Chimaeras Research Articles

Intrinsically Disordered Flanking Regions Increase the Affinity of a Transcriptional Coactivator Interaction across Vertebrates.

Interactions between two proteins are often mediated by a disordered region in one protein binding to a groove in a folded interaction domain in the other one. While the main determinants of a certain interaction are typically found within a well-defined binding interface involving the groove, recent studies show that nonspecific contacts by flanking regions may increase the affinity. One example is the coupled binding and folding underlying the interaction between the two transcriptional coactivators NCOA3 (ACTR) and CBP, where the flanking regions of an intrinsically disordered region in human NCOA3 increases the affinity for CBP. However, it is not clear whether this flanking region-mediated effect is a peculiarity of this single protein interaction or if it is of functional relevance in a broader context. To further assess the role of flanking regions in the interaction between NCOA3 and CBP, we analyzed the interaction across orthologs and paralogs (NCOA1, 2, and 3) in human, zebra fish, and ghost shark. We found that flanking regions increased the affinity 2- to 9-fold in the six interactions tested. Conservation of the amino acid sequence is a strong indicator of function. Analogously, the observed conservation of increased affinity provided by flanking regions, accompanied by moderate sequence conservation, suggests that flanking regions may be under selection to promote the affinity between NCOA transcriptional coregulators and CBP.

Sixth sense in the deep-sea: the electrosensory system in ghost shark Chimaera monstrosa

Animals that continually live in deep sea habitats face unique challenges and require adaptive specializations solutions in order to locate and identify food, predators, and conspecifics. The Ampullae of Lorenzini are specialized electroreceptors used by chondrichthyans for important biological functions. Ampullary organs of the ghost shark Chimaera monstrosa, a deep-sea species commonly captured as by-catch in the bottom trawl fishery, are here described for the first time using macroscopic, ultrastructural and histological approaches. The number of ampullary pores in C. monstrosa is about 700, distributed into the whole cephalic section of C. monstrosa, and organized in12 pore clusters and they are arranged into different configurations and form a distinct morphological pattern for this species, showing some anatomical peculiarities never described before in others cartilaginous fishes and may constitute an evolutionary adaptation of this ancient chondrichthyan species to the extreme environmental conditions of its deep sea niche.

Leopard Seals (Hydrurga leptonyx) in New Zealand Waters Predating on Chondrichthyans

Leopard seals (Hydrurga leptonyx) are top-order predators that prey on a wide variety of species including crustaceans, cephalopods, fishes, birds, and pinnipeds. While multiple diet studies have been conducted worldwide, there are no previous accounts of leopard seals predating on chondrichthyans. As part of a wider study on the diet of leopard seals in New Zealand (NZ) waters, researchers and citizen scientists recorded 39 observations of predation and collected 127 scats (166 total records) between 1942 and 2019. Predation on chondrichthyans was detected in 23.1% (n = 9) of observations of predation and 7.1% (n = 9) of scats (the latter via morphological examination and DNA sequencing). From both observations of predations and scats, three chondrichthyan species or genus were identified; elephantfish (Callorhinchus milii), ghost sharks (Hydrolagus spp.) and spiny dogfish (Squalus acanthias). While this is the first published record of leopard seals feeding on chondrichthyans, the relatively high frequency of occurrence within our NZ records, and that certain individuals appeared to target this type of prey, indicates that these species could constitute a substantial, or important, part of the diet for some leopard seals in this region. As chondrichthyans form an important part of the NZ marine ecosystems, our recognition of an additional top-order predator of these species contributes to understanding the overall health of, and future impacts of predators on, the wider NZ marine ecosystem.

