Contamination and sub-lethal toxicological effects of persistent organic pollutants in the European eel (<Emphasis Type="Italic">Anguilla anguilla</Emphasis>) in the Orbetello lagoon (Tuscany,Italy)

This paper reports on contamination levels and their sub-lethal toxicological effects in specimens of the European eel (Anguilla anguilla) in the Orbetello Lagoon, (Tuscany, Italy). Organochlorine pesticides (OC) and polychlorinated biphenyls (PCBs) were investigated as priority pollutants in muscle tissue. Phase I P450 enzymes, i.e., EROD, B(a)PMO and the two reductases (NADH ferryred and cyt c.), and cholinesterase (ChE) were assayed in liver and muscle as sensitive biological indicators of fish health. PCBs, lindane and p,p′DDE in muscles showed a wide concentration range (0.001–0.025 μg g⁻¹ wet weight) and attained the lowest levels in the eastern basin. High homogeneity and relatively low values were observed for phase I P450 enzymes, suggesting that no significant detoxification process of OC pesticides and PCBs occurred. The threat posed by organophosphate insecticides (OP) and CB compounds was also evidenced by ChE activity. The integrated response of phase I P450 enzymes and ChE activity being an indicator of potential effects of toxic contaminant levels on reproductive success and population decline of eels, can be used to assess the overall lagoon quality.     

Tolerance of acute hypercapnic acidosis by the European eel (<Emphasis Type="Italic">Anguilla anguilla</Emphasis>)

European eels ( Anguilla anguilla) were exposed sequentially to partial pressures of CO(2) in the water ( PwCO(2)) of 5, 10, 20, 40, 60 then 80 mm Hg (equivalent to 0.66-10.5 kPa), for 30 min at each level. This caused a profound drop in arterial plasma pH, from 7.9 to below 7.2, an increase in arterial PCO(2) from 3.0 mm Hg to 44 mm Hg, and a progressive decline in arterial blood O(2) content (caO(2)) from 10.0% to 1.97% volume. Gill ventilation rate increased significantly at water PwCO(2)s of 10, 20 and 40 mm Hg, followed by a decline at PwCO(2)s of 60 and 80 mm Hg, due to periodic breathing. Mean opercular pressure amplitude increased steadily throughout hypercapnic exposure and was significantly elevated at a PwCO(2) of 80 mm Hg. Hypercapnia caused a tachycardia between PwCO(2)s of 5 mmHg and 10 mm Hg, followed by a progressive decline in heart rate. Cardiac output (CO) remained unchanged throughout, as a consequence of a significant increase in stroke volume at PwCO(2)s of 40, 60 and 80 mm Hg. The eels maintained O(2) uptake at routine normocapnic levels throughout hypercapnic exposure. A comparison of the rates of blood O(2) delivery (calculated from CO and caO(2)) against O(2) consumption at PwCO(2)s of 60 mm Hg and 80 mm Hg indicated that a portion of O(2) uptake was due to cutaneous respiration. Thus, the European eel's exceptional tolerance of acute hypercapnia is probably a consequence of the tolerance of its heart to acidosis and hypoxia, and a contribution to O(2) uptake from cutaneous respiration.     

TNT biodegradation and production of dihydroxylamino-nitrotoluene by aerobic TNT degrader <Emphasis Type="Italic">Pseudomonas</Emphasis> sp. strain TM15 in an anoxic environment

Anaerobic bacteria have been used to produce 2,4-dihydroxylamino-nitrotoluene (2,4DHANT), a reductive metabolite of 2,4,6-trinitrotoluene (TNT). Here, an aerobic TNT biodegrader Pseudomonas sp. strain TM15 produced 2,4DHANT as evidenced by the molecular ion with m/z of 199 identified from LC-TOFMS analyses. TNT biodegradation with a high cell concentration (10⁹ cells/ml) led to a significant accumulation of 2,4DHANT in the culture medium, as well as hydroxylamino-dinitrotoluenes (HADNTs), although these products were not accumulated when a low cell concentration was used; also, the accumulation of diamino-nitrotoluene and of an unidentified metabolite were observed in the culture medium with the high cell concentration (10¹⁰ cells/ml). 2,4DHANT overproduction was a function of the aeration speed since cultures with low aeration speeds (30 rpm) had a 19-fold higher DHANT productivity than those aerated with high speeds (180 rpm); this indicates that molecular oxygen was related to the formation of 2,4DHANT. The quantification of dissolved oxygen (DO) in the media demonstrated that the productivity of 2,4DHANT was increased at low DO values. Moreover, supplying oxygen to the culture media produced a remarkable decrease of 2,4DHANT accumulation; these results clearly indicate that high 2,4DHANT production was a consequence of the oxygen deficit in the culture medium. This finding is useful for understanding the TNT biodegradation (bioremediation technology) in an anoxic environment. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Biological control of invasive populations of crayfish: the European eel (<Emphasis Type="Italic">Anguilla anguilla</Emphasis>) as a predator of <Emphasis Type="Italic">Procambarus clarkii</Emphasis>

