Female Sea Lampreys Research Articles

Sensory trap leads to reliable communication without a shift in nonsexual responses to the model cue.

The sensory trap model of signal evolution suggests that males manipulate females into mating using traits that mimic cues used in a nonsexual context. Despite much empirical support for sensory traps, little is known about how females evolve in response to these deceptive signals. Female sea lamprey (Petromyzon marinus) evolved to discriminate a male sex pheromone from the larval odor it mimics and orient only toward males during mate search. Larvae and males release the attractant 3-keto petromyzonol sulfate (3kPZS), but spawning females avoid larval odor using the pheromone antagonist, petromyzonol sulfate (PZS), which larvae but not males, release at higher rates than 3kPZS. We tested the hypothesis that migratory females also discriminate between larval odor and the male pheromone and orient only to larval odor during anadromous migration, when they navigate within spawning streams using larval odor before they begin mate search. In-stream behavioral assays revealed that, unlike spawning females, migratory females do not discriminate between mixtures of 3kPZS and PZS applied at ratios typical of larval versus male odorants. Our results indicate females discriminate between the sexual and nonsexual sources of 3kPZS during but not outside of mating and show sensory traps can lead to reliable sexual communication without females shifting their responses in the original context.

A review of an electric weir and fishway in a Great Lakes tributary from conception to termination

A successful management plan requires clear goals and a process for evaluation. Without them, managers risk operational shifts in which continuous changes disguised as improvements may have little beneficial effect. The conception, design, and operation of an electric barrier and fishway on the Pere Marquette River of Lake Michigan serve as an illustration. The Great Lakes Fishery Commission operated an electric weir to stop migration of adult sea lampreys (Petromyzon marinus) and a fishway to provide upstream passage of rainbow trout (Oncorhynchus mykiss) during 2000–2009. The weir and fishway were successful in blocking some of the annual spawning run of sea lampreys (trapped an annual average of 439 sea lampreys) and it allowed passage of rainbow trout (an annual average of 6,091). Even with success that yielded an estimated density of 0.03 adult female sea lampreys per 100 m2 of larval habitat upstream of the weir, lampricide treatments continued because the weir still allowed establishment of substantial densities of larval sea lampreys. Our evaluation suggests that an in-stream barrier must approach 100% blockage of sea lampreys to eliminate large recruitment events. The failure of the weir to reduce lampricide treatments was due to an informally defined purpose and measures for success at the onset, the complexity of electric weir systems (and the operational problems created by such intricacy), and lack of recognition of the reduction in larval sea lamprey recruitment needed to succeed. Control of sea lampreys in the Pere Marquette River could have benefited from an adaptive management approach.

A pheromone antagonist liberates female sea lamprey from a sensory trap to enable reliable communication

The evolution of male signals and female preferences remains a central question in the study of animal communication. The sensory trap model suggests males evolve signals that mimic cues used in nonsexual contexts and thus manipulate female behavior to generate mating opportunities. Much evidence supports the sensory trap model, but how females glean reliable information from both mimetic signals and their model cues remains unknown. We discovered a mechanism whereby a manipulative male signal guides reliable communication in sea lamprey (Petromyzon marinus). Migratory sea lamprey follow a larval cue into spawning streams; once sexually mature, males release a pheromone that mimics the larval cue and attracts females. Females conceivably benefit from the mimetic pheromone during mate search but must discriminate against the model cue to avoid orienting toward larvae in nearby nursery habitats. We tested the hypothesis that spawning females respond to petromyzonol sulfate (PZS) as a behavioral antagonist to avoid attraction to the larval cue while tracking the male pheromone despite each containing attractive 3-keto petromyzonol sulfate (3kPZS). We found 1) PZS inhibited electrophysiological responses to 3kPZS and abated preferences for 3kPZS when mixed at the same or greater concentrations, 2) larvae released more PZS than 3kPZS whereas males released more 3kPZS than PZS, and 3) mixtures of 3kPZS and PZS applied at ratios measured in larval and male odorants resulted in the discrimination observed between the natural odors. Our study elucidates how communication systems that arise via deception can facilitate reliable communication.

