biochemical differences between polar and cold-temperate notothenioids reflect the gain of important adaptive traits in both groups and the loss of traits no longer needed for life in stably cold waters in Antarctic species. This short review discusses these key differences and suggests new lines of studies, many of which are based on the new genomic technologies now becoming available for fishes, that may contribute importantly to our understanding of molecular evolution in protein-coding and gene regulatory components of the genome. 2. Characteristics of the Antarctic fish fauna: the suborder Notothenioidei Beginning in the early Miocene (2522 million years ago), the Antarctic shelf was subject to a series of tectonic and oceanographic events that undoubtedly altered faunal composition. Antarctica gradually became isolated and colder and expansion of the ice sheet led to destruction and disturbance of inshore habitat by ice, as a consequence of repeated groundings of parts of the ice sheet as far as the shelf break (Clarke and Johnston, 1996; Anderson, 1999). Loss of habitat and changes in the trophic structure of the ecosystem probably led to the local extinction of many of the Eocene components of the fish fauna. Thus, the diversity of the fauna was reduced and, as Antarctica became increasingly isolated, new niches became available to groups that were diversifying in situ (notothenioids) or immigrating into (liparids and zoarcids) this developing cold-water ecosystem (Eastman, 2005). Little is known, however, about when the fauna became modern in taxonomic composition. The first Antarctic notothenioids to be reported in the literature were collected near Kerguelen Island during the expedition of the Erebus and Terror under command of Sir James Clark Ross (18391843). Prior to these collections, it is doubtful that many believed that fishes could live in such a harsh environment. In fact, the fish fauna of the Southern Ocean is limited in both species and higher taxonomic diversity and contains only 313 species distributed among 50 families (Gon and Heemstra, 1990; Eastman, 2000). Thus, although the
Please cite this article as: Coppes Petricorena, Z.L., Somero, G.N. Biochemical adaptations of notothenioid fishes: Comparisons between cold temperate South American and New Zealand species and. Comparative Biochemistry and Physiology, Part A (2007), doi:10.1016/j.cbpa.2006.09.028
ARTICLE IN PRESS
Z.L. Coppes Petricorena, G.N. Somero / Comparative Biochemistry and Physiology, Part A xx (2007) xxxxxx 3
Southern Ocean represents approximately 10% of the world ocean, it contains only 1.3% of the world's fish fauna (Eastman and McCune, 2000). Benthic fishes are the major component of the fauna with 213 species; higher taxonomic diversity of benthic fishes is confined to 18 families (Eastman, 2000). Two perciform groups, the suborder Notothenioidei (notothenioids) and the Zoarcidae (eelpouts), and the scorpaeniform family Liparidae (snailfishes) are the most speciose taxa, accounting for 87.4% of the species (Eastman, 2000). The fish fauna as a whole is highly endemic; 88% endemism is found for species confined to water south of the APF. If only the notothenioids are considered, endemism rises to 97%, a very high percentage for a marine group, because only 10% specific endemism is enough for recognition of provinces (Eastman and McCune, 2000). The suborder Notothenioidei dominates the Antarctic fish fauna. Notothenioids account for approximately 46% of the total fish species known to occur south of the APF. More significantly, they comprise as much as 90% of the biomass of fish captures around the continent (Ekau, 1990; Eastman, 2005). The suborder Notothenioidei comprises eight families and 122 species. Five families and 96 species are Antarctic whereas three families and 26 species are non-Antarctic (Eastman, 2005). There is no fossil record of these fishes, and the origin of this group is ambiguous. According to Sidell (2000) all that can be said with certainty is that, sometime between the MidTertiary and the present, there was a massive crash of species diversity that left a sluggish, demersal stock of ancestral notothenioids to colonize the vast Southern Ocean, which at that time was not as cold as at present. The initial diversification of some of the basal families took place during fragmentation of Gondwana 6040 Mya when cooling of circumpolar waters began (Eastman, 1993). Since the Mid-Miocene, geographical isolation and a chronically cold environment have resulted in extreme stenothermality of extant species (Somero and DeVries, 1967; Cheng and Detrich, in press; Podrabsky and Somero, 2006). The collapse in species diversity may be due in part to decreases in water temperature; however, the slow geologic pace of ocean cooling has led Eastman and Clarke (1998) to conclude that the loss of shallow water habitats that accompanied Antarctica's glaciation may have been of equal, if not greater importance in causing the extinction. Whatever the causes of the initial extinctions, subsequent radiations of the fish fauna definitely are based on adaptations to the low temperatures currently found in the Southern Ocean, as discussed in the following sections of this review. 3. Non-Antarctic notothenioids To appreciate the unique biochemical features of Antarctic notothenioids, it is important to contrast our knowledge of these highly cold-adapted stenotherms with the information available about their cold-temperate relatives from South American and New Zealand waters. Although most notothenioids are endemic to the Southern Ocean, a number of species are endemic to temperate areas north of the Antarctic Polar Front such as in southern Australia, Tasmania, New Zealand, and southern South America. Three small basal families, Bovichtidae,
