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PubMed1982

Antigenic structure of sperm whale myoglobin. II. Characterization of antibodies preferentially reactive with peptides arising in response to immunization with the native protein.

The antigenicity of peptide fragments derived from CNBr-cleaved sperm whale Mb has been tested in competitive inhibition assays. These peptides appeared to be very poor inhibitors in a reaction involving radiolabeled sperm whale Mb with antipeptide antibodies, requiring about 10(4)-fold excess of peptides to achieve the same level of inhibition as cold native Mb. However, cold peptides competed much better with radiolabeled peptides for antibody binding than the native protein. These antibodies, preferentially reactive with peptides, originated from sera of animals immunized with the native molecule. This indicates heterogeneity of the antibodies produced against Mb and suggests the presence of diverse antibody subpopulations reactive with different antigenic forms of the protein. Results presented in this paper illustrate that competitive radioimmunoassays preferentially measure antibodies to the radiolabeled species, and this phenomenon can be used to demonstrate distinct populations among antibodies isolated to a single peptide.

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PubMedDate not supplied

[Study of electron transfer in hemoproteins. IV. Oxidation of sperm whale oxymoglobin catalyzed by copper ions].

The influence of cupric ions on the oxidation of sperm whale oxymoglobin and soybean oxyleghemoglobin at pH 4.8--7.5 and 10--40 degrees C has been studied. The stability constants of cupric ions to myoglobin were determined: K1 = 3.4 . 10(5) M(-1) for the more reactive center, and K2 = 2.1 . 10(3) M(-1) for the next 5--7 binding centers. The role of ionization of His-GH1 in the process of oxidation of sperm whale myoglobin is discussed. A mechanism of electron transfer from myoglobin molecule to the external acceptor is presented.

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PubMed1993

Kinetics of ligand binding to Pseudoterranova decipiens and Ascaris suum hemoglobins and to Leu-29-->Tyr sperm whale myoglobin mutant.

The kinetics of binding of O2, CO, and NO to the octameric, two-domain hemoglobins of the parasitic nematodes Pseudoterranova decipiens and Ascaris suum were determined on nanosecond and picosecond time scales using flash photolysis. The two nematode hemoglobins have very similar kinetic properties. On the picosecond time scale, they exhibit an unusual behavior in showing a geminate reaction with oxygen that is biphasic and dependent on the flash intensity. The geminate reaction with NO is also faster and more complete than for sperm whale myoglobin; however, in contrast to the O2 reaction, it is homogeneous. In addition, the oxygen dissociation rate of P. decipiens hemoglobin, 0.0035 s-1, is as low as that of A. suum hemoglobin, 0.004 s-1 (Gibson, Q. H., and Smith, M. H. (1965) Proc. R. Soc. Lond. B Biol. Sci. 163, 206-214). A mutant of sperm whale myoglobin suggested by sequence alignment of the nematode hemoglobins, Leu-29-->Tyr, did not have kinetic properties similar to them.

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PubMed1996

Microsatellites from the beluga whale Delphinapterus leucas.

Fifteen microsatellites were isolated from a beluga whale Delphinapterus leucas, genomic library. The microsatellites were amplified in 100 beluga obtained from two widely separated locations. An average of 8.6 alleles per locus were detected and the average heterozygosity was 0.65 with a range of 0.27-0.86. All microsatellites were polymorphic and 13 of the genotype distributions observed were in Hardy-Weinberg equilibrium. It was possible with these microsatellites to assign correctly individual whales to their stock-of-origin 98% of the time. Microsatellites were amplified in 15 other cetaceans with these beluga-derived primers.

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PubMed1996

Epidermal ultrastructure of the southern right whale calf (Eubalaena australis).

