Biomedical subjects

B D Sidell

Publications and source records attributed to B D Sidell.

At least 19 recordsLinked to original sources

PubMed2004

The substrate specificity of hormone-sensitive lipase from adipose tissue of the Antarctic fish Trematomus newnesi.

Antarctic fishes of the suborder Notothenioidei characteristically possess large stores of neutral lipids that have been shown to be important both in conferring buoyant lift and as a caloric resource for energy metabolism. Previous work has established that the aerobic energy metabolism of Antarctic fish is fueled predominantly by the catabolism of fatty acids, with the catabolic machinery displaying a preference for the oxidation of unsaturated fatty acids. The composition of the fatty acids released from adipose tissue of Antarctic fish during lipolysis, however, has not previously been demonstrated. Employing a substrate competition assay, we have characterized the substrate specificity of hormone-sensitive lipase (HSL) from adipose tissue of the Antarctic fish Trematomus newnesi. Rates of oleic acid release from radiolabeled triolein were quantified in the presence and absence of a nonradiolabeled cosubstrate. Polyunsaturated species of triacylglycerols (TAGs) containing 18:2 or 20:4 depressed rates of oleate release by 70-75% below control values. Most of the molecular species of TAG containing monoenoic fatty acids (i.e. those containing 14:1, 16:1 or 20:1) had no significant effect on rates of oleate release. By contrast, oleate release from triolein was actually stimulated (by 2-4-fold) by both saturated species of TAG (i.e. those containing 14:0, 16:0 and 18:0) and those possessing long-chain (22:1 and 24:1) monoenes (by 1.2-1.5-fold). Thus, the rank order of substrate preference for adipose tissue HSL was: polyunsaturates > monoenes > saturates. Degree of fatty acid unsaturation had a more marked effect on rates of hydrolysis than did fatty acid chain length. In addition, the enzyme displayed a preference for the hydrolysis of sn-1,2 rather than sn-1,3 diacylglycerols. These data indicate that the substrate specificity of adipose tissue HSL may be an important factor in determining which fatty acids are mobilized during stimulated lipolysis and which are made available for catabolism by other tissues of Antarctic fishes. Our data further suggest that TAGs containing some saturated fatty acids may be sufficiently poor substrates for catabolism by HSL to explain their disproportionate accumulation in adipose tissue. Such a mechanism could also contribute to the ontogenetic accumulation of fats that has been reported as an underlying basis for the positive correlation of buoyancy with increasing body mass in this group.

Adipose Tissue↗

PubMed2003

Muscle fine structure may maintain the function of oxidative fibres in haemoglobinless Antarctic fishes.

Muscle fine structure and metabolism were examined in four species of Antarctic fishes that vary in their expression of haemoglobin (Hb). To determine how locomotory pectoral muscles maintain function, metabolic capacity, capillary supply and fibre ultrastructure were examined in two nototheniid species that express Hb (Notothenia coriiceps and Gobionotothen gibberifrons) and two species of channichthyid icefish that lack Hb (Chaenocephalus aceratus and Chionodraco rastrospinosus). Surprisingly, icefish have higher densities of mitochondria than red-blooded species (C. aceratus, 53+/-3% of cell volume; C. rastrospinosus, 39+/-3%; N. coriiceps, 29+/-3%; G. gibberifrons, 25+/-1%). Despite higher mitochondrial densities the aerobic metabolic capacities per g wet mass, estimated from measurements of maximal activities of key metabolic enzymes, are lower in icefish compared to red-blooded species. This apparent incongruity can be explained by the significantly lower mitochondrial cristae surface area per unit mitochondrion volume in icefishes (C. aceratus, 20.8+/-1.6 microm(-1); C. rastrospinosus, 25.5+/-1.8 microm(-1)) compared to red-blooded species (N. coriiceps, 33.6+/-3.0 microm(-1); G. gibberifrons, 37.7+/-3.6 microm(-1)). Consequently, the cristae surface area per unit muscle mass is conserved at approximately 9 m(2)g(-1). Although high mitochondrial densities in icefish muscle do not enhance aerobic metabolic capacity, they may facilitate intracellular oxygen movement because oxygen is more soluble in lipid, including the hydrocarbon core of intracellular membrane systems, than in aqueous cytoplasm. This may be particularly vital in icefish, which have larger oxidative muscle fibres compared to red-blooded nototheniods (C. aceratus, 2932+/-428 microm(2); C. rastrospinosus, 9352+/-318 microm(2); N. coriiceps, 1843+/-312 microm(2); G. gibberifrons, 2103+/-194 microm(2)). These large fibres contribute to a relatively low capillary density, which is partially compensated for in icefish by a high index of tortuosity in the capillary bed (C. aceratus=1.4, N. coriiceps=1.1).