Mycobacterium chimaera genomics with regard to epidemiological and clinical investigations conducted for the open-chest post-surgical Mycobacterium chimaera infections outbreak

Post-surgical infections due to Mycobacterium chimaera appeared as a novel nosocomial threat in 2015, with a worldwide outbreak due to contaminated heater-cooler units used in open chest surgery. We report the results of investigations conducted in France including whole genome sequencing comparison of patient and HCU isolates. Methods We sought M. chimaera infection cases from 2010 onwards through national epidemiological investigations in healthcare facilities performing cardiopulmonary bypass together with a survey on good practices and systematic heater-cooler unit microbial analyses. Clinical and HCU isolates were subjected to whole genome sequencing analyzed with regards to the reference outbreak strain Zuerich-1. Results Only two clinical cases were shown to be related to the outbreak, although 23% (41/175) heater-cooler units were declared positive for M. avium complex. Specific measures to prevent infection were applied in 89% (50/56) healthcare facilities although only 14% (8/56) of them followed the manufacturer maintenance recommendations. Whole genome sequencing comparison showed that the clinical isolates and 72% (26/36) of heater-cooler unit isolates belonged to the epidemic cluster. Within clinical isolates, 5 to 9 non-synonymous single nucleotide polymorphisms were observed, among which an in vivo mutation in a putative efflux pump gene observed in a clinical isolate obtained for one patient under antimicrobial treatment. Conclusions Cases of post-surgical M. chimaera infections were declared to be rare in France, although heater-cooler units were contaminated as in other countries. Genomic analyses confirmed the connection to the outbreak and identified specific single nucleotide polymorphisms, including one suggesting fitness evolution in vivo.

Ghosts of the deep – Biodiversity, fisheries, and extinction risk of ghost sharks

AbstractGhost sharks (subclass Holocephali) remain a largely data‐poor group of cartilaginous fishes. The general paucity of attention may partially be related to identification and unresolved taxonomic issues, occurrence in the deep oceans, and their low value and interest in fisheries (which some notable exceptions). Here, we synthesize and assess the extinction risk of all known extant ghost sharks (52 species) by applying the IUCN Red List of Threatened Species Categories and Criteria. Ghost sharks have a low proportion of threatened (8%) and Near Threatened (8%) species, with most species (69%) assessed as Least Concern. The group still exhibits some data deficiency (15%), and biological information is lacking for most species. Endemism is high, with 37% of species known from only one location or one country. Species richness was highest in the Northeast Atlantic, off the northwest coast of Africa (Morocco to Mauritania), the East China Sea, New Zealand and off the northwest coast of South America (Ecuador and Peru). Ghost sharks are predominately taken as by‐catch, but some targeted fishing and/or retention for the liver oil trade occurs. Species‐specific reporting, monitoring and management are required to assess population trends, and further investigation is needed on trade and use, particularly for higher risk species including the sicklefin chimaeras (genusNeoharriotta) and the American Elephantfish (Callorhynchus callorhynchus, Callorhinidae).

Species composition and size distribution of sharks and rays caught in Bali Strait and its surrounding area and its relation to fisheries management

Muncar is one of the shark production centers in Java, where the fishing ground in Bali Strait and its surrounding area. Routine data collection was carried out from May 2018 to April 2019 at the Brak Fish Market and the Muncar Coastal Fishing Port (PPP), Banyuwangi to determine the species composition and size distribution of sharks and rays caught in Bali Strait and its surrounding area. Data collection was done by census and almost of all individuals were identified to species level and and measured. During the study, 3.551 individuals of sharks and rays were landed in this port, consisting of 75 species from 25 families (48 species of sharks, one species of ghost shark and 26 rays species). The most common types of sharks belong to Carcharhinidae and for the rays was Dasyatidae. Among these, thirteen species were listed in Appendix II CITES, i.e. Carcharhinus falciformis, Alopias pelagicus, A. superciliosus, Isurus oxyrhinchus, I. paucus, Sphyrna lewini, S. zygaena, Mobula mobular, M. tarapacana, M. thrustoni, Glaucostegus typus, Rhynchobatus australiae and Rhyna ancylostoma. Most of sharks and rays caught were juvenile to adolescent and at immature or maturing stages.

