Stickleback fishes: Bridging the gap between population biology and paleobiology

Integration of evolutionary mechanisms and phylogeny requires study of phenotypes that change in the fossil record and continue to evolve in extant populations. Pelvic reduction in the three-spined stickle-back has evolved rapidly in a Miocene fossil assemblage and in numerous extant isolated lake populations throughout its distribution. Although pelvic reduction is caused by selection, expression of reduced pelvic phenotypes is constrained by development and other factors. However, lineages with pelvis reduction rapidly go extinct while lineages that retain the fully formed pelvic girdle tend to persist. Existence of pelvic reduction since the Miocene has depended on an equilibrium between divergence and extinction. The phylogenetic topology resulting from this process differs greatly from the conventional view of evolutionary history, and could only be recognized by analysis of both extant populations and fossil material. If this phylogenetic topology is common, it may help to account for the different perceptions that population biologists and paleobiologists have of evolutionary tempo.     

Vestigial skeletal structures in dinosaurs

The existence of vestigial structures is one of the main lines of evidence for macroevolution. Here I introduce a phylogenetic bracketing approach to the identification of vestigial structures and apply it to Dinosauria. According to this approach, a structure is considered vestigial if, in comparison with its homolog in at least three successive outgroups, it is reduced to one-third or less its size relative to adjacent structures and if at least distally it has lost the specialized morphology present in the three outgroups. This approach identifies fingers IV and V as vestigial in dinosaurs in general, II–V in sauropods, III in Tyrannosauridae and Caudipteryx , II and III in Shuvuuia and I and III in modern birds. The entire forelimb distal to the elbow is vestigial in Abelisauridae. Vestigial parts of the pelvic girdle and hindlimb include the pubic shaft in Iguanodontia and Ceratopsia, the entire pubis in Ankylosauria, the first metatarsal in derived Iguanodontia, the first metatarsal shaft in Theropoda and the fifth toe in dinosaurs in general. Derived Centrosaurinae and some Chasmosaurus exhibit vestigial supraorbital horns. Some centrosaurines have a vestigial nasal horn. The antorbital cavity is vestigial in Thyreophora, Iguanodontia and Ceratopsidae. I recommend that this information be exploited to increase public awareness of the evidence for macroevolution.     

Armor morphology and reproductive output in threespine stickleback,Gasterosteus aculeatus

Synopsis The threespine stickleback, Gasterosteus aculeatus, is an extensively armored fish inhabiting both marine and fresh waters across its holarctic distribution. Marine fish nearly always possess a full complement of bony lateral plates running from just behind the head to the tail, and a robust pelvic girdle complex. These armor features appear to constrain lateral and ventral abdominal distention, and therefore clutch volume. Freshwater populations in many areas exhibit variable reduction in lateral plate number, and in some regions the pelvic girdle is also reduced or lost. Freshwater populations also vary in the degree of abdominal distention exhibited by gravid females. We tested whether reduction in armoring might be correlated with increased clutch volume using five populations from the Cook Inlet area of Alaska. The hypothesis that populations having reduced pelvic girdle complexes would have greater size-adjusted clutch volumes was not supported. In fact, our two full-pelvic populations as a group had larger volumes. Similarly, size-adjusted clutch volumes were not related to pelvic phenotype within either of our two pelvic-reduced populations, nor to lateral plate morph within a fifth population. Other factors that may explain the interpopulation differences in clutch volume in threespine stickleback include body shape, food quantity and quality, intensity of predation, and even behavior. Except for a preliminary analysis of body shape, these possibilities remain unexplored. The concept of phenotypic integration suggests that these factors should be analyzed as a suite rather than individually.     

