RESUMO
The goal of this study was to design genus-specific primers for rapid evaluation of the most abundant bacterial genera identified using amplicon-based sequencing of the 16S rRNA gene in fish-related samples and surrounding water. Efficient genus-specific primers were designed for 11 bacterial genera including Alkalimarinus, Colwellia, Enterovibrio, Marinomonas, Massilia, Oleispira, Phaeobacter, Photobacterium, Polarbacerium, Pseudomonas, and Psychrobium. The specificity of the primers was confirmed by the phylogeny of the sequenced polymerase chain reaction (PCR) amplicons that indicated primers were genus-specific except in the case of Colwellia and Phaeobacter. Copy number of the 16S rRNA gene obtained by quantitative PCR using genus-specific primers and the relative abundance obtained by 16S rRNA gene sequencing using universal primers were well correlated for the five analyzed abundant bacterial genera. Low correlations between quantitative PCR and 16S rRNA gene sequencing for Pseudomonas were explained by the higher coverage of known Pseudomonas species by the designed genus-specific primers than the universal primers used in 16S rRNA gene sequencing. The designed genus-specific primers are proposed as rapid and cost-effective tools to evaluate the most abundant bacterial genera in fish-related or potentially other metagenomics samples.
Assuntos
Metagenômica , Rhodobacteraceae , Animais , Primers do DNA/genética , Peixes , Larva , Metagenoma , Pseudomonas/genética , RNA Ribossômico 16S/genéticaRESUMO
Prostate cancer (PCa) is one of the most prevalent male tumours. Stanniocalcin-1 (STC1) is a glycoprotein and, although the role of STC1 in human cancer is poorly understood, it is suggested to be involved in the development and progression of different neoplasms. This study investigated the protein expression profile of STC1 in PCa and benign prostatic hyperplasia (BPH) samples and STC1 signalling during cell proliferation and cell death in vitro using cell lines. We found higher levels of STC1 in PCa when compared to BPH tissue and that STC1 inhibited forskolin stimulation of cAMP in PC-3 cells. A monoclonal antibody against STC1 was effective in reducing cell proliferation, in promoting cell cycle arrest, and in increasing apoptosis in the same cells. Since STC1 acts as a regulator of prostatic tissue signalling, we suggest that this protein is a novel candidate biomarker for prostate tumour clinical progression and a potential therapeutic target.
Assuntos
Biomarcadores Tumorais/metabolismo , Colforsina/farmacologia , Glicoproteínas/genética , Glicoproteínas/metabolismo , Hiperplasia Prostática/metabolismo , Neoplasias da Próstata/metabolismo , Apoptose , Pontos de Checagem do Ciclo Celular , Linhagem Celular Tumoral , Proliferação de Células , Progressão da Doença , Humanos , Masculino , Células PC-3 , Hiperplasia Prostática/genética , Neoplasias da Próstata/genética , Regulação para CimaRESUMO
The neuropeptide galanin (Gal) is a putative factor regulating puberty onset and reproduction through its actions on the pituitary. The present study investigated the pituitary responsiveness to galanin and the patterns of galanin receptors (Galrs) expression throughout the reproductive cycle of two years old male European sea bass (Dicentrarchus labrax), an important aquaculture species. Quantitative analysis of pituitary and hypothalamus transcript expression of four galr subtypes revealed differential regulation according to the testicular developmental stage, with an overall decrease in expression from the immature stage to the mid-recrudescence stage. Incubation of pituitary cells with mammalian 1-29Gal peptide induced significant changes in cAMP concentration, with sensitivities that varied according to the testicular development stages. Furthermore 1-29Gal was able to stimulate both follicle stimulating hormone (Fsh) and luteinizing hormone (Lh) release from pituitary cell suspensions. The magnitude of the effects and effective concentrations varied according to reproductive stage, with generalized induction of Fsh and Lh release in animals sampled in January (full spermiation). The differential expression of galrs in pituitary and hypothalamus across the reproductive season, together with the differential effects of Gal on gonadotropins release in vitro strongly suggests the involvement of the galaninergic system in the regulation the hypothalamus-pituitary-gonad axis of male sea bass. This is to our knowledge the first clear evidence for the involvement of galanin in the regulation of reproduction in non-mammalian vertebrates.
