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At least 19 records

Mercury concentrations in Pacific lamprey ( Entosphenus tridentatus ) and sediments in the Columbia River basin: Mercury in Columbia River Pacific lamprey

We investigated mercury accumulation in Pacific lamprey and sediments in the Columbia River basin. Mercury concentrations in larval lamprey differed significantly among sample locations (P < 0.001) and were correlated with concentrations in sediments (r 2 = 0.83), whereas adult concentrations were highly variable (range 0.1–9.5 µg/g) and unrelated to holding time after collection. The results suggest that Pacific lamprey in the Columbia River basin may be exposed to mercury levels that have adverse ecological effects.

54 ENVIRONMENTAL SCIENCES↗

Novel lamprey antibody recognizes terminal sulfated galactose epitopes on mammalian glycoproteins

The terminal galactose residues of N- and O-glycans in animal glycoproteins are often sialylated and/or fucosylated, but sulfation, such as 3-O-sulfated galactose (3-O-SGal), represents an additional, but poorly understood modification. To this end, we have developed a novel sea lamprey variable lymphocyte receptor (VLR) termed O6 to explore 3-O-SGal expression. O6 was engineered as a recombinant murine IgG chimera and its specificity and affinity to the 3-O-SGal epitope was defined using a variety of approaches, including glycan and glycoprotein microarray analyses, isothermal calorimetry, ligand-bound crystal structure, FACS, and immunohistochemistry of human tissue macroarrays. 3-O-SGal is expressed on N-glycans of many plasma and tissue glycoproteins, but recognition by O6 is often masked by sialic acid and thus exposed by treatment with neuraminidase. O6 recognizes many human tissues, consistent with expression of the cognate sulfotransferases (GAL3ST-2 and GAL3ST-3). The availability of O6 for exploring 3-O-SGal expression could lead to new biomarkers for disease and aid in understanding the functional roles of terminal modifications of glycans and relationships between terminal sulfation, sialylation and fucosylation.

59 BASIC BIOLOGICAL SCIENCES↗

Using a Customized Portable Deepwater Portable Electrofisher to Assess Larval Lamprey Populations in Irrigation Canals

Larval lamprey densities were quantified at two large-scale water diversions facilities located on the Yakima River near the city of Yakima, Washington. Water diversions are very common in the Yakima River Basin and provide water to irrigate a diverse range of valuable cropland. Out-migrating larval Pacific Lamprey are vulnerable to entrainment through fish protection screens during the withdrawal period and desiccation or predation during dewatering at the end of the season. In order to determine the impact to lamprey populations which inhabit these regions a portable deepwater electroshocking system was utilized to determine lamprey densities which could otherwise not be determined due to water depth. Surveys were conducted in the fall during the dewatering period in 2015 and 2017 and larval lamprey densities ranged from 0.8 to 8.8 fish m-2 at the Sunnyside Diversion Facility to 4.2 to 6.5 fish m-2 at the Wapato Diversion Facility. The estimates for the total lamprey numbers at each of the region using a portable deepwater electrofishing platform (PDEP) was very similar to those estimates using backpack electrofishing post-dewatering; the PDEP method was 12- 36% higher where comparisons were available. During 2017, the estimated lamprey entrainment at the Sunnyside Diversion increased by 484% from August to November. Lamprey lengths were also determined and ranged from 50 to 130 mm. Our results indicate that the use of the deepwater shocking system was very safe and effective at determining larval lamprey densities at hard to sample regions which are present near irrigation facilities.

larval lamprey, electrofishing, water diversion↗

Genetic Identification of Lamprey Genera and Anadromous Ecotypes in Watersheds of the Northeastern Pacific Ocean