Archosaurian respiration and the pelvic girdle aspiration breathing of crocodyliforms

Birds and crocodylians, the only living archosaurs, are generally believed to employ pelvic girdle movements as a component of their respiratory mechanism. This in turn provides a phylogenetic basis for inferring that extinct archosaurs, including dinosaurs, also used pelvic girdle breathing. I examined lung ventilation through cineradiography (high-speed X-ray filming) and observed that alligators indeed rotate the pubis to increase tidal volume, but did not observe pelvic girdle movement contributing to lung ventilation in guinea fowl, emus or tinamous, despite extensive soft-tissue motion. Re-examination of fossil archosaurs reveals that pubic rotation evolved in basal crocodyliforms and that pelvic girdle breathing is not a general archosaurian mechanism. The appearance of pelvic aspiration in crocodyliforms is a striking example of the ability of amniotes to increase gas exchange or circumvent constraints on respiration through the evolution of novel accessory breathing mechanisms.     

Pelvic anti-predator armour reduction in Norwegian populations of the threespine stickleback: a rare phenomenon with adaptive implications?

In order to search for armour reductions in the anti-predator armour apparatus of the threespine stickleback Gasterosteus aculeatus , we studied populations from 200 coastal lakes and 32 marine and estuarine sites in Norway. We scored the presence of reductions in the pelvic apparatus in these populations, measured the length of the second dorsal fin and pelvic spine and counted lateral plates in a subset of 96 populations. Then we looked for the relation between pelvic reduction and number of lateral plates and lengths of the second dorsal spine and pelvic spine. We also observed whether pelvic reduced fish had asymmetric development on the right or the left side of the pelvis. Pelvic reduction was not found in marine and estuarine sticklebacks (with one single exception), but in freshwater it occurred in four out of 200 lakes. The amount of pelvic reduction differed from 5% to 68% in the four populations. There appears to be a correlation between pelvic reduction and spine lengths at the population level, but not plate number. Further, it appears to be difficult to explain the occurrence of pelvic reduction in Norway with the factors proposed to be responsible for pelvic reduction in other regions.     

Implications of a fossil stickleback assemblage for Darwinian gradualism

Darwin postulated that a complete fossil record would contain numerous gradual transitions between ancestral and descendant species, but 150 years after publication of The Origin of Species, few such transitions have materialized. The fossil stickleback Gasterosteus doryssus and the deposit in which it occurs provide excellent conditions to detect such transitions. Abundant, well‐preserved fossils occur in a stratigraphic setting with fine temporal resolution. The paleoecology of G. doryssus resembles the ecology of modern lakes that harbour the phenotypically similar three‐spined stickleback Gasterosteus aculeatus. Gasterosteus aculeatus are primitively highly armoured, but G. doryssus comprised two contemporaneous biological species with relatively weak armour, including a near‐shore, benthic feeder (benthic) and an offshore planktivore (limnetic). The benthic species expanded its range into the limnetic zone of the lake, where it apparently switched to planktivory and evolved reduced armour within c. 5000 years in response to directional selection. Although gradual evolution of mean phenotypes occurred, a single major gene caused much of evolutionary change of the pelvic skeleton. Thus, Darwin's expectation that transitions between species in the fossil record would be gradual was met at a fine time scale, but for pelvic structure, a well‐studied trait, his expectation that gradual change would depend entirely on numerous, small, heritable differences among individuals was incorrect.     