Assuntos
Bass/fisiologia , Galanina/farmacologia , Hipófise/metabolismo , Reprodução/efeitos dos fármacos , Animais , Bass/genética , Colforsina/farmacologia , AMP Cíclico/biossíntese , Hormônio Foliculoestimulante/metabolismo , Gametogênese/efeitos dos fármacos , Perfilação da Expressão Gênica , Gonadotropinas/farmacologia , Hipotálamo/efeitos dos fármacos , Hipotálamo/metabolismo , Hormônio Luteinizante/metabolismo , Masculino , Hipófise/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores de Galanina/genética , Receptores de Galanina/metabolismo , Reprodução/fisiologiaRESUMO
Two hundred and seven individuals (103 females and 104 males) of bluemouth Helicolenus dactylopterus (Scorpaeniformes, Sebastidae), a commercially important deep-water species with an unusual reproductive strategy, from the eastern Atlantic Ocean ranging from 13·9 to 37·5 cm total length (LT ) were analysed from September 2011 to October 2012. The analysis included gonad maturity phases and blood-plasma levels of oestradiol-17ß (E2 ), 11-ketotestosterone (11-KT) and 17,20ß-dihydroxypregn-4-en-3-one (17,20ß-P). Results confirmed the existence of an annual reproductive cycle with asynchrony between females and males and a spawning season from January to May. A pronounced peak in 17,20ß-P in October for both sexes was associated with possible mating behaviour and recent copula. Levels of E2 increased preceding the elevation of the gonado-somatic index during ovarian growth and were lower during regression and regeneration. The frequency distribution of oocyte-embryonic stages and variation of hormone levels suggest the existence of daily rhythms. Fertilization was detected between 2000-0000 and 0800-1200 h and spawning took place throughout the day peaking between 2000 and 0000 h. The cyclic pattern of sex steroids and ovarian recruitment provides a new insight into the reproductive strategy of this species.
Assuntos
Hormônios Esteroides Gonadais/fisiologia , Perciformes/fisiologia , Animais , Oceano Atlântico , Feminino , Hormônios Esteroides Gonadais/sangue , Masculino , Oócitos/crescimento & desenvolvimento , Reprodução/fisiologia , Estações do AnoRESUMO
The present study shows that the olfactory potency of intestinal and bile fluids taken from dominant male chameleon cichlids Australoheros facetus is greater than those from subordinate males. Thus, dominant status may be communicated by odorants released in the intestinal fluid and bile acids may contribute towards this.
Assuntos
Líquidos Corporais/química , Ciclídeos/fisiologia , Fezes/química , Percepção Olfatória/fisiologia , Predomínio Social , Animais , Ácidos e Sais Biliares/química , Ácidos e Sais Biliares/metabolismo , Intestinos , MasculinoRESUMO
The copepod Calanus glacialis plays a key role in the Arctic pelagic ecosystem. Despite its ecological importance and ongoing climate changes, limited knowledge at the genomic level has hindered the understanding of the molecular processes underlying environmental stress responses and ecological adaptation. Transcriptome data was generated from an experiment with C. glacialis copepodite (CV) subjected to five different temperatures. We obtained a total of 512,352 high-quality 454 pyrosequencing reads, which were assembled into 55,562 contigs distributed in 128 KEGG pathways. Functional analysis revealed numerous genes related to diverse biological functions and processes, including members of all major conserved signaling pathways. Comparative analysis of acclimated individuals to experimental temperatures has provided information about gene variations observed in several pathways (e.g. genes involved in energy, lipid and amino acid metabolism were shown to be down-regulated with increasing temperatures). These mRNA sequence resources will facilitate further studies on genomics and physiology-driven molecular processes in C. glacialis and related species.
