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PubMed1976

Haemoglobins of the shark, Heterodontus portusjacksoni II. Amino acid sequence of the alpha-chain.

The amino acid sequence of the alpha-chain of the principal haemoglobin from the shark, H. portusjacksoni has been determined. The chain has 148 residues and is acetylated at the amino terminal. The soluble peptides obtained by tryptic and chymotryptic digestion of the protein or its cyanogen bromide fragments were isolated by gel filtration, paper ionophoresis and paper chromatography. The amino acid sequences were determined by the dansyl-Edman procedure. The insoluble "core" peptide from the tryptic digestion contained 34 residues and required cleavage by several prosteases before the sequence was established. Compared with human alpha-chain there are 88 amino acid differences including the additional seven residues which appear on the amino terminal of the shark chain. There is also one deletion and one insertion. The chain contains no tryptophan but has four cysteinyl residues which is the highest number of such residues recorded for a vertebrate globin. In the alpha1beta1 contact sites there are four changes in the oxyhaemoglobin form and six deoxy form. Nine of the 16, alpha1beta1 contact sites show variation while three of the haem contact sites have changed in comparison to the residues known to be involved in these interactions in horse haemoglobin alpha-chain. Use of the sequence data to estimate a time of divergence of the shark from the main vertebrate line yielded the value of 410 +/- 46 million years. The data, in general, support the palaeontological view that bony fishes arose before the elasmobranchs.

Amino Acid Sequence↗
PubMed1998

New host and ocean records for the copepod Ommatokoita elongata (Siphonostomatoida: Lernaeopodidae), a parasite of the eyes of sleeper sharks.

Seven of 8 Pacific sleeper sharks (Somniosus pacificus Bigelow and Schroeder, 1944) captured in Prince William Sound, Alaska, were actively infected, and all 8 had been at one time infected with the parasitic copepod Ommatokoita elongata (Grant, 1827) (Siphonostomatoida: Lernaeopodidae). Active infections consisted of adult females and chalimus larvae that had attached to the corneas of the sharks' eyes. This report documents a new host record and possibly the only reliable record of this parasite from a host other than the Greenland shark, Somniosus microcephalus (Bloch and Schneider, 1801). It also documents the first time O. elongata has been identified outside of the Atlantic Ocean or its locally adjacent straits and seas.

Animals↗
PubMed1986

Solubilized (Na+ + K+)-ATPase from shark rectal gland and ox kidney--an inactivation study.

The bi-exponential time-course of detergent inactivation at 37 degrees C of C12E8-solubilized (Na+ + K+)-ATPase from shark rectal glands and ox kidney was investigated. The data for shark enzyme, obtained at detergent/protein weight ratios between 2 and 16, are interpreted in terms of a simple model where the membrane bound enzyme is solubilized predominantly as (alpha-beta)2 diprotomers at low detergent concentrations and as alpha-beta protomers at high C12E8 (octaethyleneglycoldodecylmonoether) concentrations. It is observed that the protomers are inactivated 15-fold more rapidly than the diprotomers, and that the rate of inactivation of both oligomers is proportional to the detergent/protein ratio. Inactivation of kidney enzyme was biexponential with a very rapid inactivation of up to 40% of the enzyme activity. The observed rate of inactivation of the slower phase varied with the detergent/protein ratio, but the inactivation pattern for the kidney enzyme could not readily be accommodated within the model for inactivation of the shark enzyme. The rates of inactivation at 37 degrees C were about the same in KCl and NaCl, i.e., in the E2(K) and E1 X Na forms, for both enzymes.

Animals↗
PubMed1989

The complete amino acid sequence of growth hormone of an elasmobranch, the blue shark (Prionace glauca).