The red swamp crayfish, Procambarus clarkii, is a paradigmatic invader of freshwater systems. Several attempts have been made to mitigate its multiple impacts but none was successful. Among the different methods proposed, the use of the European eel (Anguilla anguilla) as an indigenous predator is promising but the available information about its predatory ability on crayfish is to date scanty. To fill this gap in knowledge, we ran three experiments in wetlands and irrigation ditches in Italy. The first experiment, in the laboratory, was aimed at quantifying the extent of predation by eels on crayfish, the second, in enclosures, the size classes of crayfish mainly preyed and the possible effect of the eels on P. clarkii behaviour, and the third, in the field, its ability to effectively reduce crayfish populations. Results showed that eels prey on small-sized or soft crayfish, attacking them from the back; an indirect effect was to reduce crayfish trophic activity, which in turn might increases crayfish mortality due to starvation and decreases impact on the community. However, as shown in the field, the use of eels should be appropriately calibrated to the context of application. Taken together, our results show that eels might be used as a complement to the traditional trapping method. However, additional studies are necessary to understand the adequate number of eels to be introduced and to develop appropriate methods for quantifying such effects.     

Dietary effects on fatty acid composition in muscle tissue of juvenile European eel, <Emphasis Type="Italic">Anguilla anguilla</Emphasis> (L.)

The role of intracontinental migration patterns of European eel (Anguilla anguilla) receives more and more recognition in both ecological studies of the European eel and possible management measures, but small-scale patterns proved to be challenging to study. We experimentally investigated the suitability of fatty acid trophic markers to elucidate the utilization of feeding habitats. Eight groups of juvenile European eels were fed on eight different diets in a freshwater recirculation system at 20°C for 56 days. Three groups were fed on freshwater diets (Rutilus rutilus, Chironomidae larvae, and Gammarus pulex) and four groups were reared on diets of a marine origin (Clupea harengus, Crangon crangon, Mysis spec., and Euphausia superba) and one on commercial pellets used in eel aquaculture. Fatty acid composition (FAC) of diets differed significantly with habitat. FAC of eel muscle tissue seemed to be rather insensitive to fatty acids supplied with diet, but the general pattern of lower n3:n6 and EPA:ARA ratios in freshwater prey organisms could be traced in the respective eels. Multivariate statistics of the fatty acid composition of the eels resulted in two distinct groups representing freshwater and marine treatments. Results further indicate the capability of selectively restraining certain fatty acids in eel, as e.g. the n3:n6 ratio in all treatments was <4, regardless of dietary n3:n6. In future studies on wild eel, these measures can be used to elucidate the utilization of feeding habitats of individual European eel.     

Variation between European eel <Emphasis Type="Italic">Anguilla anguilla</Emphasis> (L.) stocks in five marshes of the Thames Estuary (United Kingdom)

The European eel (Anguilla anguilla, L.) was historically widely distributed throughout the United Kingdom, in coastal waters, lakes, rivers and wetlands. Recruitment has declined in recent decades and the species is now listed as ‘Critically Endangered’ on the International Union for Conservation of Nature and Natural Resources (IUCN) Red List. Management of suitable wetland habitats may contribute to species recovery; however, little is known about the stocks in these areas. In this study, yellow (adult stage?>?300 mm) eels were sampled in ditches in five marshes bordering the Thames Estuary in England, UK. Ecological variables, including ditch characteristics, invertebrate abundance and water quality parameters were measured. Habitat features were also observed and recorded, including access, land use and water management regimes. Eels were found in all marshes, but at varying catch-per-unit-effort (CPUE). There were no significant correlations between CPUE and the ecological variables, except ditch width. However, a significant difference in CPUE was found between two of the marshes, which may be explained by variations in local habitat management. Mean lengths showed a high proportion of females and mean body condition of four of the marshes was also found to be greater than in three rivers in the same region. These findings suggest that the marshes are potentially favourable eel habitats and that factors influencing habitat quality, such as land use and water management, may affect eel abundance, production of females and body condition. Effective management of such wetlands may therefore contribute to the conservation of European eel.     

