Adult Lampreys Research Articles

Similarities and Differences for Swimming in Larval and Adult Lampreys.

The spinal locomotor networks controlling swimming behavior in larval and adult lampreys may have some important differences. As an initial step in comparing the locomotor systems in lampreys, in larval animals the relative timing of locomotor movements and muscle burst activity were determined and compared to those previously published for adults. In addition, the kinematics for free swimming in larval and adult lampreys was compared in detail for the first time. First, for swimming in larval animals, the neuromechanical phase lag between the onsets or terminations of muscle burst activity and maximum concave curvature of the body increased with increasing distance along the body, similar to that previously shown in adults. Second, in larval lampreys, but not adults, absolute swimming speed (U; mm s(-1)) increased with animal length (L). In contrast, normalized swimming speed (U'; body lengths [bl] s(-1)) did not increase with L in larval or adult animals. In both larval and adult lampreys, U' and normalized wave speed (V') increased with increasing tail-beat frequency. Wavelength and mechanical phase lag did not vary significantly with tail-beat frequency but were significantly different in larval and adult animals. Swimming in larval animals was characterized by a smaller U/V ratio, Froude efficiency, and Strouhal number than in adults, suggesting less efficient swimming for larval animals. In addition, during swimming in larval lampreys, normalized lateral head movements were larger and normalized lateral tail movements were smaller than for adults. Finally, larval animals had proportionally smaller lateral surface areas of the caudal body and fin areas than adults. These differences are well suited for larval sea lampreys that spend most of the time buried in mud/sand, in which swimming efficiency is not critical, compared to adults that would experience significant selection pressure to evolve higher-efficiency swimming to catch up to and attach to fish for feeding as well as engage in long-distance migration during spawning. Finally, the differences in swim efficiency for larval and adult lampreys are compared to other animals employing the anguilliform mode of swimming.

River Lamprey (Lampetra ayresii) Outmigrant Upstream of the John Day Dam in the Mid-Columbia River

The anadromous River Lamprey (Lampetra ayresii) is known to occur in North America from Alaska to central California, generally in association with larger estuarine systems (such as the Fraser River in British Columbia, the Sacramento River in California, and the Columbia River in Washington-Oregon; Vladykov and Follett 1958; Weitkamp and others 2015). Its occurrence in the Columbia River basin, however, has not been well documented. Although small numbers of juvenile (feeding phase) River Lamprey have been captured incidentally in the Columbia River Estuary (Bond and others 1983; Weitkamp and others 2012, 2015), there are few records of this species in freshwater (Kostow 2002). The farthest upriver report in the Columbia River basin is a March 1940 record at Bonneville (presumably at Bonneville Dam), 235 river kilometers (rkm) from the Pacific Ocean (Vladykov and Follett 1958). However, adult River Lamprey (117to 324-mm total length; Vladykov and Follett 1958; Bond and others 1983; Wydoski and Whitney 2003; Weitkamp and others 2015) are less noticeable than the larger Pacific Lamprey (Entosphenus tridentatus) (700to 800-mm total length; Clemens and others 2010) and might spawn in deep water in large river systems similar to other lamprey species (Dawson and others 2015). Thus, even when abundant, they are rarely observed in freshwater (Beamish 1980; Beamish and Youson 1987). Furthermore, during its prolonged freshwater larval stage, it is difficult or impossible to distinguish River Lamprey, either morphologically or genetically, from the closely related and widely distributed Western Brook Lamprey (Lampetra richardsoni) (Docker 2009). The nonmigratory, non-trophic adult Western Brook Lamprey (90to 180-mm total length) can be distinguished from adult River Lamprey by morphological and dentition characters (Wydoski and Whitney 2003; Docker 2009). There is a growing conservation concern for lampreys worldwide. Like other species, the River Lamprey is susceptible to various freshwater threats such as dredging and barriers to migration (Maitland and others 2015). The Columbia River Basin is currently fragmented by many mainstem hydropower, navigation, and flood-control dams, as well as numerous additional dams and irrigation diversions in tributary systems. Many of these dams do not provide fish passage and those that do are likely problematic for migratory adult lampreys (Moser and others 2015). Documentation of River Lamprey in the Columbia River basin would help inform management of this highly regulated system, as well as elucidate distribution patterns to aid conservation. A current effort is underway to increase the monitoring of outmigrating juvenile lampreys through the operation of salmonid smolt monitoring systems at mainstem Columbia River dams (Fish Passage Center 2014). Subsamples of juvenile (metamorphosed to outmigrating phase) and larval (unmetamorphosed) lampreys are collected, anesthetized, identified (to species for juvenile lampreys and to genus for larval lampreys), measured, and assessed for body condition. A putative juvenile (recently metamorphosed) River Lamprey was collected at the John Day Dam smolt bypass facility on 28 May 2013 from a sample time frame of 13:00 to 07:00. The fish was 144-mm total length and weighed 3.1 g. Its dentition pattern clearly distinguished it from the relatively more common Pacific Lamprey (Fig. 1). Pacific Lamprey have 3 large cusps on the supra-oral lamina, 4 pairs of lateral teeth, and 1 row of posterior teeth; whereas the River Lamprey has 2 cusps on the supra-oral lamina, 3 pairs of lateral teeth, and no posterior teeth (Wydoski and Whitney 2003). Its caudal pigmentation was likewise consistent with Lampetra spp. (Fig. 2, Fig. 3); lampreys in the Entosphenus genus possess a lighter caudal ridge, whereas the caudal ridge of Lampetra spp. is uniformly dark (Goodman and others GENERAL NOTES