Directional asymmetry of pelvic vestiges in threespine stickleback

Extensive reduction of the size and complexity of the pelvic skeleton (i.e., pelvic reduction) has evolved repeatedly in Gasterosteus aculeatus. Asymmetrical pelvic vestiges tend to be larger on the left side (i.e., left biased) in populations studied previously. Loss of Pitx1 expression is associated with pelvic reduction in G. aculeatus, and pelvic reduction maps to the Pitx1 locus. Pitx1 knockouts in mice have reduced hind limbs, but the left limb is larger. Thus left-biased directional asymmetry of stickleback pelvic vestiges may indicate the involvement of Pitx1 in pelvic reduction. We examined 6,356 specimens from 27 Cook Inlet populations of G. aculeatus with extensive pelvic reduction. Samples from 20 populations exhibit the left bias in asymmetrical pelvic vestiges expected if Pitx1 is involved, and three have a slight, non-significant left bias. However, samples from three populations have a significant right bias, and one large sample from another population has equal frequencies of specimens with larger vestiges on the left or right side. A sample of fossil threespine stickleback also has significantly left-biased pelvic vestiges. These results suggest that silencing of Pitx1 or the developmental pathway in which it functions in the pelvis is the usual cause of pelvic reduction in most Cook Inlet populations of G. aculeatu, and that it caused pelvic reduction at least 10 million years ago in a stickleback population. A different developmental genetic mechanism is implicated for three populations with right-biased pelvic vestiges and for the population without directional asymmetry.     

Parallel evolution of Pitx1 underlies pelvic reduction in Scottish threespine stickleback (Gasterosteus aculeatus)

Little is known about the genetic and molecular mechanisms that underlie adaptive phenotypic variation in natural populations or whether similar genetic and molecular mechanisms are utilized when similar adaptive phenotypes arise in independent populations. The threespine stickleback (Gasterosteus aculeatus) is a good model system to investigate these questions because these fish display a large amount of adaptive phenotypic variation, and similar adaptive phenotypes have arisen in multiple, independent stickleback populations. A particularly striking pattern of parallel evolution in sticklebacks is reduction of skeletal armor, which has occurred in numerous freshwater locations around the world. New genetic and genomic tools for the threespine stickleback have made it possible to identify genes that underlie loss of different elements of the skeletal armor. Previous work has shown that regulatory mutations at the Pitx1 locus are likely responsible for loss of the pelvic structures in independent stickleback populations from North America and Iceland. Here we show that the Pitx1 locus is also likely to underlie pelvic reduction in a Scottish population of threespine stickleback, which has apparently evolved pelvic reduction under a different selection regime than the North American populations.     

Variation in female life-history traits among Alaskan populations of the threespine stickleback, Gasterosteus aculeatus L. (Pisces: Gasterosteidae)

Life-history characteristics of female threespine stickleback (Gasterosteus aculeatus) were examined in 12 populations, 11 freshwater and one anadromous, within the Cook Inlet region of Alaska. Because this area has been deglaciated during the last 20 000 years, the freshwater populations are recently derived, probably independendy, from the local marine or anadromous stickleback. Freshwater threespine stickleback have undergone considerable morphological evolution within this region, apparently in response to environmental factors including predatory regimes and environmental productivity. Our freshwater study populations were selected to sample this range of morphological variation in order to determine whether life-history traits and morphologies have followed similar evolutionary trajectories. Freshwater populations could be categorized generally into one of three ecomorphotypes: those inhabiting relatively productive lakes having one or more piscivorous fishes present, and in which the stickleback exhibit a fully developed pelvic girdle; those inhabiting low-calcium lakes that lack piscivorous fishes, and in which the pelvic structures are incomplete; those living in streams with piscivorous fishes, in which the stickleback have fully developed pelvic girdles. The anadromous population constituted a fourth ecomorphotype that lives in marine waters, and is robusdy armored. The freshwater populations showed considerable variation in all life-history traits assessed, and this variation generally corresponded to our ecomorphological classifications. Nevertheless, within each ecomorphotype there was sufficient variation to suggest that morphological and life-history traits may not always respond in the same manner in response to the same selective regime.     