Aerobiosis↗

PubMed2000

The estimation of the diffusion constant and solubility of O(2)in tissue using kinetics.

The diffusion of a gas through a substance in which it is soluble is analogous to the passage of electric current through a circuit with both capacitance and resistance. We model steady-state diffusion employing this analogy, and extend the model to include a description of the kinetics of systems under circumstances of changing partial pressure, applying two physical constants from electrical circuitry to gas diffusion: capacitance (zeta) and resistance (R). We represent the substrate of the diffusion as a capacitor being charged through a resistor after the rapid imposition of a voltage change. Using the insight derived from this model we have devised an experimental system that allows us to approximate both D, the diffusion coefficient, and alpha, solubility, directly from the kinetic data. We do this by recording the exponential change in P(O(2))on one side of a sheet of material both with and without the addition of a purely resistive barrier of known resistivity. The method was used to estimate D and alpha for distilled water at a number of temperatures, olive oil, and the belly skin of Rana catesbeiana.

Animals↗

PubMed2000

Quantification of diffusion distance within the spongy myocardium of hearts from antarctic fishes.

We developed a stereological method for quantifying diffusion distance within spongy myocardium. Using this method we compared the hearts of three species of Antarctic fishes that vary in expression of oxygen-binding proteins. We examined hearts from Gobionotothen gibberifrons, a red-blooded species whose ventricle has myoglobin (Mb), and hearts of two species of icefish that lack hemoglobin (Hb) and vary in expression of cardiac Mb; Chionodraco rastrospinosus expresses Mb, Chaenocephalus aceratus does not. Average diffusion distance within ventricular tissue is greater in red-blooded Antarctic teleosts (9.82 + or - 1.37 microm) compared with icefish (C. rastrospinosus, 6.20 microm + or - 0.86; C. aceratus, 6.23 + or - 0.41 microm). Average diffusion distance to a mitochondrion parallels this trend because mitochondria are uniformly distributed within cardiac muscle. Results show that loss of Hb is correlated with increased trabeculation of heart ventricle. Loss of Mb however, is not correlated with an increase in trabeculation of ventricular tissue, despite significant differences in cellular ultrastructure compared with species that express the protein.

Animals↗

PubMed2000

Concentrations of myoglobin and myoglobin mRNA in heart ventricles from Antarctic fishes.

We used a combined immunochemical and molecular approach to ascertain the presence and concentrations of both the intracellular oxygen-binding hemoprotein myoglobin (Mb) and its messenger RNA (mRNA) in 13 of 15 known species of Antarctic channichthyid icefishes. Mb protein is present in the hearts of eight species of icefishes: Chionodraco rastrospinosus, Chionodraco hamatus, Chionodraco myersi, Chaenodraco wilsoni, Pseudochaenichthys georgianus, Cryodraco antarcticus, Chionobathyscus dewitti and Neopagetopsis ionah. Five icefish species lack detectable Mb protein: Chaenocephalus aceratus, Pagetopsis macropterus, Pagetopsis maculatus, Champsocephalus gunnari and Dacodraco hunteri. Mb concentrations range from 0.44+/-0.02 to 0.71+/-0.08 mg Mb g(-)(1 )wet mass in heart ventricle of species expressing the protein. A Mb-mRNA-specific cDNA probe was used to quantify mRNA in five Mb-expressing icefishes. Mb mRNA was found in low but detectable amounts in Champsocephalus gunnari, one of the species lacking detectable Mb. Mb mRNA concentrations in heart ventricle from Mb-expressing species ranged from 0.78+/-0.02 to 16.22+/-2.17 pg Mb mRNA microg(-)(1 )total RNA). Mb protein and Mb mRNA are absent from the oxidative skeletal muscle of all icefishes. Steady-state concentrations of Mb protein do not parallel steady-state concentrations of Mb mRNA within and among icefishes, indicating that the concentration of Mb protein is not determined by the size of its mRNA pool.