ABSTRACT Nonparasitic, nonmigratory Western Brook Lamprey (WBL; Lampetra ayresii ), and parasitic, anadromous Western River Lamprey (WRL; L. ayresii ) are sympatric lampreys that likely represent different life history variations of a single species. Novel genetic tools are critical for differentiating WBL and WRL, whose larvae preclude morphological identification (ID) and will enable comprehensive assessment of imperiled native lampreys of the Northeastern Pacific (including WBL, WRL, and Pacific Lamprey, Entosphenus tridentatus ). We developed 47 candidate single nucleotide polymorphism (SNP) markers using whole genome resequencing of WBL ( N = 24) and WRL ( N = 15) from Ksi Ts'oohl Ts'ap Creek (Nass River, British Columbia, Canada) which are likely ecotypes distinguished by few divergent SNPs across multiple chromosomes. We used five novel candidate SNPs to perform genetic ID of WBL and WRL ecotypes in collections of mixed native lampreys from lower Columbia River tributaries ( N = 1474), Ksi Ts'oohl Ts'ap Creek ( N = 352), and ocean phase WRL from the Georgia Basin (Salish Sea, British Columbia, Canada; N = 91). Two previously published SNPs were used to ID genera, Entosphenus versus Lampetra . Morphological ID utilized photographs collected from a subset of genotyped lampreys, and high concordance was demonstrated between ID methods for genera (99%) and Lampetra ecotypes (> 98%). We characterized spatial and temporal composition of lamprey genera and ecotypes surveyed across NE Pacific tributaries under the expectation these compositions would be similar across nearby sites and across years at the same site. Proportions of lamprey genera were highly variable within regions and across years; however, Lampetra ecotypic proportions were spatially and temporally stable. WRL were rare in lower Columbia tributaries (~1% average rate among Lampetra ) and common further north (> 40% of Lampetra ). Genetic ID methods are powerful monitoring tools that create the novel ability to ascertain genera and ecotypes regardless of life stage, while increasing the efficiency of surveys by eliminating time‐intensive morphological data collection.

Silver, G. S. [Columbia River Inter‐Tribal Fish Co↗

Behavior and potential threats to survival of migrating lamprey ammocoetes and macrophthalmia

Upon metamorphosis, anadromous juvenile lamprey (macrophthalmia) exhibit distinct migration behaviors that take them from larval rearing habitats in streams to the open ocean. While poorly studied, lamprey larvae (ammocoetes) also engage in downstream movement to some degree. Like migrating salmon smolts, lamprey macrophthalmia undergo behavioral changes associated with a highly synchronized metamorphosis. Unlike salmon smolts, the timing of juvenile migration in lamprey is protracted and poorly documented. Lamprey macrophthalmia and ammocoetes are not strong swimmers, attaining maximum individual speeds of less than 1 m s-1, and sustained speeds of less than 0.5 m s-1. They are chiefly nocturnal and distribute throughout the water column, but appear to concentrate near the bottom in the thalweg of deep rivers. At dams and irrigation diversions, macrophthalmia can become impinged on screens or entrained in irrigation canals, suffer increased predation, and experience physical injury that may result in direct or delayed mortality. The very structures designed to protect migrating juvenile salmonids can be harmful to juvenile lamprey. Yet at turbine intakes and spillways, lampreys, which have no swim bladder, can withstand changes in pressure and shear stress large enough to injure or kill most teleosts. Lamprey populations are in decline in many parts of the world, with some species designated as species of concern for conservation that merit legally mandated protections. Hence, provisions for safe passage of juvenile lamprey are being considered at dams and water diversions in North America and Europe.

Petromyzontiformes,Transformers, Passage, Metamorp↗

Survival, healing, and swim performance of juvenile migratory sea lamprey ( Petromyzon marinus ) implanted with a new acoustic microtransmitter designed for small eel-like fishes

Background: Little is known about the transformer stage of the parasitic lampreys, a brief but critical period that encompasses juvenile out-migration from rivers to lakes or oceans to begin parasitic feeding. Information about this life stage could have significant conservation implications for both imperiled and invasive lampreys. We investigated tag retention, survival, wound healing, and swim performance of newly transformed sea lamprey (Petromyzon marinus) implanted with a new micro-acoustic transmitter, the eel–lamprey acoustic transmitter (ELAT), in a controlled laboratory environment. Results: The 61-day survival of our tagged subjects was 71%, within the range reported in similar studies of juvenile lampreys. However, survival was significantly lower in the tagged animals (vs control), with no effect statistically attributable to measures of animal length, mass, condition, or population of origin (Great Lakes vs. Atlantic drainage). Mortality in tagged fish was concentrated in the first four days post-surgery, suggesting injury from the surgical process. An unusually long recovery time from anesthesia may have contributed to the increased mortality. In a simple burst swim assay, tagged animals swam significantly slower (– 22.5%) than untagged animals, but were not significantly different in endurance swim tests. A composite wound healing score at day four was a significant predictor of maximum burst swim speed at day 20, and wound condition was related to animal mass, but not length, at the time of tagging. Conclusions: Impairments to survival and swim performance of juvenile sea lamprey implanted with the ELAT transmitter were within currently reported ranges for telemetry studies with small, difficult to observe fishes. Our results could be improved with more refined anesthesia and surgical techniques. The ability to track migratory movements of imperiled and pest populations of parasitic lampreys will improve our ability to estimate vital rates that underlie recruitment to the adult population (growth, survival) and to investigate the environmental factors that regulate the timing and rates of movement, in wild populations.

