First evidence of attraction of adult river lamprey in the migratory phase to larval odour

Sexually immature, adult river lamprey Lampetra fluviatilis in the upstream migratory phase, were shown to be attracted to water conditioned with ammocoete larvae when tested using a two‐choice flume. Although preliminary, the data suggest that migrating adult river lamprey may be attracted to larval putative pheromone as recorded in sea lamprey Petromyzon marinus .     

An electrophoretic and immunoelectrophoretic characterization of the serum proteins of the adult lamprey, Petromyzon marinus L

1. Serum proteins of upstream migrants of Petromyzon marinus were separated electrophoretically on polyacrylamide and agarose as well as by crossed immunoelectrophoresis using anti-lamprey serum antiserum. 2. Some constituents of four of the five major bands separated by electrophoresis on polyacrylamide were located in the agarose and crossed-immunoelectrophoretic patterns. 3. Two serum proteins, SDS-1 and CB-III, were isolated. SDS-1 was found to be a glycoprotein; CB-III may be a lipoprotein. 4. Monospecific antisera were produced against SDS-1 and CB-III and were used in crossed immunoelectrophoresis to analyze lamprey serum. This method revealed that electrophoretically different forms of these two proteins exist in serum. 5. Immunoelectrophoretic methods demonstrated that three of the major bands produced by polyacrylamide gel electrophoresis of whole serum contained more than a single protein.     

An electrophoretic and immunoelectrophoretic study of serum proteins during the life cycle of the lamprey, Petromyzon marinus L

Serum proteins of upstream migrants, parasitic adults, early and late metamorphosing animals, and ammocoetes of Petromyzon marinus L. were examined by polyacrylamide gel electrophoresis and crossed-immunoelectrophoresis. The electrophoretic patterns for two proteins, SDS-1 and CB-III, were found to change during the life cycle. quantitative measurements of these two proteins showed that they continued to increase until in the adult they together constituted approx. 85% of the total serum protein. Evidence was obtained for a protein present in ammocoetes and metamorphosing animals, but not in upstream migrants.     

Population genetics of a cyclostome species pair, river lamprey (Lampetra fluviatilis L.) and brook lamprey (Lampetra planeri Bloch)

Horizontal agarose gel electrophoresis of 24 allozyme loci in four species of Central European lampreys (321 Lampetra planeri , 83 L. fluviatilis , 11 Eudontomyzon mariae and nine Petromyzon marinus ) was used to study the 'paired species' L. fluviatilis and L. planeri . The genetic differentiation of the anadromous river lamprey ( L. fluviatilis ) from the stationary brook lamprey ( L. planeri ) was within the range of ingroup differentiation of the latter, but L. fluviatilis exhibited much greater population cohesion over a more extended geographic range: G _ST = 0.0537 versus G _ST = 0.3398, N _em = 4.402 versus N _em = 0.4856, mean genetic among-stock distances D = 0.0047 versus D = 0.0257. L. planeri populations coexisting geographically with L. fluviatilis in the Rhine and Elbe river systems were genetically more cohesive than L. planeri stocks from the Danubian basin where L. fluviatilis is absent. Danubian L. planeri populations exhibit a lower degree of heterozygosity than brook lampreys from the Rhine river system, but comprise deeper genetic lineages ( G _ST = 0.4629 versus G _ST = 0.2434), despite being sampled from a much more restricted area. Isolation-by-distance is observed for L. planeri from the Danubian but not from the Atlantic drainage basins. Transspecific gene flow between L. planeri from Atlantic drainage basins and the long-distant migrating L. fluviatilis is inferred, raising doubt on the validity of two separate biospecies. E. mariae and P. marinus are clearly differentiated from Lampetra spp. at several allozyme loci.     

Variability in migration timing of adult Pacific lamprey (<Emphasis Type="Italic">Lampetra tridentata</Emphasis>) in the Columbia River,U.S.A.

