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Micronucleus test and observation of nuclear alterations in erythrocytes of Nile tilapia exposed to waters affected by refinery effluent.

Micronuclei and nuclear alterations tests were performed on erythrocytes of Oreochromis niloticus (Perciformes, Cichlidae) in order to evaluate the water quality from Paraíba do Sul river, in an area affected by effluents from an oil shale processing plant, located in the city of São José dos Campos, Brazil-SP. Water samples were collected on 2004 May and August (dry season) and on 2004 November and 2005 January (rain season), in three distinct sites, comprising 12 samples. It was possible to detect substances of clastogenic and/or aneugenic potential, as well as cytotoxic substances, chiefly at the point corresponding to the drainage of oil shale plant wastes along the river. The highest incidence of micronuclei and nuclear alterations was detected during May and August, whereas the results obtained in November and January were insignificant. This work shows that the effluent treatment provided by the oil shale plant was not fully efficient to minimize the effect of cytotoxic and mutagenic substances in the test organism surveyed.

Aneugens↗

Novel evolutionary relationship among four fish model systems.

Knowledge of the correct phylogenetic relationships among animals is crucial for the valid interpretation of evolutionary trends in biology. Zebrafish, medaka, pufferfish and cichilds are fish models for development, genomics and comparative genetics studies, although their phylogenetic relationships have not been tested rigorously. The results of phylogenomic analysis based on 20 nuclear protein-coding genes confirmed the basal placement of zebrafish in the fish phylogeny but revealed an unexpected relationship among the other three species, contrary to traditionally held systematic views based on morphology. Our analyses show that medaka (Beloniformes) and cichlids (Perciformes) appear to be more closely related to each other than either of them is to pufferfish (Tetraodontiformes), suggesting that a re-interpretation of some findings in comparative biology might be required. In addition, phylogenomic analyses show that fish typically have more copies of nuclear genes than land vertebrates, supporting the fish-specific genome duplication hypothesis.

Animals↗

Molecular characterization of rockfish (Sebastes schlegeli) gonadotropin subunits and their mRNA expression profiles during oogenesis.

In an attempt to understand the reproductive regulation in viviparous teleosts, gonadotropin (GTH) subunit cDNAs were characterized and the expression levels of GTH subunit mRNAs in the pituitary glands of the rockfish, Sebastes schlegeli (Scorpaeniformes, Scorpaenidae), were examined by Northern blot analysis. The complete sequences of rockfish GTH subunits (GTHalpha, FSHbeta, and LHbeta) were determined by rapid amplification of cDNA ends (RACE) and nucleotide sequencing. Based on the RACE analysis, the cDNAs of GTHalpha, FSHbeta, and LHbeta consisted of 655, 540, and 529 nucleotides encoding peptides of 132, 130, and 143 amino acids, respectively. The mature peptides of rockfish FSHbeta, LHbeta, and common GTHalpha showed high sequence identities (FSHbeta, 58-62%; LHbeta, 86-94%; and GTHalpha, 87-88%) to those of other perciforme (e.g., orange spotted grouper, red seabream, stripped bass, and sea bass). In a sequence alignment of the mature peptides, rockfish FSHbeta exhibited a unique feature, the lack of a conserved N-glycosylation site. This is in contrast to both LHbeta and GTHalpha which contain one and two N-glycosylation sites, respectively, and is consistent with those of other teleosts. The mRNA levels of the GTHalpha subunit increased at the vitellogenic stage and remained steady from ovulation to post-parturition. FSHbeta mRNA levels increased abruptly during the vitellogenic stages and then decreased during ovulation, embryonic development, and post-parturition. LHbeta mRNA levels were observed to rapidly increase during the vitellogenic stage, reached its highest levels during ovulation and was then followed by a decrease at post-parturition. These results suggest that in the female rockfish FSH and LH syntheses are predominant during vitellogenesis and oocyte maturation, respectively.