Animals↗

PubMed2000

The interplay among cardiac ultrastructure, metabolism and the expression of oxygen-binding proteins in Antarctic fishes.

We examined heart ventricle from three species of Antarctic fishes that vary in their expression of oxygen-binding proteins to investigate how some of these fishes maintain cardiac function despite the loss of hemoglobin (Hb) and/or myoglobin (Mb). We quantified ultrastructural features and enzymatic indices of metabolic capacity in cardiac muscle from Gobionotothen gibberifrons, which expresses both Hb and Mb, Chionodraco rastrospinosus, which lacks Hb but expresses Mb, and Chaenocephalus aceratus, which lacks both Hb and Mb. The most striking difference in cellular architecture of the heart among these species is the percentage of cell volume occupied by mitochondria, V(v)(mit,f), which is greatest in Chaenocephalus aceratus (36.53+/-2.07), intermediate in Chionodraco rastrospinosus (20.10+/-0.74) and lowest in G. gibberifrons (15.87+/-0.74). There are also differences in mitochondrial morphologies among the three species. The surface area of inner mitochondrial membrane per volume of mitochondria, S(v)(imm, mit), varies inversely with mitochondrial volume density so that S(v)(imm,mit) is greatest in G. gibberifrons (29.63+/-1.62 microm(-)(1)), lower in Chionodraco rastrospinosus (21.52+/-0.69 microm(-)(1)) and smallest in Chaenocephalus aceratus (20.04+/-0.79 microm(-)(1)). The surface area of mitochondrial cristae per gram of tissue, however, is greater in Chaenocephalus aceratus than in G. gibberifrons and Chionodraco rastrospinosus, whose surface areas are similar. Despite significant ultrastructural differences, oxidative capacities, estimated from measurements of maximal activities per gram of tissue of enzymes from aerobic metabolic pathways, are similar among the three species. The combination of ultrastructural and enzymatic data indicates that there are differences in the density of electron transport chain proteins within the inner mitochondrial membrane; proteins are less densely packed within the cristae of hearts from Chaenocephalus aceratus than in the other two species. High mitochondrial densities within hearts from species that lack oxygen-binding proteins may help maintain oxygen flux by decreasing the diffusion distance between the ventricular lumen and mitochondrial membrane. Also, high mitochondrial densities result in a high intracellular lipid content, which may enhance oxygen diffusion because of the higher solubility of oxygen in lipid compared with cytoplasm. These results indicate that features of cardiac myocyte architecture in species lacking oxygen-binding proteins may maintain oxygen flux, ensuring that aerobic metabolic capacity is not diminished and that cardiac function is maintained.

Animals↗

PubMed2000

Life at body temperatures below 0 degrees C: the physiology and biochemistry of Antarctic fishes.

Fishes of the Southern Ocean surrounding Antarctica are dominated by species of the suborder Notothenoidei. For approximately 14MY, these highly successful fishes have evolved under stable thermal conditions that result in a body temperature of ca. 0 degrees C throughout their life histories. Evolution in this chronically cold environment has led to unusual physiological and biochemical characteristics. In some cases, these characteristics are essential to survival and normal biological function at cold body temperature (e.g., development of antifreeze glycoproteins, structural modification of enzymes, cold-stable microtubules, and cardiovascular adaptations). In other instances, mutations that probably would have been lethal in warmer, less oxygen-rich environments than the Southern Ocean have been retained in Antarctic fishes (e.g., loss of hemoglobin production and variable expression of myoglobin in one notothenioid family, the Channichthyidae). These unique animals offer opportunities for insight into evolutionary processes leading to physiological and biochemical characteristics that either arise from strong selective pressure or persist through relaxation of selective pressure. After briefly describing the Antarctic marine environment, I discuss several unique aspects of the physiology and biochemistry of Antarctic fishes, specifically emphasizing our laboratory's recent studies of an unusual pattern of myoglobin expression in the Channichthyid icefishes.