Assuntos
Copépodes/genética , Regulação da Expressão Gênica/fisiologia , Temperatura , Transcriptoma , Animais , Biologia ComputacionalRESUMO
This study tested whether differences in sensitivity between the upper and lower olfactory epithelia of Solea senegalensis are associated with different odorant receptors and transduction pathways, using the electro-olfactogram. Receptor mechanisms were assessed by cross-adaptation with amino acids (L-cysteine, L-phenylalanine and 1-methyl-L-tryptophan) and bile acids (taurocholic acid and cholic acid). This suggested that relatively specific receptors exist for 1-methyl-L-tryptophan and L-phenylalanine (food-related odorants) in the lower epithelium, and for taurocholic acid (conspecific-derived odorant) in the upper. Inhibition by U73122 [a phospholipase C (PLC) inhibitor] suggested that olfactory responses to amino acids were mediated mostly, but not entirely, by PLC-mediated transduction (IC50 ; 15-55 nM), whereas bile acid responses were mediated by both PLC and adenylate cyclase-cyclic adenosine monophosphate (AC-cAMP) (using SQ-22536; an AC inhibitor). Simultaneous application of both drugs rarely inhibited responses completely, suggesting possible involvement of non-PLC and non-AC mediated mechanisms. For aromatic amino acids and bile acids, there were differences in the contribution of each transduction pathway (PLC, AC and non-PLC and non-AC) between the two epithelia. These results suggest that differences in sensitivity of the two epithelia are associated with differences in odorant receptors and transduction mechanisms.
Assuntos
Linguados/fisiologia , Receptores Odorantes/fisiologia , Transdução de Sinais/fisiologia , Olfato/fisiologia , Adenina/análogos & derivados , Adenina/farmacologia , Inibidores de Adenilil Ciclases , Adenilil Ciclases/metabolismo , Aminoácidos/metabolismo , Animais , Ácidos e Sais Biliares/metabolismo , AMP Cíclico/metabolismo , Epitélio/fisiologia , Estrenos/farmacologia , Pirrolidinonas/farmacologia , Fosfolipases Tipo C/antagonistas & inibidores , Fosfolipases Tipo C/metabolismoRESUMO
Sperm undergo maturation acquiring progressive motility and the ability to fertilize oocytes through exposure to the components of the epididymal fluid (EF). Although the establishment of a calcium (Ca(2+)) gradient along the epididymis has been described, its direct effects on epididymal function remain poorly explored. Regucalcin (RGN) is a Ca(2+)-binding protein, regulating the activity of Ca(2+)-channels and Ca(2+)-ATPase, for which a role in male reproductive function has been suggested. This study aimed at comparing the morphology, assessed by histological analysis, and function of epididymis, by analysis of sperm parameters, antioxidant potential and Ca(2+) fluxes, between transgenic rats overexpressing RGN (Tg-RGN) and their wild-type littermates. Tg-RGN animals displayed an altered morphology of epididymis and lower sperm counts and motility. Tissue incubation with (45)Ca(2+) showed also that epididymis of Tg-RGN displayed a diminished rate of Ca(2+)-influx, indicating unbalanced Ca(2+) concentrations in the epididymal lumen. Sperm viability and the frequency of normal sperm, determined by the one-step eosin-nigrosin staining technique and the Diff-Quik staining method, respectively, were higher in Tg-RGN. Moreover, sperm of Tg-RGN rats showed a diminished incidence of tail defects. Western blot analysis demonstrated the presence of RGN in EF as well as its higher expression in the corpus region. The results presented herein demonstrated the importance of maintaining Ca(2+)-levels in the epididymal lumen and suggest a role for RGN in sperm maturation. Overall, a new insight into the molecular mechanisms driving epididymal sperm maturation was obtained, which could be relevant to development of better approaches in male infertility treatment and contraception.
