RESUMO
Manganese (Mn), a cofactor for various enzyme classes, is an essential trace metal for all organisms. However, overexposure to Mn causes neurotoxicity. Here, we evaluated the effects of exposure to Mn chloride (MnCl2) on viability, morphology, synapse function (based on neurogranin expression) and behavior of zebrafish larvae. MnCl2 exposure from 2.5 h post fertilization led to reduced survival (60%) at 5 days post fertilization. Phenotypical changes affected body length, eye and olfactory organ size, and visual background adaptation. This was accompanied by a decrease in both the fluorescence intensity of neurogranin immunostaining and expression levels of the neurogranin-encoding genes nrgna and nrgnb, suggesting the presence of synaptic alterations. Furthermore, overexposure to MnCl2 resulted in larvae exhibiting postural defects, reduction in motor activity and impaired preference for light environments. Following the removal of MnCl2 from the fish water, zebrafish larvae recovered their pigmentation pattern and normalized their locomotor behavior, indicating that some aspects of Mn neurotoxicity are reversible. In summary, our results demonstrate that Mn overexposure leads to pronounced morphological alterations, changes in neurogranin expression and behavioral impairments in zebrafish larvae.
Assuntos
Comportamento Animal , Larva , Manganês , Neurogranina , Peixe-Zebra , Animais , Peixe-Zebra/metabolismo , Larva/efeitos dos fármacos , Comportamento Animal/efeitos dos fármacos , Neurogranina/metabolismo , Neurogranina/genética , Manganês/toxicidade , Cloretos/toxicidade , Compostos de ManganêsRESUMO
The torus semicircularis (TS) of teleosts is a key midbrain center of the lateral line and acoustic sensory systems. To characterize the TS in adult zebrafish, we studied their connections using the carbocyanine tracers applied to the TS and to other related nuclei and tracts. Two main TS nuclei, central and ventrolateral, were differentiable by their afferent connections. From central TS, (TSc) numerous toropetal cells were labeled bilaterally in several primary octaval nuclei (anterior, magnocellular, descending, and posterior octaval nuclei), in the secondary octaval nucleus, in the caudal octavolateralis nucleus, and in the perilemniscular region. In the midbrain, numerous toropetal cells were labeled in the contralateral TSc. In the diencephalon, toropetal cells labeled from the TSc were observed ipsilaterally in the medial prethalamic nucleus and the periventricular posterior tubercle nucleus. TSc toropetal neurons were also labeled bilaterally in the hypothalamic anterior tuberal nucleus (ATN) and ipsilaterally in the parvicellular preoptic nucleus but not in the telencephalon. Tracer application to the medial octavolateralis nucleus revealed contralateral projections to the ventrolateral TS (TSvl), whereas tracer application to the secondary octaval nucleus labeled fibers bilaterally in TSc and neurons in rostral TSc. The TSc sends ascending fibers to the ipsilateral lateral preglomerular region that, in turn, projects to the pallium. Application of DiI to the optic tectum labeled cells and fibers in the TSvl, whereas application of DiI to the ATN labeled cells and fibers in the TSc. These results reveal that the TSvl and TSc are mainly related with the mechanosensory lateral line and acoustic centers, respectively, and that they show different higher order connections.
Assuntos
Neurônios , Peixe-Zebra , Animais , Acústica , Núcleo Arqueado do Hipotálamo , Colículos SuperioresRESUMO
Articular cartilage injuries are very frequent lesions that if left untreated may degenerate into osteoarthritis. Gene transfer to mesenchymal stem cells (MSCs) provides a powerful approach to treat these lesions by promoting their chondrogenic differentiation into the appropriate cartilage phenotype. Non-viral vectors constitute the safest gene transfer tools, as they avoid important concerns of viral systems including immunogenicity and insertional mutagenesis. However, non-viral gene transfer usually led to lower transfection efficiencies when compared with their viral counterparts. Biomaterial-guided gene delivery has emerged as a promising alternative to increase non-viral gene transfer efficiency by achieving sustained delivery of the candidate gene into cellular microenvironment. In the present study, we designed hyaluronic acid-based gene-activated cryogels (HACGs) encapsulating a novel formulation of non-viral vectors based on niosomes (P80PX) to promote MSCs in situ transfection. The developed HACG P80PX systems showed suitable physicochemical properties to promote MSCs in situ transfection with very low cytotoxicity. Incorporation of a plasmid encoding for the transcription factor SOX9 (psox9) into HACG P80PX systems led to an effective MSCs chondrogenic differentiation with reduced expression of fibrocartilage and hypertrophic markers. The capacity of the developed systems to restore cartilage extracellular matrix was further confirmed in an ex vivo model of chondral defect.
