Parâmetros zootécnicos de juvenis de pacu alimentados a diferentes frequências de arraçoamento em tanques-rede

O objetivo deste trabalho foi avaliar o desempenho zootécnico e as características bromatológicas e hematológicas de pacus alimentados a diferentes frequências de arraçoamento. Três mil e duzentos peixes, com peso inicial médio de 65,9±2,36 g, foram distribuídos em 16 tanques-rede de 5 m³ de volume útil. Durante 65 dias, foram avaliadas quatro frequências de arraçoamento (tratamentos): T1, às 12:00 h; T2, às 8:00 e às 17:00 h; T3, às 8:00, 12:00 e 17:00 h; e T4, às 8:00, 11:00, 14:00 e 17:00 h. Utilizou-se o delineamento experimental inteiramente casualizado, com quatro repetições. Analisaram-se parâmetros de desempenho produtivo, índices de gordura visceral e hepatossomático, composição centesimal da carcaça e parâmetros hematológicos e bioquímicos. Quanto ao ganho de peso, as frequências T3 e T4 proporcionaram melhores resultados do que T1 e T2. Embora a proteína, o colesterol e os eritrócitos tenham apresentado diferenças entre os tratamentos, permaneceram dentro dos valores de referência para pacus cultivados em tanques-rede. A frequência de três arraçoamentos diários resultou em maior ganho de peso, em comparação aos demais tratamentos, sem interferir na saúde e na composição centesimal das carcaças dos animais.

Vitamina A em dieta de juvenis de pacu cultivados em tanques-rede

O objetivo deste trabalho foi avaliar o efeito da suplementação de vitamina A na alimentação de juvenis de pacu cultivados em tanques-rede. Foram utilizados 2.000 peixes, com peso inicial médio de 66,93±15,03 g, distribuídos em delineamento inteiramente casualizado, em 20 tanques-rede de 5,0 m³, com cinco tratamentos e quatro repetições. As rações foram formuladas com 0, 3.000, 6.000, 9.000 e 12.000 UI de vitamina A por quilograma de dieta, por meio da suplementação com acetato de retinol. O arraçoamento foi realizado às 8:30, 13:30 e 17:30 h, até a saciedade aparente dos animais. Não foram observadas diferenças quanto aos parâmetros de desempenho produtivo, rendimento e composição química da carcaça, e aos parâmetros hematológicos e lipídios no fígado dos peixes que foram alimentados com diferentes níveis de vitamina A. A suplementação de vitamina A em dietas para juvenis de pacu criados em tanques-rede não influencia o desempenho produtivo dos peixes.

Parasitofauna de cachara cultivado em tanque-rede no rio Paraguai

O objetivo deste trabalho foi descrever a fauna parasitária de cachara (Pseudoplatystoma reticulatum) cultivado em tanque-rede, no rio Paraguai. Dez peixes com peso médio de 598,0±81,3 g e comprimento total médio de 38,6±1,6 cm foram examinados. Todos os peixes necropsiados apresentaram infestação por pelo menos duas espécies de parasitos. Entre os parasitos, foram encontrados Ichthyophthirius multifiliis (Ciliophora), Myxobolus sp. e Henneguya sp. (Myxozoa), Monogenoidea, Choanoscolex abscissus e Nominoscolex sudobim (Cestoda), Dolops carvalhoi (Crustacea) e Digenea. O protozoário Ichthyophthirius multifiliis foi o parasito com maior prevalência.