Petromylidenes A⁻C: 2-Alkylidene Bile Salt Derivatives Isolated from Sea Lamprey (Petromyzon marinus).

Three novel bile acid derivatives, petromylidenes A–C (1–3), featuring uncommon alkylidene adductive scaffolds, were isolated from water conditioned with sexually mature male sea lampreys (Petromyzon marinus). Their structures were elucidated by mass spectrometry and NMR spectroscopy, and by comparison to spectral data of related structures. The identification of compounds 1–3, further illustrates the structural diversity of the 5α bile salt family. Compounds 1–3 exhibited notable biological properties as well, including high olfactory potencies in adult sea lampreys and strong behavioral attraction of ovulated female sea lampreys. Electro-olfactogram recordings indicated that the limit of detection for 1 was 10−9 M, 2 was 10−11 M, and 3 was less than 10−13 M. These results suggested 1–3 were likely male pheromones, which guide reproductive behaviors in the sea lamprey.

Three Novel Bile Alcohols of Mature Male Sea Lamprey (Petromyzon marinus) Act as Chemical Cues for Conspecifics.

Sea lamprey, Petromyzon marinus, rely heavily on chemical cues that mediate their life history events, such as migration and reproduction. Here, we describe petromyzone A-C (1-3), three novel bile alcohols that are highly oxidized and sulfated, isolated from water conditioned with spermiated male sea lamprey. Structures of these compounds were unequivocally established by spectroscopic analyses and by comparison with spectra of known compounds. Electro-olfactogram recordings showed that 1 at 10-11 M was stimulatory to the adult sea lamprey olfactory epithelium, while 2 and 3 were stimulatory at 10-13 M. Behavioral assays indicated that 1 is attractive, 2 is not attractive or repulsive, and 3 is repulsive to ovulated female sea lamprey. The results suggest that 1 and 2 may be putative pheromones that mediate chemical communication in sea lamprey. The identification of these three components enhances our understanding of the structures and functions of sex pheromone components in this species and may provide useful behavioral manipulation tools for the integrated management of sea lamprey, a destructive invader in the Laurentian Great Lakes.

Changes in sea lamprey size and fecundity through time in the Great Lakes

Sea lamprey (Petromyzon marinus) are an aquatic nuisance species that cause great harm to fishes in the Great Lakes. Control of sea lamprey is vital to protecting the Great Lakes fishery, and designing an effective control program depends on accurate knowledge of their demographics. This study compared historical and current data on female sea lamprey length, weight, fecundity, gonadosomatic index (GSI) and egg weight in the Great Lakes. Female sea lamprey captured in 2011 were compared with studies conducted in 1960 and 1981 on Lake Superior and in 1981 and 1999 on Lake Ontario, as well as available data from Lakes Huron, Michigan and Erie. In Lake Superior, mean length and weight of female sea lamprey increased by approximately 13% and 45%, respectively, while mean fecundity increased 17% and individual egg weights increased 43%. There was no difference in mean sea lamprey GSI in Lake Superior. In Lake Ontario, female sea lamprey length increased by 7%, but there were no differences in the other metrics. Across the Great Lakes, there was a general increase in fecundity from the period before the 1980s to more recent years. An increased population of larger parasitic sea lamprey will have higher feeding rates and thus a higher impact on commercially and ecologically important fishes. Managers may need to reconsider control targets and lampricide treatment frequency for lakes where sea lamprey growth and fecundity have increased.