59 BASIC BIOLOGICAL SCIENCES↗

Spatial patterns in occupancy and density of larval lampreys in freshwater habitats restored to a Stage 0 condition

Abstract We examined occupancy and density of larval lampreys ( Entosphenus tridentatus and Lampetra spp.) in two rivers in Oregon (USA) restored to a Stage 0 condition 1–5 years prior, using a multiscale occupancy model and a zero‐inflated Poisson mixture model. We sampled lampreys using backpack electrofishing in randomly distributed, paired, 1‐m 2 quadrats and recorded environmental data. Probabilities of occupancy and density were higher when water velocity was low, the substrate was noncompacted, and sediment was dominated by fines (<4 mm). At mean water depth (0.34 m) and velocity (0.09 m/s), estimated densities in occupied quadrats were 4.8 lampreys/m 2 (95%: 3.4–6.9) when the substrate was compacted, and fines were not dominant, and 21.1 lampreys/m 2 (95%: 17.7–25.3) when the substrate was noncompacted and fines were dominant. Probabilities of detecting occupancy in a 1‐m 2 quadrat sampled by backpack electrofishing were 0.76 (95%: 0.64–0.87) when captured after visual observation and 0.80 (95%: 0.71–0.88) with blind sweeps (i.e., constantly moving the net regardless of observation). The probability of capturing a single lamprey in a quadrat sampled by blind sweeps was 0.32 (95%: 0.27–0.37). Sampling in paired 1‐m 2 quadrats facilitated concurrent examination of patterns in occupancy and density while accounting for capture probability, which could aid temporal monitoring of restored habitats. To the best of our knowledge, this is the first study to document occupancy and estimate densities of larval lampreys in habitats that underwent valley floor restoration to Stage 0. We observed both lamprey genera within 5 years of restoration. Aquatic restoration that increases low‐velocity, noncompacted, fine sediment habitats could benefit lampreys.

Harris, Julianne E.↗

Detection efficiency of adult Pacific lamprey passage counts at Leaburg Dam and upstream distribution in the McKenzie River (Oregon, USA)

Abstract Adult Pacific lamprey ( Entosphenus tridentatus ) were counted using consistent methodology since 2005 with a video monitoring system as they passed Leaburg Dam (McKenzie River, Oregon, USA) en route to upstream spawning areas. In this study we evaluated the detection efficiency of the video system and upstream distribution of Pacific lamprey using the video counts of lamprey passage (herein, “dam counts”), passive integrated transponder (PIT) tags, and radio telemetry. In 2019–2020 we collected, tagged and tracked 32 adult lamprey (4 from the McKenzie River and 28 that were translocated from Willamette Falls). All fish were tagged and released into the tailrace of Leaburg Dam in June 2019. Oregon Department of Fish and Wildlife employees and volunteers from the local community conducted mobile radiotracking above and below the dam in drift boats (114 mainstem river kilometers) and on foot (several tributaries). The estimated detection efficiency for dam counts was 92% (95% confidence interval: 67%–99%). Fifty percent (16 of 32) of the tagged lamprey passed the dam, including 13 of 28 that were translocated. Thirty‐seven percent (6 of 16) of lamprey that passed Leaburg Dam were detected in a restored reach of the South Fork McKenzie, 32.6 river kilometers upstream of the dam.