Estratificação ambiental e otimização de rede de ensaios de genótipos de soja no Cerrado

O objetivo deste trabalho foi estabelecer uma estratificação ambiental consistente, para a recomendação e a avaliação de linhagens experimentais e cultivares de soja na região do Cerrado, a partir de análises da interação entre genótipos e ambientes (GxA) quanto à produtividade de grãos, além de avaliar a atual rede de ensaios de valor de cultivo e uso (VCU) para sua otimização. Os dados provieram de 559 ensaios de competição de linhagens de soja, realizados em 57 localidades, durante sete safras agrícolas (2002/2003 a 2008/2009). Realizaram-se análises conjuntas de variância, pelo modelo AMMI ("additive main effects and multiplicative interaction"), e de estratificação ambiental, pela abordagem correlata de "genótipos vencedores". A interação GxA foi sempre significativa, como resultado da resposta diferencial dos genótipos à variação ambiental. Os locais de teste se agruparam de modo diferente de acordo com os grupos de maturação. Observou-se redundância em 20% dos locais, o que indica a possibilidade de otimização da rede de ensaios, via eliminação ou substituição dessas localidades. A região-alvo deve receber estratificações distintas, congêneres a cada grupo de maturação, e pode ser dividida em 22 (ciclo precoce), 23 (ciclo médio) e 21 (ciclo tardio) estratos ambientais.

Six new loci associated with body mass index highlight a neuronal influence on body weight regulation.

Common variants at only two loci, FTO and MC4R, have been reproducibly associated with body mass index (BMI) in humans. To identify additional loci, we conducted meta-analysis of 15 genome-wide association studies for BMI (n > 32,000) and followed up top signals in 14 additional cohorts (n > 59,000). We strongly confirm FTO and MC4R and identify six additional loci (P < 5 x 10(-8)): TMEM18, KCTD15, GNPDA2, SH2B1, MTCH2 and NEGR1 (where a 45-kb deletion polymorphism is a candidate causal variant). Several of the likely causal genes are highly expressed or known to act in the central nervous system (CNS), emphasizing, as in rare monogenic forms of obesity, the role of the CNS in predisposition to obesity.

Triodothyronine enhancement of neuronal differentiation in aggregating fetal rat brain cells cultured in a chemically defined medium.

Triiodothyronine (30 nM) added to serum-free cultures of mechanically dissociated re-aggregating fetal (15-16 days gestation) rat brain cells greatly increased the enzymatic activity of choline acetyltransferase and acetylcholinesterase throughout the entire culture period (33 days), and markedly accelerated the developmental rise of glutamic acid decarboxylase specific activity. The enhancement of choline acetyltransferase and acetylcholinesterase specific activities in the presence of triiodothyronine was even more pronouned in cultures of telencephalic cells. If triiodothyronine treatment was restricted to the first 17 culture days, the level of choline acetyltransferase specific activity at day 33 was 84% of that in chronically treated cultures and 270% of that in cultures receiving triiodothyronine between days 17 and 33, indicating that relatively undifferentiated cells were more responsive to the hormone. Triiodothyronine had no apparent effect on the incorporation of [3H]thymidine at day 5 or on the total DNA content of cultures, suggesting that cellular differentiation, rather than proliferation was affected by the hormone. Our findings in vitro are in good agreement with many observations in vivo, suggesting that rotation-mediated aggregating cell cultures of fetal rat brain provide a useful model to study thyroid hormone action in the developing brain.

Neuroligin-1 links neuronal activity to sleep-wake regulation.

Maintaining wakefulness is associated with a progressive increase in the need for sleep. This phenomenon has been linked to changes in synaptic function. The synaptic adhesion molecule Neuroligin-1 (NLG1) controls the activity and synaptic localization of N-methyl-d-aspartate receptors, which activity is impaired by prolonged wakefulness. We here highlight that this pathway may underlie both the adverse effects of sleep loss on cognition and the subsequent changes in cortical synchrony. We found that the expression of specific Nlg1 transcript variants is changed by sleep deprivation in three mouse strains. These observations were associated with strain-specific changes in synaptic NLG1 protein content. Importantly, we showed that Nlg1 knockout mice are not able to sustain wakefulness and spend more time in nonrapid eye movement sleep than wild-type mice. These changes occurred with modifications in waking quality as exemplified by low theta/alpha activity during wakefulness and poor preference for social novelty, as well as altered delta synchrony during sleep. Finally, we identified a transcriptional pathway that could underlie the sleep/wake-dependent changes in Nlg1 expression and that involves clock transcription factors. We thus suggest that NLG1 is an element that contributes to the coupling of neuronal activity to sleep/wake regulation.