Calicotyle hydrolagi n. sp. (Monogenea: Monocotylidae) infecting the deep-sea Eastern Pacific black ghost shark Hydrolagus melanophasma from the Atacama Trench, with comments on host specificity of Calicotyle spp.

We describe Calicotyle hydrolagi n. sp. (Monogenea: Monocotylidae) infecting the cloaca of deep-water Eastern Pacific black ghost sharks, Hydrolagus melanophasma captured as bycatch at a local fishery for Patagonian toothfish Dissostichus eleginoides, (Nototheniidae) in the Atacama Trench using morphological and nucleotide (LSU rRNA and SSU rRNA) data. This new species is differentiated from its congeners by a number of characters, including the absence of a cecal diverticula, the size and shape of the male copulatory organ and the shape of the vagina, as well as by differences in molecular data (SSU rRNA and LSU rRNA). The suitability of some sclerotized structures such as the male copulatory organ (MCO) as a taxonomic character is discussed; specifically, we found that the relationship between MCO and total length exhibit different trends in members of Calicotyle isolated from sharks, skates and chimaeras. Additional efforts to obtain sample of Calicotyle species and further molecular studies based on ribosomal and mitochondrial genes are necessary to clarify the degree of host specificity in this genus. Additionally, this is the first report of a member of Calicotyle to be reported in the Southeastern Pacific Ocean.

Further studies of ion channels in the electroreceptor of the skate through deep sequencing, cloning and cross species comparisons

Our comparative studies seek to understand the structure and function of ion channels in cartilaginous fish that can detect very low voltage gradients in seawater. The principal channels of the electroreceptor include a calcium activated K channel whose α subunit is Kcnma1, and a voltage-dependent calcium channel, Cacna1d. It has also been suggested based on physiological and pharmacological evidence that a voltage-gated K channel is present in the basal membranes of the receptor cells which modulates synaptic transmitter release. Large conductance calcium-activated K channels (BK) are comprised of four α subunits, encoded by Kcnma1 and modulatory β subunits of the Kcnmb class. We recently cloned and published the skate Kcnma1 gene and most of Kcnmb4 using purified mRNA of homogenized electroreceptors. Bellono et al. have recently performed RNA sequencing (RNA-seq) on purified mRNA from skate electroreceptors and found several ion channels including Kcnma1. We searched the Bellono et al. RNA-seq repository for additional channels and subunits. Our most significant findings are the presence of two Shaker type voltage dependent K channel sequences which are grouped together as isoforms in the data repository. The larger of these is a skate ortholog of the voltage dependent fast potassium channel Kv1.1, which is expressed at appreciable levels. The second ortholog is similar to Kv1.5 but has fewer N-terminal amino acids than other species. The sequence for Kv1.5 in the skate is very strongly aligned with the recently reported sequence for potassium channels in the electroreceptors of the cat shark, S. retifer, which also modulate synaptic transmission. The latter channel was designated as Kv1.3 in the initial report, but we suggest that these channels are actually orthologs of each other, and that Kv1.5 is the prevailing designation. We also found a beta subunit sequence (Kcnab2) which may co-assemble with one or both of the voltage gated channels. The new channels and subunits were verified by RT-PCR and the Kv1.1 sequence was confirmed by cloning. We also searched the RNA-seq repository for accessory subunits of Kcnma1, and found a computer-generated assembly that contained a complete sequence of its β subunit, Kcnmb2. Skate Kcnmb2 has a total of 279 amino acids, with 51 novel amino acids at the N-terminus which may play a specific physiological role. This sequence was confirmed by PCR and cloning. However, skate Kcnmb2 is expressed at low levels in the electroreceptor compared to Kcnma1 and skate Kcnmb1 is absent. The evolutionary origin of the newly described K channels and their subunits was studied by alignments with mammalian sequences, including human, and also those in related fish: the whale shark (R. typus), the ghost shark (C.milii), and (S. retifer). There are also orthologous K channels of the lamprey, which has electroreceptors. Tree building and bootstrap programs were used to confirm phylogenetic inferences. Further research should focus on the subcellular locations of these channels, their gating behavior, and the effects of accessory subunits on gating.

