Evolution of variable lymphocyte receptor B antibody loci in jawless vertebrates

Three types of variable lymphocyte receptor (VLR) genes, VLRA, VLRB, and VLRC, encode antigen recognition receptors in the extant jawless vertebrates, lampreys and hagfish. The somatically diversified repertoires of these VLRs are generated by serial stepwise copying of leucine-rich repeat (LRR) sequences into an incomplete germline VLR gene. Lymphocytes that express VLRA or VLRC are T cell–like, while VLRB-expressing cells are B cell–like. Here, we analyze the composition of the VLRB locus in different jawless vertebrates to elucidate its configuration and evolutionary modification. The incomplete germline VLRB genes of two hagfish species contain short noncoding intervening sequences, whereas germline VLRB genes in six representative lamprey species have much longer intervening sequences that exhibit notable genomic variation. Genomic clusters of potential LRR cassette donors, fragments of which are copied to complete VLRB gene assembly, are identified in Japanese lamprey and sea lamprey. In the sea lamprey, 428 LRR cassettes are located in five clusters spread over a total of 1.7 Mbp of chromosomal DNA. Preferential usage of the different donor cassettes for VLRB assemblage is characterized in our analysis, which reveals evolutionary modifications of the lamprey VLRB genes, elucidates the organization of the complex VLRB locus, and provides a comprehensive catalog of donor VLRB cassettes in sea lamprey and Japanese lamprey.

Knowledge Gaps in the Definition of Threats for the Red List Assessment of European Freshwater-Dependent Fish Species

Freshwater ecosystems are disproportionally important for biodiversity conservation, as they support more than 9% of known animal species while representing less than 1% of the Earth’s surface. However, the vast majority of the threats (99%, or 826 out of 837) identified by the International Union for Conservation of Nature Red List of Threatened Species known to affect the 434 known freshwater-dependent fish and lampreys of Europe are not supported by validated published scientific knowledge. This general lack of information about freshwater-dependent fish and lamprey species may have deleterious effects on species conservation, and additional funding is required to fill baseline knowledge gaps.

New Insights into the Distribution, Physiology and Life Histories of South American Galaxiid Fishes, and Potential Threats to This Unique Fauna

South American galaxiids occupy both Patagonia and the ichthyogeographic Chilean Province, encompassing glacial Andean deep lakes, shallow plateau lakes, reservoirs, short Pacific rivers and long Atlantic rivers. The total fish fauna includes 29 species, comprising Neotropical fishes (siluriforms and characids), galaxiids, percichthyids, atherinopsids and mugilids, two lamprey species, and several exotic fishes (salmonids, Gambusia spp. and common carp). The family Galaxiidae shares a common ancestry with the Gondwanan temperate fish fauna, played a major role in the post-glacial colonization of Andean lakes and streams, and contributes key species to the food webs. Galaxiid species occupy an enormous latitudinal gradient, show a wide variety of life history patterns and are the southernmost native freshwater fishes of the world. Knowledge of South American galaxiids has improved notably, but new challenges arise due to climate change, biological invasions, damming, aquaculture and contamination. In this changing environment, the future of South American galaxiids should be carefully considered as a legacy of the old Gondwana and a unique attribute of the freshwaters of southern South America.

Hearing capabilities and behavioural response of sea lamprey (Petromyzon marinus) to low-frequency sounds

Hearing ability is well studied across teleost fishes in general, and vertebrates more broadly, but little is known about sound detection abilities of lampreys (Petromyzontiformes), a basal extant vertebrate group. The sea lamprey (Petromyzon marinus) is a destructive invader of the Laurentian Great Lakes, while numerous lamprey species (including the sea lamprey) are imperiled in their native ranges. In both management scenarios, behavioral manipulation tactics to control movement and distribution are desired. Therefore, we describe the hearing ability and behavioral responses of adult and juvenile sea lamprey to sound to reveal how hearing may have evolved in vertebrates and determine possible management applications. Based on auditory evoked potentials, sea lamprey detected tones of 50–300 Hz with equal sensitivity, but did not detect sounds above 300 Hz. In a laboratory bioassay, sea lamprey behaviorally responded to sound range of 50–200 Hz, with a general increase in swimming and a decrease in resting behaviours at both juvenile and adult stages relative to no-sound controls. To our knowledge, this is the first test of lamprey hearing, and the results support that sound may be a means to modify lamprey behaviour for management purposes.