Amino Acid Sequence↗

Sequence analysis, endocrine regulation, and signal transduction of GnRH receptors in teleost fish.

Three gonadotropin-releasing hormones (GnRHs) and three cognate receptors have been identified in vertebrates, with distinct distributions and functions. According to their sequences, the receptors can be grouped into distinct classes: types I, II, and III. One branch contains all type-I GnRH receptors (GnRH-R-I) from mammals and fish; another branch clusters mainly amphibian and human type-II GnRH receptors; and a third branch includes evolved fish, mainly perciform species, type-III GnRH receptors. Taken tilapia GnRH receptors as a model, the present study summarizes the information regarding the amino-acid residues assumed to be involved in the receptors' structure, binding, activation, and intracellular signal transduction, including arrangement of the disulfide bonds, glycosylation sites, coupling to G proteins, and protein kinase A or protein kinase C phosphorylation sites.

Animals↗

Changes in mRNA expression of grouper (Epinephelus coioides) growth hormone and insulin-like growth factor I in response to nutritional status.

Growth hormone (GH) and insulin-like growth factor-I (IGF-I) are key links to nutritional condition and growth regulation in teleost. To understand the endocrine mechanism of growth regulation in grouper, we cloned the cDNAs for grouper GH and IGF-I and examined their mRNA expression during different nutritional status. Grouper GH cDNA is 936 base pairs (bp) long excluding the poly-A tail. It contained untranslated regions of 85 and 231bp in the 5'- and 3'-ends, respectively. It has an open reading frame of 612bp coding for a signal peptide of 17 amino acids (aa) and a mature hormone of 187aa residues. Based on the aa sequence of the mature hormone, grouper GH shows higher sequence identity (>76%) to GHs of perciforms than to GHs of cyprinids and salmonids (53-69%). Grouper preproIGF-I cDNA consisted of 558bp, which codes for 186aa. This is composed of 44aa for the signal peptide, 68aa for the mature peptide comprising B, C, A, and D domains, and 74aa for the E domain. Mature grouper IGF-I shows very high sequence identity to IGF-I of teleost fishes (84-97%) compared to advanced groups of vertebrates such as chicken, pig, and human (80%). Using DNA primers specific for grouper GH and IGF-I, the changes in mRNA levels of pituitary GH and hepatic IGF-I in response to starvation and refeeding were examined by a semi-quantitative RT-PCR. Significant elevation of GH mRNA level was observed after 2 weeks of food deprivation, and increased further after 3 and 4 weeks of starvation. GH mRNA level in fed-controls did not change significantly during the same period. Hepatic IGF-I mRNA level decreased significantly starting after 1 week of starvation until the 4th week. There was no significant change in IGF-I mRNA levels in fed-controls. One week of refeeding can restore the GH and IGF-I mRNA back to its normal levels. Deprivation of food for 1-4 weeks also resulted in cessation of growth and decrease in condition factor.

Amino Acid Sequence↗

The Atlantic-Mediterranean transition: discordant genetic patterns in two seabream species, Diplodus puntazzo (Cetti) and Diplodus sargus (L.).

Sparids are a group of demersal perciform fish of high commercial value, which have experienced an extensive radiation, particularly in the Mediterranean, where they occupy a variety of different niches. The present study focuses on two species: Diplodus sargus and D. puntazzo, presenting a wide distribution from the Mediterranean to the eastern Atlantic coasts. They display similar ecological behaviour and are evolutionary closely related. Both are highly appreciated in fisheries and D. puntazzo is currently under domestication process. However, little is know on their population structure and it is an open question whether any genetic differentiation exists at the geographic level. To address this issue we examined sequence variation of a portion of the mitochondrial DNA (mtDNA) control region in population samples of each of the two species collected over a wide geographic range. In addition to the mtDNA, analysis of nuclear loci (allozymes) was included in the study to compare patterns revealed by nuclear and mitochondrial markers. The studied samples covered an area from the eastern Mediterranean to the Portuguese coasts immediately outside the Gibraltar Strait. The two species revealed a level of sequence polymorphism remarkably different for the control region with the D. puntazzo and D. sargus showing 111 and 28 haplotypes, respectively. Such a difference was not detected with allozyme markers. The two species also showed large differences in their population structure. While D. puntazzo presented a marked genetic divergence between the Atlantic and Mediterranean samples, D. sargus showed little intraspecific differentiation. These results were supported using both mtDNA and allozyme markers, and were interpreted as the consequence of differences in the history of the two species such as fluctuations in the effective population size due to bottlenecks and expansions, possibly combined with present-day differences in levels of gene flow.

