Cryofouling avoidance in the Antarctic scallop Adamussium colbecki.

The presence of supercooled water in polar regions causes anchor ice to grow on submerged objects, generating costly problems for engineered materials and life-endangering risks for benthic communities. The factors driving underwater ice accretion are poorly understood, and passive prevention mechanisms remain unknown. Here we report that the Antarctic scallop Adamussium colbecki appears to remain ice-free in shallow Antarctic marine environments where underwater ice growth is prevalent. In contrast, scallops colonized by bush sponges in the same microhabitat grow ice and are removed from the population. Characterization of the Antarctic scallop shells revealed a hierarchical micro-ridge structure with sub-micron nano-ridges which promotes directed icing. This concentrates the formation of ice on the growth rings while leaving the regions in between free of ice, and appears to reduce ice-to-shell adhesion when compared to temperate species that do not possess highly ordered surface structures. The ability to control the formation of ice may enable passive underwater anti-icing protection, with the removal of ice possibly facilitated by ocean currents or scallop movements. We term this behavior cryofouling avoidance. We posit that the evolution of natural anti-icing structures is a key trait for the survival of Antarctic scallops in anchor ice zones.

Weddell seals produce ultrasonic vocalizations.

Seals (phocids) are generally not thought to produce vocalizations having ultrasonic fundamental frequencies (≥20 kHz), although previous studies could have been biased by sampling limitations. This study characterizes common, yet, previously undescribed, ultrasonic Weddell seal (Leptonychotes weddellii) vocalizations. The vocalizations were identified in more than one year (2017-2018) of broadband acoustic data obtained by a continuously recording underwater observatory in McMurdo Sound, Antarctica. Nine recurrent call types were identified that were composed of single or multiple vocal elements whose fundamental frequencies spanned the ultrasonic range to nearly 50 kHz. Eleven vocal elements had ultrasonic center frequencies (≥20 kHz), including chirps, whistles, and trills, with two elements at >30 kHz. Six elements had fundamental frequencies always >21 kHz. The fundamental frequency of one repetitive U-shaped whistle element reached 44.2 kHz and descending chirps (≥3.6 ms duration) commenced at ≤49.8 kHz. The source amplitude of one fully ultrasonic chirp element (29.5 kHz center frequency) was 137 dB re 1 µPa-m. Harmonics of some vocalizations exceeded 200 kHz. Ultrasonic vocalizations occurred throughout the year with the usage of repetitive ultrasonic chirp-based calls appearing to dominate in winter darkness. The functional significance of these high-frequency vocalizations is unknown.

A second estrogen receptor from Japanese lamprey (Lethenteron japonicum) does not have activities for estrogen binding and transcription.

Estrogens regulate many physiological responses in vertebrates by binding to the estrogen receptor (ER), a ligand-activated transcription factor. To understand the evolution of vertebrate ERs and to investigate how estrogen acts in a jawless vertebrate, we used degenerate primer sets and PCR to isolate DNA fragments encoding two distinct ER subtypes, Esr1a and Esr1b from the Japanese lamprey, Lethenteron japonicum. Phylogenetic analysis indicates that these two ERs are the result of lineage-specific gene duplication within the jawless fishes, different from the previous duplication event of Esr1 (ERα) and Esr2 (ERß) within the jawed vertebrates. Reporter gene assays show that lamprey Esr1a displays both constitutive and estrogen-dependent activation of gene transcription. Domain swapping experiments indicate that constitutive activity resides in the A/B domain of lamprey Esr1a. Unexpectedly, lamprey Esr1b does not bind estradiol and is not stimulated by other estrogens, androgens or corticosteroids. A 3D model of lamprey Esr1b suggests that although estradiol fits into the steroid binding site, some stabilizing contacts between the ligand and side chains that are found in human Esr1 and Esr2 are missing in lamprey Esr1b.

Antifreeze protein-induced superheating of ice inside Antarctic notothenioid fishes inhibits melting during summer warming.