Romer, Jeremy D.↗

Performance Comparisons for Artificially Propagated and Wild Pacific Lamprey Juveniles and Larvae

ABSTRACT Artificially propagated Pacific lamprey ( Entosphenus tridentatus ) are produced for restoration and for use in dam passage studies to reduce the demand for wild fish. Such uses require that animals are representative of their wild counterparts. Previous work indicated that this is true for Pacific lamprey larvae and juveniles reared in the hatchery with respect to the length of sustained swimming. However, more subtle differences in behaviour and performance that lamprey need to survive have not been assessed. In this study, artificially propagated and wild fish were compared in laboratory tests under no‐flow conditions to examine light avoidance, burrowing speed, burst swim speed, volitional routine swim speed and time to come to rest. Most larvae burrowed in less than a minute, and we found highly significant differences ( p < 0.001) between artificially propagated and wild larvae burrowing times, a critical escape behaviour. This could have implications for studies of larval entrainment at irrigation diversion canals or in turbine boils at dams. Interestingly, all of the wild juveniles tested came to rest quickly after introduction to the chamber (1.5 min), while artificially propagated lamprey swam robotically near the surface and 48% did not come to rest in the first 10 min (median time to rest = 9.5 min). In contrast, wild juveniles quickly (median = 1.47 min) sought areas near the tank bottom and attached strongly with their oral disc. Such behavioural differences could have important survival consequences for artificially propagated lamprey as they approach turbine intakes, bypass screens and irrigation diversion headgates. This study highlights the need to conduct behavioural assays that examine subtleties of fish behaviour that can be missed with traditional swim tunnel comparisons.

Frick, Kinsey [Fish Ecology Division, Northwest Fi↗

Performance of a Vertical‐Slot Fish Pass for the Sea Lamprey Petromyzon marinus L. and Habitat Recolonization

Abstract In 2011, a vertical‐slot fish pass was built at the Coimbra Açude‐Ponte dam (Mondego River, Portugal), approximately 45 km upstream from the river mouth. The performance of this infrastructure for sea lamprey passage was evaluated between 2011 and 2015 using several complementary methodologies, namely radio telemetry [conventional and electromyogram (EMG)], passive integrated transponder (PIT) telemetry and electrofishing surveys. During the study period, the electrofishing revealed a 29‐fold increase in the abundance of larval sea lamprey upstream of the fish pass. Of the 20 radio‐tagged individuals released downstream from the dam, 33% managed to find and successfully surpass the obstacle in less than 2 weeks, reaching the spawning areas located in the upstream stretch of the main river and in one important tributary. Fish pass efficiency was assessed with a PIT antenna installed in the last upstream pool and revealed a 31% efficiency, with differences between and within migratory seasons. Time of day and river flow significantly influenced the attraction efficiency of the fish pass, with lampreys negotiating it mainly during the night period and when discharge was below 50 m 3 s −1 . Sea lampreys tagged with EMG transmitters took 3 h to negotiate the fish pass, during which high muscular effort was only registered during passage, or passage attempts, of the vertical slots. The use of complementary methodologies provided a comprehensive passage evaluation for sea lamprey, a species for which there is a considerable paucity of valuable data concerning behavioural, physiological and environmental influences on obstacle negotiation. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Pereira, E.↗

New technology for an ancient fish: A lamprey life cycle modeling tool with an R Shiny application

Lampreys (Petromyzontiformes) are an ancient group of fishes with complex life histories. We created a life cycle model that includes an R Shiny interactive web application interface to simulate abundance by life stage. This will allow scientists and managers to connect available demographic information in a framework that can be applied to questions regarding lamprey biology and conservation. We used Pacific lamprey ( Entosphenus tridentatus ) as a case study to highlight the utility of this model. We applied a global sensitivity analysis to explore the importance of individual life stage parameters to overall population size, and to better understand the implications of existing gaps in knowledge. We also provided example analyses of selected management scenarios (dam passage, fish translocations, and hatchery additions) influencing Pacific lamprey in fresh water. These applications illustrate how the model can be applied to inform conservation efforts. This tool will provide new capabilities for users to explore their own questions about lamprey biology and conservation. Simulations can hone hypotheses and predictions, which can then be empirically tested in the real world.

