Exposure to waterborne 4-tert-octylphenol induces vitellogenin synthesis and disrupts testis morphology in the South American freshwater fish Cichlasoma dimerus (Teleostei, Perciformes).

Exposure to environmental pollutants may disrupt endocrine functions and cause reproductive effects in human and wildlife populations. Various groups of chemicals have estrogen-like effects, including degradation products of alkylphenol polyethoxylates, such as 4-tert-octylphenol (OP). Laboratory studies have shown that exposure of male fish to xenoestrogens results in induction of circulating vitellogenin (Vtg), inhibition of testicular growth, testis abnormalities and formation of intersex gonads. In this study, the impact of the exposure to waterborne OP on reproductive aspects in the South American cichlid fish Cichlasoma dimerus was evaluated using qualitative changes in the levels of Vtg in plasma and surface mucus and histological alterations in the liver and gonads as endpoints. Adult males and females were exposed to OP via immersion during 60 days in aquaria under semi-static conditions, water changes being made every 84 h. Treatment groups were: control (ethanol 0.005%), OP 30, 150 and 300 microg/L. Using Western and Dot blot analysis, Vtg was detected in plasma and mucus of control and treated females and treated males, while no Vtg was observed in samples from control males. Morphological changes in the hepatocytes due to the accumulation of Vtg were observed in OP-exposed males. Impairment of testicular structure became apparent in males treated with the highest OP concentrations. The most salient pathological change was the alteration of lobular organization with increased testicular fibrosis and progressive disruption of spermatogenesis. No major changes were observed in ovarian architecture. Our results indicate that detection of Vtg in surface mucus may be a sensitive and non-invasive biomarker of the endocrine disrupting effects of environmental estrogens, resulting in a useful method for field monitoring.

Brain-derived neurotrophic factor (BDNF)-like immunoreactivity localization in the retina and brain of Cichlasoma dimerus (Teleostei, Perciformes).

Brain-derived neurotrophic factor (BDNF) is a neurotrophin involved in the development and maintenance of vertebrate nervous systems. Although there were several studies in classical animal models, scarce information for fish was available. The main purpose of this study was to analyze the distribution of BDNF in the brain and retina of the cichlid fish Cichlasoma dimerus. By immunohistochemistry we detected BDNF-like immunoreactive cells in the cytoplasm and the nuclei of the ganglion cell layer and the inner nuclear layer of the retina. In the optic tectum, BDNF-like immunoreactivity was detected in the nucleus of neurons of the stratum periventriculare and the stratum marginale and in neurons of the intermediate layers. In the hypothalamus we found BDNF-like immunoreactivity mainly in the cytoplasm of the nucleus lateralis tuberis and the nucleus of the lateral recess. To confirm the nuclear and cytoplasm localization of BDNF we performed subcellular fractionation, followed by Western blot, detecting a 39 kDa immunoreactive-band corresponding to a possible precursor form of BDNF in both fractions. BDNF-like immunoreactivity was distributed in areas related with photoreception (retina), the integration center of retinal projections (optic tectum) and the control center of background and stress adaptation (hypothalamus). These results provide baseline anatomical information for future research about the role of neurotrophins in the adult fish central nervous system.

Localization of Neuropeptide Y-like immunoreactive structures in the brain of the pejerrey, Odontesthes bonariensis (Teleostei, Atheriniformes).

The aim of this study was to determine the distribution of Neuropeptide-Y (NPY) immunoreactive neurons and fibres in the brain and pituitary of Odontesthes bonariensis by immunohistochemical methods. A wide distribution of immunoreactive NPY (ir-NPY) cells and fibres in the forebrain and midbrain was observed. A prominent ir-NPY nucleus was found in the ventral telencephalon and other ir-NPY cells groups were recognized at the dorso-medial telencephalon. The diencephalon showed ir-NPY cells in the Nucleus entopeduncularis, the Nucleus preopticus periventricularis and in the Nucleus lateralis tuberis. Ir-NPY fibres were conspicuous in the preoptic region and the hypothalamus. There were also numerous ir-NPY fibres at the epithalamic level running ventrally to the hypothalamus and the pituitary stalk. At the rhomboencephalic level, the ir-NPY neurons were observed in the Locus coeruleus. Double-labelled immunostaining showed a close association between ir-NPY fibres that reach the adenohypophysis and growth hormone (GH)- and gonadotropin (GtH)-expressing cells. Although our results exhibit some relevant differences when compared to other fish groups, they support the existence of a conserved NPY system in teleosts.

Ovarian follicle ultrastructure in the teleost Synbranchus marmoratus (Bloch, 1795), with special reference to the vitelline envelope development.