Antifreeze proteins (AFPs) of polar marine teleost fishes are widely recognized as an evolutionary innovation of vast adaptive value in that, by adsorbing to and inhibiting the growth of internalized environmental ice crystals, they prevent death by inoculative freezing. Paradoxically, systemic accumulation of AFP-stabilized ice could also be lethal. Whether or how fishes eliminate internal ice is unknown. To investigate if ice inside high-latitude Antarctic notothenioid fishes could melt seasonally, we measured its melting point and obtained a decadal temperature record from a shallow benthic fish habitat in McMurdo Sound, Antarctica. We found that AFP-stabilized ice resists melting at temperatures above the expected equilibrium freezing/melting point (eqFMP), both in vitro and in vivo. Superheated ice was directly observed in notothenioid serum samples and in solutions of purified AFPs, and ice was found to persist inside live fishes at temperatures more than 1 °C above their eqFMP for at least 24 h, and at a lower temperature for at least several days. Field experiments confirmed that superheated ice occurs naturally inside wild fishes. Over the long-term record (1999-2012), seawater temperature surpassed the fish eqFMP in most summers, but never exceeded the highest temperature at which ice persisted inside experimental fishes. Thus, because of the effects of AFP-induced melting inhibition, summer warming may not reliably eliminate internal ice. Our results expose a potentially antagonistic pleiotropic effect of AFPs: beneficial freezing avoidance is accompanied by melting inhibition that may contribute to lifelong accumulation of detrimental internal ice crystals.

Nonhepatic origin of notothenioid antifreeze reveals pancreatic synthesis as common mechanism in polar fish freezing avoidance.

Phylogenetically diverse polar and subpolar marine teleost fishes have evolved antifreeze proteins (AFPs) or antifreeze glycoproteins (AFGPs) to avoid inoculative freezing by internalized ice. For over three decades since the first fish antifreeze (AF) protein was discovered, many studies of teleost freezing avoidance showed hepatic AF synthesis and distribution within the circulation as pivotal in preventing the blood, and therefore the fish, from freezing. We have uncovered an important twist to this long-held paradigm: the complete absence of liver synthesis of AFGPs in any life stage of the Antarctic notothenioids, indicating that the liver plays no role in the freezing avoidance in these fishes. Instead, we found the exocrine pancreas to be the major site of AFGP synthesis and secretion in all life stages, and that pancreatic AFGPs enter the intestinal lumen via the pancreatic duct to prevent ingested ice from nucleating the hyposmotic intestinal fluids. AFGPs appear to remain undegraded in the intestinal milieu, and the composition and relative abundance of intestinal AFGP isoforms are nearly identical to serum AFGPs. Thus, the reabsorption of intact pancreas-derived intestinal AFGPs, and not the liver, is the likely source of circulatory AFGPs in notothenioid fishes. We examined diverse northern fish taxa and Antarctic eelpouts with hepatic synthesis of bloodborne AF and found that they also express secreted pancreatic AF of their respective types. The evolutionary convergence of this functional physiology underscores the hitherto largely unrecognized importance of intestinal freezing prevention in polar teleost freezing avoidance, especially in the chronically icy Antarctic waters.

Freezing resistance of antifreeze-deficient larval Antarctic fish.

Antarctic notothenioids, along with many other polar marine fishes, have evolved biological antifreeze proteins (AFPs) to survive in their icy environments. The larvae of Antarctic notothenioid fish hatch into the same frigid environment inhabited by the adults, suggesting that they must also be protected by sufficient AFPs, but this has never been verified. We have determined the contribution of AFPs to the freezing resistance of the larvae of three species: Gymnodraco acuticeps, Pagothenia borchgrevinki and Pleuragramma antarcticum. Of the three, only P. borchgrevinki larvae are protected by high, adult levels of AFPs. Hatchling G. acuticeps and P. antarcticum have drastically inadequate AFP concentrations to avoid freezing at the ambient seawater temperature (-1.91 degrees C). We raised G. acuticeps larvae and measured the AFP levels in their blood for approximately 5 months post hatching. Larval serum freezing point was -1.34+/-0.04 degrees C at the time of hatch; it began to decrease only after 30 days post hatch (d.p.h.), and finally reached the adult value (-2.61+/-0.03 degrees C) by 147 d.p.h. Additionally, AFP concentrations in their intestinal fluids were very low at hatching, and did not increase with age throughout a sampling period of 84 d.p.h. Surviving in a freezing environment without adequate AFP protection suggests that other mechanisms of larval freezing resistance exist. Accordingly, we found that G. acuticeps hatchlings survived to -3.6+/-0.1 degrees C while in contact with external ice, but only survived to -1.5+/-0.0 degrees C when ice was artificially introduced into their tissues. P. antarcticum larvae were similarly resistant to organismal freezing. The gills of all three species were found to be underdeveloped at the time of hatch, minimizing the risk of ice introduction through these delicate structures. Thus, an intact integument, underdeveloped gill structures and other physical barriers to ice propagation may contribute significantly to the freezing resistance and survival of these larval fishes in the icy conditions of the Southern Ocean.