Predation's role in repeated phenotypic and genetic divergence of armor in threespine stickleback

Predator-driven divergent selection may cause differentiation in defensive armor in threespine stickleback: (1) predatory fish and birds favor robust armor, whereas (2) predaceous aquatic insects favor armor reduction. Although (1) is well established, no direct experimental evidence exists for (2). I examined the phenotypic and genetic consequences of insect predation using F₂ families from crosses between freshwater and marine stickleback populations. I measured selection on body size, and size-adjusted spine (dorsal and pelvic) and pelvic girdle length, by splitting juvenile F₂ families between control and insect predation treatments, set in pond enclosures. I also examined the effect of insect predation on Ectodysplasin ( Eda ), a gene physically linked to quantitative trait loci for lateral plate number, spine length, and body shape. Insect predation resulted in: (1) significant selection for larger juvenile size, and shorter dorsal spine and pelvic girdle length, (2) higher mortality of individuals missing the pelvic girdle, and (3) selection in favor of the low armor Eda allele. Predatory insects favor less stickleback armor, likely contributing to the widespread reduction of armor in freshwater populations. Because size strongly influences mate choice, predator-driven divergent selection on size may play a substantial role in byproduct reproductive isolation and speciation in threespine stickleback.     

Evolutionary history shapes the association between developmental instability and population‐level genetic variation in three‐spined sticklebacks

Developmental instability (DI) is the sensitivity of a developing trait to random noise and can be measured by degrees of directionally random asymmetry [fluctuating asymmetry (FA)]. FA has been shown to increase with loss of genetic variation and inbreeding as measures of genetic stress, but associations vary among studies. Directional selection and evolutionary change of traits have been hypothesized to increase the average levels of FA of these traits and to increase the association strength between FA and population‐level genetic variation. We test these two hypotheses in three‐spined stickleback (Gasterosteus aculeatus L.) populations that recently colonized the freshwater habitat. Some traits, like lateral bone plates, length of the pelvic spine, frontal gill rakers and eye size, evolved in response to selection regimes during colonization. Other traits, like distal gill rakers and number of pelvic fin rays, did not show such phenotypic shifts. Contrary to a priori predictions, average FA did not systematically increase in traits that were under presumed directional selection, and the increases observed in a few traits were likely to be attributable to other factors. However, traits under directional selection did show a weak but significantly stronger negative association between FA and selectively neutral genetic variation at the population level compared with the traits that did not show an evolutionary change during colonization. These results support our second prediction, providing evidence that selection history can shape associations between DI and population‐level genetic variation at neutral markers, which potentially reflect genetic stress. We argue that this might explain at least some of the observed heterogeneities in the patterns of asymmetry.     

Genetic relationships and origin of two geographic groups of the freshwater threespine stickleback, 'hariyo'

'Hariyo' comprise the southernmost freshwater populations of the threespine stickleback, Gasterosteus aculeatus species complex, distributed in central Honshu, Japan. Two geographic groups (Gifu and Shiga) of the 'hariyo' have been recognized as differing from each other in some morphological and ecological features. In order to elucidate the genetic characteristics and phylogenetic position of these populations, partial sequences of the mitochondrial cytochrome b gene were compared in 123 specimens from 10 'hariyo' populations, and eight other freshwater and anadromous populations of threespine stickleback in Japan. Phylogenetic analysis resolved 22 haplotypes in a single most-parsimonious tree. In contrast to previous allozyme analyses, the haplotype tree indicated that 'hariyo' populations are monophyletic. Each of two geographic groups of the 'hariyo' was also shown to be nearly monophyletic. The two geographic groups differed from each other in sequence by an average of 0.47% and from other populations by 0.70%. Based on a molecular clock, constructed from fossil records and mtDNA genetic distances of Gasterosteus and Pungitius, it was estimated that the two groups of 'hariyo' differentiated from the other threespine stickleback populations and also from each other in the middle Pleistocene. The results suggested that each of the two groups of 'hariyo' is a distinct evolutionarily significant unit with unique genetic features, as well as morphological and ecological characteristics.     

Studies on dentin. 2. Vestigial lacteal incisor teeth of the rat.