Adaptation, Biological↗

PubMed1998

Two distinct types of fatty acid-binding protein are expressed in heart ventricle of Antarctic teleost fishes.

This report provides the first evidence for the existence of two distinct types of fatty acid-binding protein (FABP) in cardiac tissue of vertebrates. Four species of Antarctic teleost fish (Chaenocephalus aceratus, Cryodraco antarcticus, Gobionotothen gibberifrons and Notothenia coriiceps) exhibited two FABP mRNAs of 1. 0 kb and 0.8 kb, which we have termed Hh-FABP and Had-FABP (isolated from Heart tissue, with similarity to mammalian heart-type FABP or mammalian adipose-type FABP respectively). These FABP types appear to be products of distinct genes. Both FABP transcripts were abundant in cardiac and aerobic pectoral muscle. However, relative abundance of the two types varied distinctly among other tissues such as kidney, brain, spleen and white muscle. Neither FABP type was expressed in liver or intestine. The coding regions of Hh-FABP and Had-FABP cDNAs from the same species are only approximately 60% identical with one another. However, homologues of each FABP species, which exhibit >98% identity to their respective types, were isolated from three other Antarctic teleosts. Phylogenetic analysis of aligned amino-acid sequences places Hh-FABP with other vertebrate heart-type FABPs, and Had with adipose/cutaneous FABPs. Expression of two distinct FABPs in cardiac tissue of Antarctic teleosts may be related to their ability to both utilize fatty acid as the primary metabolic fuel and to store lipid intracellularly.

Amino Acid Sequence↗

PubMed1998

A novel vertebrate myoglobin gene containing three A+T-rich introns is conserved among Antarctic teleost species which differ in myoglobin expression.

This report presents the first teleost myoglobin (Mb) genomic DNA sequence, several features of which are distinct from mammalian Mb genes. We have isolated and compared genomic clones of three closely related Antarctic icefish: Chionodraco rastrospinosus, which expresses Mb mRNA and functional polypeptide; Champsocephalus gunnari, which transcribes the Mb gene but does not produce polypeptide due to a 5-base pair (bp) frameshift insertion; and Chaenocephalus aceratus, which lacks both Mb protein and mRNA. The single-copy icefish Mb gene contains three introns: two at positions identical to those found in mammalian Mb and a novel intron located in the 5' untranslated region three nucleotides upstream from the initiator codon. All three introns are shorter than those found in mammalian Mb genes and exhibit a considerably higher A+T content. The entire Mb transcriptional unit is intact in C. aceratus, indicating that the failure to express this gene is not due to aberrations in the coding region, splice junctions, polyadenylation signals, or core promoter elements. The three icefish Mb sequences display an extreme degree of identity in the transcriptional unit and putative promoter region. In contrast, sequences 65 bp downstream from the polyadenylation site bear no homology among the three species, demonstrating that rapid sequence change has occurred in the 1 million years since the divergence of these species.

Amino Acid Sequence↗

PubMed1998

Intracellular oxygen diffusion: the roles of myoglobin and lipid at cold body temperature.

Cold temperature can constrain the rate of oxygen movement through muscle cells of ectothermic animals because the kinetic energy of the solvent-solute system decreases and the viscosity of the aqueous cytoplasm increases during cooling within the physiological range of body temperatures. These factors affect the movement of both dissolved oxygen and oxymyoglobin, the two predominant routes of intracellular oxygen diffusion in vertebrate oxidative muscles. In addition, reductions in temperature have been shown to increase the affinity of myoglobin for oxygen and to slow the rate of Mb O2-dissociation, compromising the ability of this oxygen-binding protein to facilitate intracellular oxygen diffusion. Experiments with both seasonally cold-bodied fishes and polar fish species suggest that several factors combine to overcome these limitations in delivery of oxygen from the blood to the mitochondria. First, reductions in body temperature induce increases in mitochondrial density of oxidative muscle cells, reducing the mean diffusional pathlength for oxygen between capillaries and mitochondria. Second, cold body temperature in both temperate-zone and polar fishes is frequently correlated with a high content of neutral lipid in oxidative muscles, providing an enhanced diffusional pathway for oxygen through the tissue. Third, recent data indicate that myoglobins from fish species bind and release oxygen more rapidly at cold temperature than do those from mammals. Data from both oxidative skeletal muscle and cardiac muscle of fishes suggest that these factors in various combinations contribute to enhance the aerobically supported mechanical performance of the tissues at cold cellular temperatures.