Expansions, diversification, and interindividual copy number variations of AID/APOBEC family cytidine deaminase genes in lampreys

Cytidine deaminases of the AID/APOBEC family catalyze C-to-U nucleotide transitions in mRNA or DNA. Members of the APOBEC3 branch are involved in antiviral defense, whereas AID contributes to diversification of antibody repertoires in jawed vertebrates via somatic hypermutation, gene conversion, and class switch recombination. In the extant jawless vertebrate, the lamprey, two members of the AID/APOBEC family are implicated in the generation of somatic diversity of the variable lymphocyte receptors (VLRs). Expression studies linked CDA1 and CDA2 genes to the assembly of VLRA/C genes in T-like cells and the VLRB genes in B-like cells, respectively. Here, we identify and characterize several CDA1-like genes in the larvae of different lamprey species and demonstrate that these encode active cytidine deaminases. Structural comparisons of the CDA1 variants highlighted substantial differences in surface charge; this observation is supported by our finding that the enzymes require different conditions and substrates for optimal activity in vitro. Strikingly, we also found that the number of CDA-like genes present in individuals of the same species is variable. Nevertheless, irrespective of the number of different CDA1-like genes present, all lamprey larvae have at least one functional CDA1-related gene encoding an enzyme with predicted structural and chemical features generally comparable to jawed vertebrate AID. Our findings suggest that, similar to APOBEC3 branch expansion in jawed vertebrates, the AID/APOBEC family has undergone substantial diversification in lamprey, possibly indicative of multiple distinct biological roles.

First observations of spawning nests in the pouched lamprey (Geotria australis)

The pouched lamprey (Geotria australis), one of four Southern Hemisphere lamprey species, is New Zealand’s only freshwater representative of the agnathans. In contrast to that of Northern Hemisphere lampreys, the reproductive ecology of Southern Hemisphere lampreys is poorly understood, with no documented nesting sites or spawning behaviours. In the present study, we utilized passive integrated transponder tags to track migratory adult G. australis to locate spawning sites. Across 2 years, six cryptic nesting sites were discovered, each containing eggs and a single male and female lamprey. The postspawning pairs were all located underneath large boulders, with the eggs forming a coagulated cluster that adhered to the underside of the boulder. Both the male and female lamprey survived spawning for over 105 days. The observed spawning nests highlight a substantial divergence in the reproductive behaviour between Northern and Southern Hemisphere lampreys, as the present observations of G. australis are the first that support clustered cryptic egg deposition and an extended time to mortality postspawning. This study offers the first insights to the long-standing mystery of Southern Hemisphere lamprey reproduction.

Voltage-gated sodium channel gene repertoire of lampreys: gene duplications, tissue-specific expression and discovery of a long-lost gene

Studies of the voltage-gated sodium (Nav) channels of extant gnathostomes have made it possible to deduce that ancestral gnathostomes possessed four voltage-gated sodium channel genes derived from a single ancestral chordate gene following two rounds of genome duplication early in vertebrates. We investigated the Nav gene family in two species of lampreys (the Japanese lamprey Lethenteron japonicum and sea lamprey Petromyzon marinus ) (jawless vertebrates—agnatha) and compared them with those of basal vertebrates to better understand the origin of Nav genes in vertebrates. We noted six Nav genes in both lamprey species, but orthology with gnathostome (jawed vertebrate) channels was inconclusive. Surprisingly, the Nav2 gene, ubiquitously found in invertebrates and believed to have been lost in vertebrates, is present in lampreys, elephant shark ( Callorhinchus milii ) and coelacanth ( Latimeria chalumnae ). Despite repeated duplication of the Nav1 family in vertebrates, Nav2 is only in single copy in those vertebrates in which it is retained, and was independently lost in ray-finned fishes and tetrapods. Of the other five Nav channel genes, most were expressed in brain, one in brain and heart, and one exclusively in skeletal muscle. Invertebrates do not express Nav channel genes in muscle. Thus, early in the vertebrate lineage Nav channels began to diversify and different genes began to express in heart and muscle.

Evolution of the α2-adrenoreceptors in vertebrates: ADRA2D is absent in mammals and crocodiles

Evolutionary studies of genes that have been functionally characterized and whose variation has been associated with pathological conditions represent an opportunity to understand the genetic basis of pathologies. α2-adrenoreceptors (ADRA2) are a class of G protein-coupled receptors that regulate several physiological processes including blood pressure, platelet aggregation, insulin secretion, lipolysis, and neurotransmitter release. This gene family has been extensively studied from a molecular/physiological perspective, yet much less is known about its evolutionary history. Accordingly, the goal of this study was to investigate the evolutionary history of α2-adrenoreceptors (ADRA2) in vertebrates. Our results show that in addition to the three well-recognized α2-adrenoreceptor genes (ADRA2A, ADRA2B and ADRA2C), we recovered a clade that corresponds to the fourth member of the α2-adrenoreceptor gene family (ADRA2D). We also recovered a clade that possesses two ADRA2 sequences found in two lamprey species. Furthermore, our results show that mammals and crocodiles are characterized by possessing three α2-adrenoreceptor genes, whereas all other vertebrate groups possess the full repertoire of α2-adrenoreceptor genes. Among vertebrates ADRA2D seems to be a dispensable gene, as it was lost two independent times during the evolutionary history of the group. Additionally, we found that most examined species possess the most common alleles described for humans; however, there are cases in which non-human mammals possess the alternative variant.