RESUMO
Zinc oxide nanoparticles (ZnO NPs) are among the most widely used nanomaterials. They have multiple applications in cosmetics, textiles, paints, electronics and, recently, also in biomedicine. This extensive use of ZnO NPs notably increases the probability that both humans and wildlife are subjected to undesirable effects. Despite being among the most studied NPs from a toxicological point of view, much remains unknown about their ecotoxicological effects or how they may affect specific cell types, such as cells of the central nervous system. The main objective of this work was to investigate the effects of ZnO NPs on human glial cells and zebrafish embryo development and to explore the role of the released Zn2+ ions in these effects. The effects on cell viability on human A172 glial cells were assessed with an MTT assay and morphological analysis. The potential acute and developmental toxicity was assessed employing zebrafish (Danio rerio) embryos. To determine the role of Zn2+ ions in the in vitro and in vivo observed effects, we measured their release from ZnO NPs with flame atomic absorption spectrometry. Then, cells and zebrafish embryos were treated with a water-soluble salt (zinc sulfate) at concentrations that equal the number of Zn2+ ions released by the tested concentrations of ZnO NPs. Exposure to ZnO NPs induced morphological alterations and a significant decrease in cell viability depending on the concentration and duration of treatment, even after removing the overestimation due to NP interference. Although there were no signs of acute toxicity in zebrafish embryos, a decrease in hatching was detected after exposure to the highest ZnO NP concentrations tested. The ability of ZnO NPs to release Zn2+ ions into the medium in a concentration-dependent manner was confirmed. Zn2+ ions did not seem entirely responsible for the effects observed in the glial cells, but they were likely responsible for the decrease in zebrafish hatching rate. The results obtained in this work contribute to the knowledge of the toxicological potential of ZnO NPs.
Assuntos
Nanopartículas Metálicas , Nanopartículas , Óxido de Zinco , Animais , Humanos , Peixe-Zebra , Óxido de Zinco/toxicidade , Óxido de Zinco/química , Nanopartículas/toxicidade , Neuroglia , Íons , Nanopartículas Metálicas/toxicidadeRESUMO
NOP56 belongs to a C/D box small nucleolar ribonucleoprotein complex that is in charge of cleavage and modification of precursor ribosomal RNAs and assembly of the 60S ribosomal subunit. An intronic expansion in NOP56 gene causes Spinocerebellar Ataxia type 36, a typical late-onset autosomal dominant ataxia. Although vertebrate animal models were created for the intronic expansion, none was studied for the loss of function of NOP56. We studied a zebrafish loss-of-function model of the nop56 gene which shows 70% homology with the human gene. We observed a severe neurodegenerative phenotype in nop56 mutants, characterized mainly by absence of cerebellum, reduced numbers of spinal cord neurons, high levels of apoptosis in the central nervous system (CNS) and impaired movement, resulting in death before 7 days post-fertilization. Gene expression of genes related to C/D box complex, balance and CNS development was impaired in nop56 mutants. In our study, we characterized the first NOP56 loss-of-function vertebrate model, which is important to further understand the role of NOP56 in CNS function and development.
RESUMO
Neurogranin (Nrgn) is a neural protein that is enriched in the cerebral cortex and is involved in synaptic plasticity via its interaction with calmodulin. Recently we reported its expression in the brain of the adult zebrafish (Alba-González et al. J Comp Neurol 530:1569-1587, 2022). In this study we analyze the development of Nrgn-like immunoreactivity (Nrgn-like-ir) in the brain and sensory structures of zebrafish embryos and larvae, using whole mounts and sections. First Nrgn-like positive neurons appeared by 2 day post-fertilization (dpf) in restricted areas of the brain, mostly in the pallium, epiphysis and hindbrain. Nrgn-like populations increased noticeably by 3 dpf, reaching an adult-like pattern in 6 dpf. Most Nrgn-like positive neurons were observed in the olfactory organ, retina (most ganglion cells, some amacrine and bipolar cells), pallium, lateral hypothalamus, thalamus, optic tectum, torus semicircularis, octavolateralis area, and viscerosensory column. Immunoreactivity was also observed in axonal tracts originating in Nrgn-like neuronal populations, namely, the projection of Nrgn-like immunopositive primary olfactory fibers to olfactory glomeruli, that of Nrgn-like positive pallial cells to the hypothalamus, the Nrgn-like-ir optic nerve to the pretectum and optic tectum, the Nrgn-like immunolabeled lateral hypothalamus to the contralateral region via the horizontal commissure, the octavolateralis area to the midbrain via the lateral lemniscus, and the viscerosensory column to the dorsal isthmus via the secondary gustatory tract. The late expression of Nrgn in zebrafish neurons is probably related to functional maturation of higher brain centers, as reported in the mammalian telencephalon. The analysis of Nrgn expression in the zebrafish brain suggests that it may be a useful marker for specific neuronal circuitries.