The complete amino acid sequence of growth hormone (GH) from a phylogenetically ancient fish, the blue shark (Prionace glauca), was determined. The shark GH isolated from pituitary glands by U. J. Lewis, R. N. P. Singh, B. K. Seavey, R. Lasker, and G. E. Pickford (1972, Fish. Bull. 70, 933-939) was purified by reversed-phase high-performance liquid chromatography. The hormone was reduced, carboxymethylated, and subsequently cleaved in turn with cyanogen bromide and Staphylococcus aureus protease. The intact protein was also cleaved with lysyl endopeptidase and o-iodosobenzoic acid. The resulting peptide fragments were separated by rpHPLC and submitted to sequence analysis by automated and manual Edman methods. The shark GH consists of 183 amino acid residues with a calculated molecular weight of 21,081. Sequence comparisons revealed that the elasmobranch GH is considerably more similar to tetrapod GHs (e.g., 68% identity with sea turtle GH, 63% with chicken GH, and 58% with ovine GH) than teleostean GHs (e.g., 38% identities with salmon GH and 42% with bonito GH) except for eel GH (61% identity), and substantiates the earlier finding derived from the immunochemical and biological studies (Hayashida and Lewis, 1978) that the primitive fish are less diverged from the main line of vertebrate evolution leading to the tetrapod than are the modern bony fish.

Amino Acid Sequence↗
PubMed2001

Transjugular liver biopsy with use of the shark jaw needle: diagnostic yield, complications, and cost-effectiveness.

PURPOSE: Obtaining transjugular liver biopsy specimens with use of single-use needle systems is expensive, whereas biopsy specimens obtained with use of reusable needle systems are frequently associated with inadequate core specimens. The authors report their experience with the reusable Cook Shark Jaw biopsy needle, including diagnostic yield, complications, and cost-effectiveness. MATERIALS AND METHODS: A retrospective audit was performed of a cohort of 134 patients who underwent 136 transjugular liver biopsies with use of a reusable 16-gauge Shark Jaw needle during a 30-month period. Specimen adequacy and complication rates were assessed and direct costs of expendable components calculated. Cost-effectiveness was expressed as cost-per-successful biopsy. RESULTS: Biopsies were technically successful in 126 of 136 (93%) patients, with diagnostic histologic core specimens obtained in 124 of 126 (98%) patients, for an overall success rate of 91%. Complications included capsular penetration in six (4.4%) patients, cardiac arrhythmia in two (1.5%) patients, and puncture site hematoma or bleeding in 10 (7.4%) patients. Three tract embolizations were performed for capsular penetration. No instances of subcapsular hematoma, hemoperitoneum, or sepsis occurred, and no deaths were attributed to the procedure. The cost of expendable components totaled $103 per biopsy, corresponding to a cost-effectiveness of $113/successful biopsy. CONCLUSION: Transjugular liver biopsy specimens obtained with use of the Shark Jaw needle have a diagnostic yield comparable to those obtained with use of single-use biopsy systems, at a substantially lower cost with no increase in serious complications.

Adult↗
PubMed1996

Cellular and molecular biology of chloride secretion in the shark rectal gland: regulation by adenosine receptors.

The rectal gland of the dogfish shark (Squalus acanthias) is a sodium chloride secreting epithelial organ whose function was discovered in 1959 by Wendell Burger. The gland, composed of homogenous tubules of a single cell type, is an important model for secondary active chloride transport. Hormonal stimulation of chloride secretion in this system activates asymetrically arranged transport proteins (apical cAMP-activated CFTR-like Cl- channels, basolateral Na/K/2Cl cotransporters, Na/K-ATPase activity, and K+ channels). Five receptors, hormones, and membrane proteins of the shark rectal gland involved in chloride secretion have been cloned recently. Because the intact gland can be perfused via a single artery and vein, it has been possible to examine precisely the metabolic regulation of chloride transport by endogenous adenosine. Rectal gland cells have a high density of both stimulatory A2 type and inhibitory A1 type adenosine receptors. When stimulated by secretagogues, chloride secretion and venous adenosine concentrations increase in parallel, with chloride secretion increasing from approximately 150 to 2100 microEq/hr/g, and adenosine concentrations increasing from approximately 5 to approximately 890 nM. This work of ion transport is accompanied by a marked fall in intracellular ATP activity and a rise in both intracellular AMP and adenosine activity. Agents that prevent the interaction of endogenous adenosine with extracellular receptors significantly increase the chloride transport response to secretagogues. When chloride transport is inhibited by blocking the Na/K/2Cl cotransporter with bumetanide, both adenosine release and chloride secretion fall to basal values. We recently cloned a unique adenosine receptor subtype that is distinct from previously cloned mammalian adenosine receptors. Because of its highly specialized function, single cell type, and simple vascular system, the shark rectal gland is an ideal model system for examining the metabolic regulation of chloride secretion by adenosine receptors.

