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Enzyme activities support the use of liver lipid-derived ketone bodies as aerobic fuels in muscle tissues of active sharks.

Few data exist to test the hypothesis that elasmobranchs utilize ketone bodies rather than fatty acids for aerobic metabolism in muscle, especially in continuously swimming, pelagic sharks, which are expected to be more reliant on lipid fuel stores during periods between feeding bouts and due to their high aerobic metabolic rates. Therefore, to provide support for this hypothesis, biochemical indices of lipid metabolism were measured in the slow-twitch, oxidative (red) myotomal muscle, heart, and liver of several active shark species, including the endothermic shortfin mako, Isurus oxyrinchus. Tissues were assayed spectrophotometrically for indicator enzymes of fatty acid oxidation (3-hydroxy-o-acyl-CoA dehydrogenase), ketone-body catabolism (3-oxoacid-CoA transferase), and ketogenesis (hydroxy-methylglutaryl-CoA synthase). Red muscle and heart had high capacities for ketone utilization, low capacities for fatty acid oxidation, and undetectable levels of ketogenic enzymes. Liver demonstrated undetectable activities of ketone catabolic enzymes but high capacities for fatty acid oxidation and ketogenesis. Serum concentrations of the ketone beta-hydroxybutyrate varied interspecifically (means of 0.128-0.978 micromol mL(-1)) but were higher than levels previously reported for teleosts. These results are consistent with the hypothesis that aerobic metabolism in muscle tissue of active sharks utilizes ketone bodies, and not fatty acids, derived from liver lipid stores.

Aerobiosis↗

Some biological actions of alkylglycerols from shark liver oil.

Shark liver oil has been used for over 40 years as both a therapeutic and preventive agent. The active ingredients in shark liver oil have been found to be a group of ether-linked glycerols known as alkylglycerols. Initial clinical use was for treating leukemias, and later to prevent radiation sickness from cancer x-ray therapy. Studies over the last 30 years have shown that alkylglycerols are multifunctional. The level of natural alkylglycerols rises within tumor cells, apparently in an effort to control cell growth. Recent studies indicate that the activation of protein kinase C, an essential step in cell proliferation, can be inhibited by alkylglycerols. This action suggests a competitive inhibition of 1.2-diacylglycerol by alkylglycerols. Further studies on the immunostimulatory action of alkylglycerols suggest a primary action on the macrophage. The process of macrophage activation has been demonstrated with both synthetic and natural alkylglycerols. While the exact mechanism has not been found, both an autocrine and paracrine system have been suggested. Shark liver is a major natural source of alkylglycerols, which have no known side effects in dosages of 100 mg three times a day. The information presented in this article suggests that alkylglycerols may be used both as an adjunct therapy in the treatment of neoplastic disorders and as an immune booster in infectious diseases.

Animals↗

Inhibitory effect of shark serum on the Lewis lung carcinoma.

The growth and metastasis of transplanted Lewis lung carcinoma in C57BL/6J mice were inhibited by an ip injection of whole serum from Carcharhinus plumbeus, the sandbar shark. Tumors failed to develop in 69% of the animals inoculated with shark serum on days 0, 3, and 6 after tumor transplantation. Histologic examination of the tumor site at days 3 and 6 showed that tumor cells were pyknotic, and evidence of lysis of tumor cells and minor leukocytic infiltration existed. Tumor cells were not in tissue sections from day 15, and these animals still had no symptoms at day 216. The mean tumor volume of the remaining 31% of the treated animals was less than that of controls; they had a prolonged mean survival time, but ultimately they died from metastases, as did the controls. Urea- and hemoglobin-treated animals and those pretreated or intralesionally treated with shark serum were similar to the controls in both tumor kinetics and survival times.

Animals↗

Substitution rates of organelle and nuclear genes in sharks: implicating metabolic rate (again).