Assuntos
Neurogranina , Peixe-Zebra , Animais , Neurogranina/metabolismo , Encéfalo/metabolismo , Neurônios/metabolismo , Axônios/metabolismo , MamíferosRESUMO
We studied the expression of neurogranin in the brain and some sensory organs (barbel taste buds, olfactory organs, and retina) of adult zebrafish. Database analysis shows zebrafish has two paralog neurogranin genes (nrgna and nrgnb) that translate into three peptides with a conserved IQ domain, as in mammals. Western blots of zebrafish brain extracts using an anti-neurogranin antiserum revealed three separate bands, confirming the presence of three neurogranin peptides. Immunohistochemistry shows neurogranin-like expression in the brain and sensory organs (taste buds, neuromasts and olfactory epithelium), not being able to discern its three different peptides. In the retina, the most conspicuous positive cells were bipolar neurons. In the brain, immunopositive neurons were observed in all major regions (pallium, subpallium, preoptic area, hypothalamus, diencephalon, mesencephalon and rhombencephalon, including the cerebellum), a more extended distribution than in mammals. Interestingly, dendrites, cell bodies and axon terminals of some neurons were immunopositive, thus zebrafish neurogranins may play presynaptic and postsynaptic roles. Most positive neurons were found in primary sensory centers (viscerosensory column and medial octavolateral nucleus) and integrative centers (pallium, subpallium, optic tectum and cerebellum), which have complex synaptic circuitry. However, we also observed expression in areas not related to sensory or integrative functions, such as in cerebrospinal fluid-contacting cells associated with the hypothalamic recesses, which exhibited high neurogranin-like immunoreactivity. Together, these results reveal important differences with the patterns reported in mammals, suggesting divergent evolution from the common ancestor.
Assuntos
Neurogranina , Peixe-Zebra , Animais , Encéfalo/metabolismo , Diencéfalo/metabolismo , Mamíferos , Neurogranina/análise , Neurogranina/metabolismo , Rombencéfalo/química , Peixe-Zebra/metabolismoRESUMO
We studied the connections (connectome) of the adult zebrafish pallium using carbocyanine dye tracing and ancillary anatomical methods. The everted zebrafish pallium (dorsal telencephalic area, D) is composed of several major zones (medial, lateral, dorsal, central, anterior, and posterior) distinguishable by their topography, cytoarchitecture, immunohistochemistry, and genoarchitecture. Our comprehensive study reveals poor interconnectivity between these pallial areas, especially between medial (Dm), lateral/dorsal (Dl, Dd), and posterior (Dp) regions. This suggests that the zebrafish pallium has dedicated modules for different neural processes. Pallial connections with extrapallial regions also show compartmental organization. Major extratelencephalic afferents come from preglomerular nuclei (to Dl, Dd, and Dm), posterior tuberal nucleus (to Dm), and lateral recess nucleus (to Dl). The subpallial (ventral, V) zones dorsal Vv, Vd, and Vs, considered homologues of the striatum, amygdala, and pallidum, are mainly afferent to Dl/Dd and Dp. Regarding the efferent pathways, they also appear characteristic of each pallial region. Rostral Dm projects to the dorsal entopeduncular nucleus. Dp is interconnected with the olfactory bulbs. The central region (Dc) defined here receives mainly projections from Dl-Dd and projects toward the pretectum and optic tectum, connections, which help to delimiting Dc. The connectome of the adult pallium revealed here complements extant studies on the neuroanatomical organization of the brain, and may be useful for neurogenetic studies performed during early stages of development. The connectome of the zebrafish pallium was also compared with the pallial connections reported in other teleosts, a large group showing high pallial diversity.