Evidence that sea lampreys (Petromyzon marinus) complete their life cycle within a tributary of the Laurentian Great Lakes by parasitizing fishes in inland lakes

Abstract The sea lamprey ( Petromyzon marinus ) invaded the upper Laurentian Great Lakes and feeds on valued fish. The Cheboygan River, Michigan, USA, is a large sea lamprey producing tributary to Lake Huron and despite having a renovated dam 2 km from the river mouth that presumably blocks sea lamprey spawning migrations, the watershed upstream of the dam remains infested with larval sea lamprey. A navigational lock near the dam has been hypothesized as the means of escapement of adult sea lampreys from Lake Huron and source of the upper river population (H1). However, an alternative hypothesis (H2) is that some sea lampreys complete their life cycle upstream of the dam, without entering Lake Huron. To evaluate the alternative hypothesis, we gathered angler reports of lamprey wounds on game fishes upstream of the dam, and captured adult sea lampreys downstream and upstream of the dam to contrast abundance, run timing, size, and statolith microchemistry. Results indicate that a small population of adult sea lampreys (n

Potential for carbon dioxide to act as a non-physical barrier for invasive sea lamprey movement

Abstract Invasive sea lampreys have had substantial negative ecological and economic impacts on the Laurentian Great Lakes region. Control efforts, such as lampricide application and barriers, have resulted in a reduction in number of sea lampreys in the Great Lakes. Due to environmental and non-target impacts of existing control mechanisms, coupled with the fact that no non-physical barrier is completely effective at stopping fish movement, there is a critical need to develop novel control technologies to assist with the control and suppression of sea lamprey populations. Recent work has indicated that carbon dioxide gas (CO 2 ) applied to water will influence the movement and behavior of fishes, providing the potential for CO 2 to act as a non-physical barrier that can exclude fish from a target area. To date, the effectiveness of CO 2 at influencing the movement and behavior of sea lampreys has not been explored. The current study showed that CO 2 applied to water will result in behavioral agitation for both adult and transformer sea lampreys, and will eventually result in equilibrium loss. More importantly, both adult and transformer sea lampreys will ‘choose’ to avoid water with CO 2 concentrations of 85 and 160 mg/L (respectively). Together, results from this study suggest that CO 2 applied to water has the potential to act as a non-physical barrier to deter the movement of free-swimming lamprey in the wild. Carbon dioxide gas can be integrated with existing control technologies to act as a novel barrier technology and augment existing control strategies for sea lampreys.

Efficacy of a side-mounted vertically oriented bristle pass for improving upstream passage of European eel (Anguilla anguilla) and river lamprey (Lampetra fluviatilis) at an experimental Crump weir