Gomes, Dylan G. E. (ORCID:0000000226423728)↗

Methods for Implantation of a New Micro Acoustic Tag in Juvenile Pacific Lamprey and American Eel

Juvenile Pacific lamprey and American eels were used for laboratory evaluations to determine potential effects from tag implantation. Telemetry technology has been identified as a way to obtain more detailed information on movement and behavior across a broader spatial scale than is possible with other known technology. For laboratory studies using juvenile Pacific lamprey, tested fish reached the macropthalmia stage of development and ranged from 140–220 mm in length and 1.8–7.0 g in weight. Juvenile lamprey tag loss was 3% (n = 51) and testing showed no significant differences in the swimming ability of tagged lamprey compared to their un-tagged counterparts. American eels reached the yellow-phase (1.5 years post glass-stage) and were 113–175 mm in total length and 1.7–7.5 g in weight. No mortality occurred during the holding period and a tag loss of 3.8% (n = 1) was observed within the first 20 days post-tagging, which is the current projected battery life of the tag. The presence of the tag did not have any significant effect on the swimming ability of tagged eels compared to untagged controls.

Pacific lamprey, American eel, tagging↗

Pacific Lamprey Translocations to the Snake River Boost Abundance of All Life Stages

Abstract The Pacific Lamprey Entosphenus tridentatus is an anadromous fish that has severely declined in the Snake River basin. In 2007, the Nez Perce Tribe initiated translocation of adults to this region from the main‐stem Columbia River and has continued the translocations to this day. These actions are aimed at restoring larval abundance along with holistic habitat improvements. We performed parentage and sibship analyses with 260 single‐nucleotide polymorphism loci to monitor productivity of translocated lamprey over a decade (2007–2018). These results provide the first direct evidence that translocations boosted larval abundance, increased juvenile production in the interior Columbia River (~3% in 2017 and 2018), and demonstrated successful migration to the Pacific Ocean. Per capita juvenile production from Snake River adult lamprey translocations out‐performed that of their volitionally migrating counterparts in the interior Columbia River, demonstrating that translocating adults to suitable habitats increased overall productivity. We projected that the translocations may eventually return enough adult offspring to the Columbia River to replace the annual take of adults for translocations, despite many adults likely returning to other basins. These translocations are also restoring Pacific Lamprey to historical areas where they had been nearly extirpated, which has impactful benefits both culturally and ecologically. Further, this study provided an unprecedented volume of data that has redefined key biological attributes, including timing of life stage transformation (average of 6.7‐year‐old juveniles), larval growth rates (~22 mm/year for the first 4 years), ocean duration (average of 5.1 years), dispersal (~3% of Snake River‐origin adults that return to the Columbia River will divert to the Willamette River), and life span (median age of postspawn adults was 12.9 years). We even quantified a new biological phenomenon of delayed spawning, which occurs at low rates (~9%) in nature. These data provide an opportunity to monitor ongoing translocation efforts and inform adaptive management to aid species recovery.

Hess, Jon E.↗

Oxygen consumption of sexually mature adult, first-feeding larval, and yearling Pacific Lampreys

Abstract Objective The Pacific Lamprey Entosphenus tridentatus is considered a first food of Native American tribes, such as the Confederated Tribes of the Umatilla Indian Reservation. Populations of this imperiled species have declined such that harvest for traditional use is limited. As a result, propagation and culture of Pacific Lampreys have been initiated to supply animals for restoration and recovery efforts. These efforts require transport and periodic holding of both adults and larvae under static (no-flow) conditions. Hence, guidelines for ensuring an adequate oxygen supply are needed for all life stages of this species. Methods We measured oxygen consumption rates of mature adults, first-feeding larvae, and yearlings under static conditions at 12–15°C. In addition, we recorded indicators of stress during hypoxia for adults and estimated routine respiration rates during and after larval feeding. Result Adults exhibited surprisingly high metabolic rates when at rest in static chambers. At 12°C, a single adult typically reduced dissolved oxygen levels in 10 L to <2 mg/L in just 90 min (oxygen consumption rates of 100–200 mg/kg/h). Adults often started to climb the walls of open chambers when dissolved oxygen levels approached 2 mg/L, allowing them to raise their branchiopores into air. Larvae remained quiescent, even when oxygen levels dropped below 1 mg/L, and costs of feeding increased routine respiration by 22–56%. Conclusion Based on these observations, we recommend that adult Pacific Lampreys always be transported in aerated containers and with access to air at the top of the tank. Although larvae exhibited hypoxia tolerance, care should be taken to ensure adequate oxygen availability, particularly during and immediately after feeding.