Animals↗
PubMed1995

Shark, a Src homology 2, ankyrin repeat, tyrosine kinase, is expressed on the apical surfaces of ectodermal epithelia.

Tyrosine kinases, ankyrin repeats, and Src homology 2 domains play central roles in developmental processes. The cloning of a cDNA for Shark, a single protein that possesses all three domains, is described. During Drosophila embryogenesis, Shark is expressed exclusively by ectodermally derived epithelia and is localized preferentially to the apical surface of these cells. This apical localization persists, even as tissues undergo complex invaginations, moving from the external surface of embryos to form internal structures, but expression is lost when cells lose their polarity. This pattern closely mimics the expression of Crumbs, a protein necessary for proper organization of ectodermal epithelia. Shark's structure and localization pattern suggest that it functions in a signaling pathway for epithelial cell polarity, possibly transducing the Crumbs signal.

Amino Acid Sequence↗
PubMed2005

Interaction of FXYD10 (PLMS) with Na,K-ATPase from shark rectal glands. Close proximity of Cys74 of FXYD10 to Cys254 in the a domain of the alpha-subunit revealed by intermolecular thiol cross-linking.

FXYD domain-containing proteins are tissue-specific regulators of the Na,K-ATPase that have been shown to have significant physiological implications. Information about the sites of interaction between some FXYD proteins and subunits of the Na,K-ATPase is beginning to emerge. We previously identified an FXYD protein in plasma membranes from shark rectal gland cells and demonstrated that this protein (FXYD10) modulates shark Na,K-ATPase activity. The present study was undertaken to identify the location of the C-terminal domain of FXYD10 on the alpha-subunit of Na,K-ATPase, using covalent cross-linking combined with proteolytic cleavage. Treatment of Na,K-ATPase-enriched membranes with the homobifunctional thiol cross-linker 1,4-bismaleimidyl-2,3-dihydroxybutane resulted in cross-linking of FXYD10 to the alpha-subunit. Cross-linking was not affected by preincubation with sodium or potassium but was significantly reduced after pre-incubation with the non-hydrolyzable ATP analog beta,gamma-methyleneadenosine 5'-triphosphate (AMP-PCP). A peptic assay was developed, in which pepsin treatment of Na,K-ATPase at low pH resulted in extensive cleavage of the alpha-subunit while FXYD10 was left intact. Proteolytic fragments of control and cross-linked preparations were isolated by immunoprecipitation and analyzed by gel electrophoresis. A proteolytic fragment containing FXYD10 cross-linked to a fragment from the alpha-subunit could be localized on SDS gels. Sequencing of this fragment showed the presence of FXYD10 as well as a fragment within the A domain of the alpha-subunit comprising 33 amino acids, including a single Cys residue, Cys254. Thus, regulation of Na,K-ATPase by FXYD10 occurs in part via cytoplasmic interaction of FXYD10 with the A domain of the shark alpha-subunit.

Adenosine Triphosphate↗
PubMed2004

Reproduction in shark-attacked sea turtles is supported by stress-reduction mechanisms.

Vertebrates exhibit varied behavioural and physiological tactics to promote reproductive success. We examined mechanisms that could enable female loggerhead turtles to undertake nesting activities and maintain seasonal reproduction despite recent shark injuries of varying severity. We proposed that endocrinal mechanisms that regulate both a turtle's stress response and reproductive ability are modified to promote successful and continued reproduction. Irrespective of the degree of injury, females did not exhibit increased levels of the stress hormone corticosterone, nor decreased levels of the reproductive steroid testosterone; hormone responses consistent with stress. When exposed to a capture stressor, females with shark injury did not exhibit any greater corticosterone response than controls. In addition, breeding females showed a reduced corticosterone stress response compared to non-breeding females. Reduced endocrinal responses following shark injury, and during breeding in general may, in part, enable females to maintain behavioural and physiological commitment to reproduction.

Analysis of Variance↗
PubMed1991

Cl- secretion by cultured shark rectal gland cells. II. Effects of forskolin on cellular electrophysiology.