Pseudaphritidae and Eleginopidae, with 12 species are nonAntarctic. Among the other five notothenioid families, Nototheniidae, Harpagiferidae, Artedidraconidae, Bathydraconidae and Channichthyidae, 15 species occur along the cooltemperate southern coast of South America and New Zealand (Eastman and Eakin, 2000). These cold-temperate species encounter water temperatures of approximately 5 C15 C (Johnston et al., 1998; Fields and Somero, 1998). The evolutionary histories of the non-Antarctic notothenioids are not fully established, but there is compelling evidence, much of it of biochemical and molecular nature, that supports an Antarctic ancestry for some of these species. When the APF advanced 300 km to the north during the late Miocene (6.55.0 million years ago), cold water reached as far north as New Zealand (Kennett, 1982). Some notothenioids migrated along with the shifting APF and established themselves in New Zealand and cold-temperate South American waters. Other notothenioids, notably representatives of the basal lineages like the Family Bovichtidae, presumably diverged and became established in the brackish coastal water of the southern continental blocks before the isolation of Antarctica (Eastman and McCune, 2000). The lack of genes for antifreeze glycoproteins in members of the Bovichtidae, Pseudaphritidae and Eleginopidae, supports this scenario for the basal notothenioid lineages (Cheng and Detrich, in press). The presence or absence of genes encoding glycoprotein antifreezes thus provides a window albeit one that is cloudy at times (see below) into the evolutionary histories of Antarctic and non-Antarctic notothenioids. 4. Antifreeze glycoproteins We begin our comparison of Antarctic and non-Antarctic notothenioids with what is certainly the most striking difference between teleost fish that can and cannot survive in the presence of ice the occurrence in polar species of antifreeze glycoproteins or proteins that inhibit the growth of ice crystals (Cheng and DeVries, 1991; DeVries and Cheng, 2005; Cheng and Detrich, in press). Based on solute concentrations in blood and cells, notothenioids living in most regions of the Southern Ocean spend their entire lives at body temperatures below the predicted colligative freezing point of their body fluids (Cheng and Detrich, in press). What keeps these fish in liquid-state is a battery of antifreeze glycoproteins (AFGPs) that reduce the freezing point the temperature of ice crystal growth to well below ambient sea water temperatures. Recent studies by Cheng and colleagues (2006) have toppled the prevailing paradigm concerning where AFGPs are produced. They showed that hepatic synthesis, the initially proposed site of AFGP production, does not, in fact, occur; rather the production of these critical molecules is restricted to pancreatic tissue and the anterior portion of the stomach. From these sites of synthesis, AFGPs enter the entire gut cavity where the presence of ice ingested during feeding creates an acute danger of lethal ice formation (Cheng et al., 2006). How antifreezes move from the gut into the general circulation remains unknown and represents a critical question for study. Might there be a set of adaptations
ARTICLE IN PRESS
for AFGP transport that are unique to the notothenioids that have evolved freezing resistance? The occurrence of AFGPs in non-Antarctic notothenioids from South American and New Zealand waters has been examined in several studies (Cheng et al., 2003; for review, see Cheng and Detrich, in press). In two endemic New Zealand notothenioids, Notothenia angustata and Notothenia microlepidota, two or three genes encoding AFGPs are present, respectively, and low amounts of AFGPs can be detected in blood (Cheng et al., 2003). This discovery indicates that these two cold-temperate New Zealand notothenioids are derived from an Antarctic ancestor, because there is strong evidence that the gene encoding AFGP arose only once and at a time that preceded the radiation of the notothenioid families with Antarctic representatives (Eastman, 1993; Cheng, 1996; Cheng et al., 2003). Thus, the most parsimonious explanation for the presence of AFGP-containing notothenioids in a coldtemperate New Zealand habitat is that the species (or their ancestor(s)) were escapees from Antarctic waters that followed the northern-moving APF to lower latitudes when this type of oceanographic shift occurred, most recently during the late Miocene (Eastman and McCune, 2000). Two further differences between Antarctic and coldtemperate AFGP-containing notothenioids are the copy numbers of afgp genes and the numbers of AFGPs encoded by the polyprotein-encoding afgp genes. This gene family is thought to have arisen from a trypsinogen-like serine protease gene no more than 14 million years ago, a date that is consistent with the freezing of the coastal Southern Ocean (1015 million years ago; Kennett, 1977) and the diversification of the 5 Antarctic notothenioid families 715 million years ago (Bargelloni et al., 1994; Chen et al., 1997). The size of the AFGP gene family has increased in Antarctic notothenioids and the number of AFGP molecules encoded in a given antifreeze gene likewise has increased in Antarctic species (Cheng et al., 2003). Both evolutionary trends reflect the severity of the freezing threat that Antarctic notothenioids face. The small number of afgp genes in the New Zealand notothenioids and the small number of AFGPs encoded by each of these genes suggest that these species reached New Zealand waters before the production of AFGPs had reached the levels found in current Antarctic notothenioids. If this hypothesis is true, then the dating of the evolution of the expansion of the afgp gene family in terms of gene copy number and protein-encoding sites per gene can be better understood. The amino acid sequences of the AFGPs of Antarctic and New Zealand notothenioids also exhibit interesting differences. Thus, 6 of the 10 AFGPs found in N. angustata and 4 of the 11 found in N. microlepidota contain amino acid substitutions that are predicted to lead to a loss of antifreeze function. It remains unclear as to why the New Zealand notothenioids continue to express afgp genes and why expression increases in the cold. These, too, are questions for future study. It is interesting that the Patagonian tooth fish Dissostichus eleginoides (Nototheniidae), which occurs from a latitude of 40S off the coasts of South America to 60S in the Antarctic and overlaps in distribution with its strictly Antarctic congener
