Vertebrate Trait Research Articles

Computed Tomographic Measures of Funnel-Shaped Lumbar Vertebral Canal and Articular Process Dysplasia Malformations Differ Between German Shepherd and Belgian Malinois Military Working Dogs.

Researchers who study the selection and breeding program criteria for military working dogs aim to help maximize the years of active duty service. Computed tomographic (CT) quantitative phenotyping has been previously described as a method for supporting these research studies. Funnel-shaped lumbar vertebral foramen malformations have been previously described in Labrador retriever military working dogs and proposed to be risk factors for impaired arterial perfusion of nerve tissues during exercise. Articular process dysplasia malformations have been previously described in varying dog breeds and proposed to be risk factors for articular process degenerative joint disease and vertebral foramen stenosis. Aims of this retrospective, cross-sectional study were to describe quantitative CT phenotyping methods for characterizing funnel-shaped lumbar vertebral foramina and articular process dysplasia malformations and to apply these methods in a comparison between groups of German shepherd and Belgian Malinois military working dogs. A military working dog hospital's database was searched for German shepherd and Belgian Malinois dogs aged <6 years that had CT scans of the lumbosacral region during the period of 2008–2016. Observers unaware of CT findings recorded available clinical data for each of the dogs. An observer unaware of clinical data recorded CT measures of funnel-shaped lumbar vertebral foramina and articular process dysplasia malformations for each of dogs and each of the lumbar vertebrae that were available in the scans. A total of 59 dogs were sampled: 41 German shepherd and 18 Belgian Malinois. Articular process dysplasia and funnel-shaped vertebral foramen phenotypic traits were present in both breeds in this sample, with the frequency and quantitative measure of these traits being greater in German shepherd dogs and heavier dogs. Lower weight dogs had a lesser degree of a funnel-shaped foramen at all sampled vertebral locations. A consistent relationship between articular process dysplasia measures and body weight was not seen. Computed tomography measures of funnel shaped vertebral foramina were greater in German shepherd vs. Belgian Malinois dogs at the L7 vertebra (P < 0.01). The CT measures of cranial articular process dysplasia were greater in German shepherd vs. Belgian Malinois dogs at the L4 (P < 0.01) and L5 (P < 0.05) vertebrae.

Eliminating the dark matter of data deficiency by predicting the conservation status of Northeast Atlantic and Mediterranean Sea sharks and rays

Sharks and rays are threatened by overfishing, yet we have little idea of the conservation status of the hundreds of Data Deficient species. Here, we developed an ecological trait model to predict the categorical conservation status of 22 Northeast Atlantic and 13 Mediterranean Sea Data Deficient sharks and rays. We first developed an explanatory cumulative link mixed model based on regionally data-sufficient species on the International Union for Conservation of Nature (IUCN) Red List of Threatened Species™ using maximum body size, median depth, and reproductive mode, then predicted the statuses of Data Deficient species. Species exclusive to the Mediterranean were 3.8 times more likely to be threatened than species exclusive to the Northeast Atlantic. Over half of Northeast Atlantic (55%, n = 12 of 22), and two-thirds of Mediterranean (62%, n = 8 of 13) Data Deficient species were predicted to be threatened. When applied to all data-sufficient species, the mean predictive accuracy was 71% and 66% for the Northeast Atlantic and Mediterranean models, respectively. Overall, Northeast Atlantic Data Deficient species are predicted to be 1.4 times more threatened than data-sufficient species proportionally (39% assessed-threatened, n = 38 of 98), whereas threat levels in the Mediterranean Sea are similarly high for both (65% assessed-threatened, n = 39 of 60 data-sufficient). With the growing availability of vertebrate trait databases, trait-based, categorical prediction of conservation status is a cost-effective approach towards incorporating Data Deficient species into unbiased (i) estimates of lineage-wide extinction rates, (ii) protected species lists, and (iii) Red List Indices, thus preventing poorly-known species from reaching extinction unnoticed.

A Hox gene regulatory network for hindbrain segmentation.