The presence of a vestigial, lacteal incisor tooth is described in the laboratory rat. This tooth is felt to belong to the same dental generation as the other functional teeth. Accordingly, the rat is described as having a monophyodont, first dentition containing two incisor teeth in each quadrant. These vestigial teeth are then compared with other similar mammalian teeth and are defined as transient, partially formed and non-functional. As such, they are differentiated from other transient teeth. The examination of the fossil record suggests that tooth loss is a general phenomenon in rodents, but that this vestigial tooth probably represents a condition present in forms antecedent to rodents. A critical literature review strongly suggests that the teeth of the recent rat are members of the first dental generation. The presence of such a vestigial tooth and of the postincisive diastema in the rat is felt to be an example of phylogenetic reduction and progressive retardation in the sense of de Beer's concepts. These same two phenomena were analyzed with respect to the field theory of Butler and of the Zahnreihen theory of Edmund. Placed within the context of recent data on epithelioectomesenchymal interactions, both theories were supported, and both the vestigial teeth and anodontic diastema were shown to be explicable within these conceptual frameworks.     

The evolution of paired fins

Summary The Archipterygium is Gegenbaur’s most lasting contribution to the study of vertebrate limb evolution. This transformational hypothesis of gill arches to limb girdles, rays to fins, and proposal of a vertebrate fin-limb groundplan, is generally treated as a flawed alternative to the more widely accepted lateral fin-fold hypothesis of vertebrate limb evolution. When compared to the phylogenetic distribution and diversity of fins and limbs, both hypotheses fail. Dermal skeletal lateral folds, spines and keels originate repeatedly in vertebrate evolution, but paired fins with girdles originate at pectoral level and are anteroposteriorly restricted. Pelvic fins emerge later in phylogeny; pectoral and pelvic appendages primitively differ. Endoskeletal girdles never exhibit characteristics of gill arches. The emergent sequence of paired fin evolution depends upon phylogenetic hypotheses within which extant agnathan interrelationships are uncertain; positions of jawless fossil fish along the gnathostome stem are insecure; the fossil data set is patchy. However, certain features of the data set are robust. This has prompted a reconsideration of Gegenbaur’s hypothesized arch-girdle relationship, and an iterative homology between scapulocoracoid and extrabranchial cartilages is suggested. No transformation of arch to girdle is necessarily implied, but some signal of developmental relatedness is predicted.     

A comparative analysis of experimental selection on the stickleback pelvis

Mechanisms of natural selection can be identified using experimental approaches. However, such experiments often yield nonsignificant effects and imprecise estimates of selection due to low power and small sample sizes. Combining results from multiple experimental studies might produce an aggregate estimate of selection that is more revealing than individual studies. For example, bony pelvic armour varies conspicuously among stickleback populations, and predation by vertebrate and insect predators has been hypothesized to be the main driver of this variation. Yet experimental selection studies testing these hypotheses frequently fail to find a significant effect. We experimentally manipulated length of threespine stickleback (Gasterosteus aculeatus) pelvic spines in a mesocosm experiment to test whether prickly sculpin (Cottus asper), an intraguild predator of stickleback, favours longer spines. The probability of survival was greater for stickleback with unclipped pelvic spines, but this effect was noisy and not significant. We used meta‐analysis to combine the results of our mesocosm experiment with previously published experimental studies of selection on pelvic armour. We found evidence that fish predation indeed favours increased pelvic armour, with a moderate effect size. The same approach found little evidence that insect predation favours reduced pelvic armour. The causes of reduced pelvic armour in many stickleback populations remain uncertain.     

Morphological divergence of North‐European nine‐spined sticklebacks (Pungitius pungitius): signatures of parallel evolution

