Spiny-rayed Fishes Research Articles

A classic key innovation constrains oral jaw functional diversification in fishes

Abstract Modifications to the pharyngeal jaws—a prey processing system located posterior to the mouth cavity—are widely considered a key innovation that enhanced diversification within several prominent fish clades. Seen in cichlids, damselfishes, wrasses, and a few other lineages, these musculoskeletal alterations are believed to increase the evolutionary independence and, thus, the diversification of the oral and pharyngeal jaw systems. To test this classic hypothesis, we conducted comparative phylogenetic analyses to assess the effect of the pharyngeal novelty on the diversification of feeding morphology and kinematics across a taxonomically diverse sample of spiny-rayed fishes. We quantified movements of the oral jaws and other craniofacial structures from 689 suction-feeding strikes using high-speed videos collected from 228 species with and without the pharyngeal jaw novelty. Contradicting long-held predictions, we find significantly greater disparity across all traits and faster rates of oral jaw functional evolution in fishes without the specialized prey processing system. The modified pharyngeal jaw is undoubtedly a functional innovation as it enhances the strength of the prey processing system, facilitating exceptional transition rates to feeding on hard and tough prey. However, it also restricts the diversification of the feeding system, revealing that the impact of pharyngognathy is more nuanced than previously thought. In light of these and other recent findings, a reinterpretation of the macroevolutionary consequences of the pharyngeal jaw novelty is needed.

The dwarf neon rainbowfish Melanotaenia praecox, a small spiny-rayed fish with potential as a new Acanthomorpha model fish: I. Fin ray ontogeny and postembryonic staging.

Fish fins with highly variable color patterns and morphologies have many functions. In Actinopterygii, the free parts of fins are supported by "soft rays" and "spiny rays." Spiny rays have various functions and are extremely modified in some species, but they are lacking in popular model fish such as zebrafish and medaka. Additionally, some model fish with spiny rays are difficult to maintain in ordinary laboratory systems. Characteristics of the small, spiny-rayed rainbowfish Melanotaenia praecox render it useful as an experimental model species. Neither fish age nor body size correlate well with fin development during postembryonic development in this species. A four-stage developmental classification is proposed that is based on fin ray development. Melanotaenia praecox is an ideal species to rear in laboratories for developmental studies. Our classification allows for postembryonic staging of this species independent of individual age and body size. Development of each fin ray may be synchronized with dorsal fin development. We discuss the differences in mechanisms regulating soft, spiny, and procurrent ray development.

The dwarf neon rainbowfish Melanotaenia praecox, a small spiny-rayed fish with potential as a new Acanthomorpha model fish: II. Establishment of a microinjection procedure for genetic engineering.

Rainbowfish is a clade of colorful freshwater fish. Melanotaenia praecox is a small rainbowfish species with biological characteristics that make it potentially useful as an experimental model species. We anticipate that M. praecox could become a new model used in various fields, such as ecology, evolution, and developmental biology. However, few previous studies have described experimental set-ups needed to understand the molecular and genetic mechanisms within this species. We describe detailed procedures for genetic engineering in the rainbowfish M. praecox. By using these procedures, we successfully demonstrated CRISPR/Cas-mediated knockout and Tol2 transposon-mediated transgenesis in this species. Regarding the CRISPR/Cas system, we disrupted the tyrosinase gene and then showed that injected embryos lacked pigmentation over much of their body. We also demonstrated that a Tol2 construct, including a GFP gene driven by a ubiquitous promoter, was efficiently integrated into the genome of M. praecox embryos. The establishment of procedures for genetic engineering in M. praecox enables investigation of the genetic mechanisms behind a broad range of biological phenomena in this species. Thus, we suggest that M. praecox can be used as a new model species in various experimental biology fields.