Employing microelectrode techniques we have assessed the cellular electrophysiological properties of shark rectal gland (SRG) cells in primary culture. In the absence of secretagogues a 10-fold reduction in the Cl- concentration of the apical superfusate shark Ringer solution had little effect on either apical membrane electrical potential difference (Va) or fractional resistance (fRa), indicating little, if any, apical membrane Cl- conductance. Superfusing the basolateral surface with high-K+ shark Ringer solution (K+ increased 10-fold) depolarized the basolateral membrane electrical potential difference (Vb) by 43 mV, indicating that this barrier is largely K+ conductive. In addition, basolateral Ba2+ (5 mM) depolarized Vb by 12 mV and reduced fRa from 0.92 to 0.58, results consistent with a K(+)-conductive basolateral membrane in unstimulated SRG cells. Basolateral forskolin (10(-6) M) depolarized Va by 25 mV and caused a dramatic reduction in fRa from 0.97 to approximately 0.10. Under these conditions, a 10-fold decrease in apical superfusate Cl- concentration depolarized Va by 37 mV, revealing an adenosine 3',5'-cyclic monophosphate-induced apical membrane Cl- conductance. The time course of the forskolin-induced changes in Va and Vb suggests that the basolateral membrane K+ conductance increased and maintained the driving force for apical Cl- exit, as in other Cl(-)-secreting epithelia. These electrophysiological properties compare favorably with those of the perfused SRG tubule and indicate that SRG primary cultures are a suitable model for Cl(-)-secreting epithelia.

Animals↗
PubMed2002

Cyclooxygenase cloning in dogfish shark, Squalus acanthias, and its role in rectal gland Cl secretion.

The present studies were carried out with the aims to determine the cDNA sequence for cyclooxygenase (COX) in an elasmobranch species and to study its role in regulation of chloride secretion in the perfused shark rectal gland (SRG). With the use of long primers (43 bp) derived from regions of homology between zebrafish and rainbow trout COX-2 genes, a 600-bp product was amplified from SRG and was found to be almost equally homologous to mammalian COX-1 and COX-2 (65%). The full-length cDNA sequence was obtained by 5'-RACE and by analyzing an EST clone generated by the EST Project of the Mt. Desert Island Biological Laboratory Marine DNA Sequencing Center. The longest open reading frame encodes a 593-amino acid protein that has 68 and 64% homology to mammalian COX-1 and COX-2, respectively. The gene and its protein product is designated as shark COX (sCOX). The key residues in the active site (Try(385), His(388), and Ser(530)) are conserved between the shark and mammalian COX. sCOX contains Val(523) that has been shown to be a key residue determining the sensitivity to COX-2-specific inhibitors including NS-398. The mRNA of sCOX, detected by RT-PCR, was found in all tissues tested, including rectal gland, kidney, spleen, gill, liver, brain, and heart, but not in fin. In the perfused SRG, vasoactive intestinal peptide (VIP) at 5 nM induced rapid and marked Cl(-) secretion (basal: <250 microeq x h(-1) x g(-1); peak response: 3,108 +/- 479 microeq x h(-1) x g(-1)). In the presence of 50 microM NS-398, both the peak response (2,131 +/- 307 microeq x h(-1) x g(-1)) and the sustained response to VIP were significantly reduced. When NS-398 was removed, there was a prompt recovery of chloride secretion to control values. In conclusion, we have cloned the first COX in an elasmobranch species (sCOX) and shown that sCOX inhibition suppresses VIP-stimulated chloride secretion in the perfused SRG.

Amino Acid Sequence↗
PubMed1992

Distribution of tyrosine hydroxylase, serotonin, and leu-enkephalin immunoreactive cells in the brainstem of a shark, Squalus acanthias.