Assuntos
Proteínas de Ligação ao Cálcio/genética , Cálcio/metabolismo , Epididimo/fisiologia , Peptídeos e Proteínas de Sinalização Intracelular/genética , Maturação do Esperma/genética , Motilidade dos Espermatozoides/genética , Espermatozoides/metabolismo , Animais , Radioisótopos de Cálcio , Proteínas de Ligação ao Cálcio/metabolismo , Hidrolases de Éster Carboxílico , Sobrevivência Celular , Epididimo/ultraestrutura , Regulação da Expressão Gênica , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Transporte de Íons , Masculino , Ratos , Ratos Sprague-Dawley , Ratos Transgênicos , Transdução de Sinais , Contagem de Espermatozoides , Espermatozoides/citologiaRESUMO
The androgen receptor (AR) is a ligand-activated transcription factor member of the nuclear receptor superfamily. The existence of alternatively spliced variants is well recognised for several members of this superfamily, most of them having functional importance. For example, several testicular oestrogen receptor variants have been suggested to play a role in the regulation of spermatogenesis. However, information on AR variants is mostly related to cancer and androgen insensitivity syndrome (AIS) cases. The objective of this study was to investigate the expression of AR variants in the testis from humans and other vertebrates. Four AR variants [ARΔ2(Stop) , ARΔ2(23Stop) , ARΔ3 and ARΔ4(120)] were identified in human testis. ARΔ2(Stop) and ARΔ3, with exon 2 or 3 deleted, respectively, were also expressed in human liver, lung, kidney and heart. In addition, ARΔ2(Stop) was expressed in rat and gilthead seabream testis, while an ARΔ3 was detected in African clawed frog testis. This is the first report revealing the existence of AR variants in the testis of evolutionarily distant vertebrate species and in nonpathological tissues. These data suggest the functional importance of these novel AR forms and demonstrate a complexity in AR signalling that is not exclusive of pathological conditions.
Assuntos
Filogenia , Ratos/genética , Receptores Androgênicos/genética , Dourada/genética , Testículo/fisiologia , Xenopus laevis/genética , Adulto , Processamento Alternativo/genética , Animais , Variação Genética , Coração/fisiologia , Humanos , Rim/fisiologia , Fígado/fisiologia , Pulmão/fisiologia , Masculino , Especificidade de Órgãos , Fisiologia Comparada , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Especificidade da Espécie , Transcrição Gênica/genéticaRESUMO
The reproductive cycle in teleosts is timed to guarantee that eggs hatch in the right place at the right time, with environmental factors playing important roles in entraining and controlling the entire process. The effects of some environmental factors, like temperature and photoperiod, are now well understood. There are only a few studies regarding the effects of hydrostatic pressure (HP) on the reproductive cycle, in spite of its importance as a ubiquitous factor in all biological environments and affecting all living organisms. Hydrostatic pressure is of particular importance in fish because they can also experience rapid and cyclic changes in HP due to vertical movements in the water column. The aim of the present research was to investigate the effects of vertical migrations on the reproductive steroids of maturing female flounder. After a 14 day exposure to cyclic hydrostatic pressure (with a period of 12.4h and with a maximum peak of 800 kPa of absolute hydrostatic pressure), fish showed significantly lower plasmatic concentrations of "5ß,3α" steroids, metabolites of the putative maturation-inducing steroid in flounder (17α,20ß-dihydroxy-4-pregnen-3-one). Results indicate that environmentally realistic cyclic changes of hydrostatic pressure can influence the metabolism of reproductive steroids. This suggests a physiological role of tidally-associated vertical migrations, affecting oocyte maturation and retarding the reproductive cycle in this species until the spawning ground is attained.