Anatomy of the endocrine pancreas in actinopterygian fishes and its phylogenetic implications

The anatomy and organisation of the endocrine pancreas in ray-finned fishes vary widely. The two main morphoanatomical character states are diffuse versus compact pancreatic tissue. The latter are called Brockmann Bodies (BBs), or principal islets. The present study is the first comprehensive survey on the anatomy of the endocrine pancreas (diffuse versus compact) across 322 actinopterygian species in 39 orders and 135 families based on literature, specimen dissections, and Magnetic Resonance Imaging (MRI). The data obtained show that large endocrine pancreatic islets (BB) have appeared several times in teleost evolution: in some ostariophysian clades and within the Salmoniformes and Neoteleostei. Acanthomorpha (spiny-rayed fishes) is the largest clade of the Neoteleostei. Within this clade, an absence of BBs is only observed in flying fishes (Exocoetidae), parrotfishes (Scarinae), and some of the scarine relatives, the Labridae. The presence of BBs in examined jellynose fish species from the Ateleopodiformes indicates support for its sister-group relationship to the Ctenosquamata (Myctophiformes + Acanthomorpha). More investigations are still needed to corroborate the presence or absence of BBs as a putative synapomorphy for a clade comprising Ateleopodiformes and Ctenosquamata.

The Cretaceous–Paleogene transition in spiny-rayed fishes: surveying “Patterson’s Gap” in the acanthomorph skeletal record

In contrast to the rich collections of articulated spiny-rayed fishes from early Late Cretaceous and Eocene Lagerstätten, similar skeletal remains are sparse in Maastrichtian–Paleocene strata. Here we coin this poorly understood span “Patterson’s Gap” and review known articulated skeletons from it, summarizing available information on their phylogenetic affinities, age, and environmental context. Roughly fifty percent of taxa in both the Maastrichtian and Paleocene come from Europe and North America, with percomorphs representing around 60% of the skeletal acanthomorph taxa in each interval. This is higher than the only pre-Maastrichtian assemblage with a reasonable sample of percomorphs, but lower than most Eocene and younger sites. Fossils from Patterson’s Gap show a steady accumulation of the principal lineages of spiny-rayed fishes. Material from Paleocene or older strata provides evidence for most of the roughly 20 major acanthomorph divisions recovered by molecular studies. Many fossils from Patterson’s Gap remain undescribed and unnamed, and almost none have been included within formal phylogenetic analyses. Revision of existing material, combined with additional fieldwork, should be a priority for future efforts seeking to clarify this murky but significant interval in the evolutionary history of a major vertebrate radiation.

Topological analysis of vertebrate phylogenetic trees utilizing Horton’s first law

Phylogenetic trees visually represent evolution and diversification. While many studies have focused on the number and length of edges (branches), topological properties, such as edge connection patterns, are also important. In this study, the topological properties of phylogenetic trees were quantified, focusing on edge connection patterns. Horton’s first law was applied to quantify the overall, rather than local, topological properties of phylogenetic trees. The topological properties of vertebrate phylogenetic trees for spiny-rayed fishes, Amphibians, turtles, Squamata, Aves, and placental mammals were analyzed. The topological features discussed herein include the number of first-order edges, maximum order, and bifurcation ratio. The average bifurcation ratio of all trees was approximately 3, suggesting that phylogenetic trees for different taxa have a common mechanism of evolution. Vertebrate phylogenetic trees were compared with artificial branching objects created from neutral stochastic branching model simulations. The topological properties of the actual vertebrate phylogenetic trees agreed with those of the artificial branching objects. Our study suggests that evolutionary events do not change the overall topological properties of actual phylogenetic trees, even if the number and length of the edges change. Specifically, non-neutral events (e.g., environmental changes and mass extinction) are not main factors associated with topological properties. The results instead demonstrate a relationship between the bifurcation ratio and symmetricity in the context of temporal changes of topological properties. When the number of first-order edges increased and the maximum order remained constant, the bifurcation ratio increased and symmetricity decreased. When the number of first-order edges increased and the maximum order increased by one, the bifurcation ratio decreased and symmetricity increased.

A reference genome for Bluegill (Centrarchidae: Lepomis macrochirus).

North American sunfishes (Family Centrarchidae) are among the most popular sportfish throughout the United States and Canada. Despite the popularity of sunfishes, their ecological importance, and their extensive stocking and aquacultural history, few molecular studies have examined the evolutionary relationships and species boundaries among members of this group, many of which are known to hybridize. Here, we describe a chromosome-scale genome assembly representing Bluegill (Lepomis macrochirus), one of the most widespread centrarchid species. By combining long-read, Oxford Nanopore sequencing data with short-insert, whole-genome and HiC sequence reads, we produced an assembly (Lm_LA_1.1) having a total length of 889 Mb including 1,841 scaffolds and having a scaffold N50 of 36 Mb, L50 of 12, N90 of 29 Mb, and L90 of 22. We detected 99% (eukaryota_odb10) and 98% (actinopterygii_odb10) universal single-copy orthologs (BUSCOs), and ab initio gene prediction performed using this new assembly identified a set of 17,233 genes that were supported by external (OrthoDB v10) data. This new assembly provides an important addition to the growing set of assemblies already available for spiny-rayed fishes (Acanthomorpha), and it will serve as a resource for future studies that focus on the complex evolutionary history of centrarchids.