Synbranchus marmoratus is a protogynous diandric teleost fish widely distributed throughout South America. The aim of this work was to study the ultrastructure of the vitelline envelope and the relationship among oocyte and their follicular cells during oogenesis. During perinucleolar stage, the oocyte and the follicular cells form microvillar processes that project into the perivitelline space. The oocyte secretes a dense and amorphous material, which appears as the first evidence of the vitelline envelope (VE) development. The VE passes from a double to a multilayered structure during oocyte growth. In mature oocytes, the VE reach a mean thickness of 11 microm, having up to 30 layers. Oocyte microvilli are thinner than the follicular ones and were seen in contact with the follicular plasmalema, however we could not find any contact between the follicular microvilli and the oolemma. Before ovulation, microvillar processes retract and the pore canals seem to collapse. An outer electron dense layer occludes the superficial pore and forms a continuous layer. No jelly or adhesive coatings were seen at least in ovulated eggs sampled from ovarian lumen. Follicular cell and oocyte cytological characteristics do not differ from those described in other teleosts species.

Melanin-concentrating hormone system in the brain and skin of the cichlid fish Cichlasoma dimerus: anatomical localization, ontogeny and distribution in comparison to alpha-melanocyte-stimulating hormone-expressing cells.

Distribution and development of the melanin-concentrating hormone (MCH) system were examined by immunocytochemistry of the brain, pituitary gland and skin of the South American cichlid fish Cichlasoma dimerus. In adults, the most prominent group of MCH-ir perikarya was located in the nucleus lateralis tuberis (NLT). Outside the NLT, in the posterior hypothalamic region, a group of small neurons was found between the third ventricle and the lateral ventricular recess with delicate immunoreactive fibers that did not seem to contribute to the pituitary innervation. MCH-ir perikarya were identified at day 4 after hatching (AH) in a proliferating zone of the hypothalamic floor. Pituitary innervation could be detected at this stage. Another group of small MCH-ir neurons, only detected in pre-juvenile stages, originated close to the third ventricle in the medial hypothalamic region by day 6 AH. alphaMSH-ir neurons were localized in similar regions of the NLT and in the nucleus periventricularis posterior (NPP). Free MCH-ir neuromasts were detected in the ventral and dorsal skin of larval heads. These epidermal sensory organs were in close association with blood vessels and dermal melanocytes, suggesting that MCH synthesized in larval skin might act in an endocrine way reaching different targets and/or in a paracrine mode regulating melanin concentration in dermal melanocytes.

Oogenesis in the swamp eel Synbranchus marmoratus (Bloch, 1795) (Teleostei; Synbranchidae). Ovarian anatomy, stages of oocyte development and micropyle structure

Synbranchus marmoratus (Synbranchidae), commonly known as the swamp eel, is a protogynous diandric teleost fish widely distributed throughout South America. The purpose of this work was to study the ovarian anatomy and to describe oocyte developmental stages in the swamp eel, Synbranchus marmoratus. S. marmoratus has a unique sacular ovary. It is covered by a conspicuous muscular wall, probably involved in an egg-releasing system acting as a peristaltic-like mechanism. The internal ovarian anatomy shows a U-shaped ovarian lamella delimiting a dorsal ovarian lumen. The microscopic study shows evidence of the existence of a germinal epithelium in the inner surface of the lamella, which contains germinal cells, pre-follicular cells and epithelial cells. The complete oogenesis process is divided into four stages: oogonia, primary growth, cortical alveoli and vitellogenesis. Besides, the ovulated oocytes, and atretic structures were described. The structure of the micropyle was studied by scanning electron microscopy (MEB). Near the animal pole the vitelline envelope forms crests that fuse together becoming furrow-like structures with a slightly spiraled direction that converge into the micropyle pit where is located the micropylar canal. Although the sex reversal process of Synbranchids has been subject of many studies, this is the first complete description of the ovarian anatomy and oogenesis.

Oogenesis in the swamp eel Synbranchus marmoratus (Bloch, 1795) (Teleostei; Synbranchidae). Ovarian anatomy, stages of oocyte development and micropyle structure

Synbranchus marmoratus (Synbranchidae), commonly known as the swamp eel, is a protogynous diandric teleost fish widely distributed throughout South America. The purpose of this work was to study the ovarian anatomy and to describe oocyte developmental stages in the swamp eel, Synbranchus marmoratus. S. marmoratus has a unique sacular ovary. It is covered by a conspicuous muscular wall, probably involved in an egg-releasing system acting as a peristaltic-like mechanism. The internal ovarian anatomy shows a U-shaped ovarian lamella delimiting a dorsal ovarian lumen. The microscopic study shows evidence of the existence of a germinal epithelium in the inner surface of the lamella, which contains germinal cells, pre-follicular cells and epithelial cells. The complete oogenesis process is divided into four stages: oogonia, primary growth, cortical alveoli and vitellogenesis. Besides, the ovulated oocytes, and atretic structures were described. The structure of the micropyle was studied by scanning electron microscopy (MEB). Near the animal pole the vitelline envelope forms crests that fuse together becoming furrow-like structures with a slightly spiraled direction that converge into the micropyle pit where is located the micropylar canal. Although the sex reversal process of Synbranchids has been subject of many studies, this is the first complete description of the ovarian anatomy and oogenesis. (AU)

Colocalization of GnRH binding sites with gonadotropin-, somatotropin-, somatolactin-, and prolactin-expressing pituitary cells of the pejerrey, Odontesthes bonariensis, in vitro.