D. mawsoni lacks AFGPs (Cheng and Detrich, in press). It is highly unlikely that D. eleginoides diverged before the evolutionary acquisition of the afgp gene, so the most parsimonious explanation for the apparent absence of these genes in this cold-temperate nototheniid entails loss or severe mutation of this gene following entry into cold-temperate waters. Study of the mechanisms of loss of afgp genes in the cold-temperate congener of Dissostichus is clearly warranted. Puzzles also remain about the occurrence of afgp genes in the genus Patagonotothen, a large genus with 12 species, all of which are found in South American waters except for one, P. guntheri, which occurs at the northern tip of the Antarctic Peninsula. All of these species appear to lack afgp genes (Cheng and Detrich, in press). As in the case of D. eleginoides, the absence of afgp genes in this genus is unlikely to reflect the radiation of this group before the acquisition of the original afgp gene. Rather, loss of AFGP-encoding genes appears to be the likely mechanism, although this conjecture will require more analysis using genomic technologies. Perhaps remnants of afgp genes will be found in the AFGP-lacking species and a deeper understanding of gene loss during adaptation to non-freezing temperatures by AFGP-containing species will be obtained. Such discoveries would be an excellent complement to the elegant work that has shown us how the AFGP-encoding genes initially arose (Chen et al., 1997; Cheng and Detrich, in press). 5. Gene loss in stably cold waters: the heat-shock response Whereas Antarctic notothenioids are extraordinarily welladapted for life at near-freezing temperatures, they fare poorly when confronted with elevated temperatures. Upper incipient lethal temperatures for several notothenioids from McMurdo Sound acclimated to 1.9 C were near 56 C, marking these fish as extreme stenotherms (Somero and DeVries, 1967). Nonetheless, a recent study showed that some capacity for induced thermal tolerance is present in certain notothenioid species (Podrabsky and Somero, 2006). Thus, for the nototheniids Trematomus bernacchii and Trematomus pennellii, survival times at 14 C increased from approximately 20 min in 1.9 C laboratory-acclimated specimens to 60 and 140 min, respectively, in specimens acclimated for 68 weeks to 4 C. These results indicate that, although Antarctic notothenioids are among the most stenothermal species of animals known, they do have some mechanisms for increasing their resistance to acute heat stress. Unlike all other fishes so examined, however, these mechanisms fail to include the once-thought-to-be ubiquitous heat-shock response (HSR) (Hofmann et al., 2000). Cells typically respond to heat stress with the synthesis of a group of highly conserved proteins termed heat shock proteins (Hsps) belonging to several size classes (Lindsquist and Craig, 1988). Hsps, as members of a broad family of molecular chaperones, function to minimize protein denaturation and aggregation during heat stress, assist in renaturation of proteins that are partially denatured, and, in some cases, play a role in proteolytic degradation of irreversibly damaged proteins (Hochachka and Somero, 2002). Using 35S-labeled methionine and cysteine
ARTICLE IN PRESS
and SDS-PAGE separation to detect the presence of different size classes of newly synthesized proteins in T. bernacchii subjected to different levels of heat stress, Hofmann et al. (2000) showed that no size class of Hsp exhibited increased synthesis. Prior to this investigation, only a single study suggested that the HSR may not be ubiquitous: a study of congeners of Hydra showed that a species (H. oligactis) found in cool, thermally stable environments produced mRNA for Hsp70, but this message was unstable and did not get translated into protein (Bosch et al., 1988; Gellner et al., 1992). In Antarctic notothenioids, in contrast, neither mRNA for Hsp70 nor Hsp70 protein increases in abundance following heat stress (Hofmann et al., 2000; Place and Hofmann, 2004; Place et al., 2004). The lack of an HSR in Antarctic notothenioids belies the fact that constitutive expression of heat-inducible genes such as that encoding Hsp70 occurs in these fish (Place et al., 2004). Thus, unlike other fish, production of mRNA and protein appears decoupled from thermal stress or at least stress from elevated temperatures. Place and colleagues propose that the constitutive expression of Hsp-encoding genes in Antarctic notothenioids may, in fact, reflect thermal stress to protein folding that arises from constraints of low temperatures on this process. Thus, if the kinetics of protein folding are slowed at subzero temperatures, the dangers from aggregation of partially folded nascent polypeptides could be substantial. If this is the case, then high concentrations of molecular chaperones like Hsp70 would be adaptive because cold-induced aggregation would be reduced. There are examples of cold-induced induction of Hsps, so threats to protein folding from low extremes of temperature potentially exist (see Place et al., 2004, for review). Comparative study of the temperature-dependence of folding of nascent polypeptide chains is clearly warranted to determine whether reduced rates of protein maturation in extreme cold establish a need for enhanced levels of molecular chaperones. The absence of an HSR in Antarctic notothenioids contrasts with the responses of New Zealand notothenioids to heat stress (Place et al., 2004; Hofmann et al., 2005). Two New Zealand endemics, Bovichtus variegatus and N. angustata, exhibited abilities to up-regulate expression of Hsp70-encoding genes in response to heat stress of 16 C and 18.8 C, temperatures near the upper end of their environmental temperature range. The occurrence of the HSR in N. angustata, a species that probably is derived from an ancestor that escaped from Antarctic waters, helps to put a date on the timing of the loss of the HSR in Antarctic species. Thus, if the northward movement of the APF occurred during the late Miocene, 6.55 million years ago, well after the origin of the AFGP-encoding genes some 14 million years ago, the loss of the HSR would be a relatively recent event in the evolutionary histories of Antarctic notothenioids. In the case of B. variegatus, the occurrence of the HSR is consistent with the hypothesis that the Bovichtidae is a basal group that has evolved under temperate conditions. The mechanisms responsible for the lack of inducibility of Hsps in Antarctic notothenioids remain to be discovered. The transcriptional factor that is critical for expression of Hspencoding genes, heat-shock factor 1 (HSF1), is a logical