In vertebrates, the hindbrain serves as a highly conserved complex coordination center for regulating many fundamental activities of the central nervous system, such as respiratory rhythms, sleep patterns and equilibrium, and it also plays an important role in craniofacial development. The basic ground plan that underlies the diverse functions of the hindbrain and its neural crest derivatives is established and patterned by a process of segmentation. Through a dynamic series of signaling and regulatory interactions the developing hindbrain is transiently compartmentalized into a set of seven segmental units, termed rhombomeres. The nested expression of the Hox family of transcription factors is tightly coupled to the process of segmentation and provides a molecular code for specifying the unique regional properties of the hindbrain and its neural crest derived craniofacial structures. The high degree of similarity in hindbrain architecture between diverse vertebrates has enabled cross-species regulatory analysis. This has facilitated the experimental assembly of the signaling and regulatory interactions, which underlie the process of segmentation, into a Hox-dependent gene regulatory network (GRN) model. This hindbrain GRN is a key regulatory feature of head patterning, conserved to the base of vertebrate evolution. This regulatory framework also serves as a basis for comparing and contrasting GRNs that govern cranial neural crest formation and axial patterning and provide insight into regulatory mechanisms associated with the evolution of novel vertebrate traits. The purpose of this review is to discuss the majorfeatures of the GRN for hindbrain segmentation and its relationship to the broader functional role of the hindbrain in patterning head development.

3D shape analyses of extant primate and fossil hominin vertebrae support the ancestral shape hypothesis for intervertebral disc herniation

BackgroundRecently we proposed an evolutionary explanation for a spinal pathology that afflicts many people, intervertebral disc herniation (Plomp et al. [2015] BMC Evolutionary Biology 15, 68). Using 2D data, we found that the bodies and pedicles of lower vertebrae of pathological humans were more similar in shape to those of chimpanzees than were those of healthy humans. Based on this, we hypothesized that some individuals are more prone to intervertebral disc herniation because their vertebrae exhibit ancestral traits and therefore are less well adapted for the stresses associated with bipedalism. Here, we report a study in which we tested this “Ancestral Shape Hypothesis” with 3D data from the last two thoracic and first lumbar vertebrae of pathological Homo sapiens, healthy H. sapiens, Pan troglodytes, and several extinct hominins.ResultsWe found that the pathological and healthy H. sapiens vertebrae differed significantly in shape, and that the pathological H. sapiens vertebrae were closer in shape to the P. troglodytes vertebrae than were the healthy H. sapiens vertebrae. Additionally, we found that the pathological human vertebrae were generally more similar in shape to the vertebrae of the extinct hominins than were the healthy H. sapiens vertebrae. These results are consistent with the predictions of the Ancestral Shape Hypothesis. Several vertebral traits were associated with disc herniation, including a vertebral body that is both more circular and more ventrally wedged, relatively short pedicles and laminae, relatively long, more cranio-laterally projecting transverse processes, and relatively long, cranially-oriented spinous processes. We found that there are biomechanical and comparative anatomical reasons for suspecting that all of these traits are capable of predisposing individuals to intervertebral disc herniation.ConclusionsThe results of the present study add weight to the hypothesis that intervertebral disc herniation in H. sapiens is connected with vertebral shape. Specifically, they suggest that individuals whose vertebrae are towards the ancestral end of the range of shape variation within H. sapiens have a greater propensity to develop the condition than other individuals. More generally, the study shows that evolutionary thinking has the potential to shed new light on human skeletal pathologies.

The neurobiology of innate, volitional and learned vocalizations in mammals and birds.

Vocalization is an ancient vertebrate trait essential to many forms of communication, ranging from courtship calls to free verse. Vocalizations may be entirely innate and evoked by sexual cues or emotional state, as with many types of calls made in primates, rodents and birds; volitional, as with innate calls that, following extensive training, can be evoked by arbitrary sensory cues in non-human primates and corvid songbirds; or learned, acoustically flexible and complex, as with human speech and the courtship songs of oscine songbirds. This review compares and contrasts the neural mechanisms underlying innate, volitional and learned vocalizations, with an emphasis on functional studies in primates, rodents and songbirds. This comparison reveals both highly conserved and convergent mechanisms of vocal production in these different groups, despite their often vast phylogenetic separation. This similarity of central mechanisms for different forms of vocal production presents experimentalists with useful avenues for gaining detailed mechanistic insight into how vocalizations are employed for social and sexual signalling, and how they can be modified through experience to yield new vocal repertoires customized to the individual's social group. This article is part of the theme issue 'What can animal communication teach us about human language?'