Assuntos
Estradiol/sangue , Linguado/fisiologia , Testosterona/sangue , Animais , Feminino , Linguado/sangue , Pressão Hidrostática , Hidroxiprogesteronas/sangue , Hidroxiprogesteronas/urina , Ovário/citologia , Ovário/fisiologia , Progestinas/sangue , VitelogêneseRESUMO
The pituitary hormone prolactin is a pleiotropic endocrine factor that plays a major role in the regulation of ion balance in fish, with demonstrated actions mainly in the gills and kidney. The role of prolactin in intestinal ion transport remains little studied. In marine fish, which have high drinking rates, epithelial bicarbonate secretion in the intestine produces luminal carbonate aggregates believed to play a key role in water and ion homeostasis. The present study was designed to establish the putative role of prolactin in the regulation of intestinal bicarbonate secretion in a marine fish. Basolateral addition of prolactin to the anterior intestine of sea bream mounted in Ussing chambers caused a rapid (<20 min) decrease of bicarbonate secretion measured by pH-stat. A clear inhibitory dose-response curve was obtained, with a maximal inhibition of 60-65% of basal bicarbonate secretion. The threshold concentration of prolactin for a significant effect on bicarbonate secretion was 10 ng ml(-1), which is comparable with putative plasma levels in seawater fish. The effect of prolactin on apical bicarbonate secretion was independent of the generation route for bicarbonate, as shown in a preparation devoid of basolateral HCO(3)(-)/CO(2) buffer. Specific inhibitors of JAK2 (AG-490, 50 µmol l(-1)), PI3K (LY-294002, 75 µmol l(-1)) or MEK (U-012610, 10 µmol l(-1)) caused a 50-70% reduction in the effect of prolactin on bicarbonate secretion, and demonstrated the involvement of prolactin receptors. In addition to rapid effects, prolactin has actions at the genomic level. Incubation of intestinal explants of anterior intestine of the sea bream in vitro for 3 h demonstrated a specific effect of prolactin on the expression of the Slc4a4A Na(+)-HCO(3)(-) co-transporter, but not on the Slc26a6A or Slc26a3B Cl(-)/HCO(3)(-) exchanger. We propose a new role for prolactin in the regulation of bicarbonate secretion, an essential function for ion/water homeostasis in the intestine of marine fish.
Assuntos
Bicarbonatos/metabolismo , Mucosa Intestinal/metabolismo , Prolactina/fisiologia , Dourada/metabolismo , Animais , Antiportadores de Cloreto-Bicarbonato/biossíntese , Cromonas/farmacologia , Transporte de Íons , Janus Quinase 2/antagonistas & inibidores , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Morfolinas/farmacologia , Inibidores de Fosfoinositídeo-3 Quinase , Prolactina/administração & dosagem , Prolactina/farmacologia , Transdução de Sinais , Simportadores de Sódio-Bicarbonato/biossíntese , ATPase Trocadora de Sódio-Potássio/antagonistas & inibidores , Tirfostinas/farmacologia , Equilíbrio HidroeletrolíticoRESUMO
Estrogen actions are mainly mediated by specific nuclear estrogen receptors (ERs), for which different genes and a diversity of transcript variants have been identified, mainly in mammals. In this study, we investigated the presence of ER splice variants in the teleost fish gilthead sea bream (Sparus auratus), by comparison with the genomic organization of the related species Takifugu rubripes. Two exon2-deleted ERα transcript variants were isolated from liver cDNA of estradiol-treated fish. The ΔE2 variant lacks ERα exon 2, generating a premature termination codon and a putative C-terminal truncated receptor, while the ΔE2,3* variant contains an in-frame deletion of exon 2 and part of exon 3 and codes for a putative ERα protein variant lacking most of the DNA-binding domain. Both variants were expressed at very low levels in several female and male sea bream tissues, and their expression was highly inducible in liver by estradiol-17ß treatment with a strong positive correlation with the typical wild-type (wt) ERα response in this tissue. These findings identify novel estrogen responsive splice variants of fish ERα, and provide the basis for future studies to investigate possible modulation of wt-ER actions by splice variants.
Assuntos
Receptor alfa de Estrogênio/metabolismo , Estrogênios/metabolismo , Dourada/metabolismo , Processamento Alternativo/genética , Animais , Receptor alfa de Estrogênio/biossíntese , Receptor alfa de Estrogênio/genética , Éxons , Feminino , Fígado/metabolismo , Masculino , Isoformas de Proteínas/biossíntese , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Deleção de Sequência/genética , Takifugu/metabolismo , Distribuição TecidualRESUMO
The role of sex steroids in the modulation of fish immune responses has received little attention. Previous studies have demonstrated that 17ß-estradiol (E(2)) is able to alter the response of gilthead seabream leukocytes to infectious agents. We have used suppression subtractive hybridization to identify genes upregulated by E(2) (50 ng/ml) in macrophage cultures from gilthead seabream. We isolated 393 up-regulated cDNA fragments that led to the identification of 162 candidate estrogen-responsive genes. Functional analyses revealed the presence of several enriched immune processes and molecular pathways. The E(2) up-regulation of some immune-relevant genes was further confirmed by real time RT-PCR. Bioinformatics analysis revealed the ability of E(2) to orchestrate profound alterations in the macrophage expression profile, especially immune-related processes and pathways. This is the first report on E(2)-dependent modifications of fish macrophage transcriptome and lends weight to a suggested role for estrogen in the immune system, the possible significance of which is discussed.