An ultrastructural analysis by transmission and scanning electron microscopy was carried out on normal epidermis of six southern right whale (Eubalaena australis) calves which stranded over a period of several months at Peninsula Valdes, Argentina. This was undertaken to 1) provide the first normal skin ultrastructural data on this highly endangered species which is known to display skin pathology in some instances, and 2) to elucidate further the integumentary specializations which have developed in diving marine mammals. Southern right whale lipokeratinocytes demonstrated parakeratosis and numerous intracellular lipid bodies, keratin and melanosomes, as reported for other cetacean species, but showed several unique ultrastructural features as well. These included a high prevalence of intranuclear inclusion bodies resembling small fragments of cytoplasmic keratin, and close structural relationship between cytoplasmic lipid droplets and the nucleus. The subcellular morphology supported the concept of possible nuclear import of cytoplasmic keratin and lipid metabolites through enlargements of the nuclear pore complex or other disruptions of the nuclear envelope. The light microscopy and scanning electron microscopy also revealed an irregular contour of the lipokeratinocytes which comprised the thick stratum externum, and surface flaking of the outermost cells which were covered by stubby microvillous-like remnants of intercellular junctions. These results thus suggest that the long-tem aquatic evolution of this cetacean species has resulted in a number of integumentary specializations and that investigation of their submicroscopic cytology may help elucidate the general cell biology of nuclear-cytoplasmic interactions.

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PubMed1996

High-level expression and deuteration of sperm whale myoglobin. A study of its solvent structure by X-ray and neutron diffraction methods.

Neutron diffraction has become one of the best ways to study light atoms, such as hydrogens. Hydrogen however has a negative coherent scattering factor, and a large incoherent scattering factor, while deuterium has virtually no incoherent scattering, but a large positive coherent scattering factor. Beside causing high background due to its incoherent scattering, the negative coherent scattering of hydrogen tends to cancel out the positive contribution from other atoms in a neutron density map. Therefore a fully deuterated sample will yield better diffraction data with stronger density in the hydrogen position. On this basis, a sperm whale myoglobin gene modified to include part of the A c11 protein gene has been cloned into the T7 expression system. Milligram amounts of fully deuterated holo-myoglobin have been obtained and used for crystallization. The synthetic sperm whale myoglobin crystallized in P2(1) space group isomorphous with the native protein crystal. A complete X-ray diffraction dataset at 1.5A has been collected. This X-ray dataset, and a neutron data set collected previously on a protonated carbon-monoxymyoglobin crystal have been used for solvent structure studies. Both X-ray and neutron data have shown that there are ordered hydration layers around the protein surface. Solvent shell analysis on the neutron data further has shown that the first hydration layer behaves differently around polar and apolar regions of the protein surface. Finally, the structure of per-deuterated myoglobin has been refined using all reflections to a R factor of 17%.

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PubMed1978

Genetic control of immune response to sperm whale myoglobin in mice. I. T lymphocyte proliferative response under H-2-linked Ir gene control.

Studies on the genetic control of immune response to sperm whale myoglobin were initiated. As demonstrated in this paper, the T lymphocyte proliferative response to whale myoglobin is under H-2-linked Ir gene control. Mice of H-2d, H-2f, and H-2s haplotypes were high responders to the myoglobin, whereas haplotypes H-2b, H-2k, H-2p, H-2q, and H-2r were low responders. The Ir gene(s) was localized between H-2K and H2D regions, since the recombinant strain A.TL (KsIkSkDd) was a low responder and A.TH (KsIsSsDd) was a high responder. Further studies with recombinant strains revealed that the expression of the high-responder I-Ad or Ias alleles was sufficient to give a good response, since strains D2.GD (d d b b b b b b) and B10.HTT (s s s s k k k d) were high responders. The expression of the I-Cd allele in strains B10.A (k k k k k d d d) and B10.A(5R) (b b b k k d d d) also gave high response, and thus suggested a second Ir gene, derived from the H-2d haplotype. The finding that expression of the I-Cs allele in B10.S(8R) (k k ? ? s s s s) did not result in high response suggests the lack of the second Ir gene in the high-responder H-2s haplotype.

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PubMed1982

Giant neural systems in the inner retina and optic nerve of small whales.

Giant neural cell systems (dendrites, cell bodies, and axons) are present among more usual structures in the retina and optic nerve of the small whale (dolphin) Tursiops truncatus retina. Giant cell body dimensions range up to 75 microns in diameter. Nuclei of the cells are frequently larger (greater than 20 microns) than nearby ganglion, bipolar, and receptor cell bodies. The presence of the giant cell system and giant elements in the nerve fiber layer agree with the unusually broad fiber spectrum of the dolphin optic nerve where more than 6% of the axons are greater than 15 microns in diameter. Smaller axons in the size distribution are typical of dimensions found in terrestrial mammals. The axon estimate totaled 157,000 per optic nerve. The giant cell-axon systems of the whale retina may be a unique expression of the large ganglion cell-axon (transient or "Y" functional unit) systems recently identified in terrestrial mammals.