Mixtures of Two Bile Alcohol Sulfates Function as a Proximity Pheromone in Sea Lamprey

Unique mixtures of pheromone components are commonly identified in insects, and have been shown to increase attractiveness towards conspecifics when reconstructed at the natural ratio released by the signaler. In previous field studies of pheromones that attract female sea lamprey (Petromyzon marinus, L.), putative components of the male-released mating pheromone included the newly described bile alcohol 3,12-diketo-4,6-petromyzonene-24-sulfate (DkPES) and the well characterized 3-keto petromyzonol sulfate (3kPZS). Here, we show chemical evidence that unequivocally confirms the elucidated structure of DkPES, electrophysiological evidence that each component is independently detected by the olfactory epithelium, and behavioral evidence that mature female sea lamprey prefer artificial nests activated with a mixture that reconstructs the male-released component ratio of 30:1 (3kPZS:DkPES, molar:molar). In addition, we characterize search behavior (sinuosity of swim paths) of females approaching ratio treatment sources. These results suggest unique pheromone ratios may underlie reproductive isolating mechanisms in vertebrates, as well as provide utility in pheromone-integrated control of invasive sea lamprey in the Great Lakes.

Female sea lamprey shift orientation toward a conspecific chemical cue to escape a sensory trap

The sensory trap model of signal evolution hypothesizes that signalers adapt to exploit a cue used by the receiver in another context. Although exploitation of receiver biases can result in conflict between the sexes, deceptive signaling systems that are mutually beneficial drive the evolution of stable communication systems. However, female responses in the nonsexual and sexual contexts may become uncoupled if costs are associated with exhibiting a similar response to a trait in both contexts. Male sea lamprey (Petromyzon marinus) signal with a mating pheromone, 3-keto petromyzonol sulfate (3kPZS), which may be a match to a juvenile cue used by females during migration. Upstream movement of migratory lampreys is partially guided by 3kPZS, but females only move toward 3kPZS with proximal accuracy during spawning. Here, we use in-stream behavioral assays paired with gonad histology to document the transition of female preference for juvenile- and male-released 3kPZS that coincides with the functional shift of 3kPZS as a migratory cue to a mating pheromone. Females became increasingly biased toward the source of synthesized 3kPZS as their maturation progressed into the reproductive phase, at which point, a preference for juvenile odor (also containing 3kPZS naturally) ceased to exist. Uncoupling of female responses during migration and spawning makes the 3kPZS communication system a reliable means of synchronizing mate search. The present study offers a rare example of a transition in female responses to a chemical cue between nonsexual and sexual contexts, provides insights into the origins of stable communication signaling systems.

Breakdown products of heme catabolism as senescence index in sea lamprey (Petromyzon marinus Linnaeus, 1758)

Breakdown products of heme catabolism as senescence index in sea lamprey (Petromyzon marinus Linnaeus, 1758)

Investigations of novel unsaturated bile salts of male sea lamprey as potential chemical cues.

Sulfated bile salts function as chemical cues that coordinate reproduction in sea lamprey, Petromyzon marinus. 7α, 12α, 24-trihydroxy-5α-cholan-3-one 24-sulfate (3kPZS) is the most abundant known bile salt released by sexually mature male sea lampreys and attracts ovulated females. However, previous studies showed that the male-produced pheromone consists of unidentified components in addition to 3kPZS. Here, analysis of water conditioned with mature male sea lampreys indicated the presence of 4 oxidized, unsaturated compounds with molecular weights of 466Da, 468Da, and 2 of 470Da. These compounds were not detectable in water conditioned with immature male sea lampreys. By using mass spectrometry, 4 A-ring unsaturated sulfated bile salts were tentatively identified from male washings as 2 4-ene, a 1-ene, and a 1,4-diene analogs. These were synthesized to determine if they attracted ovulated female sea lampreys to spawning nests in natural streams. One of the novel synthetic bile salts, 3 keto-1-ene PZS, attracted ovulated females to the point of application at a concentration of 10(-12)M. This study reveals the structural diversity of bile salts in sea lamprey, some of which have been demonstrated to be pheromonal cues.