Induction of the unculturable state in <Emphasis Type="Italic">Escherichia coli</Emphasis> K12 with 2,4,6-trinitrotoluene

The toxic effect of high (200 mg/l) 2,4,6-trinitrotoluene (TNT) concentrations on Escherichia coli K12 cells in the absence of exogenous nutrient sources (incubation in 0.5% NaCl) was shown to manifest itself in the transfer of the culture to an unculturable but viable state; its reversal depends on the duration of culture contact with the xenobiotic and the conditions of cell recultivation. The likelihood that cell succession to death forms the basis of the physiologo-biochemical mechanism of the unculturable state in Escherichia coli K12 population under conditions of combined toxic and starvation stress is discussed.     

Comparative analysis of 2,4,6-trinitrotoluene (TNT)-induced cellular responses and proteomes in <Emphasis Type="Italic">Pseudomonas</Emphasis> sp. HK-6 in two types of media

TNT-induced cellular responses and proteomes in Pseudomonas sp. HK-6 were comparatively analyzed in two different media: basal salts (BS) and Luria broth (LB). HK-6 cells could not degrade more than 0.5 mM TNT with BS medium, while in LB medium, they exhibited the enhanced capability to degrade as much as 3.0 mM TNT. Analysis of total cellular fatty acids in HK-6 cells suggested that the relative abundance of several saturated or unsaturated fatty acids is altered under TNT-mediated stress conditions. Scanning electron microscopy showed the presence of perforations, irregular rod formations, and wrinkled extracellular surfaces in cells under TNT stress. Proteomic analysis of soluble protein fractions from HK-6 cultures grown with TNT as a substrate revealed 11 protein spots induced by TNT. Among these, seven proteins (including Alg8, AlgB, NirB, and the AhpC/Tsa family) were detected only in LB medium containing TNT. The proteins AspS, Tsf, and assimilatory nitrate reductase were increasingly expressed only in BS medium containing TNT. The protein dGTPase was found to be induced and expressed when cells were grown in either type of TNT-containing media. These results provide a better understanding of the cytotoxicity and survival mechanism used by Pseudomonas sp. HK-6 when placed under TNT stress conditions.     

First record of the tick <Emphasis Type="Italic">Ixodes</Emphasis> (<Emphasis Type="Italic">Pholeoixodes</Emphasis>) <Emphasis Type="Italic">kaiseri</Emphasis> in Turkey

Nymphs and larvae belonging to Ixodes spp. were collected from a red fox in Turkey. The ticks were identified morphologically and molecularly (16S rDNA PCR and phylogenetic analysis) as I. kaiseri. Sequence and phylogenetic analyses show that our I. kaiseri isolate is very similar to I. kaiseri isolates collected from Germany, Serbia, Romania, and Hungary. Therefore, the existence of I. kaiseri has been demonstrated for the first time in Turkey. More studies relating to the regional distribution and vectorial competence of I. kaiseri are needed.     

<Emphasis Type="Italic">Ty</Emphasis>1<Emphasis Type="Italic">/copia</Emphasis>- and <Emphasis Type="Italic">Ty</Emphasis>3<Emphasis Type="Italic">/gypsy</Emphasis>-like DNA sequences in <Emphasis Type="Italic">Helianthus </Emphasis>species