Preliminary analysis of New Zealand scampi (Metanephrops challengeri) diet using metabarcoding.

Deep sea lobsters are highly valued for seafood and provide the basis of important commercial fisheries in many parts of the world. Despite their economic significance, relatively little is known about their natural diets. Microscopic analyses of foregut content in some species have suffered from low taxonomic resolution, with many of the dietary items difficult to reliably identify as their tissue is easily digested. DNA metabarcoding has the potential to provide greater taxonomic resolution of the diet of the New Zealand scampi (Metanephrops challengeri) through the identification of gut contents, but a number of methodological concerns need to be overcome first to ensure optimum DNA metabarcoding results. In this study, a range of methodological parameters were tested to determine the optimum protocols for DNA metabarcoding, and provide a first view of M. challengeri diet. Several PCR protocols were tested, using two universal primer pairs targeting the 18S rRNA and COI genes, on DNA extracted from both frozen and ethanol preserved samples for both foregut and hindgut digesta. The selection of appropriate DNA polymerases, buffers and methods for reducing PCR inhibitors (including the use of BSA) were found to be critical. Amplification from frozen or ethanol preserved gut contents appeared similarly dependable. The COI gene was found to be more effective than 18S rRNA gene for identifying large eukaryotic taxa from the digesta; however, it was less successfully amplified. The 18S rRNA gene was more easily amplified, but identified mostly smaller marine organisms such as plankton and parasites. This preliminary analysis of the diet of M. challengeri identified a range of species (13,541 reads identified as diet), which included the ghost shark (Hydrolagus novaezealandiae), silver warehou (Seriolella punctata), tall sea pen (Funiculina quadrangularis) and the salp (Ihlea racovitzai), suggesting that they have a varied diet, with a high reliance on scavenging a diverse range of pelagic and benthic species from the seafloor.

Assessing ecological and molecular divergence between the closely related species Hydrolagus pallidus and H. affinis (Chimaeridae).

This study investigated taxonomic validity of the pale ghost shark Hydrolagus pallidus Hardy & Stehmann, 1990, which was described as a species distinct from the smalleyed rabbitfish H. affinis (de Brito Capello 1868). While few morphological characters distinguish the two taxa, a striking difference in sex ratio and fixed differences (1·1-1·6% divergence) in the cytochrome oxidase subunit I barcoding gene support the recognition of both species.

Molecular assisted identification of sharks, rays and chimaeras at landing sites in India.

Molecular assisted identification of sharks, rays and chimaeras at landing sites in India.

Illusory fears must not stifle chimaera research.

Illusory fears must not stifle chimaera research.

Searching the Evolutionary Origin of Epithelial Mucus Protein Components-Mucins and FCGBP.

The gel-forming mucins are large glycosylated proteins that are essential components of the mucus layers covering epithelial cells. Using novel methods of identifying mucins based on profile hidden Markov models, we have found a large number of such proteins in Metazoa, aiding in their classification and allowing evolutionary studies. Most vertebrates have 5–6 gel-forming mucin genes and the genomic arrangement of these genes is well conserved throughout vertebrates. An exception is the frog Xenopus tropicalis with an expanded repertoire of at least 26 mucins of this type. Furthermore, we found that the ovomucin protein, originally identified in chicken, is characteristic of reptiles, birds, and amphibians. Muc6 is absent in teleost fish, but we now show that it is present in animals such as ghost sharks, demonstrating an early origin in vertebrate evolution. Public RNA-Seq data were analyzed with respect to mucins in zebrafish, frog, and chicken, thus allowing comparison in regard of tissue and developmental specificity. Analyses of invertebrate proteins reveal that gel-forming-mucin type of proteins is widely distributed also in this group. Their presence in Cnidaria, Porifera, and in Ctenophora (comb jellies) shows that these proteins were present early in metazoan evolution. Finally, we examined the evolution of the FCGBP protein, abundant in mucus and related to gel-forming mucins in terms of structure and localization. We demonstrate that FCGBP, ubiquitous in vertebrates, has a conserved N-terminal domain. Interestingly, this domain is also present as an N-terminal sequence in a number of bacterial proteins.