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PubMed2000

Dialect change in resident killer whales: implications for vocal learning and cultural transmission.

Variation in vocal signals among populations and social groups of animals provides opportunities for the study of the mechanisms of behavioural change and their importance in generating and maintaining behavioural variation. We analysed two call types made by two matrilineal social groups of resident killer whales, Orcinus orca, over 12-13 years. We used a neural network-based index of acoustic similarity to identify mechanisms of call differentiation. A test for structural modification of the calls detected significant changes in one call type in both groups, but not in the other. For the modified call type, the rate of divergence between the two groups was significantly lower than the rate of modification within either group showing that calls were modified in a similar fashion in the two groups. An analysis of structural parameters detected no strong directionality in the change. The pattern of call modification could have been caused by maturational changes to the calls or, if killer whale dialects are learned behavioural traits, cultural drift in the structure of the calls together with horizontal transmission of modifications between the two groups. Such vocal matching between members of different matrilines would suggest that vocal learning is not limited to vertical transmission from mother to offspring, which has important implications for models of gene-culture coevolution. Copyright 2000 The Association for the Study of Animal Behaviour.

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PubMed2000

Within-pod variation in the sound production of a pod of killer whales, Orcinus orca.

Pod-specific calling behaviour of resident killer whales has been shown to include: discrete call types not shared among pods, different production rates of shared call types, and differences in the detailed structure of shared call types. To investigate the mechanisms leading to pod-specific calling, we compared the repertoire and structure of calls produced by three different matrilineal units within the same pod, and described call features encoding matrilineal-unit distinctiveness. The three matrilineal units had different production rates of shared calls, including one call type used almost exclusively by one matrilineal unit. Cross-validated discriminant function analyses revealed matrilineal-unit distinctive structure in five of the six shared call types examined, with duration of the terminal component being the most distinctive feature for all call types containing a terminal component. Calls generally consist of low- and high-frequency components that may follow different time-frequency contours. In our sample, a particular high-frequency contour was consistently paired with a particular low-frequency contour, both contours had roughly equal overall variability, and each contained independent matrilineal-unit distinctive information. The only call type that did not differ structurally between matrilineal units is reportedly used more in interpod meetings than in intrapod contexts. The differences in calling behaviour between matrilineal units were similar in form to previously described differences between pods, although more subtle. These results suggest that pod-specific calling behaviour in resident killer whales arises primarily as a consequence of accumulated drift or divergence between highly cohesive matrilineal units as they gradually separate into different pods. Copyright 2000 The Association for the Study of Animal Behaviour.

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PubMed1982

Genetic control and intersite influences on the immune response to sperm whale myoglobin.

Determination of the precise antigenic structure of sperm-whale myoglobin (Mb) has enabled us to focus our attention on the molecular and cellular factors that control and regulate the immune responses to the protein antigen. Our studies have shown that the immune responses to sperm-whale Mb are controlled by genes in the I region of the major histocompatibility complex (H-2) of mice. More importantly, the responses to the synthetic antigenic sites are each under separate genetic control. The recognition of the antigenic sites by antibodies is independent of the immunized species and of the time the antisera are obtained after the initial immunization (from nine days up to a year). The same sites are recognized by antisera raised in rabbit, goat, pig, cat, chicken and outbred and inbred mice. The same sites recognized by mouse B-cells are also recognized by mouse T-cells. No meaningful genetic control of antibody affinity was observed. Autoimmune antibody and T-lymphocyte proliferative responses were readily generated by immunizing an animal with self-Mb. With mouse Mb, the autoimmune T-lymphocyte response was under genetic control and mapped with the I-A and the H-2D end of the H-2 gene complex. In other recent studies we have shown, using several Mb variants, that the binding capacity of an antigenic site is fully accounted for by substitutions in the antigenic sites (actual contact residues) and in residues close (within 7.A) to the sites (i.e. environmental residues). The overall response to Mb is regulated by inter-site influences which can either be of a cooperative (help) nature or of a suppressive nature. Finally, genetic control of the responses to individual antigenic sites on a protein is not only determined by the genetic constitution of the host but also by the chemical properties of the individual sites. The H-2 subregions mapping the responses to given antigenic sites can also recognize other sites, which were previously unrecognizable in a homologous protein, if the chemical properties of these sites are suitably altered.

