Diel Patterns of Pheromone Release By Male Sea Lamprey

Costs to producing sexual signals can create selective pressures on males to invest signaling effort in particular contexts. When the benefits of signaling vary consistently across time, males can optimize signal investment to specific temporal contexts using biological rhythms. Sea lamprey, Petromyzon marinus, have a semelparous life history, are primarily nocturnal, and rely on pheromone communication for reproduction; however, whether male investment in pheromone transport and release matches increases in spawning activity remains unknown. By measuring 1) 3keto-petromyzonol sulfate (3kPZS, a main pheromone component) and its biosynthetic precursor petromyzonol sulfate (PZS) in holding water and tissue samples at 6 points over the course of 24 hours, and 2) 3kPZS release over the course of several days, we demonstrate that 3kPZS release exhibits a consistent diel pattern across several days with elevated pheromone release just prior to sunset and at night. Trends in hepatic concentrations and circulatory transport of PZS and 3kPZS were consistent with patterns of 3kPZS release and suggest the possibility of direct upregulation in pheromone transport and release rather than observed release patterns being solely a byproduct of increased behavioral activity. Our results suggest males evolved a signaling strategy that synchronizes elevated pheromone release with nocturnal increases in sea lamprey behavior. This may be imperative to ensure that male signaling effort is not wasted in a species having a single, reproductive event.

Bile acid production is life-stage and sex-dependent and affected by primer pheromones in the sea lamprey

Pheromonal bile salts are important for sea lampreys (Petromyzon marinus Linnaeus) to complete their life cycle. The synthesis and release of a releaser/primer pheromone 3-keto petromyzonol sulfate (3kPZS) by spermiating males have been well characterized. 3kPZS evokes sexual behaviors in ovulatory females, induces immediate 3kPZS release in spermiating males, and elicits neuroendocrine responses in prespawning adults. Another primer pheromone released by spermiating males, 3-keto allocholic acid (3kACA), antagonizes the neuroendocrine effects of 3kPZS in prespermiating males. However, the effects of 3kACA and 3kPZS on pheromone production in prespawning adults is unclear. To understand the foundation of pheromone production, we examined sea lamprey bile salt levels at different life stages. To investigate the priming effects of 3kACA and 3kPZS, we exposed prespawning adults with vehicle or synthetic 3kACA or 3kPZS. We hypothesized that endogenous bile salt levels were life-stage and sex-dependent, and differentially affected by 3kACA and 3kPZS in prespawning adults. Using ultra-performance liquid chromatography tandem mass spectrometry, we found that sea lampreys contained distinct mixtures of bile salts in the liver and plasma at different life stages. Males usually contained higher amounts of bile salts than females. Petromyzonamine disulfate was the most abundant C27 bile salt and petromyzonol sulfate was the most abundant C24 bile salt. Waterborne 3kACA and 3kPZS exerted differential effects on bile salt production in the liver and gill, their circulation and clearance in the plasma, and their release into water. We conclude that bile salt levels are life-stage and sex-dependent and differentially affected by primer pheromones.

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.

A pheromone outweighs temperature in influencing migration of sea lamprey

Organisms continuously acquire and process information from surrounding cues. While some cues complement one another in delivering more reliable information, others may provide conflicting information. How organisms extract and use reliable information from a multitude of cues is largely unknown. We examined movement decisions of sea lampreys ( Petromyzon marinus L.) exposed to a conspecific and an environmental cue during pre-spawning migration. Specifically, we predicted that the mature male-released sex pheromone 3-keto petromyzonol sulfate (3kPZS) will outweigh the locomotor inhibiting effects of cold stream temperature (less than 15°C). Using large-scale stream bioassays, we found that 3kPZS elicits an increase (more than 40%) in upstream movement of pre-spawning lampreys when the water temperatures were below 15°C. Both warming temperatures and conspecific cues increase upstream movement when the water temperature rose above 15°C. These patterns define an interaction between abiotic and conspecific cues in modulating animal decision-making, providing an example of the hierarchy of contradictory information.

Identification of putative migratory pheromones from Pacific lamprey (Lampetra tridentata)

Pacific lamprey, Lampetra tridentata , has declined precipitously throughout their range in the Columbia River basin of North America. Tribal Nations and Federal and State agencies are engaged in efforts to restore these fish. Understanding whether Pacific lamprey emit and detect migratory pheromones is particularly important for these restoration efforts. Using behavioural assays, we demonstrated that migratory adult Pacific lamprey are attracted to odors emanating from their larval conspecifics. We then identified putative pheromones released by larval Pacific lamprey. Chemical analysis of the conditioned water from larval lamprey using liquid chromatography – mass spectrometry (LC–MS) revealed that the Pacific lamprey can release petromyzonamine disulfate (PADS), petromyzosterol disulfate (PSDS), and petromyzonol sulfate (PZS). Electro-olfactogram studies further demonstrated that adult Pacific lamprey can smell those bile acid compounds. Our data strongly indicate that the Pacific lamprey employ a chemical communication system mediated by a mixture of bile acids, as evidenced by pheromonal functions of the bile acid compounds in guiding migratory adult sea lamprey to the spawning streams. Comprehensive understanding of the chemical communications involved in lamprey migratory behavior may lead to improved scientific approaches for restoration efforts.

Laboratory assessment of the role of a larval pheromone and natural stream odor in spawning stream localization by migratory sea lamprey (Petromyzon marinus)

This study used large laboratory mazes and natural stream waters to test the role of olfactory cues, including a pheromone released by larvae, in spawning stream localization by migratory sea lamprey (Petromyzon marinus). We found that migratory lamprey strongly prefer stream water over lake water and that this response is dependent upon a functional olfactory system. Responses persisted among migratory lamprey even after stream water was diluted a thousand times but were not seen among non-migratory lamprey. Experiments using waters from five streams demonstrated that a larval pheromone is a key determinant of stream attractiveness: water from streams with larval populations were consistently more attractive than those without, and adding larval odor to the latter reversed this relationship. Larval odor was attractive at low, realistic concentrations, especially when presented together with natural stream water, suggesting that streams contain other odors that synergize the actions of the pheromone. Some, but not all, of the activity of the larval pheromone could be explained by two bile acids released by larvae (petromyzonol sulfate and allocholic acid). Together, these results strongly suggest that migratory lamprey locate streams using a larval pheromone. This cue could be useful in lamprey control.