candidate for an evolutionary lesion. However, Buckley and colleagues (2004) showed that HSF1 is present in cells of T. bernacchii, so loss of this protein has not occurred. However, unlike what has been observed in other species, elevated temperature had no effect on the ability of HSF1 to bind to the gene regulatory region, the heat shock element, which modulates expression of Hsp-encoding genes. Further study will be needed to elucidate the precise alterations in the different components of the HSR that lead to constitutive expression of Hsps yet prevent heat-induced activation of the hsp genes. In addition to differing in gene-regulatory characteristics, the thermal optima of heat-shock proteins themselves differ between Antarctic and cold-temperate nototheniids. Place and Hofmann (2005) studied a constitutively expressed heat-shock protein, Hsc70 (a cognate of Hsp70), purified from N. angustata, T. bernacchii, and P. borchgrevinki. They measured abilities of Hsc70 to prevent thermal aggregation of lactate dehydrogenase (LDH) and to refold chemically denatured LDH over the temperature range of 2 C to 45 C. Hsc70 orthologs of all three species were capable of refolding chemically denatured LDH in vitro over this temperature range. However, the Hsc70 of the cold-temperate fish N. angustata outperformed the Hsc70s of the two Antarctic species at temperatures of 20 C and higher; conversely, the Hsc70s of the Antarctic species out-performed the cold-temperate ortholog at 2 C. The Hsc70 orthologs of Antarctic and cold-temperate notothenioids could thus be an excellent study system for discerning how the temperature sensitivities of molecular chaperones evolve. 6. Temperature adaptation of enzymatic proteins The discovery that orthologous Hsc70s from Antarctic and cold-temperate notothenioids differ in thermal optima complements the findings of studies of temperature adaptation of enzymatic proteins. Enzyme function is highly sensitive to temperature change, largely because of the balance that must be maintained between flexibility and stability in discrete, relatively mobile regions of the protein that are involved in catalytically important conformational changes (Fields and Somero, 1998; Hochachka and Somero, 2002; Fields and Houseman, 2004). Molecular flexibility is needed to maintain an appropriate catalytic rate, but stability is required to ensure correct active site geometry for substrate recognition. Fields and Somero (1998) compared the catalytic rate constants (kcat) and MichaelisMenten constants (Km, an index of substrate binding ability) in lactate dehydrogenase-A orthologs (LDH-As) of Antarctic and South American notothenioids. Each biogeographic group showed distinct differences. Rates of LDH-A activity, as measured by kcat values, were on average higher for the Antarctic species. Binding (Km of the PYR substrate pyruvate (Km )) was strongly conserved at normal body temperatures due to an intrinsically stronger binding PYR (lower Km at a common temperature of measurement) for the LDH-A orthologs of the South American notothenioids. PYR Conservation of Km in LDH-A orthologs of differently thermally adapted vertebrates has previously been documented
ARTICLE IN PRESS
for fishes (Yancey and Somero, 1978; Graves and Somero, 1982; Coppes and Somero, 1988, 1990; Holland et al., 1997; Johns and Somero, 2004) and other taxa (Yancey and Somero, 1978). Sequencing studies of the twelve notothenioid LDH-A orthologs revealed a number of insights into the evolution of this enzyme in the notothenioid lineages. First, only a single notothenioid species, the South American Eleginops maclovinus, had Histidine-75 in its sequence. This amino acid residue is present in all non-notothenioid teleost LDH-As sequenced to date. The deduced amino acid sequence of the LDH-A of E. maclovinus has eleven differences from the notothenioid consensus sequence, whereas other species' LDH-As differed by only one to four changes. The large divergence of the E. maclovinus sequence from other notothenioid sequences is consistent with the recent placement of this species into a new family (Eleginopidae) (see Fields and Somero, 1998). A second finding of this comparison helps to link amino acid substitutions to changes in enzyme function. The amino acid sequences of the active site regions of all of the orthologs are identical, so the temperature-adaptive differences in kinetic PYR properties (kcat and Km ) must arise from substitutions in regions of the enzyme that do not directly interact with substrate or cofactor. The majority of changes in sequence occur in regions of the enzyme that influence the conformational flexibility of the parts of the enzyme that change shape during binding events. The cold-adapted LDH-As of Antarctic notothenioids appear to have higher flexibility in these regions, which allows lower energy (enthalpy) barriers to conformaPYR tional changes and, hence, higher kcat and Km values. A number of substitutions are capable of adapting kinetic properties, such that within a biogeographic group, similar kinetic properties are found in orthologs with different primary structures. For example, although the LDH-A orthologs of two of the South American species, Patagonotothen tessellata and PYR Paranotothenia magellanica, have the same Km values, they share none of the differences from the consensus sequence. A third finding, which reflects results seen in other studies of enzymatic adaptation to temperature (e.g., Holland et al., 1997; Johns and Somero, 2004), is that only a single amino acid substitution may be sufficient to adaptively alter kinetic properties of enzymes. Fields and Houseman (2004) showed that altering one residue (number 233) from a glutamate to a methionine was sufficient to change an Antarctic LDH-A to a variant with kinetic properties identical to those of an LDH-A from a warm-temperate fish. Lastly, the kcat values for the South American species offer insights into the evolutionary histories of these species. Although on average the kcat values for these species were lower than those for the Antarctic notothenioids, the kcat of the LDH-A of P. tessellata fell into the range of values for the Antarctic species. This may reflect the retention of a coldadapted kcat in this species, which is derived from an ancestral species that escaped from cold Antarctic waters and gave rise to the 13 extant Patagonotothen species. In contrast, the low kcat of E. maclovinus may reflect the absence of cold adaptation in this species' evolutionary history.