Globally, populations of diadromous anguilliform morphotype fish, such as eel and lamprey, have experienced substantial declines, partly as a result of habitat fragmentation caused by river infrastructure. In the UK, a new configuration of hydraulically unobtrusive bristle pass (side-mounted and vertically oriented) has been developed to help upstream moving European eel (Anguilla anguilla) negotiate gauging weirs. The efficacy of vertically oriented bristle passes remains untested, despite their potential as a low-cost low-maintenance solution to improve habitat connectivity at low-head structural barriers worldwide. This study assessed the ability of small (82–320mm) and large (322–660mm) European eel and adult (291–401mm) river lamprey (Lampetra fluviatilis) to pass upstream over an experimental Crump weir installed in a large open-channel flume with (treatment) and without (control) side-mounted vertically oriented bristle passes under three different hydraulic regimes. Both species were highly motivated to explore their surroundings and move upstream during the trials. Under flooded control conditions, passage efficiency (the total number of times fish passed the structure as a percentage of total attempts) and passage success (the number of fish that passed the structure as a percentage of those that attempted) were high, delay was short, and number of failed attempts before passage was low for both species. When difference in head was at its greatest (230mm) and velocity and its variation downstream were high (maximum u and σ: 2.43ms−1 and 0.66ms−1, respectively), the upstream movement of small eel and lamprey was blocked, and passage efficiency and success for large eel low (4.6% and 17.2%, respectively). For large eel that successfully passed, delay was long, and number of failed attempts before upstream passage was high. When bristle passes were installed, passage efficiency for small (91.5%) and large eel (56.7%), and passage success for large eel (76.5%) and lamprey (36.7%) was higher, while delay and the number of attempts before passage was lower for both species. Bristle passes helped European eel and river lamprey pass a small experimental Crump weir, although interspecific variation in efficacy was evident.

Identification and characterization of a ubiquitinconjugating enzyme UBE2A gene from lamprey.

Ubiquitin-conjugating enzymes (E2s) play an important role in the mechanism of ubiquitin transfer. Although in most species many of these enzymes share high sequence and structural conservation, their existence and functions in the lamprey remain unknown. In this study, we identified and characterized a ubiquitin-conjugating enzyme (UBE2A)-like gene in lamprey. The gene, designated as LaUBE2A, contained a 456-bp open reading frame encoding a 152-amino acid protein with a typical UBC domain. Real-time PCR assay showed that LaUBE2A was expressed in various tissues of the adult lamprey, with higher levels in the leukocytes and muscle and lower levels in the skin and liver. The high conservation in amino acid sequence between LaUBE2A and UBE2As from Homo sapiens, Mus musculus, Cavia porcellus, and Alligator sinensi implied that the function of LaUBE2A may be similar to that of UBE2A.

Cell secretion from the adult lamprey supraneural body tissues possesses cytocidal activity against tumor cells.

The supraneural body was identified in the adult lamprey, and its secretions induced the death of a variety of tumor cells but had no effect on normal cells. The cell secretions from different lamprey tissues were separated, and these secretions killed human tumor cells to varying degrees. The cell secretions induced remarkable cell morphological alterations such as cell blebbing, and the plasma membrane was destroyed by the secretions. In addition, the secretions induced morphological alterations of the mitochondria, cytoskeletal structure, and endoplasmic reticulum, eventually leading to cell death. These observations suggest the presence of a novel protein in the lamprey and the possibility of new applications for the protein in the medical field.Electronic supplementary materialThe online version of this article (doi:10.1186/s40064-015-1270-6) contains supplementary material, which is available to authorized users.

Behavioural response of adult sea lamprey (Petromyzon marinus) to predator and conspecific alarm cues: evidence of additive effects

Sea lampreys Petromyzon marinus, an invasive pest in the Upper Great Lakes, avoid odours that represent danger in their habitat. These odours include conspecific alarm cues and predator cues, like 2-phenylethylamine hydrochloride (PEA HCl), which is found in the urine of mammalian predators. Whether conspecific alarm cues and predator cues function additively or synergistically when mixed together is unknown. The objectives of this experimental study were to determine if the avoidance response of sea lamprey to PEA HCl is proportional to the concentration delivered, and if the avoidance response to the combination of a predator cue (PEA HCl) and sea lamprey alarm cue is additive. To accomplish the first objective, groups of ten sea lampreys were placed in an artificial stream channel and presented with stepwise concentrations of PEA HCl ranging from 5 × 10−8 to 5 × 10−10 M and a deionized water control. Sea lampreys exhibited an increase in their avoidance behaviour in response to increasing concentrations of PEA HCl. To accomplish the second objective, sea lampreys were exposed to PEA HCl, conspecific alarm cue and a combination of the two. Sea lampreys responded to the combination of predator cue and conspecific alarm cue in an additive manner.