Mercury accumulation in sea lamprey (Petromyzon marinus) from Lake Huron

We determined whole-fish total mercury (Hg) concentrations of 40 male and 40 female adult sea lampreys (Petromyzon marinus) captured in the Cheboygan River, a tributary to Lake Huron, during May 2011. In addition, bioenergetics modeling was used to explore the effects of sex-related differences in activity and resting (standard) metabolic rate (SMR) on mercury accumulation. The grand mean for Hg concentrations was 519ng/g (standard error of the mean=46ng/g). On average, males were 16% higher in Hg concentration than females. Bioenergetics modeling results indicated that 14% higher activity and SMR in males would account for this observed sex difference in Hg concentrations. We concluded that the higher Hg concentration in males was most likely due to higher rate of energy expenditure in males, stemming from greater activity and SMR. Our findings have implications for estimating the effects of sea lamprey populations on mercury cycling within ecosystems, as well as for the proposed opening of sea lamprey fisheries. Eventually, our results may prove useful in improving control of sea lamprey, a pest responsible for substantial damage to fisheries in lakes where it is not native.

Odor-conditioned rheotaxis of the sea lamprey: modeling, analysis and validation

Mechanisms for orienting toward and locating an odor source are sought in both biology and engineering. Chemical ecology studies have demonstrated that adult female sea lamprey show rheotaxis in response to a male pheromone with dichotomous outcomes: sexually mature females locate the source of the pheromone whereas immature females swim by the source and continue moving upstream. Here we introduce a simple switching mechanism modeled after odor-conditioned rheotaxis for the sea lamprey as they search for the source of a pheromone in a one-dimensional riverine environment. In this strategy, the females move upstream only if they detect that the pheromone concentration is higher than a threshold value and drifts down (by turning off control action to save energy) otherwise. In addition, we propose various uncertainty models such as measurement noise, actuator disturbance, and a probabilistic model of a concentration field in turbulent flow. Based on the proposed model with uncertainties, a convergence analysis showed that with this simplistic switching mechanism, the lamprey converges to the source location on average in spite of all such uncertainties. Furthermore, a slightly modified model and its extensive simulation results explain the behaviors of immature female lamprey near the source location.

Polychlorinated Biphenyl Concentrations and Activity of Sea Lamprey Petromyzon marinus Vary by Sex

We determined the polychlorinated biphenyl (PCB) concentrations of 40 male and 40 female adult sea lampreys Petromyzon marinus captured in the Cheboygan River, a tributary to Lake Huron, during May 2011. In addition, we performed a laboratory experiment using passive integrated transponder tags to determine whether male adult sea lampreys were more active than female adult sea lampreys. Sex had a significant effect on PCB concentration, and PCB concentration at a given level of sea lamprey condition was approximately 25% greater in males than in females. Adjusting for the difference in condition between the sexes, males averaged a 17% greater PCB concentration compared with females. Results from the laboratory experiment indicated that males were significantly more active than females. The observed sex difference in PCB concentrations was not due to female sea lampreys releasing eggs at spawning because the sea lamprey is semelparous, and we caught the sea lampreys before spawning. Rather, we attributed the sex difference in PCB concentrations to a greater rate of energy expenditure in males compared with females. We proposed that this greater rate of energy expenditure was likely due to greater activity. Our laboratory experiment results supported this hypothesis. A greater resting metabolic rate may also have contributed to a greater rate of energy expenditure. Our findings should eventually be applicable toward improving control of sea lamprey, a pest responsible for considerable damage to fisheries in lakes where it is not native.

Effects of Sex Pheromones and Sexual Maturation on Locomotor Activity in Female Sea Lamprey (Petromyzon marinus)

Synchronization of male and female locomotor rhythmicity can play a vital role in ensuring reproductive success. Several physiological and environmental factors alter these locomotor rhythms. As sea lamprey, Petromyzon marinus, progress through their life cycle, their locomotor activity rhythm changes multiple times. The goal of this study was to elucidate the activity patterns of adult female sea lamprey during the sexual maturation process and discern the interactions of these patterns with exposure to male pheromones. During these stages, preovulated and ovulated adult females are exposed to sex pheromone compounds, which are released by spermiated males and attract ovulated females to the nest for spawning. The locomotor behavior of adult females was monitored in a natural stream with a passive integrated tag responder system as they matured, and they were exposed to a sex pheromone treatment (spermiated male washings) or a control (prespermiated male washings). Results showed that, dependent on the hour of day, male sex pheromone compounds reduce total activity (p < 0.05) and cause increases in activity during several daytime hours in preovulated and ovulated females. These results are one of the first examples of how sex pheromones modulate a locomotor rhythm in a vertebrate, and they suggest that the interaction between maturity stage and sex pheromone exposure contributes to the differential locomotor rhythms found in adult female sea lamprey. This phenomenon may contribute to the reproductive synchrony of mature adults, thus increasing reproductive success in this species.