The structurefunction analyses that could be performed through comparative studies of enzymes in cold-temperate and Antarctic notothenioids offer exciting promise for furthering our understanding of how enzymes function and evolve. In particular, the discovery that notothenioids from Antarctic and South American habitats may attain adaptation through modifying different sites in the amino acid sequence is helping to reveal the set of evolutionary options proteins possess for adaptive modification of kinetic parameters. 7. Structural adaptations of muscle fibres: relationship between diameter and number Skeletal muscle fibres are differentiated multicellular structures specialized for contraction. The functional properties of a muscle will be strongly influenced by fibre diameter and fibre number. The maximum diameter of each muscle fibre is related to ultimate body size and is probably limited by diffusional constraints that stem from metabolic demand and temperature (Archer and Johnston, 1991). Fibre number may increase during post-embryonic stages through activation of myogenic precursor cells, which proliferate before leaving the cell cycle and fuse to form new myotubes (Johnston, 2001). Antarctic notothenioids have unusually large diameter fibres, which can reach 100 m in slow muscle and 600 m in fast muscle (Battram and Johnston, 1991). Slow muscle fibres have relatively high densities of mitochondria (O'Brien et al., 2003), reaching 50% of fibre volume in some icefishes (Johnston, 1989). These mitochondria are found in the central zone of even the largest diameter slow fibres (Archer and Johnston, 1991). This localization is consistent with maintenance of adequate oxygenation at the low body temperature of this species (Egginton et al., 2002). Fast muscle fibres with diameters of 400 m have also been reported in sub-Antarctic notothenioids from the Beagle Channel, although a relatively restricted size range of fish was studied (Fernandez et al., 2000). Johnston et al. (2003) studied the number of muscle fibres and fibre diameters in 16 species of notothenioids from three geographical regions, Tierra del Fuego, South Georgia, and Antarctica, including representatives with benthic and secondarily pelagic life styles. They estimated ancestral values for body size and fibre number to explore how fibre number and size changed during the evolutionary radiation of this group. They showed that the radiation has been associated with a progressive loss in body size specific-maximum number of fast fibres in the myotomal muscle of the more derived species. An important consequence of the reduction in fibre number in the more derived lineages of notothenioid fish is an increase in their maximum diameter, which can reach 600 m in some species, depending on their final body size. The unusual evolutionary patterns in muscle fibre number and size in notothenioids can be appreciated by comparisons with other fish. Atlantic salmon with a standard length of 50 70 cm have 550,000 to 1,200,000 fast muscle fibres per trunk cross section (Johnston et al., 2000), with a fibre diameter of approximately 220 m. In contrast, the sub-Antarctic species
ARTICLE IN PRESS
Chaenocephalus aceratus with a body length of 85 cm, has only 12,700 fast muscle fibres per trunk cross section, and a fibre diameter of 600 m. The Patagonian notothenioid Eleginops maclovinus, which reaches similar size to the C. aceratus, has 164,000 fibres with a fibre diameter of approximately 490 m (Johnston et al., 2003). Thus, although data are limited, comparisons of fibre diameters in notothenioids with those of temperate and tropical fishes suggest that the large maximum diameter of muscle fibres in notothenioid fishes, especially the more derived Antarctic lineages, is exceptional. The factors that have selected for small numbers of large sized muscle fibres in derived Antarctic notothenioids, notably the Nototheniidae and Channichthyidae, are not fully understood, but may relate to the energy costs of life at near-freezing temperatures, especially the costs of producing large amounts of AFGPs (Johnston et al., 2003). The greater fibre number in muscle of two South American notothenioids lacking antifreezes, Eleginops maclovinus and Cottoperca gobio, may reflect the absence of an evolutionary period in cold Antarctic waters by these fish. According to Egginton et al. (2002) the rate of oxygen delivery to aerobic muscle fibres is a function of the fibre diameter together with factors that affect diffusion rate: temperature, distribution of mitochondria and lipid droplets within the fibre as well as the overall metabolic demand (O'Brien et al., 2003). Adequate oxygen delivery to large diameter muscle fibres is probably only possible because of the very low metabolic demand in polar fishes at low temperature (Clarke and Johnston, 1996; Steffensen, 2002). Modeling studies by Egginton et al. (2002) indicate that a low fibre number and high maximum fibre diameter do not limit adequate oxygen flux at low body temperatures in notothenioids. Further comparisons of muscle fibre size and number in cold-temperate notothenioids, including New Zealand species that arose from ancestors that escaped from Antarctica could further clarify the roles that adaptation to cold has played in the evolution of skeletal muscle in notothenioids. 8. Genetics of notothenioids: what has been lost during evolution in stably cold waters? The discovery that Antarctic notothenioids have lost genetic information that likely is essential for life in warmer waters raises a number of questions about the evolutionary histories of notothenioids and the future prospects of these species in a warming world. What other types of genetic information have been lost, in addition to genes encoding hemoglobin and myoglobin (for recent reviews of this topic, see Cheng and Detrich, in press; Sidell and O'Brien, 2006) and components of the heat-shock response (Hofmann et al., 2000; Place et al., 2004; Place and Hofmann, 2005)? Does loss of genetic information preclude acclimation to elevated temperatures such as those predicted as a result of climate change? Have cold-temperate notothenioids retained this critical genetic information? These questions about genetic lesions complement those asked by Montgomery and Clements (2000) in their analysis of loss and regain of function in numerous