Factors Influencing Capture of Invasive Sea Lamprey in Traps Baited With a Synthesized Sex Pheromone Component.

The sea lamprey, Petromyzon marinus, is emerging as a model organism for understanding how pheromones can be used for manipulating vertebrate behavior in an integrated pest management program. In a previous study, a synthetic sex pheromone component 7α,12α, 24-trihydroxy-5α-cholan-3-one 24-sulfate (3kPZS) was applied to sea lamprey traps in eight streams at a final in-stream concentration of 10(-12) M. Application of 3kPZS increased sea lamprey catch, but where and when 3kPZS had the greatest impact was not determined. Here, by applying 3kPZS to additional streams, we determined that overall increases in yearly exploitation rate (proportion of sea lampreys that were marked, released, and subsequently recaptured) were highest (20-40%) in wide streams (~40 m) with low adult sea lamprey abundance (<1000). Wide streams with low adult abundance may be representative of low-attraction systems for adult sea lamprey and, in the absence of other attractants (larval odor, sex pheromone), sea lamprey may have been more responsive to a partial sex pheromone blend emitted from traps. Furthermore, we found that the largest and most consistent responses to 3kPZS were during nights early in the trapping season, when water temperatures were increasing. This may have occurred because, during periods of increasing water temperatures, sea lamprey become more active and males at large may not have begun to release sex pheromone. In general, our results are consistent with those for pheromones of invertebrates, which are most effective when pest density is low and when pheromone competition is low.

Substance P Depolarizes Lamprey Spinal Cord Neurons by Inhibiting Background Potassium Channels.

Substance P is endogenously released in the adult lamprey spinal cord and accelerates the burst frequency of fictive locomotion. This is achieved by multiple effects on interneurons and motoneurons, including an attenuation of calcium currents, potentiation of NMDA currents and reduction of the reciprocal inhibition. While substance P also depolarizes spinal cord neurons, the underlying mechanism has not been resolved. Here we show that effects of substance P on background K+ channels are the main source for this depolarization. Hyperpolarizing steps induced inward currents during whole-cell voltage clamp that were reduced by substance P. These background K+ channels are pH sensitive and are selectively blocked by anandamide and AVE1231. These blockers counteracted the effect of substance P on these channels and the resting membrane potential depolarization in spinal cord neurons. Thus, we have shown now that substance P inhibits background K+ channels that in turn induce depolarization, which is likely to contribute to the frequency increase observed with substance P during fictive locomotion.

Tryptophan hydroxylase and serotonin receptor 1A expression in the retina of the sea lamprey

The dual development of the retina of lampreys is exceptional among vertebrates and offers an interesting EvoDevo (evolutionary developmental biology) model for understanding the origin and evolution of the vertebrate retina. Only a single type of photoreceptor, ganglion cell and bipolar cell are present in the early-differentiated central retina of lamprey prolarvae. A lateral retina appears later in medium-sized larvae (about 3 years after hatching in the sea lamprey), growing and remaining largely neuroblastic until metamorphosis. In this lateral retina, only ganglion cells and optic fibers differentiate in larvae, whereas differentiation of amacrine, horizontal, photoreceptor and bipolar cells mainly takes place during metamorphosis, which gives rise to the adult retina. Serotonin (5-hydroxytryptamine, 5-HT) is a neurotransmitter found in the retina of vertebrates whose synthesis is mediated by the rate-limiting enzyme tryptophan hydroxylase (TPH). TPH is also the first enzyme in the biosynthetic pathways of melatonin in photoreceptor cells. The serotonin 1A receptor (5-HT1A) is a major determinant of the activity of both serotonergic cells and their targets due to its pre- and post-synaptic location. Here, we report the developmental pattern of expression of tph and 5-ht1a transcripts in the sea lamprey retina by means of in situ hybridization. In larvae, strong tph mRNA signal was observed in photoreceptors and putative ganglion cells of the central retina, and in some neuroblasts of the lateral retina. In adults, strong tph expression was observed in bipolar, amacrine and ganglion cells and in photoreceptors. In the prolarval (central) retina, all the differentiated retinal cells expressed 5-ht1a transcripts, which were not observed in undifferentiated cells. In larvae, photoreceptors, bipolar cells and ganglion cells in the central retina, and neuroblasts in the lateral retina, showed 5-ht1a expression. In the adult retina, expression of 5-ht1a transcript was mainly observed in the myoid region of both short and long photoreceptors, and was also observed in bipolar, amacrine and ganglion cells. Some 5-HT-immunoreactive amacrine cells have already been reported in the adult lamprey retina. Our study supports the serotonergic phenotype of these amacrine cells of lampreys and also suggests that other retinal neurons could synthesize serotonin at levels not detectable by immunohistochemistry. The expression of the tph transcript in retinal photoreceptors of lampreys strongly suggests that they synthesize melatonin and that this pathway appeared early and has been conserved throughout evolution in vertebrates. The expression of tph and 5-ht1a in neuroblasts also indicates that serotonin might be playing developmental roles in the larval lamprey retina.