The central nervous system location of neurochemicals that are widely distributed among extant animals may give us clues to changes that occurred in the brains of these animals during evolution. We have been studying the brains of cartilaginous fishes, a heterogeneous group whose central nervous system varies considerably. Squalus acanthias, the spiny dogfish shark, was chosen to represent the squalomorphs, a group of living sharks known to possess many primitive characters. The distribution of tyrosine hydroxylase (TH+), serotonin (5-HT+), and leu-enkephalin (LENK+) positive cells within the brainstem of Squalus was determined by use of antibodies to these substances. All the major raphe groups described for mammals were found in Squalus. The 5-HT+ cells in raphe nuclei were more uniformly distributed in Squalus than in Heterodontus, the horn shark. Other nuclei that were 5-HT+ and LENK+, and that have been identified in mammals, included reticularis paragigantocellularis lateralis, a B9 cell group, and reticularis magnocellularis. The postcommissural nucleus and pretectal area contained 5-HT+ and LENK+ cells. These cells have been described in a holocephalian, in teleosts, and in reptiles but not in other elasmobranchs or in mammals. Cells that were TH+ were located in prominent A1/A2, A6 (locus coeruleus), A9 (substantia nigra), and A10 (ventral tegmental area) cell groups, and in a very small A5 group. We conclude that the variation in chondrichthian brainstems exceeds that in mammals, and we suggest that this variation is related to life-style and the long evolutionary history of these fishes.

Animals↗
PubMed1996

Heterocercal tail function in leopard sharks: a three-dimensional kinematic analysis of two models

Two different models have been proposed to explain the function of the heterocercal tail in shark locomotion. The classical model proposes that, as a result of lift generated by the tail as it beats, the net force acting on the tail is directed dorsally and anteriorly. In contrast, Thomson's model suggests that the tail generates a net force directed through the shark's center of gravity, i.e. ventrally and anteriorly. In this study, we evaluate these two models by describing the three-dimensional kinematics of the heterocercal tail in the leopard shark Triakis semifasciata during swimming. Lateral and posterior views of the tail were examined from four individuals swimming in a flow tank at 1.2 L s-1 (where L is total length) using two high-speed video cameras filming simultaneously at 250 fields s-1. These two simultaneous views allowed eight landmarks on the tail to be followed in three dimensions through time. These landmarks allowed the tail to be divided into separate surfaces whose orientation over time was calculated. Points located anteriorly on the tail go through significantly smaller excursions and reach their maximum lateral excursion significantly earlier in the beat cycle than points on the trailing edge of the tail. Three-dimensional angle calculations show that the terminal lobe leads the ventral lobe through a beat, as predicted by the classical model. Dye-stream visualizations confirmed that this pattern of movement deflects water ventrally and posteriorly to the moving tail, providing strong support for the classical model. Additionally, our results show that a three-dimensional analysis is critical to understanding the function of the heterocercal tail.

Journal Article↗
PubMed1997

Feeding mechanism and functional morphology of the jaws of the lemon shark Negaprion brevirostris (Chondrichthyes, Carcharhinidae)

This study tests the hypothesis that preparatory, expansive, compressive and recovery phases of biting behavior known for aquatically feeding anamniotes are conserved among extant elasmobranch fishes. The feeding mechanism of the lemon shark Negaprionbrevirostris is examined by anatomical dissection, electromyography and high-speed video analysis. Three types of feeding events are differentiated during feeding: (1) food ingestion primarily by ram feeding; (2) food manipulation; and (3) hydraulic transport of the food by suction. All feeding events are composed of the expansive, compressive and recovery phases common to aquatically feeding teleost fishes, salamanders and turtles. A preparatory phase is occasionally observed during ingestion bites, and there is no fast opening phase characteristic of some aquatically feeding vertebrates. During the compressive phase, palatoquadrate protrusion accounts for 26% of the gape distance during jaw closure and is concurrent with muscle activity in the dorsal and ventral preorbitalis and the levator palatoquadrati. Hydraulic transport events are shorter in duration than ram ingestion bites. Prey ingestion, manipulation and hydraulic transport events are all found to have a common series of kinematic and motor components. Individual sharks are capable of varying the duration and to a lesser extent the onset of muscle activity and, consequently, can vary their biting behavior. We propose a model for the feeding mechanism in carcharhinid sharks, including upper jaw protrusion. This study represents the first electromyographic and kinematic analysis of the feeding mechanism and behavior of an elasmobranch.

Journal Article↗
PubMed1984

[Evaluation of the protein quality of meat flour from the tollo shark (Squalus acanthias)].