Assuntos
Estrogênios/farmacologia , Perfilação da Expressão Gênica , Macrófagos/efeitos dos fármacos , Dourada/genética , Actinas/genética , Actinas/metabolismo , Animais , Células Cultivadas , Simulação por Computador , Etiquetas de Sequências Expressas , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Expressão Gênica/efeitos dos fármacos , Genes MHC Classe I , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Metaloproteinase 13 da Matriz/genética , Metaloproteinase 13 da Matriz/metabolismo , Receptores Imunológicos/genética , Receptores Imunológicos/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Transcrição Gênica/efeitos dos fármacosRESUMO
Proopiomelanocorticotrophin (POMC) in vertebrates is produced in the pituitary gland and undergoes post-translational processing to give rise to a range of biologically active peptides. Teleosts possess 2-3 different POMC transcripts which have been proposed to have originated from a whole or partial genome duplication. In the present study 2 transcripts of gilthead sea bream POMC (sbPOMC-α1 and α2) were cloned and characterised. sbPOMC-α1 is expressed principally in the melanotroph cells of the pars intermedia (PI) and sbPOMC-α2 is expressed in the corticotroph cells of the rostral pars distalis and probably also in the PI. The 2 sbPOMC transcripts have a differential tissue distribution in extra-pituitary sites. An appraisal of POMC evolution indicates sbPOMCs belong to one of the two main clades that exist in teleosts and that overall a non conservative process of gene loss occurred in this infraclass.
Assuntos
Proteínas de Peixes/genética , Duplicação Gênica , Pró-Opiomelanocortina/genética , Dourada/genética , Sequência de Aminoácidos , Animais , Encéfalo/metabolismo , Clonagem Molecular , Proteínas de Peixes/metabolismo , Dados de Sequência Molecular , Filogenia , Hipófise/metabolismo , Pró-Opiomelanocortina/metabolismo , Dourada/metabolismo , Alinhamento de Sequência , Análise de Sequência de ProteínaRESUMO
Teleosts have high olfactory sensitivity to bile salts. To assess whether this phenomenon is involved in intra-specific chemical communication alone, or is part of a more ;broad range' sensitivity to bile salts produced by heterospecifics, we investigated possible differences in the odour of bile between the sexes and among different species - the eel (Anguilla anguilla), goldfish (Carassius auratus) and Mozambique tilapia (Oreochromis mossambicus) - using the electro-olfactogram (EOG). We also identified the main bile constituents by liquid chromatography and mass spectrometry. There were marked differences in olfactory response of the eel to thin-layer chromatography fractions of bile from both sexes, and mature and immature conspecifics. Smaller differences were seen in the potency of fractions of bile from male and female goldfish and tilapia. Eels, goldfish and tilapia demonstrated similar olfactory sensitivity to bile from a range of different species, with no apparent correlation between the olfactory potency of bile and a phylogenetic closeness and/or similarity of diet of the donor to the receiver. The three species were able to detect odorants in thin-layer chromatography fractions of heterospecific bile even in the absence of activity in conspecific bile. Eels, goldfish and tilapia responded to both sulphated C(27) bile salts (5beta-scymnol-sulphate and 5alpha-cyprinol sulphate) and to taurine-conjugated C(24) bile salts (taurochenodeoxycholic acid, taurolithocholic acid and taurocholic acid), irrespective of whether these bile salts were present in conspecific bile. Together, these results suggest that teleosts have a broad-range olfactory sensitivity to bile salts, with potential roles in both intra-specific chemical communication and in inter-specific interactions.