We examined the effects of river environment on the timing of spawning migrations by anadromous Pacific lamprey, Lampetra tridentata, in the Columbia River (U.S.A.). In a 41-year time series of adult lamprey counts, migration timing was earliest in warm, low-discharge years and latest in cold, high-flow years. Threshold temperatures associated with run timing were similar throughout the dataset despite significant impoundment-related warming, suggesting that temperature-dependent migration cues have been temporally stable. Within each year, migration rates of PIT-tagged lampreys were positively correlated with temperature and negatively correlated with discharge through multiple river reaches, offering additional evidence for environmental control of upstream movement. Both visual count and PIT-tag data indicated that there may be population-based differences in migration timing within the aggregate Columbia River lamprey run. These life history and behavioral results have potentially far-reaching implications for management of lamprey species.     

Profiles of the female-specific serum protein in the Japanese river lamprey, Lampetra japonica (Martens), and the sand lamprey, Lampetra reissneri (Dybowski), in relation to sexual maturation

The occurrence of female-specific serum protein (FSSP) was examined in the anadromous lamprey, Lampetra japonica, and a freshwater lamprey, Lampetra reissneri, by an immunodiffusion technique using antiserum raised against the serum of sexually mature female lamprey. A considerable amount of FSSP was consistently detected in the blood of vitellogenic, but not in that of nonvitellogenic, females of both species of lampreys. The relative amount of the FSSP tended to become larger in female lampreys at the spawning period. The FSSP may correspond to the yolk precursor protein which is incorporated into ovarian oocytes during the period of exogenous vitellogenesis in the lampreys.     

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The investigation is a light microscopical study of the structural development of the lamprey labyrinth, especially in Lampetra fluviatilis. Some comparisons have been made with Lampetra planeri and Petromyzon marinus. The following parts of the ear have been investigated; macula communis, its subdivision and innervation; the semicircular ducts and their ampullae; the two endolymphatic ducts and the ciliated chambers. Particular attention has been paid to the origin of these parts, the function of the ciliated chambers and the ampullae. On the basis of the present knowledge of the labyrinth an attempt has been made to see how the structural changes in it during the life cycle of the river lamprey harmonize with its way of living. Also comparative and phylogenetic aspects of the lamprey labyrinth have been made.     

HPLC and ELISA analyses of larval bile acids from Pacific and western brook lampreys

Comparative studies were performed on two native lamprey species, Pacific lamprey (Lampetra tridentata) and western brook lamprey (Lampetra richardsoni) from the Pacific coast along with sea lamprey (Petromyzon marinus) from the Great Lakes, to investigate their bile acid production and release. HPLC and ELISA analyses of the gall bladders and liver extract revealed that the major bile acid compound from Pacific and western brook larval lampreys was petromyzonol sulfate (PZS), previously identified as a migratory pheromone in larval sea lamprey. An ELISA for PZS has been developed in a working range of 20 pg-10 ng per well. The tissue concentrations of PZS in gall bladder were 127.40, 145.86, and 276.96 micro g/g body mass in sea lamprey, Pacific lamprey, and western brook lamprey, respectively. Releasing rates for PZS in the three species were measured using ELISA to find that western brook and sea lamprey released PZS 20 times higher than Pacific lamprey did. Further studies are required to determine whether PZS is a chemical cue in Pacific and western brook lampreys.     

Sea lamprey Petromyzon marinus: an exception to the rule of homing in anadromous fishes

Anadromous fishes are believed to make regular circuits of migration in the sea before homing to their natal rivers. Sea lamprey Petromyzon marinus is an anadromous fish that is an exception to this life-history pattern. It also differs from other anadromous fishes in that its adult phase is parasitic, a feeding strategy that should make homing problematic for lamprey cohorts that become widely dispersed through transport by the diverse hosts they parasitize. We sequenced a portion of the mitochondrial DNA control region from sea lampreys collected from 11 North American east coast rivers to test for genetic evidence of homing. There were no significant differences (chi2=235.1, p=0.401) in haplotype frequencies among them, with almost 99 per cent of haplotypic diversity occurring within populations. These findings, together with concordant genetic results from other geographical regions and ancillary information on pheromonal communication, suggest that sea lamprey does not home but rather exhibits regional panmixia while using a novel 'suitable river' strategy to complete its life cycle.     