Prolonged morphological expansion of spiny-rayed fishes following the end-Cretaceous.

Spiny-rayed fishes (Acanthomorpha) dominate modern marine habitats and account for more than a quarter of all living vertebrate species. Previous time-calibrated phylogenies and patterns from the fossil record explain this dominance by correlating the origin of major acanthomorph lineages with the Cretaceous-Palaeogene mass extinction. Here we infer a time-calibrated phylogeny using ultraconserved elements that samples 91.4% of all acanthomorph families and investigate patterns of body shape disparity. Our results show that acanthomorph lineages steadily accumulated throughout the Cenozoic and underwent a significant expansion of among-clade morphological disparity several million years after the end-Cretaceous. These acanthomorph lineages radiated into and diversified within distinct regions of morphospace that characterize iconic lineages, including fast-swimming open-ocean predators, laterally compressed reef fishes, bottom-dwelling flatfishes, seahorses and pufferfishes. The evolutionary success of spiny-rayed fishes is the culmination of multiple species-rich and phenotypically disparate lineages independently diversifying across the globe under a wide range of ecological conditions.

Spiny and soft-rayed fin domains in acanthomorph fish are established through a BMP-gremlin-shh signaling network

With over 18,000 species, the Acanthomorpha, or spiny-rayed fishes, form the largest and arguably most diverse radiation of vertebrates. One of the key novelties that contributed to their evolutionary success are the spiny rays in their fins that serve as a defense mechanism. We investigated the patterning mechanisms underlying the differentiation of median fin Anlagen into discrete spiny and soft-rayed domains during the ontogeny of the direct-developing cichlid fish Astatotilapia burtoni Distinct transcription factor signatures characterize these two fin domains, whereby mutually exclusive expression of hoxa13a/b with alx4a/b and tbx2b marks the spine to soft-ray boundary. The soft-ray domain is established by BMP inhibition via gremlin1b, which synergizes in the posterior fin with shh secreted from a zone of polarizing activity. Modulation of BMP signaling by chemical inhibition or gremlin1b CRISPR/Cas9 knockout induces homeotic transformations of spines into soft rays and vice versa. The expression of spine and soft-ray genes in nonacanthomorph fins indicates that a combination of exaptation and posterior expansion of an ancestral developmental program for the anterior fin margin allowed the evolution of robustly individuated spiny and soft-rayed domains. We propose that a repeated exaptation of such pattern might underly the convergent evolution of anterior spiny-fin elements across fishes.

Large-bodied sabre-toothed anchovies reveal unanticipated ecological diversity in early Palaeogene teleosts.

Many modern groups of marine fishes first appear in the fossil record during the early Palaeogene (66–40 Ma), including iconic predatory lineages of spiny-rayed fishes that appear to have originated in response to ecological roles left empty after the Cretaceous/Palaeogene extinction. The hypothesis of extinction-mediated ecological release likewise predicts that other fish groups have adopted novel predatory ecologies. Here, we report remarkable trophic innovation in early Palaeogene clupeiforms (herrings and allies), a group whose modern representatives are generally small-bodied planktivores. Two forms, the early Eocene (Ypresian) †Clupeopsis from Belgium and a new genus from the middle Eocene (Lutetian) of Pakistan, bear conspicuous features indicative of predatory ecology, including large size, long gapes and caniniform dentition. Most remarkable is the presence of a single, massive vomerine fang offset from the midline in both. Numerous features of the neurocranium, suspensorium and branchial skeleton place these taxa on the engraulid (anchovy) stem as the earliest known representatives of the clade. The identification of large-bodied, piscivorous anchovies contributes to an emerging picture of a phylogenetically diverse guild of predatory ray-finned fishes in early Palaeogene marine settings, which include completely extinct lineages alongside members of modern marine groups and taxa that are today restricted to freshwater or deep-sea environments.