Fine‐Scale Pathways Used By Adult Sea Lampreys during Riverine Spawning Migrations

AbstractBetter knowledge of upstream migratory patterns of spawning Sea Lampreys Petromyzon marinus, an invasive species in the Great Lakes, is needed to improve trapping for population control and assessment. Although trapping of adult Sea Lampreys provides the basis for estimates of lake‐wide abundance that are used to evaluate the Sea Lamprey control program, traps have only been operated at dams due to insufficient knowledge of Sea Lamprey behavior in unobstructed channels. Acoustic telemetry and radiotelemetry were used to obtain movement tracks for 23 Sea Lampreys in 2008 and 18 Sea Lampreys in 2009 at two locations in the Mississagi River, Ontario. Cabled hydrophone arrays provided two‐dimensional geographic positions from acoustic transmitters at 3‐s intervals; depth‐encoded radio tag detections provided depths. Upstream movements occurred at dusk or during the night (2015–0318 hours). Sea Lampreys were closely associated with the river bottom and showed some preference to move near banks in shallow glide habitats, suggesting that bottom‐oriented gears could selectively target adult Sea Lampreys in some habitats. However, Sea Lampreys were broadly distributed across the river channel, suggesting that the capture efficiency of nets and traps in open channels would depend heavily on the proportion of the channel width covered. Lack of vertical movements into the water column may have reflected lamprey preference for low water velocities, suggesting that energy conservation was more beneficial for lampreys than was vertical searching in rivers. Improved understanding of Sea Lamprey movement will assist in the development of improved capture strategies for their assessment and control in the Great Lakes.

Expression patterns of Eph genes in the "dual visual development" of the lamprey and their significance in the evolution of vision in vertebrates.

Image-forming vision is crucial to animals for recognizing objects in their environment. In vertebrates, this type of vision is achieved with paired camera eyes and topographic projection of the optic nerve. Topographic projection is established by an orthogonal gradient of axon guidance molecules, such as Ephs. To explore the evolution of image-forming vision in vertebrates, lampreys, which belong to the basal lineage of vertebrates, are key animals because they show unique "dual visual development." In the embryonic and pre-ammocoete larval stage (the "primary" phase), photoreceptive "ocellus-like" eyes develop, but there is no retinotectal optic nerve projection. In the late ammocoete larval stage (the "secondary" phase), the eyes grow and form into camera eyes, and retinotectal projection is newly formed. After metamorphosis, this retinotectal projection in adult lampreys is topographic, similar to that of gnathostomes. In this study, we explored the involvement of Ephs in lamprey "dual visual development" and establishment of the image-form vision. We found that gnathostome-like orthogonal gradient expression was present in the retina during the "secondary" phase; i.e., EphB showed a gradient of expression along the dorsoventral axis, while EphC was expressed along the anteroposterior axis. However, no orthogonal gradient expression was observed during the "primary" phase. These observations suggest that Ephs are likely recruited de novo for the guidance of topographical "second" optic nerve projection. Transformations during lamprey "dual visual development" may represent "recapitulation" from a protochordate-like ancestor to a gnathostome-like vertebrate ancestor.

