Hydrophobicity patterns and biological adaptation: an exemplary case from fish hemoglobins.

The dissection of phylogenetic and environmental components in biological evolution is one of the main themes of general biology. Here we propose an approach to this theme relying upon the comparison between a phylogenetic oriented metrics spanning the hemoglobin beta chains of different fishes and a more physiologically oriented metrics defining the same sequences in terms of the dynamical features of their hydrophobic distributions. By analyzing the set of sequences more similar to the Gadus morhua (Atlantic cod) hemoglobin beta chain, we were able to give a proof of concept of the possibility to discriminate the phylogenetic and environmental (evolutive convergence) components by the comparative analysis of the Clustal W (phylogenetics first) and Recurrence Quantification Analysis (physiology first) metrics in which the sequences were embedded. The use of a molecular system like hemoglobin playing a crucial role in fishes adaptation to environmental cues allowed us to span different levels of biological variability by means of the same paradigm. Starting from the reconstruction of the general taxonomy of vertebrate groups we went down to the exploitation of the peculiar role played by Met55Val and Lys62Ala polymorphisms in the beta1 hemoglobin chain of the Atlantic cod able to influence the geographical distribution of its various stocks.

Whether depositing fat or losing weight, fish maintain a balance.

In fish, the relative amount of tissues of different densities changes significantly over short periods throughout the year, depending on the availability of food, nutrition and their developmental status, such as sexual maturation. If a land-living animal accumulates fat it influences not only its general state of health, but also markedly increases its energy expenditure for locomotion owing to the force of gravity. On a body submerged in water, this force, which acts on the centre of gravity (COG), is counterbalanced by a lifting force that is negligible in air and which acts on the centre of buoyancy (COB). Any difference in the longitudinal positions of the two centres will therefore result in pitching moments that must be counteracted by body or fin movements. The displacement of the COG away from the COB is a result of tissues of different density (e.g. bones and fat) not being distributed homogeneously along the body axis. Moreover, the proportions of tissues of different densities change significantly with feeding status. It is still unknown whether these changes produce a displacement of the COG and thus affect the hydrostatic stability of fish. Analysis of computed tomography and magnetic resonance imaging images of Atlantic herring, Atlantic salmon and Atlantic mackerel reveals that the COG is fairly constant in each species, although we recorded major interspecies differences in the relative amount of fat, muscle and bone. We conclude that the distribution of different tissues along the body axis is very closely adjusted to the swimming mode of the fish by keeping the COG constant, independent of the body fat status, and that fish can cope with large variations in energy intake without jeopardizing their COG and thus their swimming performance.

Haemoglobin polymorphisms affect the oxygen-binding properties in Atlantic cod populations.

A major challenge in evolutionary biology is to identify the genes underlying adaptation. The oxygen-transporting haemoglobins directly link external conditions with metabolic needs and therefore represent a unique system for studying environmental effects on molecular evolution. We have discovered two haemoglobin polymorphisms in Atlantic cod populations inhabiting varying temperature and oxygen regimes in the North Atlantic. Three-dimensional modelling of the tetrameric haemoglobin structure demonstrated that the two amino acid replacements Met55beta1Val and Lys62beta1Ala are located at crucial positions of the alpha1beta1 subunit interface and haem pocket, respectively. The replacements are proposed to affect the oxygen-binding properties by modifying the haemoglobin quaternary structure and electrostatic feature. Intriguingly, the same molecular mechanism for facilitating oxygen binding is found in avian species adapted to high altitudes, illustrating convergent evolution in water- and air-breathing vertebrates to reduction in environmental oxygen availability. Cod populations inhabiting the cold Arctic waters and the low-oxygen Baltic Sea seem well adapted to these conditions by possessing the high oxygen affinity Val55-Ala62 haplotype, while the temperature-insensitive Met55-Lys62 haplotype predominates in the southern populations. The distinct distributions of the functionally different haemoglobin variants indicate that the present biogeography of this ecologically and economically important species might be seriously affected by global warming.

Temperature acclimation modulates the oxygen binding properties of the Atlantic cod (Gadus morhua L.) genotypes-HbI*1/1, HbI*1/2, and HbI*2/2-by changing the concentrations of their major hemoglobin components (results from growth studies at different temperatures).

The influence of long-term acclimation temperatures in Atlantic cod (Gadus morhua) was studied by growth experiments carried out over a total of 272 individuals. The attention focused on the structural and functional modulation of the five electrophoretically distinguishable genotypes of cod hemoglobin (HbI*1/1, HbI*1/2, HbI*2/2, HbI*1/2b, and HbI*2/2b) and on the correlation with body length/weight. The main results can be summarized as follows. (1) Acclimation to lower (4 and 8 degrees C) and higher (12 and 15 degrees C) temperatures favors the expression of, respectively, more anodic and more cathodic hemoglobin components. (2) The optimal O(2) transporting features are observed at 12 degrees C, as well as a saturation-dependent temperature dependence of O(2) binding, which furthermore is strongly dependent upon the acclimation background. (3) The optimal growth condition for the three main genotypes (HbI*1/1, HbI*1/2, and HbI*2/2) is associated with T=12 degrees C. The overall results are consistent with the idea that environmental temperatures constitute a primary factor in the aggregation of individuals physiologically more than genetically homogeneous. This is fully confirmed by careful statistical analysis carried out over a subset of individuals for which the full set of structural (isoelectric focusing), functional (O(2) binding), and growth data was available.