Feeding ecology underlies the evolution of cichlid jaw mobility.

The fish feeding apparatus is among the most diverse functional systems in vertebrates. While morphological and mechanical variations of feeding systems are well studied, we know far less about the diversity of the motions that they produce. We explored patterns of feeding movements in African cichlids from Lakes Malawi and Tanganyika, asking whether the degree of kinesis is associated with dietary habits of species. We used geometric morphometrics to measure feeding kinesis as trajectories of shape change, based on 326 high-speed videos in 56 species. Cranial morphology was significantly related to feeding movements, both of which were distributed along a dietary axis associated with prey evasiveness. Small-mouthed cichlids that feed by scraping algae and detritus from rocks had low kinesis strikes, while large-mouthed species that eat large, evasive prey (fishes and shrimps) generated the greatest kinesis. Despite having higher overall kinesis, comparisons of trajectory shape (linearity) revealed that cichlids that eat mobile prey also displayed more kinematically conserved, or efficient, feeding motions. Our work indicates that prey evasiveness is strongly related to the evolution of cichlid jaw mobility, suggesting that this same relationship may explain the origins and diversity of highly kinetic jaws that characterize the super-radiation of spiny-rayed fishes.

Histology of the endothermic opah (Lampris sp.) suggests a new structure-function relationship in teleost fish bone.

Endothermy, production and retention of heat by the body, appeared convergently in mammals, birds and four spiny-rayed teleost fish lineages. Of these, red-muscle endothermy over most or all of the body has only appeared in two groups: tunas and the opah (Lampris). Hitherto, tunas have been the only spiny-rayed fishes known to have bones containing embedded osteocyte cells; others have acellular bone. We examined bone histology in Lampris for the first time, demonstrating the presence of cellular bone very similar to that of tunas. This contrasts with the acellular condition of its ectothermic close relatives. The distribution of this character suggests that it co-evolved with red-muscle endothermy, hinting at a common physiological mechanism that would link bone histology to endothermy in these distantly related teleost lineages.

Exceptional preservation of a Cretaceous intestine provides a glimpse of the early ecological diversity of spiny-rayed fishes (Acanthomorpha, Teleostei)

Acanthomorph teleosts (spiny-rayed fishes) account for approximately a third of extant vertebrate species. They appeared during the Late Cretaceous and have been a major component of aquatic biodiversity since the early Cenozoic. They occupy today most trophic levels and ecological niches in aquatic environments, however very little is known about those that were adopted by the earliest representatives of the group. Here, we report on an exceptional glimpse into the ecological diversity of early spiny-rayed fishes provided by the unusual preservation of a newly discovered specimen of the freshwater acanthomorph Spinocaudichthys from the Upper Cretaceous of Morocco. A combination of major-to-trace elemental mapping methods reveals that the gross morphology of the specimen’s intestine has been remarkably preserved owing to the rapid mineralization of iron hydroxides around it. Differing with the typically short and straight intestinal tract of carnivorous teleosts, the intestine in Spinocaudichthys is long and highly convoluted, indicating a probable herbivorous diet. Acanthomorphs would therefore have conquered various ecological niches in their early evolutionary history, prior to their subsequent phylogenetic diversification in both marine and freshwater environments that followed the K-Pg extinction event.

Explosive diversification of marine fishes at the Cretaceous-Palaeogene boundary.

The Cretaceous-Palaeogene (K-Pg) mass extinction is linked to the rapid emergence of ecologically divergent higher taxa (for example, families and orders) across terrestrial vertebrates, but its impact on the diversification of marine vertebrates is less clear. Spiny-rayed fishes (Acanthomorpha) provide an ideal system for exploring the effects of the K-Pg on fish diversification, yet despite decades of morphological and molecular phylogenetic efforts, resolution of both early diverging lineages and enormously diverse subclades remains problematic. Recent multilocus studies have provided the first resolved phylogenetic backbone for acanthomorphs and suggested novel relationships among major lineages. However, these new relationships and associated timescales have not been interrogated using phylogenomic approaches. Here, we use targeted enrichment of >1,000 ultraconserved elements in conjunction with a divergence time analysis to resolve relationships among 120 major acanthomorph lineages and provide a new timescale for acanthomorph radiation. Our results include a well-supported topology that strongly resolves relationships along the acanthomorph backbone and the recovery of several new relationships within six major percomorph subclades. Divergence time analyses also reveal that crown ages for five of these subclades, and for the bulk of the species diversity in the sixth, coincide with the K-Pg boundary, with divergences between anatomically and ecologically distinctive suprafamilial clades concentrated in the first 10 million years of the Cenozoic.