Rates of nucleotide substitution for nuclear genes are thought to be governed primarily by the number of germ line replication events (the so-called "generation time" hypothesis). In contrast, rates of mitochondrial DNA evolution appear to be set primarily by DNA damage pathways of mutation mediated by mutagenic by-products of oxidative phosphorylation (the so-called "metabolic-rate" hypothesis). Comparison of synonymous substitution rates estimated for the mitochondrial cytochrome b gene and nuclear-encoded dlx, hsp70, and RAG-1 genes in mammals and sharks shows that rates of molecular evolution for sharks are approximately an order of magnitude slower than those for mammals for both nuclear and mitochondrial genes. In addition, there is significant positive covariation of substitution rate for mitochondrial and nuclear genes within sharks. These results, interpreted in light of the pervasiveness of DNA damage by mutagenic by-products of oxygen metabolism to both nuclear and mitochondrial genes and coupled with increasing evidence for cross-genome activity of DNA repair enzymes, suggest that molecular clocks for mitochondrial and nuclear genes may be set primarily by common mutational mechanisms.

Animals↗

Mitochondrial DNA sequence evolution in sharks: rates, patterns, and phylogenetic inferences.

Abundant representation of sharks in the fossil record makes this group a superb system in which to investigate rates and patterns of molecular evolution and to explore the strengths and weaknesses of phylogenetic inferences from molecular data. In this report, the molecular evolution of the cytochrome b gene in sharks is described and the information related to results from phylogenetic analysis of the data evaluated in the light of a phylogeny derived independently of the molecular data. Across divergent lineages of sharks there is evidence for significant substitution rate variation, departure from compositional equilibrium, and substantial homoplasy; nevertheless, the signal of evolutionary history is evident in patterns of shared transversions and amino acid replacements.

Animals↗

Isolation and genetic diversity of endangered grey nurse shark (Carcharias taurus) populations.

Anthropogenic impacts are believed to be the primary threats to the eastern Australian population of grey nurse sharks (Carcharias taurus), which is listed as critically endangered, and the most threatened population globally. Analyses of 235 polymorphic amplified fragment length polymorphisms (AFLP) loci and 700 base pairs of mitochondrial DNA control region provide the first account of genetic variation and geographical partitioning (east and west coasts of Australia, South Africa) in C. taurus. Assignment tests, analysis of relatedness and Fst values all indicate that the Australian populations are isolated from South Africa, with negligible migration between the east and west Australian coasts. There are significant differences in levels of genetic variation among regions. Australian C. taurus, particularly the eastern population, has significantly less AFLP variation than the other sampling localities. Further, the eastern Australian sharks possess only a single mitochondrial haplotype, also suggesting a small number of founding individuals. Therefore, historical, rather than anthropogenic processes most likely account for their depauperate genetic variation. These findings have implications for the viability of the eastern Australian population of grey nurse sharks.

Animal Migration↗

Discrimination of sounds based on the phase difference between particle motion and acoustic pressure in the shark Chiloscyllium griseum.

The reported psychophysical study indicates that a shark not only detects particle motion in a sound field (f = 84 Hz) but moreover shows a sensitivity to acoustic pressure. These results are remarkable because sharks lack a gas bladder that can act as a pressure-to-displacement transformer enabling the hair cells in the labyrinths to respond to acoustic pressure indirectly. The probable significance of the observed phenomena for acoustic localization in fish is discussed and a possible pressure-to-displacement transformer in shark is proposed.

Animals↗

Structure of a shark IgNAR antibody variable domain and modeling of an early-developmental isotype.

The new antigen receptor (IgNAR) antibodies from sharks are disulphide bonded dimers of two protein chains, each containing one variable and five constant domains. Three types of IgNAR variable domains have been discovered, with Type 3 appearing early in shark development and being overtaken by the antigen-driven affinity-matured Type 1 and 2 response. Here, we have determined the first structure of a naturally occurring Type 2 IgNAR variable domain, and identified the disulphide bond that links and stabilizes the CDR1 and CDR3 loops. This disulphide bridge locks the CDR3 loop in an "upright" conformation in contrast to other shark antibody structures, where a more lateral configuration is observed. Further, we sought to model the Type 3 isotype based on the crystallographic structure reported here. This modeling indicates (1) that internal Type 3-specific residues combine to pack into a compact immunoglobulin core that supports the CDR loop regions, and (2) that despite apparent low-sequence variability, there is sufficient plasticity in the CDR3 loop to form a conformationally diverse antigen-binding surface.