Assuntos
Telencéfalo , Peixe-Zebra , Animais , Córtex Cerebral , Vias Eferentes , Bulbo OlfatórioRESUMO
Ruthenium(II) complexes are currently considered attractive alternatives to the widely used platinum-based drugs. We present herein the synthesis and characterization of half-sandwich ruthenium compounds formulated as [Ru(p-cymene)(L)Cl][CF3SO3] (L = 1,1-bis(methylenediphenylphosphano)ethylene, 1; L = 1,1-bis(diphenylphosphano)ethylene, 2), which were characterized by elemental analysis, mass spectrometry, 1H and 31P{1H} NMR, UV-vis and IR spectroscopy, conductivity measurements and cyclic voltammetry. The molecular structures for both complexes were determined by single-crystal X-ray diffraction. Their cytotoxic activity was evaluated using the MTT assay against human tumor cells, namely ovarian (A2780) and breast (MCF7 and MDA-MB-231). Both complexes were active against breast adenocarcinoma cells, with complex 1 exhibiting a quite remarkable cytotoxicity in the submicromolar range. Interestingly, at concentrations equivalent to the IC50 values in the MCF7 cancer cells, complexes 1 and 2 presented lower cytotoxicity in normal human primary fibroblasts. The antiproliferative effects of 1 and 2 in MCF7 cells might be associated with the induction of reactive oxygen species (ROS), leading to a combined cell death mechanism via apoptosis and autophagy. Despite the fact that in vitro a partial intercalation between complexes and DNA was observed, no MCF7 cell cycle delay or arrest was observed, indicating that DNA might not be a direct target. Complexes 1 and 2 both exhibited a moderate to strong interaction with human serum albumin, suggesting that protein targets may be involved in their mode of action. Their acute toxicity was evaluated in the zebrafish model. Complex 1 (the most toxic of the two) exhibited a lethal toxicity LC50 value about 1 order of magnitude higher than any IC50 concentrations found for the cancer cell models used, highlighting its therapeutic relevance as a drug candidate in cancer chemotherapy.
Assuntos
Antineoplásicos/farmacologia , Complexos de Coordenação/farmacologia , Fosfinas/farmacologia , Animais , Antineoplásicos/síntese química , Antineoplásicos/metabolismo , Antineoplásicos/toxicidade , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Complexos de Coordenação/síntese química , Complexos de Coordenação/metabolismo , Complexos de Coordenação/toxicidade , DNA/metabolismo , Ensaios de Seleção de Medicamentos Antitumorais , Fibroblastos/efeitos dos fármacos , Humanos , Fosfinas/síntese química , Fosfinas/metabolismo , Fosfinas/toxicidade , Ligação Proteica , Espécies Reativas de Oxigênio/metabolismo , Rutênio/química , Albumina Sérica Humana/metabolismo , Peixe-ZebraRESUMO
This study describes the cytoarchitecture of the torus longitudinalis (TL) in adult zebrafish by using light and electron microscopy, as well as its main connections as revealed by DiI tract tracing. In addition, by using high resolution confocal imaging followed by digital tracing, we describe the morphology of tectal pyramidal cells (type I cells) that are GFP positive in the transgenic line Tg(1.4dlx5a-dlx6a:GFP)ot1. The TL consists of numerous small and medium-sized neurons located in a longitudinal eminence attached to the medial optic tectum. A small proportion of these neurons are GABAergic. The neuropil shows three types of synaptic terminals and numerous dendrites. Tracing experiments revealed that the main efference of the TL is formed of parallel-like fibers that course within the marginal layer of the optic tectum. A toral projection to the thalamic nucleus rostrolateralis is also observed. Afferents to the TL come from visual and cerebellum-related nuclei in the pretectum, namely the central, intercalated and the paracommissural pretectal nuclei, as well as from the subvalvular nucleus in the isthmus. Additional afferents to the TL may come from the cerebellum but their origins could not be confirmed. The tectal afferent projection to the TL originates from cells similar to the type X cells described in other cyprinids. Tectal pyramidal neurons show round or piriform cell bodies, with spiny apical dendritic trees in the marginal layer. This anatomical study provides a basis for future functional and developmental studies focused on this cerebellum-like circuit in zebrafish.
