Differential expression of zinc transporters accompanies the differentiation of C2C12 myoblasts

Zinc transporters facilitate metal mobilization and compartmentalization, playing a key role in cellular development. Little is known about the mechanisms and pathways of Zn movement between Zn transporters and metalloproteins during myoblast differentiation. We analyzed the differential expression of ZIP and ZnT transporters during C2C12 myoblast differentiation. Zn transporters account for a transient decrease of intracellular Zn upon myogenesis induction followed by a gradual increase of Zn in myotubes. Considering the subcellular localization and function of each of the Zn transporters, our findings indicate that a fine regulation is necessary to maintain correct metal concentrations in the cytosol and subcellular compartments to avoid toxicity, maintain homeostasis, and for loading metalloproteins needed during myogenesis. This study advances our basic understanding of the complex Zn transport network during muscle differentiation

Noise analysis to evaluate chick thermal comfort

A relação entre o ambiente térmico e o desempenho de pintinhos tem sido estudada, entretanto a consideração de que a medida de conforto térmico possa ser estimada pela interpretação da amplitude e da freqüência da vocalização, em condições tropicais de alojamento é um conceito novo. Esta pesquisa teve como objetivo estimar uma medida de bem-estar térmico para pintinhos em fase de aquecimento, pela medida de amplitude e freqüência do ruído emitido pelo grupo alojado. Inicialmente foi feito um experimento em câmara climática (A), para se estabelecer padrões de comportamento a partir de limites de temperatura ambiental. Foram alojados 45 pintos em um box de 2,3 m². Uma câmera foi colocada a 2,0 m acima das aves e a captura das imagens foi registrada em um computador. A partir dos padrões determinados no primeiro experimento, foi feita uma avaliação desses resultados em uma granja comercial (experimento B) utilizando densidade de aves semelhante, de maneira a possibilitar uma validação. Foram medidas continuamente: a temperatura ambiental e a amplitude e freqüência de ruídos, do grupo de pintos alojados durante a fase de aquecimento em ambos experimentos. Houve correlação entre um padrão de comportamento de agrupamento e a vocalização, medida não somente pela amplitude do ruído do grupo, mas também pelo espectro da sua freqüência. Na etapa de validação observou-se que, mantida a inércia térmica pelo uso adequado de cortinas, as aves vocalizam menos, coincidentemente com a baixa variação súbita de temperatura. Foi possível estimar o bem-estar térmico de pintinhos na fase de aquecimento pelo registro da amplitude e freqüência de ruído do grupo alojado.The relationship between thermal environment and chick performance has widely been evaluated, however the consideration that the assessment of the comfort may be estimated by interpreting both amplitude and frequency of bird vocalization under tropical housing conditions is a new concept. This research had as objective of estimating thermal comfort for chicks during the heating phase using this new concept. An experiment was carried out inside a climate controlled chamber (A) for establishing the behavioral pattern related to environmental temperature limits. Forty five chicks were reared inside a 2.3 m² box. A video camera was placed 2.0 m above the birds and the images were captured and registered in a computer. From the pattern determined in the controlled chamber an evaluation of the results was proceeded in a commercial broiler farm (experiment B) using similar bird density in order to validate the data. Environmental temperature, and both amplitude and frequency of the vocalizations of the chick group reared under heating were continuously recorded in both experiments. A correlation between group behavioral pattern and their vocalization, was found, evaluated not only by the noise amplitude but also by the noise frequency spectrum. When the thermal inertia is maintained by adequate use of curtains the birds vocalized less which is coincident with the low sudden variation of the temperature of the environmental. It was possible to estimate the thermal comfort for chicks at the heating stage by recording the amplitude and the frequency of the noise emitted by the reared group

MTF1, a classic metal sensing transcription factor, promotes myogenesis in response to copper [preprint]

MTF1 is a conserved metal-binding transcription factor in eukaryotes that binds to conserved DNA sequence motifs, termed metal response elements (MREs). MTF1 responds to metal excess and deprivation, protects cells from oxidative and hypoxic stresses, and is required for embryonic development in vertebrates. We used multiple strategies to identify an unappreciated role for MTF1 and copper (Cu) in cell differentiation. Upon initiation of myogenesis from primary myoblasts, MTF1 expression increased, as did nuclear localization. Mtf1 knockdown impaired differentiation, while addition of non-toxic concentrations of Cu+ enhanced MTF1 expression and promoted myogenesis. Cu+ bound stoichiometrically to a C-terminus tetra-cysteine of MTF1. MTF1 bound to chromatin at the promoter regions of myogenic genes and binding was stimulated by copper. MTF1 formed a complex with MyoD at myogenic promoters, the master transcriptional regulator of the myogenic lineage. These studies establish novel mechanisms by which copper and MTF1 regulate gene expression in myoblast differentiation