Amino Acid Sequence↗

When bigger is not better: selection against large size, high condition and fast growth in juvenile lemon sharks.

Selection acting on large marine vertebrates may be qualitatively different from that acting on terrestrial or freshwater organisms, but logistical constraints have thus far precluded selection estimates for the former. We overcame these constraints by exhaustively sampling and repeatedly recapturing individuals in six cohorts of juvenile lemon sharks (450 age-0 and 255 age-1 fish) at an enclosed nursery site (Bimini, Bahamas). Data on individual size, condition factor, growth rate and inter-annual survival were used to test the 'bigger is better', 'fatter is better' and 'faster is better' hypotheses of life-history theory. For age-0 sharks, selection on all measured traits was weak, and generally acted against large size and high condition. For age-1 sharks, selection was much stronger, and consistently acted against large size and fast growth. These results suggest that selective pressures at Bimini may be constraining the evolution of large size and fast growth, an observation that fits well with the observed small size and low growth rate of juveniles at this site. Our results support those of some other recent studies in suggesting that bigger/fatter/faster is not always better, and may often be worse.

Age Factors↗

Plasma biochemistry reference values of wild bonnethead sharks, Sphyrna tiburo.

BACKGROUND: Elasmobranchs (sharks, skates, and rays) are of commercial, sport, research, and exhibit importance, however, blood chemistry reference values have been determined for few of these species. OBJECTIVE: The purpose of this study was to establish plasma biochemistry and PCV reference values for wild bonnethead sharks (Sphyrna tiburo). METHODS: Heparinized blood samples were collected from 24 bonnethead sharks at the time of capture in trawl nets off the coast of South Carolina and Georgia. Weight, length, PCV, total solids (TS, by refractometry), and plasma biochemical analyses were done using standard techniques. Wilcoxon rank-sum and Kendall tau b tests were used to compare values by animal size, boat and sex; 1-way ANOVA was used to compare TS and total protein (TP) concentrations. RESULTS: Median (quartile; minimum-maximum) values were as follows: PCV 22% (22%, 26%; 17-28%), TS 6.3 (6.0, 6.8; 5.8-7.5) g/dL, total protein 2.9 (2.7, 3.4; 2.2-4.3) g/dL, albumin 0.4 (0.4, 0.4; 0.3-0.5) g/dL, globulins 2.6 (2.3, 3.0;1.9-3.8) g/dL, sodium 282 (279, 285;273-292) mmol/L, potassium 7.3 (6.4, 7.9; 5.7-9.2) mmol/L, chloride 290 (285, 296; 277-304) mmol/L, total CO2 3 (2, 4; 0-5) mmol/L, calcium 16.8 (16.2, 17.4;15.8-18.2) mg/dL, phosphorus 8.8 (7.5, 10.0; 5.9-12.7) mg/dL, urea nitrogen 1004 (986, 1028; 944-1068) mg/dL, creatinine <0.1 mg/dL, glucose 184 (165, 191; 155-218) mg/dL, aspartate aminotransferase 42 (33, 66; 15-132) U/L, lactate dehydrogenase <5 U/L, creatine kinase 82 (47, 233; 18-725) U/L, and osmolality 1094 (1078, 1111; 1056-1139) mOsm/kg. No differences based on sex were detected. TS and TP values were related by the fitted line TS = (1.006 X TP) + 3.318. CONCLUSION: Values reported here will be useful for evaluating the health status of bonnetheads in wild and captive research conditions and in exhibits.

Animals↗

Hematology of the sandbar shark, Carcharhinus plumbeus: standardization of complete blood count techniques for elasmobranchs.