Assuntos
Ácidos e Sais Biliares , Bile , Enguias/fisiologia , Carpa Dourada/fisiologia , Olfato , Tilápia/fisiologia , Animais , Feminino , Masculino , Caracteres Sexuais , Especificidade da EspécieRESUMO
The effect of nest aggregation in courtship behaviour was tested experimentally in an ecologically constrained, sex-role reversed population of the peacock blenny Salaria pavo. Mixed sex groups of eight males and eight females were tested in experimental tanks, containing eight potential nests either aggregated or dispersed. In the aggregated treatment, males spent more time inside their nests and monopolized other potential nests, causing a female-biased operational sex ratio (OSR). In the aggregated treatment, females also expressed more courtship behaviour. The results in general support the prediction that the aggregation of nests promotes male monopolization of potential nests, resulting in fewer nest-holding males and therefore a female-biased OSR that leads to the reversal of sex roles.
Assuntos
Comportamento de Nidação/fisiologia , Perciformes/fisiologia , Comportamento Sexual Animal/fisiologia , Comportamento Agonístico/fisiologia , Animais , Feminino , MasculinoRESUMO
Calcium mobilization from internal stores, such as scales, induced by 17beta-estradiol during sexual maturation in salmonids is well documented. This calcium mobilization from scales is proposed to be mediated by the estrogen receptor (ER). However, the ER subtypes involved and signaling mechanisms responsible for this effect remain to be fully characterized. In the present study, we have localized ERalpha, ERbetaa and ERbetab proteins in juvenile and adult sea bream (Sparus auratus) and Mozambique tilapia (Oreochromis mossambicus) scales by immunohistochemistry with sea bream ER subtype specific antibodies. The three ERs were detected in isolated or small groups of round cells, in the basal layer of the scales of both juvenile and adult fish and the localization and signal intensity varied with the species and age of the animals. The ERs may be co-localized in cells of the scale posterior region that expressed tartrate-resistant acid phosphatase (TRAP), a marker for osteoclasts. These results suggest that the calcium mobilizing action of 17beta-estradiol on fish scales is via its direct action on ERs localized in osteoclasts.
Assuntos
Imuno-Histoquímica/métodos , Receptores de Estrogênio/metabolismo , Pele/metabolismo , Fosfatase Ácida/metabolismo , Animais , Cálcio/metabolismo , Estradiol/farmacologia , Receptor beta de Estrogênio/metabolismo , Isoenzimas/metabolismo , Osteoclastos/efeitos dos fármacos , Osteoclastos/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Dourada , Fosfatase Ácida Resistente a Tartarato , TilápiaRESUMO
Gilthead sea bream (Sparus auratus L.) were fed a vitamin D-deficient diet for 22 weeks. Growth rate, whole body mineral pools and calcium balance were determined. Plasma parathyroid hormone-related protein (PTHrP) and calcitriol levels were assessed. Expression of mRNA for pthrp and pth1r was quantified in gills and hypophysis. Fish on vitamin D-deficient diet (D- fish) showed reduced growth and lower calcium turnover (calcium influx, efflux and accumulation rates decreased) and unaltered plasma calcium levels. Plasma calcitriol levels became undetectable, PTHrP levels decreased in the D- fish. In controls, a significant increase in plasma PTHrP level over time was seen, i.e. it increased with body mass. Relationships were found between plasma PTHrP and the whole body pools of calcium, phosphorus and magnesium, indicative of a role for PTHrP in bone development. Expression of pthrp and pth1r mRNA was down-regulated in the hypophysis of D-fish, whereas in gill tissue, pthrp and pth1r mRNA were up-regulated. We conclude that lower pthrp mRNA expression and plasma values in D- fish reflect lower turnover of PTHrP under conditions of hampered growth; up-regulation of pthrp mRNA in gills indicate compensatory paracrine activity of PTHrP during calcitriol deficiency to guarantee well-regulated branchial calcium uptake. This is the first report to document a relation between PTHrP and calcitriol in fish.