A brief natural history of the orgasm

Why the sexual climax, in humans, results in a pleasurable experience remains an important biological question. Analysis of evolutionary traits in numerous Vertebrates suggests that orgasm evolved through three phylogenetic stages during the transition from external to internal fertilization and viviparity. First, orgasm is directly dependent on ejaculation in males and the expulsion of fluids from the ovarian and urethral glands (Skene’s) in females. I propose that sexual orgasm could come from the primitive reflex of discharging gametes to ensure reproduction. Thus, the understanding of orgasm should not be reduced to a penis- or a clitoris-centred paradigm. Secondly, orgasm has evolved to stimulate sexual activity because the evolutionary transition from external fertilization to internal fertilization has been accompanied in numerous species with a lessening in reproductive rates. Because sexual activity encourages reproduction, it can be argued that orgasm has evolved to increase sexual activity, particularly in viviparous species with low reproductive rates. Third, internal fertilization in the genital tract of females weakens the visibility of the putative success of fertilization. Female sexual fluids and proteins can bias fertilization in favour of preferred males. Because orgasm could promote a better choice of partner, I argue that female orgasm may have evolved as a post-copulatory selection tactic by which females can increase their control of mates.

The underlying mechanisms of vertebrate seasonal reproduction.

Animals make use of changes in photoperiod to adapt their physiology to the forthcoming breeding season. Comparative studies have contributed to our understanding of the mechanisms of seasonal reproduction in vertebrates. Birds are excellent models for studying these phenomena because of their rapid and dramatic responses to changes in photoperiod. Deep brain photoreceptors in birds perceive and transmit light information to the pars tuberalis (PT) in the pituitary gland, where the thyroid-stimulating hormone (TSH) is produced. This PT-TSH locally increases the level of the bioactive thyroid hormone T3 via the induction of type 2 deiodinase production in the mediobasal hypothalamus, and an increased T3 level, in turn, controls seasonal gonadotropin-releasing hormone secretion. In mammals, the eyes are the only photoreceptive structure, and nocturnal melatonin secretion encodes day-length information and regulates the PT-TSH signaling cascade. In Salmonidae, the saccus vasculosus plays a pivotal role as a photoperiodic sensor. Together, these studies have uncovered the universality and diversity of fundamental traits in vertebrates.