BACKGROUND: Standardized hematologic methods and reference intervals have not been established for cartilaginous fishes (sharks, skates, and rays) despite the large number of animals displayed in zoos and aquariums worldwide. OBJECTIVE: The focus of this study was to validate CBC methods for sandbar shark (Carcharhinus plumbeus) blood, based on criteria established in human medicine, for the following tests: RBC count, total WBC count, PCV, hemoglobin (Hgb) concentration, and WBC differential percentages. METHODS: Replicate CBCs were performed using blood samples from 5 captive sandbar sharks. Three protocols for RBC and total WBC counts were compared, as were different centrifugation times for PCV determination, and 2 methods for Hgb concentration. Means, minimum and maximum values, and CVs were compared to CAP and CLIA performance guidelines for human tests. RESULTS: Total WBC counts in a diluent modified for elasmobranch blood, Hgb concentration by the cyanmethemoglobin method after removal of nuclei, and WBC differential percentages showed acceptable performance. PCV results were acceptable when tubes were centrifuged for at least 5 minutes. Total RBC counts by all 3 methods exceeded the acceptable error for manual counts of human cells. CONCLUSIONS: Standardized CBC tests can be used as health assessment tools for elasmobranchs. Total RBC counts should be viewed as estimates.

Animals↗

Shark cartilage contains inhibitors of tumor angiogenesis.

Shark cartilage contains a substance that strongly inhibits the growth of new blood vessels toward solid tumors, thereby restricting tumor growth. The abundance of this factor in shark cartilage, in contrast to cartilage from mammalian sources, may make sharks an ideal source of the inhibitor and may help to explain the rarity of neoplasms in these animals.

Animals↗

Regulation of MRP2-mediated transport in shark rectal salt gland tubules.

We examined endothelin-1 (ET-1) regulation of the xenobiotic efflux pump, multidrug resistance-associated protein isoform 2 (MRP2), in intact dogfish shark rectal salt gland tubules using a fluorescent substrate sulforhodamine 101 and confocal microscopy. Subnanomolar to nanomolar concentrations of ET-1 rapidly reduced the cell-to-lumen transport of sulforhodamine 101. These effects were prevented by an ET(B) receptor antagonist but not by an ET(A) receptor antagonist. Immunostaining with an antibody to mammalian ET(B) receptors showed specific localization to the basolateral membrane of the shark rectal gland epithelial cells. ET-1 effects on transport were blocked by a protein kinase C (PKC)-selective inhibitor, implicating PKC in ET-1 signaling. A protein kinase A (PKA)-selective inhibitor had no effect. Forskolin reduced luminal accumulation of sulforhodamine 101, but inhibition of PKA did not block the forskolin effect. Consistent with this observation, a cAMP analog that does not activate PKA reduced luminal accumulation of sulforhodamine 101. These results indicate that shark rectal gland transport on MRP2 is regulated by ET acting through an ET(B) receptor and PKC. In addition, cAMP affects transporter function through a PKA-independent mechanism, possibly by competition for transport.

Animals↗

Characteristics of renal Na(+)-D-glucose cotransport in the skate (Raja erinacea) and shark (Squalus acanthias).

We have investigated the properties of the skate (Raja erinacea) and shark (Squalus acanthias) kidney Na(+)-D-glucose cotransporters (SGLT) in uptake studies of radiolabeled substrates into isolated renal brush-border membrane vesicles (BBMV). Scatchard plot analysis of the substrate dependence revealed that the Na(+)-D-glucose cotransporter population is homogenous within each species. Skate BBMV showed a relatively high affinity for D-glucose [Michaelis constant (K(m)) = 0.12 mM] with an apparent coupling ratio of approximately 2 Na+ to 1 D-glucose, whereas the shark transporter was much lower in affinity (K(m) = 1.90 mM) and had a lower coupling ratio, more like 1 Na+ to 1 D-glucose. These characteristics resemble the properties of SGLT1 and SGLT2, which are known to coexist in the mammalian kidney. Inhibitor studies using sugar analogs and glucosides suggested structural differences of the D-glucose binding site among these transporters, whereas the hydrophobic transporter domains in the vicinity of the D-glucose binding site appeared to be similar. In the high-affinity skate system, D-glucose was recognized by hydrogen bonds to the hydroxy groups at C-2, C-3, and C-4 and by hydrophobic interaction with the C-6 methylene group. In contrast, the low-affinity shark system seemed to lack the hydrophobic recognition motif for the C-6 methylene group of D-glucose.