The behavioural response of migratory sea lamprey (Petromyzon marinus) to potential damage-released larval and migratory chemical alarm cues

We investigated the presence of damage-released alarm cues and the reactions they may cause in landlocked migratory sea lamprey (Petromyzon marinus) using semi-natural laboratory conditions during the day. In two separate experiments, migratory sea lampreys were exposed to stimuli prepared from the skinless carcasses and skin tissue of larval sea lamprey and from the skin tissue and muscle tissue of migratory sea lamprey. Migratory female sea lamprey swam significantly longer after being exposed to the stimulus prepared from the skinless carcass of larval sea lamprey. No significant changes were seen in the behaviour of the migratory female lamprey in response to larval skin extract or in the behaviour of male sea lamprey to any experimental extract in the larval extract experiment. In the second experiment that utilised migratory lamprey stimuli, neither male and nor female migratory sea lamprey showed a significant difference in their behavioural response among different treatments and controls. Our findings indicate that adult female sea lamprey respond strongly to damage-released alarm cues from larval sea lamprey; hence, the latter holds promise for sea lamprey behavioural manipulations for control purposes during the day. Further research is needed to examine responses to damage-released alarm cues at night, when migratory sea lampreys are more active.

Expression of three GnRH receptors in specific tissues in male and female sea lampreys Petromyzon marinus at three distinct life stages

Two recently cloned gonadotropin-releasing hormone (GnRH) receptors (lamprey GnRH-R-2 and lamprey GnRH-R-3) along with lamprey (l) GnRH-R-1 were shown to share similar structural features and amino acid motifs common to other vertebrate receptors. Here we report on our findings of RNA expression of these three GnRH receptors in the three major life stages (larval, parasitic, and adult phases) of the sea lamprey, Petromyzon marinus, a basal vertebrate. For each stage, we examined the expression of messenger RNA encoding the receptors in the brain, pituitary, gonad, heart, muscle, liver, eye, intestine, kidney, skin, thyroid, gill, and endostyle by RT-PCR. In adult lampreys, the spatial expression of the three receptors in the brain and pituitary was investigated by in situ hybridization. In general, the receptors were more widely expressed in adult tissues as compared to parasitic-phase tissues and least widely expressed in the larval tissues. There were noted differences in male and female lampreys in the adult and parasitic phases for all three receptors. The data showed the presence of all three receptor transcripts in brain tissues for adult and parasitic phases and all three receptor transcripts were expressed in the adult pituitaries, but not in the parasitic pituitaries. However, in the larval phase, only lGnRH-R-1 was expressed in the larval brain and pituitary. In situ hybridization revealed that lGnRH-R-2 and -3 were expressed in the pineal tissue of adult female lampreys while lGnRH-R-1 was expressed in the pineal in adult male lampreys, all restricted to the pineal pellucida. In summary, these data provide an initial comparative analysis of expression of three lamprey GnRH receptors suggesting differential regulation within males and females at three different life/reproductive stages.

A synthesized mating pheromone component increases adult sea lamprey (Petromyzon marinus) trap capture in management scenarios

Application of chemical cues to manipulate adult sea lamprey (Petromyzon marinus) behavior is among the options considered for new sea lamprey control techniques in the Laurentian Great Lakes. A male mating pheromone component, 7α,12α,24-trihydroxy-3-one-5α-cholan-24-sulfate (3kPZS), lures ovulated female sea lamprey upstream into baited traps in experimental contexts with no odorant competition. A critical knowledge gap is whether this single pheromone component influences adult sea lamprey behavior in management contexts containing free-ranging sea lampreys. A solution of 3kPZS to reach a final in-stream concentration of 10−12 mol·L−1 was applied to eight Michigan streams at existing sea lamprey traps over 3 years, and catch rates were compared between paired 3kPZS-baited and unbaited traps. 3kPZS-baited traps captured significantly more sexually immature and mature sea lampreys, and overall yearly trapping efficiency within a stream averaged 10% higher during years when 3kPZS was applied. Video analysis of a trap funnel showed that the likelihood of sea lamprey trap entry after trap encounter was higher when the trap was 3kPZS baited. Our approach serves as a model for the development of similar control tools for sea lamprey and other aquatic invaders.