The estuarine biology of the River lamprey, Lampetra fluviatilis, in the Firth of Forth, Scotland, with particular reference to size composition and feeding

Samples collected regularly during 1979, 1980 and 1981 from the intake screens of power stations in the estuary of the River Forth produced data on the estuarine phase in the life cycle of the River lamprey, Lampetra fluviatilis. Recently metamorphosed animals were common during the spring while sexually maturing adults were abundant in the late summer and autumn. These were assumed to be lampreys at the end of their downstream and start of their upstream migrations respectively. The respective sizes of the presumed downstream and upstream migrants were 69–135 mm (0-2-2-8 g) and 200–361 mm (7-0-93-2 g). Smaller numbers of animals of intermediate size were collected during the summer and late autumn. Males were usually the predominant sex with an average of 56–5% of the population in 1980–1981. In all months, the mean weight and with one exception also the mean length, was greater for females than for males. The intestine frequently contained fish remains, especially muscle, bone and scales of clupeids and thus the River lamprey seems to feed mainly on teleosts in this estuary. An analysis of the size of scales in the intestines of lampreys of various body lengths indicates that there is a relationship between the size of host and predator. Comparisons with other studies of Lampetra fluviatilis emphasize the variability that exists in aspects of the biology of this species within and among populations.     

The structure of Mordacia mordax insulin supports the monophyly of the Petromyzontiformes and an ancient divergence of Mordaciidae and Geotriidae

The phylogenetic relationships between the two southern hemisphere lamprey families (Geotriidae and Mordaciidae) and their northern hemisphere counterparts (Petromyzontidae) are unresolved. Insulin was isolated from an extract of islet-containing intestinal tissue from ammocoetes of the Australian lamprey, Mordacia mordax. Its primary structure was established as A-chain: GIVEQCCHRK10CSIYDMENYC20N and B-chain: SALMGTGGTH10LCGSHLVEAL20YVVCGQRGFF30 YTP[SKG]. Although the residues in parentheses are only tentatively assigned, mass spectrometry supports the proposed sequence and demonstrates that Mordacia proinsulin, unlike proinsulin from Geotria australis, is fully processed to mature insulin. Insulins from M. mordax and G. australis and from the northern hemisphere lampreys Petromyzon marinus and Lampetra fluviatilis share a pentapeptide extension to N-terminus of the B-chain (Ser-Ala-Leu-Xaa-Gly) that has never been found in the insulins of any other vertebrate class. This observation provides support for the claim that the Petromyzontiformes constitute a monophyletic group. M. mordax insulin differs from that of G. australis by 18 amino acid residues but by only four residues from the common sequence of P. marinus and L. fluviatilis insulin. These data are consistent with the view that Geotriidae and Mordaciidae have been separated for a long period and suggest that G. australis insulin has undergone an accelerated rate of molecular evolution.     

Choline acetyltransferase immunoreactivity in the hypothalamoneurohypophysial system of the lamprey.

The cholinergic innervation of the neurohypophysis of the lampreys Petromyzon marinus and Lampetra fluviatilis was studied by means of immunocytochemical techniques with antibodies directed against the enzyme choline acetyltransferase (ChAT). The results obtained in both species were basically similar. A rich innervation by ChAT-immunoreactive fibres was found throughout the neurohypophysis. These fibres originate from cholinergic neurons located in the preoptic region and the paraventricular nucleus. Some of these cholinergic neurons are in contact with the cerebrospinal fluid. Numerous axonal swellings were evident in the tuberal region of the sea lamprey, but not in the river lamprey. The possible pathways of cholinergic release in the lamprey hypophysis are discussed.     

Conservation status of Northern Hemisphere lampreys (Petromyzontidae)

Among the 34 nominal lamprey species in the Northern Hemisphere, ten are endangered; nine are vulnerable at least in part of their range, and one is extinct. The major cause is habitat degradation through pollution and stream regulation. Four conservation priorities are recommended: 1. The protection of all lamprey species or populations thereof listed as endangered; 2. where needed, the rehabilitation of the spawning streams of non-parasitic species and the removal or circumvention of any barriers preventing access to the spawning sites; 3. the protection of permanent freshwater resident populations of anadromous species, specifically of Entosphenus tridentatus, Lampetra ayresi, L. fluviatilis, Lethenteron japonicum and Petromyzon marinus : 4. the study of the conservation status of Asian lampreys.     