anatomical, physiological and behavioral capacities of Antarctic notothenioids. Although at present we can provide, at best, only preliminary answers to any of these questions about genetic losses during evolution in stably cold waters, the exploitation of genomic technologies in the study of notothenioids may soon allow us to examine in fine detail the genetic tool kits of Antarctic and non-Antarctic notothenioids (Peck et al., 2005). Sequencing and annotating of Antarctic notothenioid genomes is under way, and we may soon know what protein-encoding genes and regulatory loci have been lost (or rendered dysfunctional) during evolution in the Southern Ocean. Complementary comparative studies of the genomes of cold-temperate notothenioids could reveal whether escapees from Antarctica migrated to temperate waters before some (or all) of these genetic lesions occurred. Better defining the acclimatory capacities of Antarctic and coldtemperate notothenioids will allow firmer predictions to be made about the potential effects of climate change on this suborder of fish. Analyses of gene expression using complementary DNA (cDNA) microarrays (gene chips) offer another promising experimental strategy for elucidating the genetic capabilities of notothenioids fishes. Studies of eurythermal fishes have revealed that hundreds of genes may shift expression during thermal acclimation (Gracey et al., 2004; Podrabsky and Somero, 2004; Buckley et al., 2006). Microarray analysis of temperature-induced changes in gene expression of Antarctic notothenioids is underway and is beginning to show similarities and differences between these extreme stenotherms and the more eurythermal fishes recently examined with this technology (Buckley and Somero, in preparation). Absence of induction of Hsp-encoding genes is confirmed, but other stress-related genes show qualitatively similar expression patterns to those seen in temperate species. Thus, not all of the acclimatory abilities of Antarctic fishes have been lost during millions of years of evolution at cold, stable temperatures. The finding that notothenioids have the capacity to increase their resistance to acute heat shock suggests that some temperature-dependent gene regulatory abilities related to stress tolerance remain in the tool kits of Antarctic notothenioids, even though the HSR has been lost (Podrabsky and Somero, 2006). How the genetic repertoires of Antarctic notothenioids have changed and how they differ from those of cold-temperate notothenioids represents an exciting frontier in the study of this fascinating suborder of fishes. Acknowledgements We thank Dr. Bradley Buckley for his critical reading of this manuscript. References
Anderson, J.B., 1999. Antarctic Marine Geology. Cambridge University Press, Cambridge. Archer, S.D., Johnston, I.A., 1991. Density of cristae and distribution of mitochondria in the slow muscles of Antarctic fish. Physiol. Zool. 62, 242258.
ARTICLE IN PRESS
8 Z.L. Coppes Petricorena, G.N. Somero / Comparative Biochemistry and Physiology, Part A xx (2007) xxxxxx Egginton, S., Stilbeck, C., Hoofd, L., Calvo, J., Johnston, I.A., 2002. Peripheral oxygen transport in skeletal muscle of Antarctic and sub-Antarctic notothenioid fish. J. Exp. Biol. 205, 769779. Ekau, W., 1990. Demersal fish fauna of the Weddell Sea, Antarctica. Antarct. Sci. 2, 129137. Fernandez, D.A., Calvo, J., Franklin, C.E., Johnston, I.A., 2000. Muscle fibre types and size distribution in sub-Antarctic notothenioid fishes. J. Fish Biol. 56, 12951311. Fields, P.A., Houseman, D.E., 2004. Decreases in activation energy and substrate affinity in cold-adapted A4-LDH Dehydrogenase: evidence from the Antarctic notothenioid fish Chaenocephalus aceratus. Mol. Biol. Evol. 21, 22462255. Fields, P.A., Somero, G.N., 1998. Hot spots in cold adaptation: localized increases in conformational flexibility in lactate dehydrogenase A4 orthologs of Antarctic notothenioid fishes. Proc. Natl. Acad. Sci. U. S. A. 95, 1147611481. Gellner, K., Praetzel, G.A., Bosch, T.C.G., 1992. Cloning and expression of a heat-inducible hsp70 gene in two species of Hydra which differ in their stress response. Eur. J. Biochem. 210, 683691. Gon, O., Heemstra, P.C., 1990. Fishes of the Southern Ocean. J. L. B. Smith Institute of Ichthyology, Grahamstown, South Africa. Gordon, A.L., 1988. The Southern Ocean and global climate. Oceanus 31, 3946. Gordon, A.L., 1999. The Southern Ocean. Current 15, 46. Gordon, A.L., 2003. Oceanography the brawniest retroflection. Nature 421, 904905. Gracey, A.Y., Fraser, E.J., Li, W., Fang, Y., Taylor, R.R., Rogers, J., Brass, A., Cossins, A.R., 2004. Coping with cold: an integrative, multitissue analysis of the transcriptome of a poikilothermic vertebrate. Proc. Natl. Acad. Sci. U. S. A. 101, 1697016975. Graves, J.E., Somero, G.N., 1982. Electrophoretic and functional enzymic evolution in 4 species of eastern Pacific barracudas from different thermal environments. Evolution 36, 97106. Hochachka, P.W., Somero, G.N., 2002. Biochemical Adaptation. Mechanism and Process in Physiological Evolution. Oxford University Press, New York. 466 pp. Hofmann, G.E., Buckley, B.A., Airaksinen, S., Keen, J.E., Somero, G.N., 2000. Heat-shock protein expression is absent in the Antarctic fish Trematomus bernacchii (Family Nototheniidae). J. Exp. Biol. 203, 23312339. Hofmann, G.E., Lund, S.G., Place, S.P., Whitmer, A.C., 2005. Some like it hot, some like it cold: the heat shock response is found in New Zealand but not Antarctic notothenioid fishes. J. Exp. Mar. Biol. Ecol. 316, 7989. Holland, L.Z., McFall-Ngai, M., Somero, G.N., 1997. Evolution of lactate dehydrogenase-A homologues of barracuda fishes (genus Sphyraena) from different thermal environments: differences in kinetic properties and thermal stability are due to amino acid substitutions outside the active site. Biochemistry 36, 32073215. Hunt, B.M., Hoefling, K., Cheng, C.