Effects of FGF-2 and EGF removal on the differentiation of mouse neural precursor cells

Cell therapy for neurological disorders has advanced, and neural precursor cells (NPC) may become the ideal candidates for neural transplantation in a wide range of diseases. However, additional work has to be done to determine either the ideal culture environment for NPC expansion in vitro, without altering their plasticity, or the FGF-2 and EGF mechanisms of cell signaling in neurospheres growth, survival and differentiation. In this work we evaluated mouse neurospheres cultured with and without FGF-2 and EGF containing medium and showed that those growth factors are responsible for NPC proliferation. It is also demonstrated that endogenous production of growth factors shifts from FGF-2 to IGF-1/PDGFb upon EGF and FGF-2 withdrawal. Mouse NPC cultured in suspension showed different patterns of neuronal localization (core versus shell) for both EGF and FGF-2 withdrawal and control groups. Taken together, these results show that EGF and FGF-2 removal play an important role in NPC differentiation and may contribute to a better understanding of mechanisms of NPC differentiation. Our findings suggest that depriving NPC of growth factors prior to grafting might enhance their chance to effectively integrate into the host.As terapias celulares para doenças neurológicas têm avançado e células precursoras neurais (NPC) surgem como candidatas ideais para o transplante de células neurais em muitas doenças. No entanto, trabalhos adicionais devem ser feitos para determinar o ambiente de cultivo ideal para a expansão in vitro das NPC, sem alterar sua plasticidade, e os mecanismos de sinalização celular do fator de crescimento epidérmico (EGF) e fator de crescimento de fibroblasto 2 (FGF-2) no crescimento, sobrevivência e diferenciação da neuroesfera. Nesse trabalho avaliamosNPCcultivadas na presença e na ausência de FGF-2 e EGF e mostramos que esses fatores de crescimento são responsáveis pela proliferação das NPC. Também foi demonstrado que a produção endógena de fatores de crescimento alterna de FGF-2 a fator de crescimento de insulina 1 (IGF-1) e fator de crescimento derivado de plaquetas b (PDGFb) após remoção de EGF e FGF-2. NPC de camundongo cultivadas em suspensão mostraram padrões de localização neuronal distintos (centro versus borda) tanto no grupo controle como no grupo sem EGF e FGF-2. Juntos, esses resultados mostram que a remoção de EGF e FGF-2 exerce importante ação na diferenciação de NPC e possivelmente contribui para melhor compreensão dos mecanismos envolvidos na diferenciação. Nossos achados sugerem que, privando as NPC de fatores de crescimento antes do transplante, talvez aumente as chances de que as células efetivamente se integrem ao hospedeiro.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo (UNIFESP) Departamento de FisiologiaUniversidade Federal de São Paulo (UNIFESP) Departamento de BiofísicaUNIFESP, Depto. de FisiologiaUNIFESP, Depto. de BiofísicaSciEL

A review of the Cis-Andean species of Hemibrycon Günther (Teleostei: Characiformes: Characidae: Stevardiinae), with description of two new species