An Intact Retroviral Gene Conserved in Spiny-Rayed Fishes for over 100 My

We have identified a retroviral envelope gene with a complete, intact open reading frame (ORF) in 20 species of spiny-rayed fishes (Acanthomorpha). The taxonomic distribution of the gene, “percomORF”, indicates insertion into the ancestral lineage >110 Ma, making it the oldest known conserved gene of viral origin in a vertebrate genome. Underscoring its ancient provenence, percomORF exists as an isolated ORF within the intron of a widely conserved host gene, with no discernible proviral sequence nearby. Despite its remarkable age, percomORF retains canonical features of a retroviral glycoprotein, and tests for selection strongly suggest cooption for a host function. Retroviral envelope genes have been coopted for a role in placentogenesis by numerous lineages of mammals, including eutherians and marsupials, representing a variety of placental structures. Therefore percomORF’s presence within the group Percomorpha—unique among spiny-finned fishes in having evolved placentation and live birth—is especially intriguing.

The Phylogenetic Intrarelationships of Spiny-Rayed Fishes (Acanthomorpha, Teleostei, Actinopterygii): Fossil Taxa Increase the Congruence of Morphology with Molecular Data

Acanthomorpha (spiny-rayed fishes) is a clade of teleosts that includes more than 15 000 extant species. Their deep phylogenetic intrarelationships, first reconstructed using morphological characters, have been extensively revised with molecular data. Moreover, the deep branches of the acanthomorph tree are still largely unresolved, with strong disagreement between studies. Here, we review the historical propositions for acanthomorph deep intrarelationships and attempt to resolve their earliest branching patterns using a new morphological data matrix compiling and revising characters from previous studies. The taxon sampling we use constitutes a first attempt to test all previous hypotheses (molecular and morphological alike) with morphological data only. Our sampling also includes Late Cretaceous fossil taxa, which yield new character state combinations that are absent in extant taxa. Analysis of the complete morphological data matrix yields a new topology that shows remarkable congruence with the well-supported molecular results. Lampridiformes (oarfishes and allies) are the sister to all other acanthomorphs. Gadiformes (cods and allies) and Zeiformes (dories) form a clade with Percopsiformes (trout-perches) and the enigmatic Polymixia (beardfish) and Stylephorus (tube-eye). Ophidiiformes (cusk-eels and allies) and Batrachoidiformes (toadfishes) are nested within Percomorpha, the clade that includes most of modern acanthomorph diversity. These results provide morphological synapomorphies and independent corroboration of clades previously only recovered from molecular data, thereby suggesting the emergence of a congruent picture of acanthomorph deep intrarelationships. Fossil taxa play a critical role in achieving this congruence, since a very different topology is found when they are excluded from the analysis.

Evolution of Venomous Cartilaginous and Ray-Finned Fishes.

Venom and its associated delivery systems have evolved in numerous animal groups ranging from jellyfishes to spiders, lizards, shrews, and the male platypus. Building off new data and previously published anatomical and molecular studies, we explore the evolution of and variation within venomous fishes. We show the results of the first multi-locus, ordinal-level phylogenetic analysis of cartilaginous (Chondrichthyes) and ray-finned (Actinopterygii) fishes that hypothesizes 18 independent evolutions of this specialization. Ancestral-states reconstruction indicates that among the 2386-2962 extant venomous fishes, envenomed structures have evolved four times in cartilaginous fishes, once in eels (Anguilliformes), once in catfishes (Siluriformes), and 12 times in spiny-rayed fishes (Acanthomorpha). From our anatomical studies and phylogenetic reconstruction, we show that dorsal spines are the most common envenomed structures (∼95% of venomous fish species and 15 independent evolutions). In addition to envenomed spines, fishes have also evolved venomous fangs (2% of venomous fish species, two independent evolutions), cleithral spines (2% of venomous fish species, one independent evolution), and opercular or subopercular spines (1% of venomous fish species, three independent evolutions).