Animals↗

Vitellogenin detection in surface mucus of the South American cichlid fish Cichlasoma dimerus (Heckel, 1840) induced by estradiol-17beta. Effects on liver and gonads.

During the last decade, special attention has been focused on the consequences of exposure to environmental estrogens on reproduction in wild fish species. For this reason, characterization of biomarkers of such exposures could result in useful tools for these studies. The detection of vitellogenin (Vtg), a precursor of yolk proteins, is being intensely studied since its synthesis in the liver is regulated by the estradiol-17beta and is influenced by other estrogenic compounds. The aim of this work was to assess the presence of Vtg in the surface mucus of males of Cichlasoma dimerus (Teleostei, Perciformes), a typical South American freshwater cichlid, after hormonal treatment with estradiol-17beta (intraperitoneal injections of 10 microg E(2)/g fish). Plasma and mucus from females and treated males analyzed by Western blot revealed that different heterologous antisera against Vtg bind to putative protein of 180 & 120 kDa and 120 & 110 kDa, respectively, whereas no reaction was found in samples of untreated males. The same profile was observed in mucus samples using Dot blot, a very easy and direct technique. Using immunocytochemistry techniques, immunoreactive Vtg (ir-Vtg) producing cells in the liver of females and treated males were detected. Testes and liver tissues were also assessed by histological techniques. Marked alterations in both organs were observed, such as lower sperm production, presence of immature germ cells in the lobular lumen of testes, and some morphology changes in the hepatocytes due to the accumulation of Vtg. This is the first report about the effects of an estrogen in the Vtg synthesis and their consequences on liver and gonads of a South American fresh water cichlid. These results also support the possibility of using Vtg from surface mucus as a potential biomarker for estrogenic compounds through a noninvasive, useful and easy assay to monitor the presence of endocrine disrupting chemicals in the environment.

Animals↗

Mercury in fish from three rift valley lakes (Turkana, Naivasha and Baringo), Kenya, East Africa.

Total mercury (THg) concentrations were measured for various fish species from Lakes Turkana, Naivasha and Baringo in the rift valley of Kenya. The highest THg concentration (636 ng g(-1) wet weight) was measured for a piscivorous tigerfish Hydrocynus forskahlii from Lake Turkana. THg concentrations for the Perciformes species, the Nile perch Lates niloticus from Lake Turkana and the largemouth bass Micropterus salmoides from Lake Naivasha ranged between 4 and 95 ng g(-1). The tilapiine species in all lakes, including the Nile tilapia Oreochromis niloticus, had consistently low THg concentrations ranging between 2 and 25 ng g(-1). In Lake Naivasha, the crayfish species, Procambrus clarkii, had THg concentrations similar to those for the tilapiine species from the same lake, which is consistent with their shared detritivore diet. THg concentrations in all fish species were usually consistent with their known trophic position, with highest concentrations in piscivores and declining in omnivores, insectivores and detritivores. One exception is the detritivore Labeo cylindricus from Lake Baringo, which had surprisingly elevated THg concentrations (mean=75 ng g(-1)), which was similar to those for the top trophic species (Clarias and Protopterus) in the same lake. Except for two Hydrocynus forskahlii individuals from Lake Turkana, which had THg concentrations near or above the international marketing limit of 500 ng g(-1), THg concentrations in the fish were generally below those of World Health Organization's recommended limit of 200 ng g(-1) for at-risk groups.