As espécies de Hemibrycon que ocorrem a leste das Cordilheiras dos Andes são revisadas com base na análise do material tipo e exemplares adicionais. Nove espécies são redescritas: H. beni da bacia do río Beni, Bolívia; H. helleri do alto río Ucayali, Peru; H. huambonicus para as bacias dos ríos Huallaga e Marañon, Peru; H. jeslkii para as porções superiores das bacias dos rios Marañon, Ucayali e Madeira, Bolívia, Brasil e Peru; H. metae para a bacia do río Orinoco, Colômbia e Venezuela, e bacias costeiras do Caribe na Venezuela; H. polyodon (espécie-tipo) para a bacia do río Pastaza, Equador; H. surinamensis para as bacia costeiras da Guiana Francesa e Suriname, e porção inferior das bacias dos rios Tapajós, Tocantins e Xingu, Brasil; H. taeniurus para os rios da ilha de Trinidad, Trinidad and Tobago; H. tridens para a bacia do alto río Ucayali, Peru. Duas espécies novas são descritas: H. inambari para a bacia do alto río Madre de Dios, Peru e H. mikrostiktos para a bacia do río Ucayali, Peru. Estas espécies distinguem-se principalmente pelo padrão de colorido e caracteres merísticos. A área de distribuição do gênero é ampliada para as porções inferiores das bacias do Tapajós, Tocantins e Xingu, Brasil, com o primeiro registro de H. surinamensis para este país. Uma nova diagnose e descrição são fornecidas para a espécie-tipo do gênero, apesar de ter sido recentemente redescrita. A série-tipo de H. helleri previamente considerada questionável é encontrada e descrita. Hemibrycon coxeyi e H. pautensis são considerados sinônimos juniores de H. polyodon. Tetragonopterus (Hemibrycon) trinitatis, anteriormente considerada como species inquirenda em Characidae, e Hemibrycon guppyi são sinônimos juniores de H. taeniurus. Hemibrycon orcesi é transferida para Boehlkea. É apresentada uma chave taxonômica para as espécies do gênero a leste das Cordilheiras dos Andes.The species of Hemibrycon occurring in the east of the Andean Cordilleras are reviewed based on their type series and additional specimens. Nine species are redescribed: H. beni from río Beni basin, Bolivia; H. helleri from the upper rio Ucayali, Peru; H. huambonicus from the ríos Huallaga and Marañon basins, Peru; H. jeslkii from the upper portions of ríos Marañon, Ucayali and Madeira basins, Bolivia, Brazil, and Peru; H. metae from río Orinoco basin, Colombia and Venezuela, and Caribbean coastal basins of Venezuela; H. polyodon (type species) from río Pastaza basin, Ecuador; H. surinamensis from coastal basins of French Guiana and Suriname, and from lower rios Tapajós, Tocantins and Xingu basins, Brazil; H. taeniurus from river basins from Trinidad Island, Trinidad and Tobago, and H. tridens from upper río Ucayali basin, Peru. Two new species are described: H. inambari from the upper río Madre de Dios basin, Peru and H. mikrostiktos from río Ucayali basin, Peru. These species are distinguished among themselves mainly by the color pattern, and meristic characters. The distribution area of the genus is enlarged reaching the lower Tapajós, Tocantins and Xingu river basins, Brazil, with the first record of the occurrence of H. surinamensis in this country. A new description and diagnosis is provided for the type species of the genus, regardless its recent redescription. The type series of H. helleri that was previously considered dubious is found and described. Hemibrycon coxeyi and H. pautensis are considered junior synonyms of H. polyodon. Tetragonopterus (Hemibrycon) trinitatis, previously considered species inquirenda in Characidae, and Hemibrycon guppyi are junior synonyms of H. taeniurus. Hemibrycon orcesi is transferred to Boehlkea. A taxonomic key for the species of the genus to the east of the Andean Cordilleras is presented

Chemically-Induced RAT Mesenchymal Stem Cells Adopt Molecular Properties of Neuronal-Like Cells but Do Not Have Basic Neuronal Functional Properties

Induction of adult rat bone marrow mesenchymal stem cells (MSC) by means of chemical compounds (β-mercaptoethanol, dimethyl sulfoxide and butylated hydroxyanizole) has been proposed to lead to neuronal transdifferentiation, and this protocol has been broadly used by several laboratories worldwide. Only a few hours of MSC chemical induction using this protocol is sufficient for the acquisition of neuronal-like morphology and neuronal protein expression. However, given that cell death is abundant, we hypothesize that, rather than true neuronal differentiation, this particular protocol leads to cellular toxic effects. We confirm that the induced cells with neuronal-like morphology positively stained for NF-200, S100, β-tubulin III, NSE and MAP-2 proteins. However, the morphological and molecular changes after chemical induction are also associated with an increase in the apoptosis of over 50% of the plated cells after 24 h. Moreover, increased intracellular cysteine after treatment indicates an impairment of redox circuitry during chemical induction, and in vitro electrophysiological recordings (patch-clamp) of the chemically induced MSC did not indicate neuronal properties as these cells do not exhibit Na+ or K+ currents and do not fire action potentials. Our findings suggest that a disruption of redox circuitry plays an important role in this specific chemical induction protocol, which might result in cytoskeletal alterations and loss of functional ion-gated channels followed by cell death. Despite the neuronal-like morphology and neural protein expression, induced rat bone marrow MSC do not have basic functional neuronal properties, although it is still plausible that other methods of induction and/or sources of MSC can achieve a successful neuronal differentiation in vitro