-H.C., 2003. Annual warming episodes in seawater temperatures in McMurdo Sound in relationship to endogenous ice in notothenioid fish. Antarct. Sci. 15, 333338. Johns, G.C., Somero, G.N., 2004. Evolutionary convergence in adaptation of proteins to temperature: A4-lactate dehydrogenases of Pacific damselfishes (Chromis spp.). Mol. Biol. Evol. 21, 314320. Johnston, I.A., 1989. Antarctic fish musclesstructure, function and physiology. Antarct. Sci. 1, 97108. Johnston, I.A., 2001. Genetic and environmental determination of muscle growth patterns. In: Johnston, I.A. (Ed.), Muscle Development and Growth. . Fish Physiol., vol. 18. Academic Press, San Diego, pp. 141148. Johnston, I.A., Calvo, J., Guderly, H., Fernandez, D.A., Palmer, L., 1998. Latitudinal variation in the abundance and oxidative capacities of muscle mitochondria in Perciform fishes. J. Exp. Biol. 210, 112. Johnston, I.A., Manthri, S., Robertson, B., Campbell, P., Mitchell, D., Anderson, R., 2000. Family and population differences in muscle fibre recruitment in farmed Atlantic salmon (Salmo salar). Basic Appl. Myol. 10, 291296. Johnston, I.A., Fernandez, D.A., Calvo, J., Vieira, V.L.A., North, A.W., Abercromby, M., Garland Jr., T., 2003. Reduction in muscle fibre number during the adaptive radiation of notothenioid fishes: a phylogenetic perspective. J. Exp. Biol. 206, 25952609. Bargelloni, L., Ritchie, P.A., Patarnello, T., Battaglia, B., Lambert, D.M., Meyer, A., 1994. Molecular evolution at subzero temperatures: mitochondrial and nuclear phylogenies of fishes from Antarctica (suborder Notothenioidei), and the evolution of antifreeze glycopeptides. Mol. Biol. Evol. 11, 854863. Battram, J.C., Johnston, I.A., 1991. Muscle growth in the Antarctic teleost, Notothenia neglecta (Nybelin). Antarct. Sci. 3, 2933. Bosch, T.C.G., Krylow, S.M., Bode, H.R., Steele, R.E., 1988. Thermotolerance and synthesis of heat shock proteins: these responses are present in Hydra attenuata but absent in Hydra oligactis. Proc. Natl. Acad. Sci. U. S. A. 85, 70317927. Buckley, B.A., Place, S.P., Hofmann, G.E., 2004. Regulation of heat shock genes in isolated hepatocytes from an Antarctic fish, Trematomus bernacchii. J. Exp. Biol. 207, 36493656. Buckley, B.A., Gracey, A.Y., Somero, G.N., 2006. The cellular response to heat stress in the goby Gillichthys mirabilis: a cDNA microarray and proteinlevel analysis. J. Exp. Biol. 209, 26602677. Chen, L., De Vries, A.L., Cheng, C.-H.C., 1997. Convergent evolution of antifreeze glycoproteins in Antarctic notothenioid fish and Arctic cod. Proc. Natl. Acad. Sci. U. S. A. 94, 38173822. Cheng, C.-H.C., 1996. Genomic basis for antifreeze glycopeptide heterogeneity and abundance in Antarctic fishes. In: Ennion, S., Goldspink, G. (Eds.), Gene Expression and Manipulation in Aquatic Organisms. Cambridge University Press, Cambridge, pp. 120. Cheng, C.-H.C., DeVries, A.L., 1991. Evolution of an antifreeze glycoprotein. Nature 401, 443444. Cheng, C-H.C., Detrich, H.W. III., in press. Molecular ecophysiology of Antarctic notothenioid fishes. Phil. Trans. Roy. Soc. Lond. Series B. Cheng, C.-H.C., Chen, L., Near, T., Jin, Y., 2003. Functional antifreeze glycoprotein genes in temperate-water New Zealand nototheniid fish infer an Antarctic evolutionary origin. Mol. Biol. Evol. 20 (11), 18971908. Cheng, C.-H.C., Cziko, P.A., Evans, C.W., 2006. Non-hepatic origin of notothenioid antifreeze reveals pancreatic synthesis as common mechanism in polar fish freezing avoidance. Proc. Natl. Acad. Sci. U. S. A. 103, 1049110496. Clarke, A., Johnston, I.A., 1996. Evolution and adaptive radiation of Antarctic fishes. Trends Ecol. Evol. 11, 212218. Clarke, A., Barnes, D.K.A., Hodgsone, D.A., 2005. How isolated is Antarctica? Trends Ecol. Evol. 20, 13. Coppes, Z.L., Somero, G.N., 1988. Temperature adaptation of Lactate Dehydrogenase (LDH-A4) from white skeletal muscle of Sciaenops ocellatus (Sciaenidae). Contr. Mar. Sci. 30, 147152. Coppes, Z.L., Somero, G.N., 1990. Temperature-adaptive differences between the M4-lactate dehydrogenases of stenothermal and eurythermal Sciaenid fishes. J. Exp. Zool. 254, 127131. DeVries, A.L., Cheng, C.-H.C., 2005. Antifreeze proteins and organismal freezing avoidance in polar fishes. In: Farrell, A.P., Steffensen, J.F. (Eds.), The Physiology of Polar Fishes. . Fish Physiol., vol. 22. Elsevier Academic Press, San Diego, pp. 155201. Dewitt, H.H., 1970. A revision of the fishes of the genus Notothenia from the New Zealand region, including Macquarie Island. Proc. Calif. Acad. Sci. 38, 299340. Eastman, J.T., 1993. Antarctic Fish Biology Evolution in a Unique Environment. Academic Press, San Diego. 322 pp. Eastman, J.T., 2000. Antarctic notothenioid fishes as subjects for research in evolutionary biology. Antarct. Sci. 12, 276287. Eastman, J.T., 2005. The nature of the diversity of Antarctic fishes. Polar Biol. 28, 94107. Eastman, J.T., Clarke, A., 1998. Radiations of Antarctic and non-Antarctic fish. In: DiPrisco, G., Pisano, E., Clarke, A. (Eds.), Fishes of Antarctica: A Biological Overview. Springer-Verlag, Milan, Italy, pp. 326. Eastman, J.T., Eakin, R.R., 2000. An updated species list for notothenioid fish (Perciformes: Notothenioidei), with comments on Antarctic species. Arch. Fish. Mar. Res. 48, 1120. Eastman, J.T., McCune, A.R., 2000. Fishes on the Antarctic continental shelf: evolution of a marine species flock? J. Fish Biol. 57 (Supplement A), 84102.