Animals↗

Tilapia glycoprotein hormone alpha subunit: cDNA cloning and hypothalamic regulation.

The cDNA encoding the glycoprotein alpha (GPalpha) subunit of tilapia (Oreochromis mossambicus) was partially cloned using RACE-polymerase chain reaction (PCR) technique. The amplified cDNA was found to be 583 bases long, and to consist of a portion of the signal peptide, the full sequence encoding the mature peptide (94 amino acids) and the 3' untranslated region. Northern blot analysis revealed a single band of approximately 600 bp. Alignment of the deduced amino acids of the mature protein showed that the tilapia GPalpha subunit shares more than 80% identity with that of other perciform fish (i.e. striped bass, sea bream and yellowfin porgy) and less than 70% with that of more taxonomically remote fish and other vertebrates. Exposure of dispersed tilapia pituitary cells to salmon gonadotropin-releasing hormone (sGnRH) elevated GPalpha mRNA levels via both PKC and cAMP-protein kinase A (PKA) pathways. The transcript levels were also regulated by pituitary adenylate cyclase activating polypeptide (PACAP) and neuropeptide Y (NPY), both acting through PKC and PKA pathways. Moreover, a combined treatment of PACAP or NPY with GnRH seems to have an additive effect on the GPalpha subunit gene transcription. These results suggest that in tilapia the expression of GPalpha subunit is regulated by GnRH mainly via PKC and PKA pathways. Furthermore, PACAP and NPY can elevate the GnRH-stimulated GPalpha subunit transcription and can directly affect the subunit mRNA levels, via the same transduction pathways.

Animals↗

CDNA cloning and sequence of European sea bass (Dicentrarchus labrax) somatolactin.

Three cDNA clones encoding for European sea bass somatolactin (SL) were obtained by RT-PCR and 3' RACE of RNA of pituitary origin. Clone 1 was 582 bp in length, and included a part of the signal peptide and the 5' end of the mature protein. Clone 2 (1075 bp) included a fragment of the coding sequence and the 3' untranslated region, which was 888 bp in length and contained two putative polyadenylation signals (AATAAA) at 12-17, and 202-207 nucleotides upstream of the poly (A) tail. Clone 3 was 624 bp in length and its nucleotide sequence encoding for the entire mature protein confirmed the sequence already determined from the first two clones. The size of SL mRNA transcripts was estimated by Northern blot analysis and a single band of approximately 1.6 kb was observed with pituitary RNAs. No band was found with RNAs of brain and liver origin. Alignment of the deduced amino acid sequence revealed that European sea bass SL shared 90-84% identity with perciform, pleuronectiform and scorpaeniform fish SLs, and 77-57% with other SLs of more distant fish orders, with a strict conservation of Cys residues and the N-glycosylation site (Asn-Lys-Thr) at 121-123 amino acid positions. The reconstruction of the phylogenetic tree based on SL nucleotide sequences. and analyzed by maximum likelihood distances, showed the same clustering as the present hierarchy of fish. When comparisons were made among SL, prolactin and growth hormone of European sea bass, the overall amino identity was relatively low (22-23%). However, a high degree of amino acid homology was found at the C-terminus, which contains three of the four Cys residues strictly conserved in all the members of GH/PRL family.

3' Untranslated Regions↗

Developmental expression of three different prepro-GnRH (gonadotrophin-releasing hormone) messengers in the brain of the European sea bass (Dicentrarchus labrax).