Animals↗

PubMed1997

Variable expression of myoglobin among the hemoglobinless Antarctic icefishes.

The important intracellular oxygen-binding protein, myoglobin (Mb), is thought to be absent from oxidative muscle tissues of the family of hemoglobinless Antarctic icefishes, Channichthyidae. Within this family of fishes, which is endemic to the Southern Ocean surrounding Antarctica, there exist 15 known species and 11 genera. To date, we have examined eight species of icefish (representing seven genera) using immunoblot analyses. Results indicate that Mb is present in heart ventricles from five of these species of icefish. Mb is absent from heart auricle and oxidative skeletal muscle of all species. We have identified a 0.9-kb mRNA in Mb-expressing species that hybridizes with a Mb cDNA probe from the closely related red-blooded Antarctic nototheniid fish, Notothenia coriiceps. In confirmation that the 0.9-kb mRNA encodes Mb, we report the full-length Mb cDNA sequence of the ocellated icefish, Chionodraco rastrospinosus. Of the eight icefish species examined, three lack Mb polypeptide in heart ventricle, although one of these expresses the Mb mRNA. All species of icefish retain the Mb gene in their genomic DNA. Based on phylogeny of the icefishes, loss of Mb expression has occurred independently at least three times and by at least two distinct molecular mechanisms during speciation of the family.

Amino Acid Sequence↗

PubMed1997

Kinetic characterization of myoglobins from vertebrates with vastly different body temperatures.

Fish myoglobins are structurally distinct from the previously characterized mammalian myoglobins. Teleost fishes express generally lower levels of myoglobin than those found in mammals. Although the oxygen binding affinity is essentially the same as mammalian myoglobins, oxygen dissociation rates and carbon monoxide combination rates of the teleost myoglobins studied are significantly faster. Thus, the kinetic parameters of myoglobin from two Antarctic teleost species, measured close to their body temperature of -1 degree C, are comparable to those of mammalian myoglobins with higher body temperatures. These data suggest myoglobins from Antarctic teleosts may function at extreme environmental temperatures.

Animals↗

PubMed1997

Myoglobin enhances cardiac performance in antarctic icefish species that express the protein.

Channichthyid icefishes of Antarctica are unique among adult vertebrates. All icefish species lack hemoglobin and red blood cells in their circulating blood. All icefishes examined to date also lack the intracellular oxygen-binding protein myoglobin (Mb) in their oxidative skeletal muscles. However, some icefish species do express Mb in their heart ventricles. It is unknown whether Mb in those species in which it is present represents an evolutionary relic or has functional significance. To address this problem, we compared mechanical performance of isolated, perfused hearts from two species of icefish in which Mb is either present (Chionodraco rastrospinosus) or is absent (Chaenocephalus aceratus). Hearts were challenged with increasing afterload (2.5-4.0 kPa) under conditions of defined basal flow (approximately 100 ml.min-1.kg-1), in both the presence and absence of 5 mM sodium nitrite, a Mb poison. Unlike hearts from C. aceratus, which were unable to maintain a constant cardiac output under pressure loading, those from C. rastrospinosus retained a constant flow up to 3.5 kPa afterload. At the upper range of power outputs, hearts of Mb-lacking C. aceratus display greater oxygen utilization than those of Mb-containing C. rastrospinosus. Poisoning of Mb significantly impaired the ability of C. rastrospinosus hearts to face pressure loading without reduction in flow, whereas those of C. aceratus were refractory to the treatment. The results strongly support a functional role for Mb in the former species.

Animals↗

PubMed1997

Structural and biochemical analyses of cardiac ventricular enlargement in cold-acclimated striped bass.