The protein quality of tollo flour evaluated by various biological methods, including digestibility, is reported. The product contained 91.52% crude protein and 69.86% true protein, the difference being non-protein nitrogen, mainly urea. Due to this reason the biological evaluations were carried out with different shark flour concentrations and evaluated at different lengths of time with each method. Diets containing 0, 3, 6, 9 and 12% shark flour were prepared and evaluated by the PER, NPR, and NGI methods. Casein was used as reference protein. The protein digestibility was 91.2%, not corrected for non-protein nitrogen. On this same basis, PER and NPR--at a protein level of 9.97% in the diet--were 2.58 and 2.05, respectively. The casein control diet gave a PER of 2.93 and an NPR of 4.52. NGI was calculated by using the non-protein diet, or without it, with values of 3.32 and 2.95, respectively. As expected, all values corrected for non-protein nitrogen were higher for protein digestibility. The differences were significant at the 5% level. Apparently, urea influenced negatively the biological evaluations. It can be concluded, therefore, that the tollo shark meat is of excellent quality.

Animals↗
PubMed1964

ELECTROENCEPHALOGRAMS OF SHARKS.

Patterns of electrical potentials recorded from the brains of sharks exhibit definite relationships to chemical and visual stimuli. Forebrain potentials reflect olfactory processes. Both restrained and free-swimming sharks exhibit mesencephalic responses to light and neural triggering of respiratory reflexes from the medulla. Early evolution of typical vertebrate brain functions, with emphasis upon chemoreception, is indicated

Animals↗
PubMed1998

Morphological and cytological ontogenesis of the ampullae of lorenzini and lateral line canals in the Oman shark, Iago omanensis Norman 1939 (Triakidae), from the Gulf of Aqaba, Red Sea.

The Oman shark, Iago omanensis, is a small, placental viviparous species encountered in great numbers in the deeper waters of the Gulf of Aqaba, Red Sea. It reproduces year-round, providing an opportunity to study ontogenesis of organ systems at various stages of development. This study examines the morphological and cytological development of the mechanoreceptive lateral line (LL) system and the electrosensory Ampullae of Lorenzini. Female I. omanensis were collected bimonthly from the Gulf of Aqaba at depths of 300-800 m and sacrificed with an overdose of MS222. Their uteri were dissected and the embryos separated and fixed for light and electron microscopy. A total of 260 embryos of varying dimensions were studied. The first primordia of neuroectodermal LL neuromasts are seen in embryos of 18 mm TL. These then sink into the dermis, ripen, and develop tubuli that join to form the LL canal systems, especially developed on the head. In contrast, the primordia of Ampullae of Lorenzini start out as groups of embryonic cells situated subdermally. In embryos of 24-26 mm TL initially they develop into tubuli. With growth, the ampullar alveoli gradually widen at their ends to form the sensory epithelium. The ampullar tubuli elongate, bringing the alveoli to sites over the rostrum and head, where the ampullar capsules are formed. The presynaptic electrosensory cells are attached to afferent neural extensions forming sensory rami which extend, as in adult sharks, to the dorsal nucleus in the medulla. In preterm juveniles of 150-160 mm TL, the LL system and the Ampullae of Lorenzini are fully developed cytologically. The results of this study support the hypothesis that the LL system and electrosensory Ampullae of Lorenzini develop as separate modalities and that their structural similarity is due to the origin from the embryonic neuroectoderm. The dichotomy of their evolution occurred in very early ancestry as an ecomorphological adaptation to different sensory functions.

Animals↗
PubMed1981

Two visual pathways to the telencephalon in the nurse shark (Ginglymostoma cirratum). I. Retinal projections.

The central projections of the retina in the nurse shark were studied by anterograde transport of horseradish peroxidase and tritiated proline. With regard to efferent retinal fibers, both techniques gave completely identical results. Projections were found to pretectal area, dorsal thalamus, basal optic nucleus, and optic tectum, all at the contralateral side. The retinal target cells in the dorsal thalamus are restricted to the ventrolateral optic nucleus and the posterior optic nucleus. No evidence was found for an earlier-reported projection to the lateral geniculate nucleus. The present findings show that the ventrolateral optic nucleus exhibits homological features of the dorsal lateral geniculate nucleus in other vertebrate groups, whereas the lateral geniculate nucleus of the nurse shark is much more comparable to the nucleus rotundus of teleosts and birds and would be more appropriately so named. The application of the HRP technique also allowed us to study afferents to the retina by retrograde transport of tracer. Retrogradely labeled cells were observed in the contralateral optic tectum and are apparently similar to those reported for teleosts and birds.

Animals↗