In this study, we have analyzed the ontogenic expression of three gonadotrophin-releasing hormones (GnRH) systems expressed in the brain of a perciform fish, the European sea bass, using in situ hybridization. The riboprobes used correspond to the GnRH-associated peptide (GAP) coding regions of the three prepro-GnRH cDNAs cloned from the same species: prepro-salmon GnRH, prepro-seabream GnRH and prepro-chicken GnRH II. On day 4 after hatching, the first prepro-chicken GnRH-II mRNA-expressing cells appeared in the germinal zone of the third ventricle. They increased in number and size from 10 to 21 days, reaching at day 30 their adult final position, within the synencephalic area, at the transitional zone between the diencephalon and the mesencephalon. First prepro-salmon GnRH mRNA-expressing cells became evident on day 7 arising from the olfactory placode and migrating towards the olfactory nerve. On day 10, this cell group reached the olfactory bulb, being evident in the ventral telencephalon and preoptic area from days 15 and 45, respectively. Weakly labeled prepro-seabream GnRH mRNA-expressing cells were first detected at 30 days in the olfactory area and ventral telencephalon. On day 45, prepro-seabream GnRH mRNA-expressing cells were also present in the preoptic region reaching the ventrolateral hypothalamus on day 60. The results obtained in sea bass indicate that sGnRH and sbGnRH cells have a common origin in an olfactory primordium suggesting that both forms might arise from a duplication of a single ancestral gene, while cGnRH-II cells develop from a synencephalic primordium.

Animals↗

Olfactory sensitivity of the gilthead seabream (Sparus auratus L) to conspecific body fluids.

The potential for intraspecific chemical communication in the gilthead seabream (a marine perciform) was investigated by assessing the olfactory sensitivity to conspecific body-fluids (water occupied by conspecifics, intestinal fluid, urine, semen, egg fluid) by multiunit electrophysiological recording from the olfactory nerve. The olfactory system was responsive to water previously occupied by conspecifics, and the active compound(s) could be extracted by solid-phase extraction. The olfactory system was extremely sensitive to body fluids of sexually mature conspecifics: thresholds of detection were 1:10(7.4) (intestinal fluid), 1:10(6.1) (gametes), and 1:10(4.2) (urine). The olfactory system was also sensitive to amino acids with thresholds of detection from 10(-8.1) M (L-leucine) to 10(-6.1) M (L-phenylalanine). However, a range of other known fish odorants (steroids, bile acids, and prostaglandins) failed to evoke significant responses. Given the high olfactory sensitivity to intestinal fluid and the low urine release rates of marine compared with freshwater fish, we suggest that chemical communication is likely to be mediated via compounds present in the intestinal fluid rather than urine. Furthermore, the types of chemicals involved are likely to be different from those of freshwater fish. Their exact chemical identity and biological roles remain to be established.

Animal Communication↗

The conus arteriosus of the adult gilthead seabream (Sparus auratus).

This paper reports on the presence of the conus arteriosus in the heart of the adult gilthead seabream, Sparus auratus (Perciformes, Teleostei). The junctional region between the single ventricle and the bulbus arteriosus has been studied by conventional light microscopy, and by scanning and transmission electron microscopy. In addition, fluorescent phalloidin and antibodies against the muscle myosin heavy chains, laminin and collagen type IV have been used. The conus arteriosus is a distinct muscular segment interposed between the ventricle and the bulbus arteriosus. It is clearly different from the bulbus arteriosus due to its myocardial nature. It can also be distinguished from the ventricular myocardium because: (1) it has a conus shape; (2) it is formed by compact, well-vascularized myocardium; (3) it is surrounded on its inner and outer faces by fibrous layers rich in collagen and elastin; (4) it constitutes the anatomical support of the so-termed conus valves; (5) it shows intense staining for laminin and type-IV collagen; and (6) the myocardial cells located close to the inner fibrous layer are helicoidally arranged. By contrast, the ventricular myocardium is highly trabecular, lacks a compacta, shows no vessels, and presents barely detectable amounts of laminin and collagen type IV. The presence of a distinct conus arteriosus in the heart of an evolutionary advanced teleost species indicates that the conus is not a vestigial segment from the evolutionary or embryological points of view. The characteristic spatial arrangement of the conus myocytes strongly suggests that the conus is implicated in the mechanical performance of the conus valves.

Animals↗

Consequences of a catadromous life-strategy for levels of mitochondrial DNA differentiation among populations of the Australian bass, Macquaria novemaculeata.