NaturalHeritage: Bridging Belgian natural history collections

The Royal Belgian Institute of Natural Sciences (RBINS), the Royal Museum for Central Africa (RMCA) and Meise Botanic Garden house more than 50 million specimens covering all fields of natural history. While many different research topics have their own specificities, throughout the years it became apparent that with regards to collection data management, data publication and exchange via community standards, collection holding institutions face similar challenges (James et al. 2018, Rocha et al. 2014). In the past, these have been tackled in different ways by Belgian natural history institutions. In addition to local and national collaborations, there is a great need for a joint structure to share data between scientific institutions in Europe and beyond. It is the aim of large networks and infrastructures such as the Global Biodiversity Information Facility (GBIF), the Biodiversity Information Standards (TDWG), the Distributed System of Scientific collections (DiSSCo) and the Consortium of European Taxonomic Facilities (CETAF) to further implement and improve these efforts, thereby gaining ever increasing efficiencies. In this context, the three institutions mentioned above, submitted the NaturalHeritage project (http://www.belspo.be/belspo/brain-be/themes_3_HebrHistoScien_en.stm) granted in 2017 by the Belgian Science Policy Service, which runs from 2017 to 2020. The project provides links among databases and services. The unique qualities of each database are maintained, while the information can be concentrated and exposed in a structured way via one access point. This approach aims also to link data that are unconnected at present (e.g. relationship between soil/substrate, vegetation and associated fauna) and to improve the cross-validation of data. (1) The NaturalHeritage prototype (http://www.naturalheritage.be) is a shared research portal with an open access infrastructure, which is still in the development phase. Its backbone is an ElasticSearch catalogue, with Kibana, and a Python aggregator gathering several types of (re)sources: relational databases, REpresentational State Transfer (REST) services of objects databases and bibliographical data, collections metadata and the GBIF Internet Publishing Toolkit (IPT) for observational and taxonomical data. Semi-structured data in English are semantically analysed and linked to a rich autocomplete mechanism. Keywords and identifiers are indexed and grouped in four categories (“what”, “who”, “where”, “when”). The portal can act also as an Open Archives Initiatives Protocol for Metadata Harvesting (OAI-PMH) service and ease indexing of the original webpage on the internet with microdata enrichment. (2) The collection data management system of DaRWIN (Data Research Warehouse Information Network) of RBINS and RMCA has been improved as well. External (meta)data requirements, i.e. foremost publication into or according to the practices and standards of GBIF and OBIS (Ocean Biogeographic Information System: https://obis.org) for biodiversity data, and INSPIRE (https://inspire.ec.europa.eu) for geological data, have been identified and evaluated. New and extended data structures have been created to be compliant with these standards, as well as the necessary procedures developed to expose the data. Quality control tools for taxonomic and geographic names have been developed. Geographic names can be hard to confirm as their lack of context often requires human validation. To address this a similarity measure is used to help map the result. Species, locations, sampling devices and other properties have been mapped to the World Register of Marine Species and DarwinCore (http://www.marinespecies.org), Marine Regions and GeoNames, the AGRO Agronomy and Vertebrate trait ontologies and the British Oceanographic Data Centre (BODC) vocabularies (http://www.obofoundry.org/ontology/agro.html). Extensive mapping is necessary to make use of the ExtendedMeasurementOrFact Extension of DarwinCore (https://tools.gbif.org/dwca-validator/extensions.do). External (meta)data requirements, i.e. foremost publication into or according to the practices and standards of GBIF and OBIS (Ocean Biogeographic Information System: https://obis.org) for biodiversity data, and INSPIRE (https://inspire.ec.europa.eu) for geological data, have been identified and evaluated. New and extended data structures have been created to be compliant with these standards, as well as the necessary procedures developed to expose the data. Quality control tools for taxonomic and geographic names have been developed. Geographic names can be hard to confirm as their lack of context often requires human validation. To address this a similarity measure is used to help map the result. Species, locations, sampling devices and other properties have been mapped to the World Register of Marine Species and DarwinCore (http://www.marinespecies.org), Marine Regions and GeoNames, the AGRO Agronomy and Vertebrate trait ontologies and the British Oceanographic Data Centre (BODC) vocabularies (http://www.obofoundry.org/ontology/agro.html). Extensive mapping is necessary to make use of the ExtendedMeasurementOrFact Extension of DarwinCore (https://tools.gbif.org/dwca-validator/extensions.do).

A cognitive map in a poison frog.

A fundamental question in cognitive science is whether an animal can use a cognitive map. A cognitive map is a mental representation of the external world, and knowledge of one's place in this world, that can be used to determine efficient routes to any destination. Many birds and mammals are known to employ a cognitive map, but whether other vertebrates can create a cognitive map is less clear. Amphibians are capable of using beacons, gradients and landmarks when navigating, and many are proficient at homing. Yet only one prior study directly tested for a cognitive map in amphibians, with negative results. Poison frogs exhibit unusually complex social and spatial behaviors and are capable of long-distance homing after displacement, suggesting that they may be using complex spatial navigation strategies in nature. Here, we trained the poison frog Dendrobates auratus in a modified Morris water maze that was designed to suppress thigmotaxis to the maze wall, promoting exploration of the arena. In our moat maze, the poison frogs were able to use a configuration of visual cues to find the hidden platform. Moreover, we demonstrate that they chose direct paths to the goal from multiple random initial positions, a hallmark of a cognitive map. The performance of the frogs in the maze was qualitatively similar to that of rodents, suggesting that the potential to evolve a cognitive map is an evolutionarily conserved trait of vertebrates.