Seasonal Migration Behaviors and Distribution of Adult Pacific Lampreys in Unimpounded Reaches of the Snake River Basin

AbstractComplex life histories render anadromous fishes particularly susceptible to environmental and anthropogenic change. Adult Pacific Lampreys Entosphenus tridentatus migrating in the Columbia River and its tributaries must ascend a series of dams to reach interior spawning sites. While considerable research has focused on improving dam passage for lampreys, little is known about adult Pacific Lamprey behavior and distribution patterns within free‐flowing environments, particularly within the interior portions of their distribution. In this 3‐year study, we monitored the movements of 146 adult Pacific Lampreys in the Snake River and its tributaries upstream from Lower Granite Dam, the eighth dam from the Pacific Ocean. Our objectives were to characterize migration and test several hypotheses about adult upstream movement after dam passage. A majority of radio‐tagged adults, released above Lower Granite Dam, migrated upstream after release and many moved hundreds of kilometers upstream into Snake River tributaries. Of those with telemetry records after release, 59–70% were recorded in the Clearwater River, 16–25% were in the Snake River, and 13–16% were in the Salmon River. Lampreys that passed the Snake River–Clearwater River confluence were significantly more likely, in most years, to enter the lower‐discharge Clearwater River. Adults moved primarily at night during the summer–fall migration and did not exhibit a consistent response to changes in water temperature or discharge. These findings highlight the importance of the Clearwater River to Pacific Lampreys in the lower Snake River basin and indicate that adults that successfully pass through the Columbia–Snake hydrosystem can continue upstream migration into many Snake River subbasins. This distribution suggests that improved passage efficiency at dams may increase the number of adult Pacific Lampreys available for spawning within the interior portions of their distribution.Received May 12, 2014; accepted November 5, 2014

Sea lamprey mark type, marking rate, and parasite–host relationships for lake trout and other species in Lake Ontario

We examined how attack frequency by sea lampreys on fishes in Lake Ontario varied in response to sea lamprey abundance and preferred host abundance (lake trout >433mm). For this analysis we used two gill net assessment surveys, one angler creel survey, three salmonid spawning run datasets, one adult sea lamprey assessment, and a bottom trawl assessment of dead lake trout. The frequency of fresh sea lamprey marks observed on lake trout from assessment surveys was strongly related to the frequency of sea lamprey attacks observed on salmon and trout from the creel survey and spawning migrations. Attack frequencies on all salmonids examined were related to the ratio between the abundances of adult sea lampreys and lake trout. Reanalysis of the susceptibility to sea lamprey attack for lake trout strains stocked into Lake Ontario reaffirmed that Lake Superior strain lake trout were among the most and Seneca Lake strain among the least susceptible and that Lewis Lake strain lake trout were even more susceptible than the Superior strain. Seasonal attack frequencies indicated that as the number of observed sea lamprey attacks decreased during June–September, the ratio of healing to fresh marks also decreased. Simulation of the ratios of healing to fresh marks indicated that increased lethality of attacks by growing sea lampreys contributed to the decline in the ratios and supported laboratory studies about wound healing duration.

Life stage dependent responses to the lampricide, 3-trifluoromethyl-4-nitrophenol (TFM), provide insight into glucose homeostasis and metabolism in the sea lamprey (Petromyzon marinus)

The primary method of sea lamprey (Petromyzon marinus) control in the Great Lakes is the treatment of streams and rivers with the pesticide 3-trifluoromethyl-4-nitrophenol (TFM), which targets larval sea lamprey. However, less is known about the effects of TFM on other stages of the sea lamprey's complex life cycle. The goal of this study was to determine how TFM affected internal energy stores, metabolites, and ion balance in larval, juvenile (parasitic) and adult sea lamprey. The larvae were more tolerant to TFM than the adults, with a 2-fold higher 12h TFM LC50 and a 1.5-fold higher LC99.9. Acute (3h) exposure of the larvae, parasites and adults to their respective 12h TFM LC99.9 led to marked reductions in glycogen and phosphocreatine in the adult brain, with lesser or no effect in the larvae and parasites. Increased lactate in the brain, at less than the expected stoichiometry, suggested that it was exported to the blood. Kidney glycogen declined after TFM exposure, suggesting that this organ plays an important role in glucose homeostasis. TFM-induced disturbances to ion balance were minimal. In conclusion, TFM perturbs energy metabolism in all major stages of the sea lamprey life cycle in a similar fashion, but the adults appear to be the most sensitive. Thus, the adult stage could be a viable and effective target for TFM treatment, particularly when used in combination with other existing and emerging strategies of sea lamprey control.

The Sea Lamprey as an Etiological Model for Biliary Atresia.