The influence of a catadromous life-strategy on levels of spatial genetic structuring in fish is poorly understood. In an effort to gain a better appreciation of how this specialized life-strategy determines population genetic structuring, we assessed variation in the mitochondrial DNA (mtDNA) control region in a catadromous perciform, the Australian bass Macquaria novemaculeata. Nineteen putative haplotypes were resolved using temperature gradient gel electrophoresis from 10 geographically distinct populations. Significant heterogeneity was revealed in haplotype frequencies and their spatial distributions among many locales. Gene partitioning statistics (AMOVA) for both raw haplotype frequency data and frequency data with sequence divergences were concordant, indicating that M. novemaculeata populations were moderately genetically structured (phi ST = 0.05, 0.06; P < 0.001, respectively). Isolation by distance seems to be a strong structuring force in M. novemaculeata, culminating in no detectable phylogeographic structuring among haplotypes. Low sequence divergences were observed among many haplotypes and it is suggested that these are the result of pruning of maternal lineages by cyclical variations in female reproductive success. This study highlights the importance of life-history patterns and, in particular, spawning locality, in determining spatial structuring of mtDNA variation in catadromous species.

Animals↗

Functional antifreeze glycoprotein genes in temperate-water New Zealand nototheniid fish infer an Antarctic evolutionary origin.

The fish fauna of the Antarctic Ocean is dominated by five endemic families of the Perciform suborder Notothenioidei, thought to have arisen in situ within the Antarctic through adaptive radiation of an ancestral stock that evolved antifreeze glycoproteins (AFGPs) enabling survival as the ocean chilled to subzero temperatures. The endemism results from geographic confinement imposed by a massive oceanographic barrier, the Antarctic Circumpolar Current, which also thermally isolated Antarctica over geologic time, leading to its current frigid condition. Despite this voluminous barrier to fish dispersal, a number of species from the Antarctic family Nototheniidae now inhabit the nonfreezing cool temperate coasts of the southern continents. The origin of these temperate-water nototheniids is not completely understood. Since the AFGP gene apparently evolved only once, before the Antarctic notothenioid radiation, the presence of AFGP genes in extant temperate-water nototheniids can be used to infer an Antarctic evolutionary origin. Genomic Southern analysis, PCR amplification of AFGP genes, and sequencing showed that Notothenia angustata and Notothenia microlepidota endemic to southern New Zealand have two to three AFGP genes, structurally the same as those of the Antarctic nototheniids. At least one of these genes is still functional, as AFGP cDNAs were obtained and low levels of mature AFGPs were detected in the blood. A phylogenetic tree based on complete ND2 coding sequences showed monophyly of these two New Zealand nototheniids and their inclusion in the monophyletic Nototheniidae consisted of mostly AFGP-bearing taxa. These analyses support an Antarctic ancestry for the New Zealand nototheniids. A divergence time of approximately 11 Myr was estimated for the two New Zealand nototheniids, approximating the upper Miocene northern advance of the Antarctic Convergence over New Zealand, which might have served as the vicariant event that lead to the northward dispersal of their most recent common ancestor. Similar secondary northward dispersal likely applies to the South American nototheniid Paranotothenia magellanica, which has four AFGP genes in its DNA, but not to the sympatric nototheniid Patagonotothen tessellata, which does not appear to have any AFGP sequences in its genome at all.

Amino Acid Sequence↗

Primary structures of sardaines Z1 and Z2, protamines isolated from striped bonito (Sarda orientalis).