We examined effects of temperature acclimation on ultrastructural characteristics of cardiac myocytes and maximal activities of metabolic enzymes in cardiac tissue of striped bass (Morone saxatilis). Ventricular mass and ventricular mass divided by body weight were significantly increased (29% and 40%, respectively) in animals acclimated to cold (5 degrees C) vs. warm temperatures (25 degrees C). Mean myocyte diameter was increased at cold temperature (3.47 +/- 0.14 vs. 2.98 +/- 0.08 microns), which is sufficient to explain the increase in ventricular mass. Ventricular enlargement did not alter volume densities of mitochondria, myofibrils, protein concentration, or citrate synthase activity. Thus total volume of mitochondria and myofibrils increased proportionately with cardiac mass in cold animals. Activities of hexokinase (34%) and carnitine palmitoyltransferase (42%) increased in cold animals, suggesting positive compensation and increased aerobic capacity for utilization of glucose and fatty acids for energy production. Enlargement of the ventricle and an increased capacity for ATP production in striped bass may help compensate for kinetic constraints at cold temperatures and maintain circulatory support to oxidative axial musculature for swimming activity.

Acclimatization↗

PubMed1997

Conservation of the myoglobin gene among Antarctic notothenioid fishes.

We determined the myoglobin cDNA sequence for seven Antarctic notothenioid fish species. These data identify mutations in the myoglobin gene for Champsocephalus gunnari and Pagetopsis macropterus, two icefish species that lack detectable quantities of the polypeptide but express myoglobin mRNA. a third species lacking myoglobin polypeptide, Chaenocephalus aceratus, is devoid of myoglobin mRNA and accordingly failed to produce myoglobin products on polymerase chain reaction (PCR) amplification. Myoglobin cDNA sequences were highly conserved among the species the express the protein, particularly in the coding region. Sequence variation among the myoglobin-expressing channichthyid species was 2.0% to 2.9% in the coding region and 2.6% to 3.3% over the entire cDNA. The same extent of variation, 1.6% to 3.2% in the coding sequence and 2.8% to 3.7% overall, was observed between the icefishes and more distantly related, red-blooded nototheniid species. The two species expressing mutant myoglobin mRNA, C. gunnari and P. macropterus, exhibited the highest degree of sequence variation among the fish myoglobins examined. Drift in the myoglobin sequence in these two species, and conservation of myoglobin cDNA among fishes from two distinct families, suggest that a selective pressure operates to maintain myoglobin in the species that express the protein.

Amino Acid Sequence↗

PubMed1996

Binding site polarity and ligand affinity of homologous fatty acid-binding proteins from animals with different body temperatures.

Binding affinity and binding-pocket polarity is determined for intracellular fatty acid-binding protein (FABP) from aerobic muscle of Chaenocephalus aceratus, the Antarctic icefish, and from rat heart. FABPs bind fatty acids via weak-bond forces (both ionic and hydrophobic), and these bond forces are temperature sensitive, yet FABPs are present in animals whose body temperatures range over nearly 40 degrees C. To investigate FABP's sensitivity to body temperature, fatty acid binding affinity (Kd) was determined for both rat heart-FABP and icefish heart-FABP at two physiological temperatures (0 degrees C or 37 degrees C). Saturated and unsaturated fatty acids (16:0 and 16:1), delivered in model membranes (liposomes) whose composition is typical of either Antarctic fish (16:0/22:6 phosphatidylcholine) or mammals (bovine-heart phosphatidylcholine) were examined. Incubation at 0 degree C or 37 degrees C dose not significantly affect Kd for rat heart FABP, regardless of liposome composition or fatty acid ligand (Kd = 0.686 +/- 0.127 - 1.129 +/- 0.356 microM at 0 degree C, 0.775 +/- 0.307 - 1.605 +/- 0.427 microM at 37 degrees C). Incubation temperature significantly affects icefish FABP's affinity for 16:1 (0.626 +/- 0.093 microM at 37 degrees C vs. 1.896 +/- 0.343 microM at 0 degree C for fatty acid presented in Antarctic fish liposomes; 0.331 +/- 0.101 microM at 37 degrees C vs. 0.949 +/- 0.121 microM at 0 degree C for bovine heart liposomes) but not 16:0. Kd is not significantly different between FABPs under any set of conditions (with one exception: Kd is significantly lower in rat FABP vs. icefish FABP for 16:0 at 0 degree C for fatty acids delivered in bovine heart liposomes). Although Kd values are largely equivalent between the two FABPs, relative contributions from ionic vs. hydrophobic weak-bond forces are different between the two animals. Rat heart FABP has a binding pocket that is significantly more nonpolar than that of icefish FABP (as measured by quantum yield of the bound fluorescent fatty-acid analogue (PA-DPH); Q = 0.067 +/- 0.008 vs. 0.034 +/- 0.005 at 0 degree C, 0.030 +/- 0.003 vs. 0.019 +/- 0.002 at 37 degrees C). This suggests that rat-heart FABP realizes a micromolar Kd with a greater reliance upon hydrophobic interactions than does icefish FABP.