Biliary atresia (BA) is a progressive, inflammatory, and fibrosclerosing cholangiopathy in infants that results in obstruction of both extrahepatic and intrahepatic bile ducts. It is the most common cause for pediatric liver transplantation. In contrast, the sea lamprey undergoes developmental BA with transient cholestasis and fibrosis during metamorphosis, but emerges as a fecund adult with steatohepatitis and fibrosis in the liver. In this paper, we present new histological evidence and compare the sea lamprey to existing animal models to highlight the advantages and possible limitations of using the sea lamprey to study the etiology and compensatory mechanisms of BA and other liver diseases. Understanding the signaling factors and genetic networks underlying lamprey BA can provide insights into BA etiology and possible targets to prevent biliary degeneration and to clear fibrosis. In addition, information from lamprey BA can be used to develop adjunct treatments for patients awaiting or receiving surgical treatments. Furthermore, the cholestatic adult lamprey has unique adaptive mechanisms that can be used to explore potential treatments for cholestasis and nonalcoholic steatohepatitis (NASH).

Complete Spinal Cord Injury and Brain Dissection Protocol for Subsequent Wholemount &lt;em&gt;In Situ&lt;/em&gt; Hybridization in Larval Sea Lamprey

After a complete spinal cord injury, sea lampreys at first are paralyzed below the level of transection. However, they recover locomotion after several weeks, and this is accompanied by short distance regeneration (a few mm) of propriospinal axons and spinal-projecting axons from the brainstem. Among the 36 large identifiable spinal-projecting neurons, some are good regenerators and others are bad regenerators. These neurons can most easily be identified in wholemount CNS preparations. In order to understand the neuron-intrinsic mechanisms that favor or inhibit axon regeneration after injury in the vertebrates CNS, we determine differences in gene expression between the good and bad regenerators, and how expression is influenced by spinal cord transection. This paper illustrates the techniques for housing larval and recently transformed adult sea lampreys in fresh water tanks, producing complete spinal cord transections under microscopic vision, and preparing brain and spinal cord wholemounts for in situ hybridization. Briefly, animals are kept at 16°C and anesthetized in 1% Benzocaine in lamprey Ringer. The spinal cord is transected with iridectomy scissors via a dorsal approach and the animal is allowed to recover in fresh water tanks at 23 °C. For in situ hybridization, animals are reanesthetized and the brain and cord removed via a dorsal approach.

Genes predict long distance migration and large body size in a migratory fish, Pacific lamprey.

Elucidation of genetic mechanisms underpinning migratory behavior could help predict how changes in genetic diversity may affect future spatiotemporal distribution of a migratory species. This ability would benefit conservation of one such declining species, anadromous Pacific lamprey (Entosphenus tridentatus). Nonphilopatric migration of adult Pacific lamprey has homogenized population-level neutral variation but has maintained adaptive variation that differentiates groups based on geography, run-timing and adult body form. To investigate causes for this adaptive divergence, we examined 647 adult lamprey sampled at a fixed location on the Columbia River and radiotracked during their subsequent upstream migration. We tested whether genetic variation [94 neutral and adaptive single nucleotide polymorphisms (SNPs) previously identified from a genomewide association study] was associated with phenotypes of migration distance, migration timing, or morphology. Three adaptive markers were strongly associated with morphology, and one marker also correlated with upstream migration distance and timing. Genes physically linked with these markers plausibly influence differences in body size, which is also consistently associated with migration distance in Pacific lamprey. Pacific lamprey conservation implications include the potential to predict an individual's upstream destination based on its genotype. More broadly, the results suggest a genetic basis for intrapopulation variation in migration distance in migratory species.

Neural mechanisms underlying respiratory rhythm generation in the lamprey

The isolated brainstem of the adult lamprey spontaneously generates respiratory activity. The paratrigeminal respiratory group (pTRG), the proposed respiratory central pattern generator, has been anatomically and functionally characterized. It is sensitive to opioids, neurokinins and acetylcholine. Excitatory amino acids, but not GABA and glycine, play a crucial role in the respiratory rhythmogenesis. These results are corroborated by immunohistochemical data. While only GABA exerts an important modulatory control on the pTRG, both GABA and glycine markedly influence the respiratory frequency via neurons projecting from the vagal motoneuron region to the pTRG. Noticeably, the removal of GABAergic transmission within the pTRG causes the resumption of rhythmic activity during apnea induced by blockade of glutamatergic transmission. The same result is obtained by microinjections of substance P or nicotine into the pTRG during apnea. The results prompted us to present some considerations on the phylogenesis of respiratory pattern generation. They may also encourage comparative studies on the basic mechanisms underlying respiratory rhythmogenesis of vertebrates.