Assuntos
Colículos Superiores/anatomia & histologia , Colículos Superiores/ultraestrutura , Vias Visuais/anatomia & histologia , Vias Visuais/ultraestrutura , Peixe-Zebra/anatomia & histologia , Fatores Etários , Animais , Animais Geneticamente Modificados , Microscopia/métodos , Microscopia Eletrônica/métodos , Colículos Superiores/química , Vias Visuais/químicaRESUMO
Ruthenium(II) complexes are currently considered a viable alternative to the widely used platinum complexes as efficient anticancer agents. We herein present the synthesis and characterization of half-sandwich ruthenium compounds with the general formula [Ru( p-cymene)(L-N,N)Cl][CF3SO3] (L = 3,6-di-2-pyridyl-1,2,4,5-tetrazine (1) 6,7-dimethyl-2,3-bis(pyridin-2-yl)quinoxaline (2)), which have been synthesized by substitution reactions from the precursor dimer [Ru( p-cymene)(Cl)(µ-Cl)]2 and were characterized by elemental analysis, mass spectrometry, 1H NMR, UV-vis, and IR spectroscopy, conductivity measurements, and cyclic voltammetry. The molecular structure for complex 2 was determined by single-crystal X-ray diffraction. The cytotoxic activity of these compounds was evaluated against human tumor cells, namely ovarian carcinoma A2780 and breast MCF7 and MDAMB231 adenocarcinoma cells, and against normal primary fibroblasts. Whereas the cytotoxic activity of 1 is moderate, IC50 values found for 2 are among the lowest previously reported for Ru( p-cymene) complexes. Both compounds present no cytotoxic effect in normal human primary fibroblasts when they are used at the IC50 concentration in A2780 and MCF7 cancer cells. Their antiproliferative capacity is associated with a combined mechanism of apoptosis and autophagy. A strong interaction with DNA was observed for both with a binding constant value of the same magnitude as that of the classical intercalator [Ru(phen)2(dppz)]2+. Both complexes bind to human serum albumin with moderate to strong affinity, with conditional binding constants (log Kb) of 4.88 for complex 2 and 5.18 for complex 1 in 2% DMSO/10 mM Hepes pH7.0 medium. The acute toxicity was evaluated in zebrafish embryo model using the fish embryo acute toxicity test (FET). Remarkably, our results show that compounds 1 and 2 are not toxic/lethal even at extremely high concentrations. The novel compounds reported herein are highly relevant antitumor metallodrug candidates, given their in vitro cytotoxicity toward cancer cells and the lack of in vivo toxicity.
Assuntos
Antineoplásicos/farmacologia , Complexos de Coordenação/farmacologia , Monoterpenos/farmacologia , Rutênio/farmacologia , Animais , Antineoplásicos/efeitos adversos , Antineoplásicos/síntese química , Antineoplásicos/química , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Complexos de Coordenação/efeitos adversos , Complexos de Coordenação/síntese química , Complexos de Coordenação/química , Cimenos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Estrutura Molecular , Monoterpenos/efeitos adversos , Monoterpenos/química , Rutênio/efeitos adversos , Rutênio/química , Relação Estrutura-Atividade , Peixe-ZebraRESUMO
The pretectum is a complex region of the caudal diencephalon which in adult zebrafish comprises both retinorecipient (parvocellular superficial, central, intercalated, paracommissural, and periventricular) and non-retinorecipient (magnocellular superficial, posterior, and accessory) pretectal nuclei distributed from periventricular to superficial regions. We conducted a comprehensive study of the connections of pretectal nuclei by using neuronal tracing with fluorescent carbocyanine dyes. This study reveals specialization of efferent connections of the various pretectal nuclei, with nuclei projecting to the optic tectum (paracommissural, central, and periventricular pretectal nuclei), the torus longitudinalis and the cerebellar corpus (paracommissural, central, and intercalated pretectal nuclei), the lateral hypothalamus (magnocellular superficial, posterior, and central pretectal nuclei), and the tegmental regions (accessory and superficial pretectal nuclei). With regard to major central afferents to the pretectum, we observed projections from the telencephalon to the paracommissural and central pretectal nuclei, from the optic tectum to the paracommissural, central, accessory and parvocellular superficial pretectal nuclei, from the cerebellum to the paracommissural and periventricular pretectal nuclei and from the nucleus isthmi to the parvocellular superficial and accessory pretectal nuclei. The parvocellular superficial pretectal nucleus sends conspicuous projections to the contralateral magnocellular superficial pretectal nucleus. The composite figure of results reveals large differences in connections of neighbor pretectal nuclei, indicating high degree of nuclear specialization. Our results will have important bearings in functional studies that analyze the relationship between specific circuits and behaviors in zebrafish. Comparison with results available in other species also reveals differences in the organization and connections of the pretectum in vertebrates.
Assuntos
Rede Nervosa/fisiologia , Vias Neurais/fisiologia , Neurônios/fisiologia , Área Pré-Tectal/citologia , Peixe-Zebra/anatomia & histologia , Aminoácidos/metabolismo , Animais , Feminino , MasculinoRESUMO
Resumen Los leiomyosarcomas pueden originarse en la mayor parte de los órganos desarrollados fuera del sistema nervioso central. Se han documentado casos de leiomyosarcomas de origen: intestinal, mesentérico, uterino, retroperitoneal, de tejidos blandos y vascular. Sin embargo, los casos de leiomyosarcomas primarios de ovario son extremadamente infrecuentes y pocos casos han sido descritos en la literatura. Se cree que estos leiomyosarcomas se originan en el músculo liso alrededor de los folículos o a partir de remanentes del conducto de Wolff a partir de las paredes de la vasculatura existente en el parénquima ovárico. La mayor parte de los casos se presentan en pacientes postmenopáusicas y con un pronóstico a corto plazo desfavo rable. El pilar del tratamiento continúa siendo la citorreducción tumoral completa obteniendo márgenes quirúrgicos negativos con miras a disminuir el potencial de recidiva. El beneficio de la utilización de quimioterapia adyuvante como parte del tratamiento de esta patología sigue siendo incierto.