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PubMed1994

Immunocytochemical demonstration of astrocytes and microglia in the whale brain.

Whale brains have attracted the attention of neuroscientists but there are only sparse studies on whale glial cells. Here we report on immunolabeling of astrocytes by antibodies to glial fibrillary acidic protein (GFAP) or protein S-100 beta (both by the streptavidin/biotin technique), and labeling of microglial cells by Griffonia simplicifolia agglutinin (GSA I-B4, coupled to horseradish peroxidase), in the neocortex of a harbour porpoise Phocoena phocoena L. Many subpial and perivascular astrocytes were stained; they differed greatly in thickness and length of their processes. Subpial astrocytes were coarse with a few stout stem processes, whereas perivascular astrocytes deeper in the brain had many long and slender processes. Additionally, some long radial astrocytes were observed. Microglia were labeled throughout the brain, and showed similar features as 'resting' (ramified) microglia in the brain of other mammals.

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PubMed1997

Extractable organohalogens in tissues of beluga whales from the Canadian Arctic and the St. Lawrence estuary.

Samples of blubber, brain, kidney, liver, and muscle of individual beluga whales were extracted and analyzed for extractable organic Cl (EOCl), Br (EOBr), and I (EOI) by neutron activation analysis. The highest levels of EOCl (554 microg g(-1) lipid) were found in the kidney of Arctic beluga and the lowest (24 microg g(-1) lipid) in the blubber. Levels in brain and liver lipids were on average of equal magnitude. The unequal distribution of EOCl in tissue lipids is in marked contrast to the equal distribution, in lipid, demonstrated for organochlorines such as DDTs and PCBs by a number of investigations. The distribution of brominated (EOBr) and iodinated (EOI) compounds was similar and markedly different from that of EOCl. The tissue distribution of EOCl seems to be related to the proportion to polar lipids in the tissues. Beluga from St. Lawrence had significantly higher brain EOCl levels than the Arctic samples whereas the levels of EOCl in liver and kidney were higher in the Arctic whales.

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PubMed2000

Distribution of mercury in the organs and tissues of five toothed-whale species of the Mediterranean.

Mercury levels were determined in the tissues and organs (lung, liver, kidney, skin, muscle, bone) of five toothed-whales stranded along the Corsican coast between November 1993 and February 1996. The species taken into consideration were the bottlenose dolphin Tursiops truncatus, the common dolphin Delphinus delphis, the striped dolphin Stenella coeruleoalba, the pilot whale Globicephala melas and the Risso's dolphin Grampus griseus. The variation in mercury levels between the different tissues and organs (lung, liver, kidney, skin, muscle, bone) of the cetacean species are discussed as regards storage, biotransformation and elimination. In all cases, the liver appears to be the preferential organ for mercury accumulation (with concentrations as high as 4250 microg Hg/g dw and 3298 microg Hg/g in the livers of Tursiops truncatus and Grampus griseus, respectively). The kidney and lung are the next organs in terms of mercury uptake followed by the muscle, bone and skin. The stomach contents of Grampus griseus and D. delphis were determined and consisted of cephalopods for Grampus griseus, and of sardines Sardina pilchardus and mackerels Trachurus sp. for D. delphis. Cephalopods had higher mercury concentrations (25.4 microg Hg/g dw) than fish (ca 1 microg Hg/g). These contents represent only one meal and mercury levels found in livers may integrate mercury uptake having occurred during the whole life span of animals.

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PubMed2000

Cellular localization of CYP3A proteins in various tissues from pilot whale (Globicephala melas).