Sea lamprey orient toward a source of a synthesized pheromone using odor-conditioned rheotaxis

Characterization of vertebrate chemo-orientation strategies over long distances is difficult because it is often not feasible to conduct highly controlled hypothesis-based experiments in natural environments. To overcome the challenge, we couple in-stream behavioral observations of female sea lampreys (Petromyzon marinus) orienting to plumes of a synthesized mating pheromone, 7α,12α,24-trihydroxy-5α-cholan-3-one-24-sulfate (3kPZS), and engineering algorithms to systematically test chemo-orientation hypotheses. In-stream field observations and simulated movements of female sea lampreys according to control algorithms support that odor-conditioned rheotaxis is a component of the mechanism used to track plumes of 3kPZS over hundreds of meters in flowing water. Simulated movements of female sea lampreys do not support that rheotaxis or klinotaxis alone is sufficient to enable the movement patterns displayed by females in locating 3kPZS sources in the experimental stream. Odor-conditioned rheotaxis may not only be effective at small spatial scales as previous described in crustaceans, but may also be effectively used by fishes over hundreds of meters. These results may prove useful for developing management strategies for the control of invasive species that exploit the odor-conditioned tracking behavior and for developing biologically inspired navigation strategies for robotic fish.

Multiple functions of a multi‐component mating pheromone in sea lamprey Petromyzon marinus

The role of the C24 sulphate in the mating pheromone component, 7α,12α,24-trihydroxy-5α-cholan-3-one 24-sulphate (3kPZS), to specifically induce upstream movement in ovulated female sea lampreys Petromyzon marinus was investigated. 7α,12α-dihydroxy-5α-cholan-3-one 24-oic acid (3kACA), a structurally similar bile acid released by spermiated males, but lacking the C24 sulphate ester, was tested in bioassays at concentrations between 10(-11) and 10(-14) molar (M). 3kACA did not induce upstream movement in females or additional reproductive behaviours. In contrast, spermiated male washings induced upstream movement, prolonged retention on a nest and induced an array of nesting behaviours. Differential extraction and elution by solid-phase extraction resins showed that components other than 3kPZS + 3kACA are necessary to retain females on nests and induce nest cleaning behaviours. All pheromone components, including components in addition to 3kPZS + 3kACA that retain females and induce nest cleaning behaviours were released from the anterior region of the males, as had been reported for 3kPZS. It is concluded that the sea lamprey male mating pheromone has multiple functions and is composed of multiple components.

The efficacy of two synthesized sea lamprey sex pheromone components as a trap lure when placed in direct competition with natural male odors

The female sea lamprey (Petromyzon marinus), a devastating invasive fish of the Laurentian Great Lakes, locates potential mates by tracking a sex pheromone emitted by nesting males. We tested whether combinations of two putative components of the sex pheromone, 3-keto-petromyzonol sulfate (3kPZS) and 3-keto-allocholic acid (3kACA), were sufficiently attractive to function as a trap-bait when placed in direct competition with male odors. Ovulating females successfully located point sources of 3kPZS both in the presence and absence of a competing odor emitted by mature males placed upstream. However, 3kPZS was not able to retain females in the vicinity of a trap longer than two minutes, and retention time was reduced by 57% when competing male odors were present. 3kACA failed to elicit a response on its own and did not improve attraction to, or retention near, a source of 3kPZS. Application of an incomplete pheromone in trapping-for-control scenarios will require devices configured to minimize the effort necessary to enter a trap, features to offset the probable decrease in trap retention, and deployment into favorable habitats where competition with spawning males is minimal.