Abstract Leiomyosarcomas may originate in most of the organs developed outside the central nervous system. There are documented cases of leiomyosarcomas of intestinal, mesenteric, ute rine, retroperitoneal, and of soft and vascular tissue origin. However, cases of primary ovarian leiomyosarcoma are extremely rare, with few cases reported in the international literature. Leiomyosarcomas are believed to be those that originate from the walls of existing vasculature in ovarian parenchyma, in the smooth muscle around the follicles, or from remnants of the Wolff duct. Most cases occur in post-menopausal patients, and have an unfavourable prognosis in the short term. The mainstay of treatment remains the complete tumour debulking, with negative surgical margins in order to reduce the potential for recurrence. The benefit of the use of adjuvant chemotherapy as part of treatment of this condition remains uncertain.
Assuntos
Humanos , Leiomiossarcoma , Quimioterapia Adjuvante , Miócitos de Músculo Liso , OvárioRESUMO
The central connections of the gustatory/general visceral system of the adult zebrafish (Danio rerio) were examined by means of carbocyanine dye tracing. Main primary gustatory centers (facial and vagal lobes) received sensory projections from the facial and vagal nerves, respectively. The vagal nerve also projects to the commissural nucleus of Cajal, a general visceral sensory center. These primary centers mainly project on a prominent secondary gustatory and general visceral nucleus (SGN/V) located in the isthmic region. Secondary projections on the SGN/V were topographically organized, those of the facial lobe mainly ending medially to those of the vagal lobe, and those from the commissural nucleus ventrolaterally. Descending facial lobe projections to the medial funicular nucleus were also noted. Ascending fibers originating from the SGN/V mainly projected to the posterior thalamic nucleus and the lateral hypothalamus (lateral torus, lateral recess nucleus, hypothalamic inferior lobe diffuse nucleus) and an intermediate cell- and fiber-rich region termed here the tertiary gustatory nucleus proper, but not to a nucleus formerly considered as the zebrafish tertiary gustatory nucleus. The posterior thalamic nucleus, tertiary gustatory nucleus proper, and nucleus of the lateral recess gave rise to descending projections to the SGN/V and the vagal lobe. The connectivity between diencephalic gustatory centers and the telencephalon was also investigated. The present results showed that the gustatory connections of the adult zebrafish are rather similar to those reported in other cyprinids, excepting the tertiary gustatory nucleus. Similarities between the gustatory systems of zebrafish and other fishes are also discussed. J. Comp. Neurol. 525:333-362, 2017. © 2016 Wiley Periodicals, Inc.
Assuntos
Vias Aferentes/citologia , Encéfalo/citologia , Peixe-Zebra/anatomia & histologia , Vias Aferentes/fisiologia , Animais , Encéfalo/fisiologia , Carbocianinas , Coloração e Rotulagem , Percepção Gustatória/fisiologia , Vísceras/inervação , Peixe-Zebra/fisiologiaRESUMO
The habenulae are bilateral nuclei located in the dorsal diencephalon that are conserved across vertebrates. Here we describe the main afferents to the habenulae in larval and adult zebrafish. We observe afferents from the subpallium, nucleus rostrolateralis, posterior tuberculum, posterior hypothalamic lobe, median raphe; we also see asymmetric afferents from olfactory bulb to the right habenula, and from the parapineal to the left habenula. In addition, we find afferents from a ventrolateral telencephalic nucleus that neurochemical and hodological data identify as the ventral entopeduncular nucleus (vENT), confirming and extending observations of Amo et al. (2014). Fate map and marker studies suggest that vENT originates from the diencephalic prethalamic eminence and extends into the lateral telencephalon from 48 to 120 hour post-fertilization (hpf). No afferents to the habenula were observed from the dorsal entopeduncular nucleus (dENT). Consequently, we confirm that the vENT (and not the dENT) should be considered as the entopeduncular nucleus "proper" in zebrafish. Furthermore, comparison with data in other vertebrates suggests that the vENT is a conserved basal ganglia nucleus, being homologous to the entopeduncular nucleus of mammals (internal segment of the globus pallidus of primates) by both embryonic origin and projections, as previously suggested by Amo et al. (2014).