Previous studies in the pejerrey, Odontesthes bonariensis, have demonstrated that fibers with immunoreactivity to gonadotropin-releasing hormone (ir-GnRH) reach all areas of the pituitary gland, the rostral pars distalis (RPD), the proximal pars distalis (PPD), and the pars intemedia (PI). A close association was shown between ir-GnRH fibers and gonadotropin (GtH)-, growth hormone (GH)-, somatolactin (SL)-, and prolactin (PRL)-expressing cells. The presence of only one GnRH variant, suspected to be a novel form, has been shown in pituitary extracts of this fish. In addition, GnRH may stimulate GtHs, GH, SL, and PRL levels in different fish species. The objective of the present study was to seek GnRH receptors and therefore colocalization with GtHs, GH, SL, and PRL cells in O. bonariensis using a pituitary primary cell culture system. GnRH binding sites were revealed by autoradiography of an iodinated superactive GnRH agonist ([(125)I]GnRH-A) and pituitary cells were identified by immunocytochemistry using piscine antisera. Following autoradiography, silver grains representing specific [(125)I]GnRH-A binding were associated with anti GtH, GH, SL, and PRL positive cells. These results demonstrate the presence of GnRH binding sites on these cells. It is suggested that GnRH may play a wide role in the neuroendocrine control of different pituitary hormones in addition to the GtHs.

Concentration and seasonal variations of purine bases, nucleotides and nucleotide-sugars in liver of Bufo arenarum. Comparison with early embryogenesis.

A comparative study of the bases, nucleotides and nucleotide-sugars was performed in liver of Bufo arenarum. Samples were obtained at different seasons of the year, according to the periods of active (December-March) and passive metabolism (April-July). It was found that guanine and hypoxanthine increase significantly in spring. A remarkable decrease in the levels of the nucleosides triphosphates adenosine 5'-triphosphate (ATP) and guanosine 5'-triphosphate (GTP) was found in spring; correlatively, an increase in the levels of adenosine 5'-monophosphate (AMP) and guanosine 5'-monophosphate (GMP) and adenosine 5'-pyrophosphate (ADP) and guanosine 5'-pyrophosphate (GDP was observed. A notable increase in the amounts of uridine 5'-diphosphate- D - glucose (UDP-glucose) was detected at the end of the period of low metabolism, which is coincident wtih a decrease in the level of glycogen, observed by other authors.

Concentration and seasonal variations of purine bases, nucleotides and nucleotide-sugars in liver of Bufo arenarum. Comparison with early embryogenesis

A comparative study of the bases, nucleotides and nucleotide-sugars was performed in liver of Bufo arenarum. Samples were obtained at different seasons of the year, according to the periods of active (December-March) and passive metabolism (April-July). It was found that guanine and hypoxanthine increase significantly in spring. A remarkable decrease in the levels of the nucleosides triphosphates adenosine 5’-triphosphate (ATP) and guanosine 5’-triphosphate (GTP) was found in spring; correlatively, an increase in the levels of adenosine 5’-monophosphate (AMP) and guanosine 5’-monophosphate (GMP) and adenosine 5’-pyrophosphate (ADP) and guanosine 5’-pyrophosphate (GDP was observed. A notable increase in the amounts of uridine 5’-diphosphate- D - glucose (UDP-glucose) was detected at the end of the period of low metabolism, which is coincident wtih a decrease in the level of glycogen, observed by other authors.

Concentration and seasonal variations of purine bases, nucleotides and nucleotide-sugars in liver of Bufo arenarum. Comparison with early embryogenesis.

A comparative study of the bases, nucleotides and nucleotide-sugars was performed in liver of Bufo arenarum. Samples were obtained at different seasons of the year, according to the periods of active (December-March) and passive metabolism (April-July). It was found that guanine and hypoxanthine increase significantly in spring. A remarkable decrease in the levels of the nucleosides triphosphates adenosine 5-triphosphate (ATP) and guanosine 5-triphosphate (GTP) was found in spring; correlatively, an increase in the levels of adenosine 5-monophosphate (AMP) and guanosine 5-monophosphate (GMP) and adenosine 5-pyrophosphate (ADP) and guanosine 5-pyrophosphate (GDP was observed. A notable increase in the amounts of uridine 5-diphosphate- D - glucose (UDP-glucose) was detected at the end of the period of low metabolism, which is coincident wtih a decrease in the level of glycogen, observed by other authors.