Earliest axial fossils from the genus Australopithecus

Australopitheus anamensis fossils demonstrate that craniodentally and postcranially the taxon was more primitive than its evolutionary successor Australopithecus afarensis. Postcranial evidence suggests habitual bipedality combined with primitive upper limbs and an inferred significant arboreal adaptation. Here we report on A. anamensis fossils from the Assa Issie locality in Ethiopia's Middle Awash area dated to ∼4.2 Ma, constituting the oldest known Australopithecus axial remains. Because the spine is the interface between major body segments, these fossils can be informative on the adaptation, behavior and our evolutionary understanding of A. anamensis. The atlas, or first cervical vertebra (C1), is similar in size to Homo sapiens, with synapomorphies in the articular facets and transverse processes. Absence of a retroglenoid tubercle suggests that, like humans, A. anamensis lacked the atlantoclavicularis muscle, resulting in reduced capacity for climbing relative to the great apes. The retroflexed C2 odontoid process and long C6 spinous process are reciprocates of facial prognathism, a long clivus and retroflexed foramen magnum, rather than indications of locomotor or postural behaviors. The T1 is derived in shape and size as in Homo with an enlarged vertebral body epiphyseal surfaces for mitigating the high-magnitude compressive loads of full-time bipedality. The full costal facet is unlike the extant great ape demifacet pattern and represents the oldest evidence for the derived univertebral pattern in hominins. These fossils augment other lines of evidence in A. anamensis indicating habitual bipedality despite some plesiomorphic vertebral traits related to craniofacial morphology independent of locomotor or postural behaviors (i.e., a long clivus and a retroflexed foramen magnum). Yet in contrast to craniodental lines of evidence, some aspects of vertebral morphology in A. anamensis appear more derived than its descendant A. afarensis.

An atlas of anterior hox gene expression in the embryonic sea lamprey head: Hox-code evolution in vertebrates

In the hindbrain and the adjacent cranial neural crest (NC) cells of jawed vertebrates (gnathostomes), nested and segmentally-restricted domains of Hox gene expression provide a combinatorial Hox-code for specifying regional properties during head development. Extant jawless vertebrates, such as the sea lamprey (Petromyzon marinus), can provide insights into the evolution and diversification of this Hox-code in vertebrates. There is evidence for gnathostome-like spatial patterns of Hox expression in lamprey; however, the expression domains of the majority of lamprey hox genes from paralogy groups (PG) 1-4 are yet to be characterized, so it is unknown whether they are coupled to hindbrain segments (rhombomeres) and NC. In this study, we systematically describe the spatiotemporal expression of all 14 sea lamprey hox genes from PG1-PG4 in the developing hindbrain and pharynx to investigate the extent to which their expression conforms to the archetypal gnathostome hindbrain and pharyngeal hox-codes. We find many similarities in Hox expression between lamprey and gnathostome species, particularly in rhombomeric domains during hindbrain segmentation and in the cranial neural crest, enabling inference of aspects of Hox expression in the ancestral vertebrate embryonic head. These data are consistent with the idea that a Hox regulatory network underlying hindbrain segmentation is a pan vertebrate trait. We also reveal differences in hindbrain domains at later stages, as well as expression in the endostyle and in pharyngeal arch (PA) 1 mesoderm. Our analysis suggests that many Hox expression domains that are observed in extant gnathostomes were present in ancestral vertebrates but have been partitioned differently across Hox clusters in gnathostome and cyclostome lineages after duplication.