Assuntos
Vias Aferentes/fisiologia , Mapeamento Encefálico , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Habenula/fisiologia , Aminoácidos/metabolismo , Animais , Animais Geneticamente Modificados , Gânglios da Base , Calbindina 2/genética , Calbindina 2/metabolismo , Embrião não Mamífero , Lateralidade Funcional , Regulação da Expressão Gênica no Desenvolvimento/genética , Ácido Glutâmico/metabolismo , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Habenula/citologia , Habenula/crescimento & desenvolvimento , Larva , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Neurônios/metabolismo , Bulbo Olfatório , Peixe-Zebra , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismoRESUMO
Los tumores derivados de células germinales representan el 20-25% de tumores ováricos, siendo sólo 3% malignos. La degeneración maligna se presenta únicamente en 1-2% de los casos. La edad de presentación promedio de la transformación maligna es a los 50 años, siendo los más frecuentes el carcinoma de células escamosas y el adenocarcinoma (1). Dado que la supervivencia a largo plazo se encuentra en relación con la detección temprana y la completa resección quirúrgica, se hace necesario determinar las características clínico-patológicas que sugieran una posible transformación maligna de estos tumores. A continuación, se presenta el caso de una paciente perimenopáusica, con hallazgos histopatológicos indicativos de transformación maligna de un teratoma quístico maduro a cistoadenocarcinoma mucinoso bien diferenciado de tipo endocervical e intestinal.
The germ cell-derived tumors account for 20-25% of ovarian tumors, with 3% being malignant. The degeneration occurs in 1-2% of cases. The age of presentation is menopause, around the age 50, and the most common malignant histological types in this context are squamous cell carcinoma and adenocarcinoma. It is important to determine the clinical-pathological characteristics of malignant transformation (although currently not well established), as because early detection and complete surgical resection are important for long-term survival. A case is presented on a peri-menopausal woman with histopathological findings of malignant transformation of a mature cystic teratoma to well-differentiated mucinous cystadenocarcinoma endocervical and intestinal type.
Assuntos
Humanos , Feminino , Ovário , Teratoma , Carcinoma de Células Escamosas , Cistadenocarcinoma Mucinoso , Adenocarcinoma Mucinoso , Mulheres , Sobrevivência , Células GerminativasRESUMO
Polypteriform fishes are believed to be basal to other living ray-finned bony fishes, and they may be useful for providing information of the neural organization that existed in the brain of the earliest ray-finned fishes. The calcium-binding proteins calretinin (CR) and calbindin-D28k (CB) have been widely used to characterize neuronal populations in vertebrate brains. Here, the distribution of the immunoreactivity against CR and CB was investigated in the olfactory organ and brain of Polypterus senegalus and compared to the distribution of these molecules in other ray-finned fishes. In general, CB-immunoreactive (ir) neurons were less abundant than CR-ir cells. CR immunohistochemistry revealed segregation of CR-ir olfactory receptor neurons in the olfactory mucosa and their bulbar projections. Our results confirmed important differences between pallial regions in terms of CR immunoreactivity of cell populations and afferent fibers. In the habenula, these calcium-binding proteins revealed right-left asymmetry of habenular subpopulations and segregation of their interpeduncular projections. CR immunohistochemistry distinguished among some thalamic, pretectal, and posterior tubercle-derived populations. Abundant CR-ir populations were observed in the midbrain, including the tectum. CR immunoreactivity was also useful for characterizing a putative secondary gustatory/visceral nucleus in the isthmus, and for distinguishing territories in the primary viscerosensory column and octavolateral region. Comparison of the data obtained within a segmental neuromeric context indicates that some CB-ir and CR-ir populations in polypteriform fishes are shared with other ray-finned fishes, but other positive structures appear to have evolved following the separation between polypterids and other ray-finned fishes.