Neofunctionalization of Androgen Receptor by Gain-of-Function Mutations in Teleost Fish Lineage.

Steroid hormone receptor family provides an example of evolution of diverse transcription factors through whole-genome duplication (WGD). However, little is known about how their functions have been evolved after the duplication. Teleosts present a good model to investigate an accurate evolutionary history of protein function after WGD, because a teleost-specific WGD (TSGD) resulted in a variety of duplicated genes in modern fishes. This study focused on the evolution of androgen receptor (AR) gene, as two distinct paralogs, ARα and ARβ, have evolved in teleost lineage after TSGD. ARα showed a unique intracellular localization with a higher transactivation response than that of ARβ. Using site-directed mutagenesis and computational prediction of protein-ligand interactions, we identified two key substitutions generating a new functionality of euteleost ARα. The substitution in the hinge region contributes to the unique intracellular localization of ARα. The substitution on helices 10/11 in the ligand-binding domain possibly modulates hydrogen bonds that stabilize the receptor-ligand complex leading to the higher transactivation response of ARα. These substitutions were conserved in Acanthomorpha (spiny-rayed fish) ARαs, but not in an earlier branching lineage among teleosts, Japanese eel. Insertion of these substitutions into ARs from Japanese eel recapitulates the evolutionary novelty of euteleost ARα. These findings together indicate that the substitutions generating a new functionality of teleost ARα were fixed in teleost genome after the divergence of the Elopomorpha lineage. Our findings provide a molecular explanation for an adaptation process leading to generation of the hyperactive AR subtype after TSGD.

The Rise of Jaw Protrusion in Spiny-Rayed Fishes Closes the Gap on Elusive Prey

Jaw protrusion is one of the most important innovations in vertebrate feeding over the last 400 million years [1, 2]. Protrusion enables a fish to rapidly decrease the distance between itself and its prey [2, 3]. We assessed the evolution and functional implications of jaw protrusion in teleost fish assemblages from shallow coastal seas since the Cretaceous. By examining extant teleost fishes, we identified a robust morphological predictor of jaw protrusion that enabled us to predict the extent of jaw protrusion in fossil fishes. Our analyses revealed increases in both average and maximum jaw protrusion over the last 100 million years, with a progressive increase in the potential impact of fish predation on elusive prey. Over this period, the increase in jaw protrusion was initially driven by a taxonomic restructuring of fish assemblages, with an increase in the proportion of spiny-rayed fishes (Acanthomorpha), followed by an increase in the extent of protrusion within this clade. By increasing the ability of fishes to catch elusive prey [2, 4], jaw protrusion is likely to have fundamentally changed the nature of predator-prey interactions and may have contributed to the success of the spiny-rayed fishes, the dominant fish clade in modern oceans [5].

Phylogeny and tempo of diversification in the superradiation of spiny-rayed fishes

Spiny-rayed fishes, or acanthomorphs, comprise nearly one-third of all living vertebrates. Despite their dominant role in aquatic ecosystems, the evolutionary history and tempo of acanthomorph diversification is poorly understood. We investigate the pattern of lineage diversification in acanthomorphs by using a well-resolved time-calibrated phylogeny inferred from a nuclear gene supermatrix that includes 520 acanthomorph species and 37 fossil age constraints. This phylogeny provides resolution for what has been classically referred to as the "bush at the top" of the teleost tree, and indicates acanthomorphs originated in the Early Cretaceous. Paleontological evidence suggests acanthomorphs exhibit a pulse of morphological diversification following the end Cretaceous mass extinction; however, the role of this event on the accumulation of living acanthomorph diversity remains unclear. Lineage diversification rates through time exhibit no shifts associated with the end Cretaceous mass extinction, but there is a global decrease in lineage diversification rates 50 Ma that occurs during a period when morphological disparity among fossil acanthomorphs increases sharply. Analysis of clade-specific shifts in diversification rates reveal that the hyperdiversity of living acanthomorphs is highlighted by several rapidly radiating lineages including tunas, gobies, blennies, snailfishes, and Afro-American cichlids. These lineages with high diversification rates are not associated with a single habitat type, such as coral reefs, indicating there is no single explanation for the success of acanthomorphs, as exceptional bouts of diversification have occurred across a wide array of marine and freshwater habitats.