Quantitative morphometric analysis of adult teleost fish by X-ray computed tomography

Vertebrate models provide indispensable paradigms to study development and disease. Their analysis requires a quantitative morphometric study of the body, organs and tissues. This is often impeded by pigmentation and sample size. X-ray micro-computed tomography (micro-CT) allows high-resolution volumetric tissue analysis, largely independent of sample size and transparency to visual light. Importantly, micro-CT data are inherently quantitative. We report a complete pipeline of high-throughput 3D data acquisition and image analysis, including tissue preparation and contrast enhancement for micro-CT imaging down to cellular resolution, automated data processing and organ or tissue segmentation that is applicable to comparative 3D morphometrics of small vertebrates. Applied to medaka fish, we first create an annotated anatomical atlas of the entire body, including inner organs as a quantitative morphological description of an adult individual. This atlas serves as a reference model for comparative studies. Using isogenic medaka strains we show that comparative 3D morphometrics of individuals permits identification of quantitative strain-specific traits. Thus, our pipeline enables high resolution morphological analysis as a basis for genotype-phenotype association studies of complex genetic traits in vertebrates.

GABAergic modulation of olfactomotor transmission in lampreys.

Odor-guided behaviors, including homing, predator avoidance, or food and mate searching, are ubiquitous in animals. It is only recently that the neural substrate underlying olfactomotor behaviors in vertebrates was uncovered in lampreys. It consists of a neural pathway extending from the medial part of the olfactory bulb (medOB) to locomotor control centers in the brainstem via a single relay in the caudal diencephalon. This hardwired olfactomotor pathway is present throughout life and may be responsible for the olfactory-induced motor behaviors seen at all life stages. We investigated modulatory mechanisms acting on this pathway by conducting anatomical (tract tracing and immunohistochemistry) and physiological (intracellular recordings and calcium imaging) experiments on lamprey brain preparations. We show that the GABAergic circuitry of the olfactory bulb (OB) acts as a gatekeeper of this hardwired sensorimotor pathway. We also demonstrate the presence of a novel olfactomotor pathway that originates in the non-medOB and consists of a projection to the lateral pallium (LPal) that, in turn, projects to the caudal diencephalon and to the mesencephalic locomotor region (MLR). Our results indicate that olfactory inputs can induce behavioral responses by activating brain locomotor centers via two distinct pathways that are strongly modulated by GABA in the OB. The existence of segregated olfactory subsystems in lampreys suggests that the organization of the olfactory system in functional clusters may be a common ancestral trait of vertebrates.

Expression profiles of MicroRNAs from multiple lumbar spine in sheep

The formation of the spine is a critical stage of mammalian development. The increase of the number of individual axons affects its performance, especially in meat production. To understand the role of miRNAs in sheep vertebrae development, the purpose of this article is to screen candidate microRNAs (miRNAs) associated with sheep spine development. MicroRNAs (miRNAs) are a rich family of small regulatory RNAs that negatively regulate gene expression at the post-transcriptional level. In this study, we used high-throughput sequencing techniques to analyze the microRNAs (miRNAs) expression profiles of L6 (6 lumbar vertebrae) and L7 (7 lumbar vertebrae) in sheep. A total number of 223 miRNAs were detected in the two libraries, and a total of 150 and 148 conserved miRNAs were obtained in L6 and L7, respectively. A total of 5 miRNAs expression differences in L6 compared to L7 (P < 0.05). Of the five obviously differently expressed miRNAs, four miRNAs were down-regulated in the L6 of sheep, and one was up-regulated. In order to further explore the functions of these miRNAs, we predicted the target genes of these differently expressed miRNAs, and obtained 1298 target genes. At the same time, NDRG2 gene, targeted by novel miR-391, which possible plays an important role in the development of the spine. Linkage-integration analysis method was used to construct the interaction network of spinal-associated miRNA and its hypothesized target. In summary, this study provides valuable resources for the transcriptome of multiple vertebral traits in sheep.

An ascending visual pathway to the dorsal telencephalon through the optic tectum and nucleus prethalamicus in the yellowfin goby Acanthogobius flavimanus (Temminck & Schlegel, 1845).