Animals↗

PubMed1996

High lipid content enhances the rate of oxygen diffusion through fish skeletal muscle.

The diffusion coefficient for O2 (Do2) and the solubility constant for O2 (alpha O2) were measured at 15 degrees C in oxidative muscle from striped bass (Morone saxatilis) that had been acclimated to 5 degrees and 25 degrees C. This design allowed us to test the hypothesis that changes in composition of the tissue that are known to occur during thermal acclimation may affect O2 movement. Our measurements permitted calculation of the diffusion constant for O2 (Ko2) through the tissue, which is a primary determinant of capacity for O2 flux. Under isothermal conditions, alpha O2 was 3.59 +/- 0.20 x 10(-2) and 6.64 +/- 0.27 x 10(-2) ml O2.cm-3.atm-1 in tissues from 25 degrees- and 5 degrees C-acclimated animals, respectively. Because O2 is more soluble in lipid than aqueous phase, higher alpha O2 in tissues from cold-acclimated animals can be accounted for by the 13-fold increase in lipid content that is known to occur in oxidative muscle of striped bass during acclimation from 25 degrees to 5 degrees C. When measured under similar isothermal conditions, Do2 showed no significant difference between animals acclimated to warm or cold temperature; Do2 through tissues from 25 degrees- and 5 degrees C-acclimated animals was 2.50 +/- 0.18 and 2.57 +/- 0.40 cm2/s, respectively. Because alpha O2 increases, the calculated KO2 (DO2. alpha O2) is greater in tissue from cold- than from warm-acclimated fish. At physiological temperature, elevated lipid content in oxidative muscle of cold-acclimated striped bass should result in enhanced intracellular movement of O2 and at least partially offset the expected decrease in DO2 at cold temperature.

Acclimatization↗

PubMed1995

Purification and characterization of fatty acid-binding protein from aerobic muscle of the Antarctic icefish Chaenocephalus aceratus.

Intracellular fatty acid-binding protein is purified and characterized from aerobic skeletal muscle of the Antarctic icefish Chaenocephalus aceratus. Molecular mass of C. aceratus FABP (CA-FABP) is 14,936 Da as estimated by electrospray mass spectrometry. CA-FABP is expressed at an intracellular concentration of 0.984 +/- 0.115 mg CA-FABP g-1 wet weight aerobic muscle and binds 0.859 +/- 0.013 moles oleic acid per mole of protein at a physiological temperature of 0 degrees C. Dissociation constants (KdS for various fatty acid ligands range from 1.38 to 2.71 microM; KdS are not significantly different among palmitic acid (16:0), palmitoleic acid (16:1), and oleic acid (18:1). Competition assays reveal that CA-FABP does not have preferential affinity for the very-long-chain, polyunsaturated fatty acids that are common in Antarctic fish (e.g., docosahexaenoic acid; 22:6). Partial amino acid sequence from CA-FABP aligns with mammalian heart-type FABPs with as high as 74% identity. These data are strikingly similar to mammalian values, yet they are derived from an organism that is distant from mammals in terms of phylogeny, body temperature, and physiology. This suggests that the FABP family is conserved not only in primary sequence, but also in its physiological properties.

Amino Acid Sequence↗