Two repeated DNA sequences isolated from a partial genomic DNA library of Helianthus annuus, p HaS13 and p HaS211, were shown to represent portions of the int gene of a Ty3 /gypsy retroelement and of the RNase-Hgene of a Ty1 /copia retroelement, respectively. Southern blotting patterns obtained by hybridizing the two probes to BglII- or DraI-digested genomic DNA from different Helianthus species showed p HaS13 and p HaS211 were parts of dispersed repeats at least 8 and 7 kb in length, respectively, that were conserved in all species studied. Comparable hybridization patterns were obtained in all species with p HaS13. By contrast, the patterns obtained by hybridizing p HaS211 clearly differentiated annual species from perennials. The frequencies of p HaS13- and p HaS211-related sequences in different species were 4.3x10(4)-1.3x10(5) copies and 9.9x10(2)-8.1x10(3) copies per picogram of DNA, respectively. The frequency of p HaS13-related sequences varied widely within annual species, while no significant difference was observed among perennial species. Conversely, the frequency variation of p HaS211-related sequences was as large within annual species as within perennials. Sequences of both families were found to be dispersed along the length of all chromosomes in all species studied. However, Ty3 /gypsy-like sequences were localized preferentially at the centromeric regions, whereas Ty1/ copia-like sequences were less represented or absent around the centromeres and plentiful at the chromosome ends. These findings suggest that the two sequence families played a role in Helianthusgenome evolution and species divergence, evolved independently in the same genomic backgrounds and in annual or perennial species, and acquired different possible functions in the host genomes.     

New combinations and typifications in <Emphasis Type="Italic">Bistorta</Emphasis>, <Emphasis Type="Italic">Persicaria</Emphasis>, <Emphasis Type="Italic">Polygonum,</Emphasis> and <Emphasis Type="Italic">Rumex</Emphasis> (Polygonaceae)

New combinations are proposed in anticipation of the Polygonaceae treatment in the forthcoming volume of Intermountain Flora: Polygonum kelloggii var. esotericum, P. kelloggii var. watsonii , Rumex densiflorus var. pycnanthus , R. salicifolius var. utahensis, and R. occidentalis var. tomentellus. Typifications are proposed to facilitate ongoing studies in Polygonaceae and to maintain current usage.     

Size-dependent foraging niches of European Perch <Emphasis Type="Italic">Perca fluviatilis</Emphasis> (Linnaeus, 1758) and North American Yellow Perch <Emphasis Type="Italic">Perca flavescens</Emphasis> (Mitchill, 1814)

Body size of consumer species is a fundamental trait that influences the trophic ecology of individuals and their contribution to the functioning of freshwater ecosystems. However, the relationship between body size and trophic ecology can be highly variable both within and between closely-related and similarly-sized species. In this study we compared the intra- and interspecific relationship between body size and trophic position for North American Yellow Perch Perca flavescens and European Perch Perca fluviatilis, which share similarities in morphology, life history traits and trophic requirements. We used stable isotope ratios (δ¹⁵N and δ¹³C) to characterize differences in size-dependency of trophic position and to trace consumer foraging history of Yellow Perch in lakes in the Northwestern United States and European Perch in lakes in Germany. The trophic position and stable isotope ratios of Yellow Perch and European Perch steadily increased with total body length, but European Perch were consistently feeding at higher trophic positions than Yellow Perch at a given length. European Perch occupied considerably higher trophic positions (mean trophic position = 3.9) than Yellow Perch (mean trophic position = 2.8). Large European Perch were increasingly piscivorous, whereas large Yellow Perch were more opportunistic and omnivorous predators of invertebrate prey. Overall, the trophic position among individual Yellow Perch varied more strongly than in European Perch. We conclude that both species similarly increase in trophic position with size, but the specific size-dependency of both trophic position and resource use varies with taxonomy and local ecological conditions. Thus, body size as a sole measure of trophic position should be considered cautiously when generalizing across populations and species.     

Genetic population diversity of European eel <Emphasis Type="Italic">Anguilla anguilla</Emphasis> elvers in two Egyptian water bodies,Rosetta (Rachid) estuary and Burullus Lake

The European eel Anguilla anguilla is a species of great economic importance for Egypt. Its long trans-Atlantic migrations for semelparous spawning then larval homing provides a possible way for worldwide mixing of populations. Its natural habitats in Egypt, as in other Mediterranean countries, are under heavy load of pollution due to anthropogenic activities. Hence, a major priority should be given to assess the genetic diversity within/among eels’ populations as a step towards revealing populations’ status, identification of possible conservation units, and, subsequently, implementation of proper conservation and management strategies for them. For this, A. anguilla elvers were collected from Rosetta estuary and Burullus coastal lagoon in the North of Egypt. Sequencing of the barcode region of the cytochrome c oxidase subunit 1 gene (COI) was carried out for all collected elvers. The sequences were analyzed using various genetic, phylogenetic, and population analyses to assess their variability. A total of 33 haplotypes were identified, 14 of which were unique to Burullus Lake, and 11 to Rosetta estuary. Very low fixation (F _st ?=?0–0.04) was found for A. anguilla populations in Egypt and worldwide, except for the Turkish and Czech ones (F _st ?=?0.47–0.62 and 0.18–0.41, respectively). Both Egyptian populations showed signs of recent populations’ expansion, besides low, non-significant raggedness values. The results suggested Burullus Lake and Rosetta estuary as separate conservation units for A. anguilla in Egypt. These units require special attention for proper protection and management of the existing eel diversity and permitting sustainability in the activities dealing with eel’s fishing and aquaculture.     