Dual visual pathways reaching the telencephalon appear to be an ancient vertebrate trait, but some teleost fish seem to possess only one pathway via the optic tectum. We undertook the present study to determine if and when this loss occurred during evolution. Tracer injection experiments to the optic nerve, the optic tectum, and the dorsal telencephalon were performed in the present study, to investigate ascending visual pathways to the dorsal telencephalon in an acanthopterygian teleost, the yellowfin goby Acanthogobius flavimanus (Temminck & Schlegel, 1845). We confirmed the presence of a nucleus prethalamicus (PTh) in the goby, which has been convincingly identified only in holocentrids, suggesting that this nucleus is present in other acanthopterygians. We found that the optic tectum projects to the PTh bilaterally. The PTh projects in turn to the dorsal telencephalon, ipsilaterally. These results suggest that the yellowfin goby possesses only an extrageniculate-like pathway, while a geniculate-like pathway could not be identified. This situation is common with that of holocentrids and may be a character common in acanthopterygians. It is possible that a geniculate-like system was lost in the common ancestor of acanthopterygians, although the scenario for the evolution of ascending visual systems in actinopterygians remains uncertain due to the lack of precise knowledge in a number of actinopterygian taxons.

Evolutionarily‐Conserved Mechanisms of Nutrient Acquisition in the Primordial Vertebrate, the Pacific Hagfish

Hagfish occupy the basal position of vertebrate phylogeny and are thus of interest for studies examining the evolution of vertebrate traits. Despite a morphologically basic digestive tract, the carnivorous hagfish can rapidly and effectively acquire nutrients from which they can persist off of for upwards of 11 months. Previous studies indicate that hagfish preferentially utilize carbohydrates and transition into fat‐fuelled energy sources as fasting progresses, yet few studies examine the mechanisms by which hagfish acquire nutrients. The aim of the present study was to examine and characterize the mechanisms of nutrient acquisition in the hindgut of the Pacific hagfish (Eptatretus stoutii), focusing on one molecule from each category of macronutrients: carbohydrates (glucose), fats (oleic acid) and proteins (alanyl‐alanine). Radiotracer fluxes across intestinal tissue demonstrated concentration‐dependent uptake indicative of regulated transport, for glucose (Km 0.37 mM, Jmax 8.48 nmol/cm2/h), oleic acid (Km 55 μM, Jmax 1131 pmol/cm2/h) and alanyl‐alanine (Km 1071 μM, Jmax 70 nmol/cm2/h). Pharmacological inhibition of glucose transporters (34% decrease with 200 μM cytochalasin‐b) and sodium‐glucose linked transporters (52–83% decrease with 0.0001–1μM phlorizin) suggest mechanisms akin to those in mammals exist in the hagfish hindgut. Further similarity in transport arises in that dipeptide transport is dependent upon sodium‐proton exchange, wherein a reduced sodium environment yields a 48% decrease in transport rates. While specific isoforms of mammalian fatty acid transporters demonstrate insulin sensitivity, hagfish oleic acid transport rates were unaffected by insulin application (average rate 2741± 262 pmol/cm2/h, N=10), despite a noted 71% decrease in plasma glucose concentration. Finally, feeding resulted in significant remodeling of the intestinal mucosa with hindgut absorptive microvilli lengthening by 125% post‐feeding. Correspondingly, an increased rate of transport was noted post‐feeding, with the degree of change dependent upon the substrate type. This study reveals evolutionarily‐conserved mechanisms of nutrient transport over a broad range of substrates in the oldest extant vertebrate and solidifies the applicability of hagfish as a model for studies of early vertebrate evolution.Support or Funding InformationSupported by NSERC Discovery Grants to GG (203736) and CG (251083)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Paw preferences in the Asian small-clawed otter – using an inexpensive, video-based protocol to study laterality of rare species in the zoo

ABSTRACTIt is still debated whether limb-use preferences represent a common trait in vertebrates, which is based on a shared phylogenetic history. Unravelling the evolutionary origin and pattern of paw preferences in vertebrates requires the analysis of a larger number of species within an ecologically relevant setting. We therefore investigated whether observations in a zoo enable the collection of reliable data sets by quantifying paw use in two independent groups of Asian small-clawed otters (Amblonyx cinerea). Employing a continuous focal animal sampling method, each day one of the ten individuals was video recorded from outside of the enclosure during usual activity. We selected four types of unimanual behaviour (reaching for food, reaching for non-food, reaching into a hole, carrying an object) and quantified paw use for each animal. Our study provides first evidences for individual paw preferences in otters, which were in line with previously reported forelimb use pattern in carnivoran species. Preferences differed between motor acts but for “reaching into a hole” a population-level right paw bias was detected. These data support that observations in a zoological setting are useful to explore task-dependent paw preferences and may facilitate future studies investigating paw preferences under experimentally controlled conditions.