Animals↗

The microanatomy of the rectal salt gland of the Port Jackson Shark, Heterodontus portusjacksoni (Meyer) (Heterodontidae): suggestions for a counter-current exchange system.

A comprehensive anatomical study was undertaken to examine the rectal salt gland in the Port Jackson shark, Heterodontus portusjacksoni, a shark known to invade estuarine environments. The microstructure and vascular organisation of the rectal salt gland was investigated using histological observation and scanning electron microscopy of vascular corrosion casts. Cellular specialisation was observed in the lining of the central lumen of this gland. This may indicate that there is some modification of the principal product of the gland prior to its secretion. The rectal salt gland has a complex structure related to its function. Contrary to previous reports, the flow in secretory tubules is in the opposite direction to that of the capillaries and thus constitutes a counter-current arrangement. The similarity in the organisation of the counter-current and lobulate arrangement of salt-secreting glands through phylogenetically diverse organisms, such as sharks and birds, suggests that this arrangement is important in achieving efficient salt secretion.

Animals↗

Mesencephalic trigeminal nucleus in sharks. A light microscopic study.

In 22 adult sharks representing eight species, the mesencephalic trigeminal nucleus (Mes V) was examined employing the Romanes reduced silved method. Morphological evidence is presented of extensive development of neurites and intranuclear communication among Mes V cells of sharks, as well as their apparent innervation by extranuclear neurons. Species differences and the possible significance of the morphology of the Mes V in sharks to certain aspects of their behavior are discussed. A brief comparison of the Mes V of elasmobranchs and those of reptiles and mammals is included.

Animals↗

Cerebrospinal fluid contacting and supraependymal mesencephalic trigeminal cells in the blue and mako sharks. A scanning electron microscopic study.

Scanning electron microscopic observations reveal cerebrospinal fluid contacting and supraependymal cell bodies of the mesencephalic trigeminal (Mes V) nucleus in adult specimens of mako and blue sharks, but not in very young specimens of the smooth dogfish shark. Possible functional roles are discussed in terms of participation of cerebrospinal fluid contacting Mes V cells in a neuroendocrine regulatory system. Comparisons are made with light microscopic observations of the relationship of Mes V cells with the ventricular wall. General features of the scanning electron microscopic appearance of the roof of the ventricle of the optic tectum in sharks are described and compared to similar general features found in the third ventricle of a variety of vertebrates.

Animals↗

Perfusion of isolated tubules of the shark rectal gland. Electrical characteristics and response to hormones.

Both the mammalian thick ascending limb of Henle's loop and the shark rectal gland actively transport Cl against an electrochemical gradient by mechanisms involving hormone-sensitive NaCl transport. In contrast to mammalian renal tubules, individual tubules of the shark rectal gland previously have not been perfused in vitro. Using a combination of renal slice and microdissection techniques we were able to isolate and perfuse single rectal gland tubules without the use of enzyme treatment. Single tubules consistently generated lumen-negative transepithelial voltages (Vt) of -1.8 mV when perfused and bathed with identical shark Ringer's solution. The addition of cyclic AMP, vasoactive intestinal peptide (VIP), and adenosine to the bath increased Vt to -7.5, -9.0, and -4.3 mV, respectively (all P less than 0.02 compared with paired controls). Each stimulation could be reversed by addition by furosemide to the bath. The adenosine response was inhibited by theophylline, a specific inhibitor of adenosine receptors. The tubules had a low transepithelial electrical resistance of 12-26 omega X cm2 and exhibited a transepithelial permselectivity for small cations. These results indicate that tubules of the rectal gland can be perfused in vitro and have receptors for VIP and adenosine. Cyclic AMP and secretagogues hyperpolarize the membrane consistent with electrogenic chloride transport, and these effects are reversed by furosemide, an inhibitor of coupled sodium-potassium-chloride co-transport. The response of Vt to cyclic AMP and furosemide, the transepithelial electrical resistance, and the cation selective permeability of tubules are remarkably similar to measurements in perfused mammalian thick ascending limbs.

Adenosine↗