Calcium activated K+ channels in the electroreceptor of the skate confirmed by cloning. Details of subunits and splicing

Elasmobranchs detect small potentials using excitable cells of the ampulla of Lorenzini which have calcium-activated K+ channels, first described in 1974. A distinctive feature of the outward current in voltage clamped ampullae is its apparent insensitivity to voltage. The sequence of a BK channel α isoform expressed in the ampulla of the skate was characterized. A signal peptide is present at the beginning of the gene. When compared to human isoform 1 (the canonical sequence), the largest difference was absence of a 59 amino acid region from the S8–S9 intra-cellular linker that contains the strex regulatory domain. The ampulla isoform was also compared with the isoform predicted in late skate embryos where strex was also absent. The BK voltage sensors were conserved in both skate isoforms. Differences between the skate and human BK channel included alternative splicing. Alternative splicing occurs at seven previously defined sites that are characteristic for BK channels in general and hair cells in particular. Skate BK sequences were highly similar to the Australian ghost shark and several other vertebrate species. Based on alignment of known BK sequences with the skate genome and transcriptome, there are at least two isoforms of Kcnma1α expressed in the skate. One of the β subunits (β4), which is known to decrease voltage sensitivity, was also identified in the skate genome and transcriptome and in the ampulla. These studies advance our knowledge of BK channels and suggest further studies in the ampulla and other excitable tissues.

The Evolution of the Scavenger Receptor Cysteine-Rich Domain of the Class A Scavenger Receptors.

The class A scavenger receptor (cA-SR) family is a group of five evolutionarily related innate immune receptors. The cA-SRs are known for their promiscuous ligand binding; as they have been shown to bind bacteria, such as Streptococcus pneumoniae and Escherichia coli, as well as different modified forms of low-density lipoprotein. Three of the five family members possess a scavenger receptor cysteine-rich (SRCR) domain while the remaining two receptors lack the domain. Previous work has suggested that the macrophage-associated receptor with collagenous structure (MARCO) shares a recent common ancestor with the non-SRCR-containing receptors; however, the origin of the SRCR domain within the cA-SRs remains unknown. We hypothesize that the SRCR domains of the cA-SRs have a common origin that predates teleost fish. Using the newly available sequence data from sea lamprey and ghost shark genome projects, we have shown that MARCO shares a common ancestor with the SRCR-containing proteins. In addition, we explored the evolutionary relationships within the SRCR domain by reconstructing the ancestral SRCR domains of the cA-SRs. We identified a motif that is highly conserved between the cA-SR SRCR domains and the ancestral SRCR domain that consist of WGTVCDD. We also show that the GRAEVYY motif, a functionally important motif within MARCO, is poorly conserved in the other cA-SRs and in the reconstructed ancestral domain. Further, we identified three sites within MARCO’s SRCR domain, which are under positive selection. Two of these sites lie adjacent to the conserved WGTVCDD motif, and may indicate a potential biological function for these sites. Together, these findings indicate a common origin of the SRCR domain within the cA-SRs; however, different selective pressures between the proteins may have caused MARCOs SRCR domain to evolve to contain different functional motifs when compared to the other SRCR-containing cA-SRs.