Cryptic Diversity of African Tigerfish (Genus Hydrocynus) Reveals Palaeogeographic Signatures of Linked Neogene Geotectonic Events

The geobiotic history of landscapes can exhibit controls by tectonics over biotic evolution. This causal relationship positions ecologically specialized species as biotic indicators to decipher details of landscape evolution. Phylogeographic statistics that reconstruct spatio-temporal details of evolutionary histories of aquatic species, including fishes, can reveal key events of drainage evolution, notably where geochronological resolution is insufficient. Where geochronological resolution is insufficient, phylogeographic statistics that reconstruct spatio-temporal details of evolutionary histories of aquatic species, notably fishes, can reveal key events of drainage evolution. This study evaluates paleo-environmental causes of mitochondrial DNA (mtDNA) based phylogeographic records of tigerfishes, genus Hydrocynus, in order to reconstruct their evolutionary history in relation to landscape evolution across Africa. Strong geographical structuring in a cytochrome b (cyt-b) gene phylogeny confirms the established morphological diversity of Hydrocynus and reveals the existence of five previously unknown lineages, with Hydrocynus tanzaniae sister to a clade comprising three previously unknown lineages (Groups B, C and D) and H. vittatus. The dated phylogeny constrains the principal cladogenic events that have structured Hydrocynus diversity from the late Miocene to the Plio-Pleistocene (ca. 0–16 Ma). Phylogeographic tests reveal that the diversity and distribution of Hydrocynus reflects a complex history of vicariance and dispersals, whereby range expansions in particular species testify to changes to drainage basins. Principal divergence events in Hydrocynus have interfaced closely with evolving drainage systems across tropical Africa. Tigerfish evolution is attributed to dominant control by pulses of geotectonism across the African plate. Phylogenetic relationships and divergence estimates among the ten mtDNA lineages illustrates where and when local tectonic events modified Africa's Neogene drainage. Haplotypes shared amongst extant Hydrocynus populations across northern Africa testify to recent dispersals that were facilitated by late Neogene connections across the Nilo-Sahelian drainage. These events in tigerfish evolution concur broadly with available geological evidence and reveal prominent control by the African Rift System, evident in the formative events archived in phylogeographic records of tigerfish

Spermiogenesis and spermatozoa ultrastructure in the Serrasalminae (Ostariophysi: Characiformes) with further evidence on the relationships of the piranhas and pacus

A ultraestrutura dos espermatozoides e espermiogênese foram estudadas nas espécies Piaractus mesopotamicus, Mylossoma duriventre, Serrasalmus maculatus e Metynnis mola de Serrasalminae, sendo dois padrões distintos reconhecidos: um comum a Mylossoma, Serrasalmus e Metynnis, e o segundo característico de Piaractus. Este último é mais similar ao observado em Salminus e Brycon. Por outro lado, o espermatozoide dos serrasalmíneos compartilha algumas características com espécies da superfamília Anostomoidea. O significado filogenético destes caracteres é discutido.Spermatozoa and spermiogenesis ultrastructure were studied in the serrasalmine species Piaractus mesopotamicus, Mylossoma duriventre, Serrasalmus maculatus, and Metynnis mola and two distinct patterns may be recognized: the first common to Mylossoma, Serrasalmus and Metynnis, and the other, characteristic of Piaractus. The latter pattern is more similar to the conditions found in Salminus and Brycon. On the other hand, serrasalmine spermatozoa also share characteristics with the spermatozoa of species of the superfamily Anostomoidea. The phylogenetic significance of these characters is discussed