A comprehensive mitochondrial DNA mixed-stock analysis clarifies the composition of loggerhead turtle aggregates in the Adriatic Sea

Migration is one of several marine vertebrate traits increasingly affected by human encroachment. The Adriatic Sea is an important foraging and wintering site for loggerhead turtles (Caretta caretta), and one of the Mediterranean regions where they are most heavily impacted, particularly by fisheries bycatch. Conservation measures concern foraging and wintering areas and nesting grounds, and must, therefore, be informed by the natal origin of individuals in these mixed aggregates. Genetic diversity was investigated among 488 loggerheads found stranded or incidentally captured in fishing gear across the Adriatic and the origin of individuals was assessed through mixed-stock analysis based on comparison of 755 bp of mitochondrial DNA control region sequence. In addition, we highlight the importance of the Gulf of Manfredonia (Apulia, Italy), and of the northeastern Adriatic—a previously genetically undescribed foraging ground. No significant genetic divergence was recorded among sampling areas, between turtles sampled in cold and warm months or between adults and juveniles. The distribution of turtles across the Adriatic Sea appeared not to depend on individual origin. Rookeries in western Greece and Crete provided the most important contributions to the Adriatic mixed stock. In particular, the Greek populations were the most abundant locally in the Gulf of Manfredonia, so they are likely to experience an even greater impact than previously thought because of the severe fishery bycatch levels in this area. This study also provides an example of how substantial increases in sample sizes permit a relatively comprehensive testing of genetic structure across groupings in foraging aggregations.

A Notch-regulated proliferative stem cell zone in the developing spinal cord is an ancestral vertebrate trait.

Vertebrates have evolved the most sophisticated nervous systems we know. These differ from the nervous systems of invertebrates in several ways, including the evolution of new cell types, and the emergence and elaboration of patterning mechanisms to organise cells in time and space. Vertebrates also generally have many more cells in their central nervous systems than invertebrates, and an increase in neural cell number may have contributed to the sophisticated anatomy of the brain and spinal cord. Here, we study how increased cell number evolved in the vertebrate central nervous system, investigating the regulation of cell proliferation in the lamprey spinal cord. Markers of proliferation show that a ventricular progenitor zone is found throughout the lamprey spinal cord. We show that inhibition of Notch signalling disrupts the maintenance of this zone. When Notch is blocked, progenitor cells differentiate precociously, the proliferative ventricular zone is lost and differentiation markers become expressed throughout the spinal cord. Comparison with other chordates suggests that the emergence of a persistent Notch-regulated proliferative progenitor zone was a crucial step for the evolution of vertebrate spinal cord complexity.

The Genome and Adult Somatic Transcriptome of the Mormyrid Electric Fish Paramormyrops kingsleyae.

Several studies have begun to elucidate the genetic and developmental processes underlying major vertebrate traits. Few of these traits have evolved repeatedly in vertebrates, preventing the analysis of molecular mechanisms underlying these traits comparatively. Electric organs have evolved multiple times among vertebrates, presenting a unique opportunity to understand the degree of constraint and repeatability of the evolutionary processes underlying novel vertebrate traits. As there is now a completed genome sequence representing South American electric eels, we were motivated to obtain genomic sequence from a linage that independently evolved electric organs to facilitate future comparative analyses of the evolution and development of electric organs. We report here the sequencing and de novo assembly of the genome of the mormyrid Paramormyrops kingsleyae using short-read sequencing. In addition, we have completed a somatic transcriptome from 11 tissues to construct a gene expression atlas of predicted genes from this assembly, enabling us to identify candidate housekeeping genes as well as genes differentially expressed in the major somatic tissues of the mormyrid electric fish. We anticipate that this resource will greatly facilitate comparative studies on the evolution and development of electric organs and electroreceptors.