Moser, Mary↗

Evaluation of survival estimates generated from tracking downstream migrating juvenile sea lamprey ( Petromyzon marinus ) with a miniature acoustic telemetry tag

Fish that undertake juvenile migration pass through habitats that vary in mortality risk. The ability to identify regions of persistently low survival would improve fishery management. We conducted a field study combined with predictive modeling of survival in downstream migrating juvenile sea lamprey using a new micro-acoustic telemetry tag designed for implantation into small, slender-bodied fishes. Detection data were collected from eight receivers placed across a coastal riverine–wetland–lake complex. Juvenile sea lamprey initiated downstream movement near nautical twilight, stopped frequently, and were more likely to move during high river discharge. Estimated survival was highest in the riverine reach, declined through the river–wetland complex, and dropped precipitously in the drowned rivermouth lake. However, the high transmission rate and resulting short battery life of the microtransmitters (as configured) likely resulted in missed detections in the lower reaches. Simulation analyses suggested survival estimation could be improved by increasing the number of tagged lamprey and staggering release locations. We offer practical recommendations for the use of this new transmitter in field studies with small anguilliform fish.

59 BASIC BIOLOGICAL SCIENCES↗

Lamprey Tagging

Pacific Northwest National Laboratory has developed a super-small acoustic tracking tag designed just for juvenile lamprey. In this video, PNNL researcher Alison Colotelo describes how she and her colleague Kate Deters inject young lamprey with the PNNL tag.

59 BASIC BIOLOGICAL SCIENCES↗

Id expression in amphioxus and lamprey highlights the role of gene cooption during neural crest evolution

Neural crest cells are unique to vertebrates and generate many of the adult structures that differentiate them from their closest invertebrate relatives, the cephalochordates. Id genes are robust markers of neural crest cells at all stages of development. We compared Id gene expression in amphioxus and lamprey to ask if cephalochordates deploy Id genes at the neural plate border and dorsal neural tube in a manner similar to vertebrates. Furthermore, we examined whether Id expression in these cells is a basal vertebrate trait or a derived feature of gnathostomes. We found that while expression of Id genes in the mesoderm and endoderm is conserved between amphioxus and vertebrates, expression in the lateral neural plate border and dorsal neural tube is a vertebrate novelty. Furthermore, expression of lamprey Id implies that recruitment of Id genes to these cells occurred very early in the vertebrate lineage. Based on expression in amphioxus we postulate that Id cooption conferred sensory cell progenitor-like properties upon the lateral neurectoderm, and pharyngeal mesoderm-like properties upon cranial neural crest. Amphioxus Id expression is also consistent with homology between the anterior neurectoderm of amphioxus and the presumptive placodal ectoderm of vertebrates. These observations support the idea that neural crest evolution was driven in large part by cooption of multipurpose transcriptional regulators from other tissues and cell types.

NASA Program Fundamental Space Biology↗

Neural crest contributions to the lamprey head

The neural crest is a vertebrate-specific cell population that contributes to the facial skeleton and other derivatives. We have performed focal DiI injection into the cranial neural tube of the developing lamprey in order to follow the migratory pathways of discrete groups of cells from origin to destination and to compare neural crest migratory pathways in a basal vertebrate to those of gnathostomes. The results show that the general pathways of cranial neural crest migration are conserved throughout the vertebrates, with cells migrating in streams analogous to the mandibular and hyoid streams. Caudal branchial neural crest cells migrate ventrally as a sheet of cells from the hindbrain and super-pharyngeal region of the neural tube and form a cylinder surrounding a core of mesoderm in each pharyngeal arch, similar to that seen in zebrafish and axolotl. In addition to these similarities, we also uncovered important differences. Migration into the presumptive caudal branchial arches of the lamprey involves both rostral and caudal movements of neural crest cells that have not been described in gnathostomes, suggesting that barriers that constrain rostrocaudal movement of cranial neural crest cells may have arisen after the agnathan/gnathostome split. Accordingly, neural crest cells from a single axial level contributed to multiple arches and there was extensive mixing between populations. There was no apparent filling of neural crest derivatives in a ventral-to-dorsal order, as has been observed in higher vertebrates, nor did we find evidence of a neural crest contribution to cranial sensory ganglia. These results suggest that migratory constraints and additional neural crest derivatives arose later in gnathostome evolution.

Non-NASA Center↗