The in situ expression of cytochrome P450 3A- (CYP3A) like proteins in hepatic and extrahepatic tissues from a marine mammal, pilot whale (Globicephala melas), was investigated. Polyclonal antibodies (PAb) raised against either rat CYP3A1 or trout CYP3A27 both recognized a microsomal protein band in liver, lung, kidney and heart. The protein band observed in liver and lung had slightly lower molecular weight than that observed in kidney and heart, suggesting the existence of two CYP3A forms in pilot whale. Immunohistochemical analyses showed strong CYP3A-staining in hepatocytes, bile duct epithelial cells, bronchial epithelial cells, in primordial- and primary follicles and their surrounding zona glomerulosa. Moderate to strong CYP3A staining was seen in smooth muscle-like cells of large arteries and arterioles in all organs examined. Mild to moderate staining was evident in alveolar epithelial cells and in kidney tubular epithelial cells. Weak staining was seen in glomerular epithelial cells and in seminiferous tubular epithelial cells.

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PubMed1965

The immunological activity of some of the chymotryptic peptides of sperm-whale myoglobin.

1. Sperm-whale apomyoglobin was digested with chymotrypsin in a dialysis sac. The ultrafiltrate contained incompletely hydrolysed fragments which partially inhibited the precipitation of metmyoglobin and apomyoglobin by some antisera produced against metmyoglobin. The inhibitory activity was stable to heating at 100 degrees and depended on the peptide structure. 2. The fragments were fractionated according to molecular size and were purified by ion-exchange chromatography. Six pure peptides and two peptides which contained a minor impurity were isolated. Their amino acid compositions and N-terminal amino acid sequences were determined and their entire amino acid sequences deduced from the known amino acid sequence of sperm-whale myoglobin. 3. The peptides formed no detectable precipitates with the antisera. Five of the eight peptides partially inhibited the precipitation of apomyoglobin and/or metmyoglobin by one antiserum. Six of the peptides inhibited the precipitation of apomyoglobin by one or other of two antisera; at least two of these peptides inhibited both antisera. One peptide failed to inhibit the precipitation of either antigen by either antiserum. Two of the peptides possessed the same serological specificity. 4. The molar ratios of inhibitors to antigen for 50% of the maximum inhibition decreased as the molecular size of the inhibitor increased. With one antiserum and with apomyoglobin as the antigen, molar ratios 12 and 80 were obtained for peptides with molecular weights 2051 and 793 respectively. 5. The size and structure of an antigenic site is discussed in relation to the known steric configuration of myoglobin.

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PubMed1966

Absorption spectra of sperm-whale ferrimyoglobin.

1. Sperm-whale ferrimyoglobin was found to contain 0.308% of Fe, on a dry weight basis, corresponding to a molecular weight of 18130. The solid takes up moisture to an equilibrium state, and was then assayed to contain 0.280% of Fe. 2. Absorption spectra are presented for acidic ferrimyoglobin, Fe(+)(H(2)O), and its conjugate base, Fe-OH, as well as for the fluoride and cyanide complexes, within the range 200-2500mmu. Data for ferromyoglobin-carbon monoxide, Fe(II)-CO, in the visible region are also included. 3. Minor spectral differences have been found between whale and horse myoglobins, particularly in the effect of temperature on the visible-absorption spectrum of Fe-OH.

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PubMed2000

Population structure of long-finned pilot whales in the North Atlantic: a correlation with sea surface temperature?

The long-finned pilot whale, Globicephala melas, is a social, pelagic odontocete distributed widely in the cold temperate waters of the North Atlantic. Despite genetic, morphometric, physiological and observational studies, it remains unclear whether any population substructure exists. We have used eight highly polymorphic microsatellite loci to analyse samples from four disparate sampling sites: USA East Coast (Cape Cod), West Greenland, the Faeroe Islands and the UK. Our results indicate that substructure does exist, and is particularly pronounced between West Greenland and other sites. The magnitudes of the various pairwise comparisons do not support a simple isolation-by-distance model. Instead, the patterns of genetic differentiation suggest that population isolation occurs between areas of the ocean which differ in sea surface temperature. Such a mechanism is supported by the observation that temperature is a primary factor determining the relative distributions of two short-finned pilot whale (G. macrorhynchus) populations off the Pacific coast of Japan.

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