Heart Ratios at Different Stages in the Life Cycle of Lampreys

Heart ratios (heart weight/body weight. ×100) have been obtained for freshly killed and preserved (10% formalin) specimens of the lampreys Petromyzon marinus, Lampetra fluviatilis and L. planeri. Values for comparable stages were significantly higher in the preserved material, probably as a result of retention and fixation of blood in the heart chambers. The regression slope for the logarithmic relationship between heart and body weights was close to one in all stages of the three species investigated. A marked increase was observed in the heart ratio during metamorphosis, apparently attributable to an increase in the number and density of myocardial fibres. Heart ratios in the lamprey were generally higher than for most other fishes, with values of 0.1 and 0.3 being obtained for example in the case of freshly killed ammocoete and sexually mature adult stages of L. fluviatilis.     

Ultrastructural changes in the liver of the sand lamprey,Lampetra reissneri,during sexual maturation

Ultrastructural changes of hepatocytes were examined in the sand lamprey,Lampetra reissneri, during various phases of the life cycle. In hepatocytes of ammocoetes, the rough endoplasmic reticulum was composed of short cisternae and the Golgi apparatus were scarcely developed, showing no sexual differences at this stage of life cycle. In hepatocytes of female lampreys at the metamorphic stages 4 to 5, the rough endoplasmic reticulum was developed to form long parallel cisternae and the Golgi apparatus were well-developed. The rough endoplasmic reticulum developed further to form stacks of long parallel cisternae extending over the cytoplasm in hepatocytes of females at the young adult stage, and became composed of both long parallel and vesicular cisternae in the cells of females at the adult stage. The Golgi apparatus were invariably welldeveloped in hepatocytes of young adult and adult females. No consipcuous development was observed in profiles of the rough endoplasmic reticulum and the Golgi apparatus in hepatocytes of males during and after metamorphosis. The ultrastructural changes of the rough endoplasmic reticulum and the Golgi apparatus observed in hepatocytes of female sand lampreys are considered to have an intimate relation to the activity of vitellogenin synthesis in the liver, and it is suggested that the hepatocytes begin to rapidly synthesize vitellogenin in the sand lamprey at the metamorphic stages 4 to 5.     

Availability of and access to critical habitats in regulated rivers: effects of low-head barriers on threatened lampreys

1.  Conservation of freshwater animal populations requires their access to, as well as sufficient availability of, critical habitats, such as those for reproduction. Abundant small-scale barriers may cause extensive fragmentation of freshwater habitat but, by comparison to larger structures their effects are rarely considered by catchment managers. The relationship between the distribution of, and access to, spawning habitat in a regulated river, characterized by abundant small barriers, was examined for river lamprey Lampetra fluviatilis , a threatened migratory fish.
2.  Telemetry of adult lamprey in the River Derwent, North East England was used to quantify upriver migration and access to spawning habitat, together with surveys of spawning habitat availability and spawning activity between 2002 and 2007.
3.  Access in to the Derwent appeared severely restricted by a tidal barrage, beyond which lamprey migrated rapidly in unobstructed reaches. Of all lamprey tagged in the lower 4 km of river, or ascending the barrage, 64% and 17% passed the first and second weirs respectively, with high flows crucial for this. Although over 98% of lamprey spawning habitat occurred more than 51 km upstream, on average just 1.8% of river lamprey spawners were recorded there.
4.  In order to protect or rehabilitate species or species assemblages, greater attention needs to be paid to the relative spatial distribution of low-head barriers and the resultant availability of key habitats within individual catchments. This is particularly important given the renewed emphasis internationally on low-head hydropower solutions as a source of renewable energy, and the rapid growth in numbers of low-head barriers in many catchments.     