Análise de ruído para a avaliação do conforto térmico de pintinhos

The relationship between thermal environment and chick performance has widely been evaluated, however the consideration that the assessment of the comfort may be estimated by interpreting both amplitude and frequency of bird vocalization under tropical housing conditions is a new concept. This research had as objective of estimating thermal comfort for chicks during the heating phase using this new concept. An experiment was carried out inside a climate controlled chamber (A) for establishing the behavioral pattern related to environmental temperature limits. Forty five chicks were reared inside a 2.3 m² box. A video camera was placed 2.0 m above the birds and the images were captured and registered in a computer. From the pattern determined in the controlled chamber an evaluation of the results was proceeded in a commercial broiler farm (experiment B) using similar bird density in order to validate the data. Environmental temperature, and both amplitude and frequency of the vocalizations of the chick group reared under heating were continuously recorded in both experiments. A correlation between group behavioral pattern and their vocalization, was found, evaluated not only by the noise amplitude but also by the noise frequency spectrum. When the thermal inertia is maintained by adequate use of curtains the birds vocalized less which is coincident with the low sudden variation of the temperature of the environmental. It was possible to estimate the thermal comfort for chicks at the heating stage by recording the amplitude and the frequency of the noise emitted by the reared group.A relação entre o ambiente térmico e o desempenho de pintinhos tem sido estudada, entretanto a consideração de que a medida de conforto térmico possa ser estimada pela interpretação da amplitude e da freqüência da vocalização, em condições tropicais de alojamento é um conceito novo. Esta pesquisa teve como objetivo estimar uma medida de bem-estar térmico para pintinhos em fase de aquecimento, pela medida de amplitude e freqüência do ruído emitido pelo grupo alojado. Inicialmente foi feito um experimento em câmara climática (A), para se estabelecer padrões de comportamento a partir de limites de temperatura ambiental. Foram alojados 45 pintos em um box de 2,3 m². Uma câmera foi colocada a 2,0 m acima das aves e a captura das imagens foi registrada em um computador. A partir dos padrões determinados no primeiro experimento, foi feita uma avaliação desses resultados em uma granja comercial (experimento B) utilizando densidade de aves semelhante, de maneira a possibilitar uma validação. Foram medidas continuamente: a temperatura ambiental e a amplitude e freqüência de ruídos, do grupo de pintos alojados durante a fase de aquecimento em ambos experimentos. Houve correlação entre um padrão de comportamento de agrupamento e a vocalização, medida não somente pela amplitude do ruído do grupo, mas também pelo espectro da sua freqüência. Na etapa de validação observou-se que, mantida a inércia térmica pelo uso adequado de cortinas, as aves vocalizam menos, coincidentemente com a baixa variação súbita de temperatura. Foi possível estimar o bem-estar térmico de pintinhos na fase de aquecimento pelo registro da amplitude e freqüência de ruído do grupo alojado.438443Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

The classic metal-sensing transcription factor MTF1 promotes myogenesis in response to copper

Metal-regulatory transcription factor 1 (MTF1) is a conserved metal-binding transcription factor in eukaryotes that binds to conserved DNA sequence motifs, termed metal response elements. MTF1 responds to both metal excess and deprivation, protects cells from oxidative and hypoxic stresses, and is required for embryonic development in vertebrates. To examine the role for MTF1 in cell differentiation, we use multiple experimental strategies [including gene knockdown (KD) mediated by small hairpin RNA and clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9), immunofluorescence, chromatin immunopreciptation sequencing, subcellular fractionation, and atomic absorbance spectroscopy] and report a previously unappreciated role for MTF1 and copper (Cu) in cell differentiation. Upon initiation of myogenesis from primary myoblasts, both MTF1 expression and nuclear localization increased. Mtf1 KD impaired differentiation, whereas addition of nontoxic concentrations of Cu(+)-enhanced MTF1 expression and promoted myogenesis. Furthermore, we observed that Cu(+) binds stoichiometrically to a C terminus tetra-cysteine of MTF1. MTF1 bound to chromatin at the promoter regions of myogenic genes, and Cu addition stimulated this binding. Of note, MTF1 formed a complex with myogenic differentiation (MYOD)1, the master transcriptional regulator of the myogenic lineage, at myogenic promoters. These findings uncover unexpected mechanisms by which Cu and MTF1 regulate gene expression during myoblast differentiation