A case of sectorial chimaera obtained after irradiation of Petunia seed

A case of sectorial chimaera obtained after irradiation of Petunia seed

SHARKS AND RAYS IN INDONESIA

Indonesia is considered to have a high diversity of sharks and rays due in part to its position in the equatorial line, between two continents and two oceans. According to recent studies and various publications, the number of chondrichthyan species known to date is 207 species of 44 families, comprised of 109 species of sharks, 96 species of batoids, and two species of ghost sharks (chimaera). The Dasyatidae (stingrays) is the most speciose family of chondrichthyans occurring in Indonesia, followed by the Carcharhinidae (whaler sharks). Most species of elasmobranchs commonly occur on the continental or insular shelves and rays are more commonly found than sharks. This type of habitat can be found mostly in the western part of Indonesia (the Java Sea, Karimata Strait, South China Sea, Malacca Strait, and Macassar Strait). Due to the separation by the Wallace Line, there are some differences in species diversity between western and eastern region. The shark and ray fauna of the eastern region is more similar to the Australian fauna, while the fauna in the western region is more similar to the Asian fauna. There are also some endemic species known to occur in Indonesian waters. The endemicity of some species can be restricted in a certain area or in a regional area such as the Indo-West Pacific.

First record of Hydrolagus melanophasma James, Ebert, Long & Didier, 2009 (Chondrichthyes, Chimaeriformes, Holocephali) from the southeastern Pacific Ocean

The eastern Pacific black ghost shark, Hydrolagus melanophasma is reported from deep waters off Chile and is described from specimens collected off Valdivia at depths of 1150 to 1720 m. This species is distinguished from all other members of the genus by its large, curved dorsal fin spine, which extends beyond the dorsal fin apex; a second dorsal fin of uniform height along the caudal peduncle; large pectoral fins extending beyond the pelvic fin insertion; and a uniform black color of the body. Catch records of this species in Chile suggest a discontinuous distribution along the continental slope of the eastern Pacific Ocean, restricted to the Middle America and Atacama trenches. These observations elevate the number of chimaeroid fishes inhabiting Chile to five species.

Phylogenetic analysis reveals wide distribution of globin X

The vertebrate globin gene repertoire consists of seven members that differ in terms of structure, function and phyletic distribution. While hemoglobin, myoglobin, cytoglobin, and neuroglobin are present in almost all gnathostomes examined so far, other globin genes, like globin X, are much more restricted in their phyletic distribution. Till today, globin X has only been found in teleost fish and Xenopus. Here, we report that globin X is also present in the genomes of the sea lamprey, ghost shark and reptiles. Moreover, the identification of orthologs of globin X in crustacean, insects, platyhelminthes, and hemichordates confirms its ancient origin.

Glial architecture of the ghost shark (Callorhinchus milii, Holocephali, Chondrichthyes) as revealed by different immunohistochemical markers

This article presents the first study on the glial architecture of a representative species of Holocephali, Callorhinchus milii (ghost shark). Holocephali are a small subclass of Chondrichthyes, with only a few extant genera, and those are considered to have a brain organization more similar to squalomorph sharks than to galeomorph sharks, skates, and rays. Three different astroglial markers--glial fibrillary acidic protein, S-100 protein, and glutamine synthetase (GS)--were investigated by immunohistochemical methods, applying both diaminobenzidine (DAB) and fluorescent techniques. They revealed similar glial structures, although most of them were detected by immunohistochemical reaction against GS and visualized by DAB. The predominant elements were radial ependymoglia spanning the area between the ventricular and meningeal surfaces, as in squalomorph sharks. Other similar features were the light appearance of myelinated neural tracts devoid of immunoreactivity, and the glial architecture of the reticular formation of the brain stem, cerebellum, and tectum, the latter with recognizable layers. The immunoreactivity of the vascular walls was similar; however, it is believed that different cell types form the blood-brain barrier in chimeras and in elasmobranchs. Some glial structures, however, resembled those of skates, rays, and galeomorph sharks. In C. milii astrocyte-like elements were observed in the telencephalon, using GS and S-100, although typical astrocyte-rich regions were not found. In some areas, especially the telencephalon, not only endfeet but also cell bodies were observed to be attached to the meningeal surface, with processes extending into the brain substance.