The role of BAP in somatic embryogenesis induction from seed explants of <Emphasis Type="Italic">Arachis</Emphasis> species from Sections <Emphasis Type="Italic">Erectoides</Emphasis> and <Emphasis Type="Italic">Procumbentes</Emphasis>

Somatic embryogenesis was induced from seed explants of Arachis archeri, A. porphyrocalix (Section Erectoides) and A. appressipila (Section Procumbentes) in response to 6-benzylaminopurine (BAP). Embryo axes first developed into single shoots in response to 4.4 μM BAP. Friable embryogenic calluses were produced from the hypocotyl region of these explants in response to different BAP concentrations. Embryonic leaflets also gave rise to friable calluses, but somatic embryos were only observed in explants of A. archeri and A. appressipila. Histological analyses revealed the presence of heart-shaped, torpedo and cotyledonary stages embryos, both as isolated and fused structures. A low frequency of embryo-to-plant conversion was achieved by inducing shoot development on medium solidified with 0.5% phytagel and supplemented with 1.5% or 3% sucrose. Rooting was induced on MS supplemented with indole-3-acetic acid (IAA).     

Floral development and systematic position of <Emphasis Type="Italic">Arillastrum</Emphasis>, <Emphasis Type="Italic">Allosyncarpia</Emphasis>, <Emphasis Type="Italic">Stockwellia</Emphasis> and <Emphasis Type="Italic">Eucalyptopsis</Emphasis> (Myrtaceae)

We have investigated the floral ontogeny of Arillastrum, Allosyncarpia, Stockwellia and Eucalyptopsis (of the eucalypt group, Myrtaceae) using scanning electron microscopy and light microscopy. Several critical characters for establishing relationships between these genera and to the eucalypts have been determined. The absence of compound petaline primordia in Arillastrum, Allosyncarpia, Stockwellia and Eucalyptopsis excludes these taxa from the eucalypt clade. Post-anthesis circumscissile abscission of the hypanthium above the ovary in Stockwellia, Eucalyptopsis and Allosyncarpia is evidence that these three taxa form a monophyletic group; undifferentiated perianth parts and elongated fusiform buds are characters that unite Stockwellia and Eucalyptopsis as sister taxa. No floral characters clearly associate Arillastrum with either the eucalypt clade or the clade of Stockwellia, Eucalyptopsis and Allosyncarpia.We gratefully acknowledge Clyde Dunlop and Bob Harwood (Northern Territory Herbarium) for collecting specimens of Allosyncarpia, and Bruce Gray (Atherton) for collecting specimens of Stockwellia. The Australian National Herbarium (CANB) kindly lent herbarium specimens of Eucalyptopsis for examination. This research was supported by a University of Melbourne Research Development Grant to Andrew Drinnan.     

<Emphasis Type="Italic">Quinua</Emphasis> and Relatives (<Emphasis Type="Italic">Chenopodium</Emphasis> sect.<Emphasis Type="Italic">Chenopodium</Emphasis> subsect.<Emphasis Type="Italic">Celluloid</Emphasis>)