ARTICLE IN PRESS
Z.L. Coppes Petricorena, G.N. Somero / Comparative Biochemistry and Physiology, Part A xx (2007) xxxxxx Kennett, J.P., 1977. Cenozoic evolution of Antarctic glaciation. The circumAntarctic Ocean, and the impact of global paleoceanography. J. Geophys. Res. 82, 38433860. Kennett, J.P., 1982. Marine Geology. Prentice-Hall, Englewood Cliffs, N.J. Lindsquist, S., Craig, E.A., 1988. The heat shock proteins. Annu. Rev. Genet. 22, 631678. Littlepage, J.L., 1965. Oceanographic investigation in McMurdo Sound, Antarctica. In: Llano, G.A. (Ed.), Biology of the Antarctic Seas, vol. II. American Geophysical Union, Washington, DC, pp. 137. Montgomery, J., Clements, K., 2000. Disaptation and recovery in the evolution of Antarctic fishes. Trends Ecol. Evol. 15, 267271. O'Brien, K.M., Skilbeck, C., Sidell, B.D., Egginton, S., 2003. Muscle fine structure may maintain the function of oxidative fibers in hemoglobin Antarctic fishes. J. Exp. Biol. 206, 411421. Peck, L.S., Clark, M.S., Clarke, A., Cockell, C.S., Convey, P., Detrich III, H.W., Fraser, K.P.P., Johnston, I.A., Methe, B.A., Murray, A.E., Rmisch, K., Rogers, A.D., 2005. Genomics: applications to Antarctic ecosystems. Polar Biol. 28, 361365. Place, S.P., Hofmann, G.E., 2004. Temperature interactions of the molecular chaperone Hsc70 from the eurythermal marine goby Gillichthys mirabilis. J. Exp. Biol. 204, 26752682. Place, S.P., Hofmann, G.E., 2005. Comparison of Hsc70 orthologues from polar and temperate notothenioid fishes: differences in the prevention of aggregation and refolding of denatured proteins. Am. J. Physiol.: Regul., Integr. Comp. Physiol. 288, R1195R1202. 9 Place, S.P., Zippay, M.L., Hofmann, G.E., 2004. Constitutive roles for inducible genes: evidence for the alteration in expression of the inducible hsp70 gene in Antarctic notothenioid species. Am. J. Physiol.: Regul., Integr. Comp. Physiol. 287, R429R436. Podrabsky, J.E., Somero, G.N., 2004. Changes in gene expression associated with acclimation to constant temperatures and fluctuating daily temperatures in an annual killifish, Austrofundulus limnaeus. J. Exp. Biol. 207, 22372254. Podrabsky, J.E., Somero, G.N., 2006. Inducible heat tolerance in Antarctic notothenioid fishes. Polar Biol., doi:10.1007/s00300-006-0157-y Scher, H.D., Martin, E.E., 2006. Timing and climatic consequences of the opening of the Drake Passage. Science 312, 428430. Sidell, B.D., 2000. Life at body temperatures below 0 C: the physiology and biochemistry of Antarctic fishes. Gravit. Space Biol. Bull. 13, 2534. Sidell, B.D., O'Brien, K.M., 2006. When bad things happen to good fish: the loss of hemoglobin and myoglobin in Antarctic icefishes. J. Exp. Biol. 209, 17911802. Somero, G.N., DeVries, A.L., 1967. Temperature tolerance of some Antarctic fishes. Science 156, 257258. Steffensen, J.F., 2002. Metabolic cold adaptation of polar fish based on measurement of aerobic oxygen consumption: fact or artefact? Artefact! Comp. Biochem. Physiol., Part A: Mol. Integr. Physiol. 132, 789795. Yancey, P.H., Somero, G.N., 1978. Temperature dependence of intracellular pH: its role in the conservation of pyruvate apparent Km values of vertebrate lactate dehydrogenases. J. Comp. Physiol., B 125, 129134.

Coppes and Somero CBP 2007

Uploaded by

Document Information

Original Description:

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Coppes and Somero CBP 2007

Uploaded by

Copyright:

Available Formats

CBA-08049; No.

Comparative Biochemistry and Physiology, Part A xx (2007) xxx xxx www.elsevier.com/locate/cbpa

You might also like