Assuntos
Calbindina 2/metabolismo , Calbindinas/metabolismo , Rajidae/fisiologia , Animais , Especificidade de Anticorpos , Química Encefálica , Imunofluorescência , Imuno-Histoquímica , Mesencéfalo/química , Mesencéfalo/metabolismo , Neurônios/metabolismo , Neurônios/fisiologia , Bulbo Olfatório/metabolismo , Mucosa Olfatória/citologia , Mucosa Olfatória/inervação , Mucosa Olfatória/fisiologia , Nervo Olfatório/citologia , Nervo Olfatório/fisiologia , Neurônios Receptores Olfatórios/metabolismo , Rombencéfalo/citologia , Rombencéfalo/metabolismo , Colículos Superiores/citologia , Colículos Superiores/fisiologia , Núcleo Supraquiasmático/citologia , Núcleo Supraquiasmático/fisiologia , Tálamo/citologia , Tálamo/fisiologiaRESUMO
Chondrosteans represent an ancient lineage in ray-finned bony fishes and hence in jawed vertebrates. This immunohistochemical study in the brain of the Siberian sturgeon reports the neuronal distribution of three cytosolic calcium-binding proteins: calbindin-D28k (CB), calretinin (CR), and parvalbumin (PV). CB and CR are widely expressed in different neuron subsets distributed throughout the sturgeon brain. Studies using double immunofluorescence reveal a wide co-distribution of CB and CR in the brain nuclei but scarce co-localization at cellular level. In the forebrain, CR- and CB-immunoreactive (ir) populations were observed in the olfactory bulbs, in pallial and subpallial telencephalic areas, and in some diencephalic nuclei. CR-ir cells were also observed in the posterior tubercle and CB-ir cells in the preglomerular complex. At midbrain and hindbrain basal levels, CB-ir and CR-ir cell bodies were mainly distributed in periventricular areas. In the cerebellum, CB and CR cells were co-localized in some granular cell subsets in laterodorsal and dorsolateral regions, and in some Purkinje-like cells. CB-ir and CR-ir fibers were mainly observed in the olfactory bulbs, hypothalamus, and habenula, and in fiber tracts that coursed in the optic tectum and through the mesencephalic and rhombencephalic basal areas. With regard to PV, the sturgeon brain showed a rather limited distribution of PV-ir perikarya and fibers. Thus, CR, CB, and PV allowed the identification of subpopulations of neurons not distinguished on the basis of cytoarchitecture alone, which provided a better understanding of the anatomical organization of the sturgeon brain. These results reveal numerous shared features with teleosts, but also important differences.
Assuntos
Encéfalo/metabolismo , Proteínas de Ligação ao Cálcio/metabolismo , Rajidae/anatomia & histologia , Animais , Encéfalo/anatomia & histologia , Calbindina 2 , Calbindinas , Parvalbuminas/metabolismo , Proteína G de Ligação ao Cálcio S100/metabolismoRESUMO
The pineal organ of fish is a photosensitive structure that receives light information from the environment and transduces it into hormonal (rhythmic melatonin secretion) and neural (efferent projections/neurotransmitters) signals. In this study, we focused on this neural output. Thus, we performed a tract-tracing study using 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI), a fluorescent carbocyanine dye, in order to elucidate the efferent and afferent connections of the pineal organ in the European sea bass. The axonal transport of DiI revealed extensive bilateral projections in the sea bass brain. The efferent projections of the sea bass pineal organ reach the habenula, ventral thalamus, periventricular pretectum, central pretectal area, posterior tubercle and medial and dorsal tegmental areas. In addition, in this study we also examined the pinealopetal system in sea bass. This analysis demonstrated that the sea bass pineal organ receives central projections from neurons located, to a large extent, in brain areas innervated by pineal efferent projections, i.e. the thalamic eminence, habenula, ventral thalamus, dorsal thalamus, periventricular pretectum, posterior commissure, posterior tubercle and medial tegmental area. This study is the first description of pinealofugal projections in a representative of Perciformes, which constitutes a derived order within teleosts. Moreover, it represents the first evidence for the presence of pinealopetal neurons in the brain of a teleost species. Our findings, together with the analysis of retinal connections, represent a step forward in the understanding of the integration of photoperiodic signals into the central nervous system of sea bass.
Assuntos
Vias Aferentes/citologia , Bass/anatomia & histologia , Vias Eferentes/citologia , Glândula Pineal/inervação , Animais , CarbocianinasRESUMO
In elasmobranchs the terminal nerve courses separately from the olfactory nerve. This characteristic makes elasmobranchs excellent models to study the anatomy and function of these two systems. Here we study the neural connections of the terminal nerve and olfactory system in two sharks by experimental tracing methods using carbocyanine dyes. The main projections from the terminal nerve system (consisting of three ganglia in Scyliorhinus canicula) course ipsilaterally to the medial septal nucleus and bilaterally to the ventromedial telencephalic pallial region. Minor terminal nerve projections were also traced ipsilaterally to diencephalic and mesencephalic levels. With regard to the olfactory connections, our results show that in sharks, unlike ray-finned fishes, the primary olfactory projections are mainly restricted to the olfactory bulb. We also performed tracer application to the olfactory bulb in order to analyze the possible central neuroanatomical relationship between the projections of the terminal nerve and the olfactory bulb. In these experiments labeled neurons and fibers were observed from telencephalic to caudal mesencephalic regions. However, we observe almost no overlap between the two systems at central levels. The afferent and the putatively efferent connections of the dogfish olfactory bulb are compared with those previously reported in other elasmobranchs. The significance of the extratelencephalic secondary olfactory projections is also discussed in a comparative context.