Parallel evolution is characterised by repeated, independent occurrences of similar phenotypes in a given habitat type, in different parts of the species distribution area. We studied body shape and body armour divergence between five marine, four lake, and ten pond populations of nine‐spined sticklebacks [Pungitius pungitius (Linnaeus, 1758)] in Fennoscandia. We hypothesized that marine and lake populations (large water bodies, diverse fish fauna) would be similar, whereas sticklebacks in isolated ponds (small water bodies, simple fish fauna) would be divergent. We found that pond fish had deeper bodies, shorter caudal peduncles, and less body armour (viz. shorter/absent pelvic spines, reduced/absent pelvic girdle, and reduced number of lateral plates) than marine fish. Lake fish were intermediate, but more similar to marine than to pond fish. Results of our common garden experiment concurred with these patterns, suggesting a genetic basis for the observed divergence. We also found large variation among populations within habitat types, indicating that environmental variables other than those related to gross habitat characteristics might also influence nine‐spined stickleback morphology. Apart from suggesting parallel evolution of morphological characteristics of nine‐spined sticklebacks in different habitats, the results also show a number of similarities to the evolution of three‐spined stickleback (Gasterosteus aculeatus Linnaeus, 1758) morphology. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 101 , 403–416.     

Phylogeny, divergence times and species limits of spiny lizards (Sceloporus magister species group) in western North American deserts and Baja California

The broad distribution of the Sceloporus magister species group (squamata: phrynosomatidae) throughout western North America provides an appropriate model for testing biogeographical hypotheses explaining the timing and origins of diversity across mainland deserts and the Baja California Peninsula. We inferred concordant phylogenetic trees describing the higher-level relationships within the magister group using 1.6 kb of mitochondrial DNA (mtDNA) and 1.7 kb of nuclear DNA data. These data provide strong support for the parallel divergence of lineages endemic to the Baja California Peninsula (S. zosteromus and the orcutti complex) in the form of two sequential divergence events at the base of the magister group phylogeny. A relaxed phylogenetic analysis of the mtDNA data using one fossil and one biogeographical constraint provides a chronology of these divergence events and evidence that further diversification within the Baja California clades occurred simultaneously, although patterns of geographical variation and speciation between clades differ. We resolved four major phylogeographical clades within S. magister that (i) provide a novel phylogenetic placement of the Chihuahuan Desert populations sister to the Mojave Desert; (ii) illustrate a mixed history for the Colorado Plateau that includes Mojave and Sonoran Desert components; and (iii) identify an area of overlap between the Mojave and Sonoran Desert clades near Yuma, Arizona. Estimates of bidirectional migration rates among populations of S. magister using four nuclear loci support strong asymmetries in gene flow among the major mtDNA clades. Based on the nonexclusivity of mtDNA haplotypes, nuclear gene flow among populations and wide zones of phenotypic intergradation, S. magister appears to represent a single geographically variable and widespread species.     

What is bred in the bone: Ecomorphological associations of pelvic girdle form in greater Antillean Anolis lizards

Ecological niche partitioning of Anolis lizards of the Greater Antillean islands has been the focus of many comparative studies, and much is known about external morphological convergence that characterizes anole ecomorphs. Their internal anatomy, however, has rarely been explored in an ecomorphological context, and it remains unknown to what degree skeletal morphology tracks the diversity and ecological adaptation of these lizards. Herein, we employ CT scanning techniques to visualise the skeleton of the pelvic girdle in situ, and 3D geometric morphometrics to compare the form of the ilium, ischium, and pubis within and between ecomorphs. We examine 26 species of anoles representing four ecomorphs (trunk‐ground, trunk‐crown, crown‐giant, twig) from three islands (Jamaica, Hispaniola, and Puerto Rico). The subtle variations in pelvic girdle morphology discovered are directly associable with all three parameters that we set out to focus on: phylogenetic relationship, specimen size, and assigned ecomorph category. Morphometric variation that correlates with size and/or phylogenetic signal varies between species and cannot be eliminated from the data set without markedly reducing its overall variability. The discovered patterns of skeletal variation are consistent with the demands of locomotor mechanics pertinent to the structural configuration of the microhabitat of three of the four ecomorphs, with the fourth having no discernible distinctive features. This manifests itself chiefly in the relative anteroposterior extent and anteroventral inclination of the ilium and pubis, which differ between ecomorphs and are postulated to reflect optimization of the direction of muscle vectors of the femoral protractors and retractors. Our investigation of the form of the pelvic girdle of anoles allows us to generalize our findings to entire ecomorph categories within a broad phylogenetic and biogeographic context. Differences in the form and configuration of the postcranial skeleton are directly related to ecological patterns.     