Traditionally viewed as an Andean grain crop,Chenopodium quinoa Willd. includes domesticated populations that are not Andean, and Andean populations that are not domesticated. Comparative analysis of leaf morphology and allozyme frequencies have demonstrated that Andean populations, both domesticated(quinua) and free-living(ajara), represent an exceptionally homogeneous unit that is well differentiated from allied domesticates of coastal Chile(quingua) and freeliving populations of the Argentine lowlands(C. hircinum). This pattern of relationships indicates that Andean populations represent a monophyletic crop/weed system that has possibly developed through cyclic differentiation (natural vs. human selection) and introgressive hybridization. Relative levels of variation suggest that this complex originated in the southern Andes, possibly from wild types allied withC. hircinum, with subsequent dispersal north to Colombia and south to the Chilean coast. Coastal populations were apparently isolated from post-dispersal differentiation and homogenization that occurred in the Andes. Other data point toward a center of origin in the northern Andes with secondary centers of genetic diversity subsequently developing in the southern Andes and the plains of Argentina. Comparative linkage of South American taxa, all tetraploid, with North American tetraploids of the subsection will eventually clarify this problem. While the possibility of a direct phyletic connection betweenC. quinoa and the Mexican domesticate(C. berlandieri subsp. nuttalliae,) cannot be excluded, available evidence indicates that the latter represents an autonomous lineage that is associated with the basal tetraploid, C. b. subsp.berlandieri, through var.sinuatum, whereas South American taxa show possible affinities to either var. zschackei or var.berlandieri. An extinct domesticate of eastern North America,C. b. subsp.jonesianum, represents either another instance of independent domestication, possibly from subsp. b. var.zschackei, or a northeastern outlier of subsp.nuttalliae.     

Revision of the <Emphasis Type="Italic">Serrulati</Emphasis> species of <Emphasis Type="Italic">Penstemon</Emphasis> section <Emphasis Type="Italic">Saccanthera</Emphasis> subsection <Emphasis Type="Italic">Serrulati</Emphasis>

A revision of Penstemon sect. Saccanthera subsect. Serrulati includes a new species (P. salmonensis), a new variety (P. triphyllus var. infernalis), and the elevation of a subspecies to species (P. curtiflorus), bringing the total number of species to eight, which are keyed and described, complete with nomenclature and type citations.     

<Emphasis Type="Italic">Galleria</Emphasis><Emphasis Type="Italic">mellonella</Emphasis> are Resistant to <Emphasis Type="Italic">Pneumocystis</Emphasis><Emphasis Type="Italic">murina</Emphasis> Infection

Studying Pneumocystis has proven to be a challenge from the perspective of propagating a significant amount of the pathogen in a facile manner. The study of several fungal pathogens has been aided by the use of invertebrate model hosts. Our efforts to infect the invertebrate larvae Galleria mellonella with Pneumocystis proved futile since P. murina neither caused disease nor was able to proliferate within G. mellonella. It did, however, show that the pathogen could be rapidly cleared from the host.     

<Emphasis Type="Italic">In silico</Emphasis> and <Emphasis Type="Italic">in situ</Emphasis> characterization of the zebrafish (<Emphasis Type="Italic">Danio rerio</Emphasis>) <Emphasis Type="Italic">gnrh3</Emphasis> (sGnRH) gene

Background

Gonadotropin releasing hormone (GnRH) is responsible for stimulation of gonadotropic hormone (GtH) in the hypothalamus-pituitary-gonadal axis (HPG). The regulatory mechanisms responsible for brain specificity make the promoter attractive for in silico analysis and reporter gene studies in zebrafish (Danio rerio).

Results

We have characterized a zebrafish [Trp⁷, Leu⁸] or salmon (s) GnRH variant, gnrh 3. The gene includes a 1.6 Kb upstream regulatory region and displays the conserved structure of 4 exons and 3 introns, as seen in other species. An in silico defined enhancer at -976 in the zebrafish promoter, containing adjacent binding sites for Oct-1, CREB and Sp1, was predicted in 2 mammalian and 5 teleost GnRH promoters. Reporter gene studies confirmed the importance of this enhancer for cell specific expression in zebrafish. Interestingly the promoter of human GnRH-I, known as mammalian GnRH (mGnRH), was shown capable of driving cell specific reporter gene expression in transgenic zebrafish.

Conclusions

The characterized zebrafish Gnrh3 decapeptide exhibits complete homology to the Atlantic salmon (Salmo salar) GnRH-III variant. In silico analysis of mammalian and teleost GnRH promoters revealed a conserved enhancer possessing binding sites for Oct-1, CREB and Sp1. Transgenic and transient reporter gene expression in zebrafish larvae, confirmed the importance of the in silico defined zebrafish enhancer at -976. The capability of the human GnRH-I promoter of directing cell specific reporter gene expression in zebrafish supports orthology between GnRH-I and GnRH-III.