Novel neutrophil inhibitory factor homologue in the buccal gland secretion of Lampetra japonica

Abstract To identify the functional gene fragment, a neutrophil inhibitory factor (NIF) like protein was found in the buccal gland of Lampetra japonica, suggesting that this related lamprey protein represents a novel class of integrin receptor antagonists. The recombinant Lampetra japonica-NIF like (rLj-NIF) was identified by SDS-PAGE and purified by using His·Bind affinity chromatography. Effect of rLj-NIF on neutrophil migration suggested that rLj-NIF can act as a neutrophil inhibitory factor. Besides that, oxidative burst activity of neutriphil was tested by flow cytometry using dihydrorhodamine (DHR123) as a fluorogenic substrate, and the data suggested that the mean fluorescence intensity significantly decreased compared with positive controls (p<0.01). All above results indicated that rLj-NIF could also prevent the binding of β2 integrins to the surface of PMN and its FITC-labeled monoclonal antibodies (p<0.05). These data suggest that Lampetra japonica-NIF like protein is secreted by the stage of the parasite at the site of attachment. rLj-NIF plays an essential role in physiological reaction of neutrophil by a novel class of β2 integrin receptor antagonists. The activity of immunosuppressant of L. japonica-NIF could have potential medicinal value in anti-inflammation and therapy of autoimmune diseases.     

Riverscape genetics in brook lamprey: genetic diversity is less influenced by river fragmentation than by gene flow with the anadromous ecotype

Understanding the effect of human-induced landscape fragmentation on gene flow and evolutionary potential of wild populations has become a major concern. Here, we investigated the effect of riverscape fragmentation on patterns of genetic diversity in the freshwater resident European brook lamprey (Lampetra planeri) that has a low ability to pass obstacles to migration. We tested the hypotheses of (i) asymmetric gene flow following water current and (ii) an effect of gene flow with the closely related anadromous river lamprey (L. fluviatilis) ecotype on L. planeri genetic diversity. We genotyped 2472 individuals, including 225 L. fluviatilis, sampled from 81 sites upstream and downstream barriers to migration, in 29 western European rivers. Linear modelling revealed a strong positive relationship between genetic diversity and the distance from the river source, consistent with expected patterns of decreased gene flow into upstream populations. However, the presence of anthropogenic barriers had a moderate effect on spatial genetic structure. Accordingly, we found evidence for downstream-directed gene flow, supporting the hypothesis that barriers do not limit dispersal mediated by water flow. Downstream L. planeri populations in sympatry with L. fluviatilis displayed consistently higher genetic diversity. We conclude that genetic drift and slight downstream gene flow drive the genetic make-up of upstream L. planeri populations whereas gene flow between ecotypes maintains higher levels of genetic diversity in L. planeri populations sympatric with L. fluviatilis. We discuss the implications of these results for the design of conservation strategies of lamprey, and other freshwater organisms with several ecotypes, in fragmented dendritic river networks.Subject terms: Conservation biology, Ecological genetics, Evolutionary genetics, Genetic variation     

Expression of foreign genes in lamprey embryos: an approach to study evolutionary changes in gene regulation

Evolution in development can be viewed as a sequence of changes in gene regulation. To investigate the cross-species compatibility of 5' upstream regulatory regions, we introduced exogenous gene constructs derived from a gnathostome genome into fertilized eggs of the Japanese lamprey, Lampetra japonica, a sister group of the gnathostomes. Eggs were injected with gene constructs in which a sequence encoding the green fluorescent protein (GFP) had been located downstream of either a virus promoter or 5' regulatory regions of medaka actin genes. Reporter gene expression was recorded for more than a month starting two days after injection. Although the expression patterns were highly mosaic and differed among individuals, GFP was expressed predominantly in the striated muscles of lamprey embryos when driven by the 5' upstream regions of the medaka muscle actin genes. This implies that a pan-vertebrate muscle-specific gene regulatory mechanism may have evolved before the agnathan/gnathostome divergence. This gene-transfer technique potentially facilitates the visualization of cells in various differentiating tissues throughout development. The introduction of developmental genes of the lamprey or other animals into lamprey embryos is another potentially important application, one that could provide us with information on the evolutionary changes in functions of genes or gene cascades.