Striped bonito protamine, sardaine, was isolated from the sperm of striped bonito (Sarda orientalis) by extraction with sulfuric acid followed by ion-exchange chromatography. The preparation gave a single band upon polyacrylamide gel electrophoresis. Sardaine consists of 34 amino acid residues, and its sequence is: Pro-Arg-Arg-Arg-Arg-Arg-Ser(Ala)-Ser-Arg-Pro-Val-Arg-Arg-Arg-Arg-Arg-Tyr -Arg- Arg-Ser-Thr-Ala-Ala-Arg-Arg-Arg-Arg-Arg-Val-Val-Arg-Arg-Arg-Arg. At position 7, serine (sardaine Z1) is partially replaced by alanine (sardaine Z2). The ion spray mass spectrum shows that sardaines Z1 and Z2 have molecular masses of 4,612.49 and 4,596.09 Da, respectively. The sequence of sardaine Z1 is 100% identical with that of thynnine Z2 from tuna fish (both fish belong to Scombridae, Perciformes).

Amino Acid Sequence↗

Molecular evolution at subzero temperatures: mitochondrial and nuclear phylogenies of fishes from Antarctica (suborder Notothenioidei), and the evolution of antifreeze glycopeptides.

Most fishes of the perciform suborder Notothenioidei are endemic to the subzero marine waters of Antarctica. A number of remarkable physiological attributes allow them to inhabit this extreme environment; for example, the blood of almost all notothenioid species contains antifreeze glycopeptides. To establish a molecular phylogenetic hypothesis for these fishes, DNA sequences from two mitochondrial genes, portions of the 12S and 16S ribosomal genes (928 base pairs [bp]), were determined for 18 species. These belong to 15 genera in five families of the suborder. The DNA data suggest that two of these families are unnatural groups and consequently that the classification and phylogeny of this suborder is in need of revision. In terms of DNA variation, the Bovichtidae are a distantly related sister group to the other families of the suborder that includes the icefishes, the only vertebrates without hemoglobin. The fishes of the suborder (except the Bovichtidae) seem to have speciated rapidly, forming an adaptive radiation in the Antarctic waters. A phylogenetic analysis of published hemoglobin amino acid sequences for other notothenioid fishes supports these results from mtDNA. On the basis of molecular phylogeny, the evolution of antifreeze glycopeptides was studied. The age of the radiation of notothenioid fishes had been estimated to be at least 38 Mya. However, the level of mtDNA variation detected in notothenioid fishes appears to be too low to agree with this date of origin and might instead suggest a younger age (10-15 Mya). Alternatively, the low level of detected mtDNA variation would agree with the traditional old-age estimate if an extremely slow rate of mtDNA evolution is postulated for this group. This slow-rate hypothesis, if true, could be explained by decreased metabolic rates slowing down the tempo of molecular evolution.

Animals↗

Changes in plasma gonadotropin II and sex steroid hormones, and sperm production of striped bass after treatment with controlled-release gonadotropin-releasing hormone agonist-delivery systems.

Spermiating striped bass, a perciform fish, were treated with two controlled-release gonadotropin-releasing hormone agonist (GnRHa)-delivery systems, and the resulting changes in plasma gonadotropin II (GTH II), testosterone (T), 11-ketotestosterone (11-KT), 17,20beta-dihydroxy-4-pregnen-3-one (17,20beta-P), and 17,20beta,21-trihydroxy-4-pregnen-3-one (17,20beta,21-P) were correlated with changes in milt production and sperm density. GnRHa-delivery systems induced a sustained elevation of plasma GnRHa and GTH II for 14 days. Plasma T levels were unchanged after GnRHa treatment, while 11-KT levels increased significantly. Plasma 17,20beta-P also increased after GnRHa treatment and remained elevated compared to levels in controls, while plasma 17,20beta,21-P levels were unaffected. Both GnRHa-delivery systems induced many-fold increases in total expressible milt, lasting throughout the 14 days of the study. Sperm density decreased 2 days after GnRHa treatment, with a subsequent increase by Day 7. This study demonstrates that GnRHa-delivery systems induce a sustained elevation of plasma GTH II levels in striped bass, resulting in a long-term enhancement of milt production. The endocrine changes observed suggest that 11-KT and 17,20beta-P, but not 17,20beta,21 -P, are mediating the effects of GTH II on spermiation.

Animals↗