Assuntos
Cálcio/metabolismo , Regulação da Expressão Gênica , Proteína Relacionada ao Hormônio Paratireóideo/sangue , Dourada/metabolismo , Deficiência de Vitamina D/metabolismo , Ração Animal , Animais , Calcitriol/sangue , Brânquias/metabolismo , Proteína Relacionada ao Hormônio Paratireóideo/genética , Hipófise/metabolismo , RNA Mensageiro/análise , Receptor Tipo 1 de Hormônio Paratireóideo/genética , Receptor Tipo 1 de Hormônio Paratireóideo/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Dourada/crescimento & desenvolvimentoRESUMO
Testicular development and plasma levels of sex steroids (11-ketotestosterone (11-KT), testosterone (T) and 17,20beta-dihydroxy-4-pregnen-3-one (17,20beta-P)) were investigated for the first time in cultured male Senegalese sole Solea senegalensis. The germ cell dynamics and gonadosomatic index (IG) were monitored. Based on the relative abundance of the different types of germ cells present, the spermatogenetic cycle was divided into five stages: early (I; spermatogonia (SPG)), mid (II; SPG, spermatocytes (SPC) and spermatids (SPD)), and late spermatogenesis (III; SPC, SPD, and spermatozoa (SPZ)), functional maturation (IV; SPD and SPZ), and recovery (V; SPD, SPZ, and SPG). During summer, fish had stage I and V testes and the lowest values in plasma levels of sex steroids and IG. Testicular recrudescence seemed to begin in autumn, as denoted by the first increase in IG and in the levels of 11-KT and T, and the appearance of testes at stage II and III. During winter, the levels of 11-KT and T peaked and soon began to decrease, the IG slightly declined and the proportion of running males (RM) gradually increased. In spring, levels of 11-KT and T continued to decline, the IG slightly increased and the proportion of RM peaked concomitantly with the occurrence of stage IV testes. Plasma levels of 17,20beta-P did not change significantly throughout testicular development. Transformation of SPD into SPZ followed a group-synchronous fashion, a phenomenon which parallels asynchronous oocyte development reported in females. This mechanism would be consistent with the observed small quantity of sperm that can be manually stripped at any one time and other aspects of S. senegalensis reproductive biology.
Assuntos
Linguados/sangue , Linguados/crescimento & desenvolvimento , Hormônios Esteroides Gonadais/sangue , Espermatogênese/fisiologia , Testículo/crescimento & desenvolvimento , Animais , Aquicultura , Feminino , Hidroxiprogesteronas/sangue , Masculino , Reprodução/fisiologia , Testículo/citologia , Testosterona/análogos & derivados , Testosterona/sangueRESUMO
Whole animal studies have indicated that Ca(2+) uptake by the gastrointestinal tract is regulated by the action of parathyroid hormone-related peptide (PTHrP) in teleost fish. We have characterised PTH receptors (PTHR) in piscine enterocytes and established, by using amino-terminal PTHrP peptides, the amino acid residues important for receptor activation and for stabilising the ligand/receptor complex. Ligand binding of (125)I-(1-35(tyr)) PTHrP to the membrane fraction of isolated sea bream enterocytes revealed the existence of a single saturable high-affinity receptor (K (D)=2.59 nM; B (max)=71 fmol/mg protein). Reverse transcription/polymerase chain reaction with specific primers for sea bream PTH1R and PTH3R confirmed the mRNA expression of only the later receptor. Fugu (1-34)PTHrP increased cAMP levels in enterocytes but had no effect on total inositol phosphate accumulation. The amino-terminal peptides (2-34)PTHrP, (3-34)PTHrP and (7-34)PTHrP bound efficiently to the receptor but were severely defective in stimulating cAMP in enterocyte cells indicating that the first six residues of piscine (1-34)PTHrP, although not important for receptor binding, are essential for activation of the adenylate cyclase/phosphokinase A (AC-PKA)-receptor-coupled intracellular signalling pathway. Therefore, PTHrP in teleosts acts on the gastrointestinal tract through PTH3R and the AC-PKA intracellular signalling pathway and might regulate Ca(2+) uptake at this site. Ligand-receptor binding and activity throughout the vertebrates appears to be allocated to the same amino acid residues of the amino-terminal domain of the PTHrP molecule.