Osteology of Priocharax and remarkable developmental truncation in a miniature Amazonian fish (Teleostei: Characiformes: Characidae)

Establishing phylogenetic relationships of miniature fishes is challenging in taxa with developmental truncation. Within the Characiformes, developmental truncation appears to be relatively rare, with the Neotropical genus Priocharax being an example. Priocharax includes three miniature species among the smallest of the order and has been hypothesized to belong to the Heterocharacinae. The pronounced reduction in its skeleton, however, prevented a clearer evaluation of its relationships. The present detailed osteological study was designed to address this question and revealed that 21 bones are absent and nine other skeletal structures are simplified in Priocharax when compared to other characids. Comparison of the skeleton of adult Priocharax with early developmental stages of other characids demonstrated that most of the absences and simplifications can be interpreted as developmental truncations. The most striking developmental truncations are in the pectoral girdle, in which the endoskeleton remains entirely cartilaginous. Other interesting truncations are in the ethmoid region of the skull, infraorbital series, and Weberian apparatus, in which the claustrum is absent. Our study also revealed some unusual sexual dimorphisms in the pelvic girdle. Two cladistic analyses were performed to assess the relationships of Priocharax within the Heterocharacinae. The first consisted of a traditional analysis in which all absences and reductions of Priocharax were coded in the same way as in the remaining taxa. This resulted in three equally most parsimonious topologies, all of which have Priocharax as the most basal taxon of the Heterocharacinae. The second analysis incorporated ontogenetic information, and most absences and reductions of Priocharax were reinterpreted as apomorphic conditions and thus, coded differently from similar conditions in outgroups. This resulted in a single phylogenetic hypothesis with Priocharax and Gnathocharax as sister groups based on seven synapomorphies. Our approach demonstrates the importance of developmental studies to better understand morphological evolution of miniaturized, truncated taxa, and to generate hypotheses of their relationships. J. Morphol. 277:65–85, 2016. © 2015 Wiley Periodicals, Inc.     

Skeletal and muscular pelvic morphology of hillstream loaches (Cypriniformes: Balitoridae)

The rheophilic hillstream loaches (Balitoridae) of South and Southeast Asia possess a range of pelvic girdle morphologies, which may be attributed to adaptations for locomotion against rapidly flowing water. Specifically, the connectivity of the pelvic plate (basipterygium) to the vertebral column via a sacral rib, and the relative size and shape of the sacral rib, fall within a spectrum of three discrete morphotypes: long, narrow rib that meets the basipterygium; thicker, slightly curved rib meeting the basipterygium; and robust crested rib interlocking with the basipterygium. Species in this third category with more robust sacral rib connections between the basipterygium and vertebral column are capable of walking out of water with a tetrapod-like lateral-sequence, diagonal-couplet gait. This behavior has not been observed in species lacking direct skeletal connection between the vertebrae and the pelvis. The phylogenetic positions of the morphotypes were visualized by matching the morphological features onto a novel hypothesis of relationships for the family Balitoridae. The morphotypes determined through skeletal morphology were correlated with patterns observed in the pelvic muscle morphology of these fishes. Transitions towards increasingly robust pelvic girdle attachment were coincident with a more anterior origin on the basipterygium and more lateral insertion of the muscles on the fin rays, along with a reduction of the superficial abductors and adductors with more posterior insertions. These modifications are expected to provide a mechanical advantage for generating force against the ground. Inclusion of the enigmatic cave-adapted balitorid Cryptotora thamicola into the most data-rich balitorid phylogeny reveals its closest relatives, providing insight into the origin of the skeletal connection between the axial skeleton and basipterygium.