Transfer entropy reconstruction and labeling of neuronal connections from simulated calcium imaging

Neuronal dynamics are fundamentally constrained by the underlying structural network architecture, yet much of the details of this synaptic connectivity are still unknown even in neuronal cultures in vitro. Here we extend a previous approach based on information theory, the Generalized Transfer Entropy, to the reconstruction of connectivity of simulated neuronal networks of both excitatory and inhibitory neurons. We show that, due to the model-free nature of the developed measure, both kinds of connections can be reliably inferred if the average firing rate between synchronous burst events exceeds a small minimum frequency. Furthermore, we suggest, based on systematic simulations, that even lower spontaneous inter-burst rates could be raised to meet the requirements of our reconstruction algorithm by applying a weak spatially homogeneous stimulation to the entire network. By combining multiple recordings of the same in silico network before and after pharmacologically blocking inhibitory synaptic transmission, we show then how it becomes possible to infer with high confidence the excitatory or inhibitory nature of each individual neuron.

Neuronal survival induced by neurotrophins requires calmodulin

It has been reported that phosphoinositide 3-kinase (PI 3-kinase) and its downstream target, protein kinase B (PKB), play a central role in the signaling of cell survival triggered by neurotrophins (NTs). In this report, we have analyzed the involvement of Ca2+ and calmodulin (CaM) in the activation of the PKB induced by NTs. We have found that reduction of intracellular Ca2+ concentration or functional blockade of CaM abolished NGF-induced activation of PKB in PC12 cells. Similar results were obtained in cultures of chicken spinal cord motoneurons treated with brain-derived neurotrophic factor (BDNF). Moreover, CaM inhibition prevented the cell survival triggered by NGF or BDNF. This effect was counteracted by the transient expression of constitutive active forms of the PKB, indicating that CaM regulates NT-induced cell survival through the activation of the PKB. We have investigated the mechanisms whereby CaM regulates the activation of the PKB, and we have found that CaM was necessary for the proper generation and/or accumulation of the products of the PI 3-kinase in intact cells.

Lack of GDAP1 induces neuronal calcium and mitochondrial defects in a knockout mouse model of charcot-marie-tooth neuropathy.

Mutations in GDAP1, which encodes protein located in the mitochondrial outer membrane, cause axonal recessive (AR-CMT2), axonal dominant (CMT2K) and demyelinating recessive (CMT4A) forms of Charcot-Marie-Tooth (CMT) neuropathy. Loss of function recessive mutations in GDAP1 are associated with decreased mitochondrial fission activity, while dominant mutations result in impairment of mitochondrial fusion with increased production of reactive oxygen species and susceptibility to apoptotic stimuli. GDAP1 silencing in vitro reduces Ca2+ inflow through store-operated Ca2+ entry (SOCE) upon mobilization of endoplasmic reticulum (ER) Ca2+, likely in association with an abnormal distribution of the mitochondrial network. To investigate the functional consequences of lack of GDAP1 in vivo, we generated a Gdap1 knockout mouse. The affected animals presented abnormal motor behavior starting at the age of 3 months. Electrophysiological and biochemical studies confirmed the axonal nature of the neuropathy whereas histopathological studies over time showed progressive loss of motor neurons (MNs) in the anterior horn of the spinal cord and defects in neuromuscular junctions. Analyses of cultured embryonic MNs and adult dorsal root ganglia neurons from affected animals demonstrated large and defective mitochondria, changes in the ER cisternae, reduced acetylation of cytoskeletal α-tubulin and increased autophagy vesicles. Importantly, MNs showed reduced cytosolic calcium and SOCE response. The development and characterization of the GDAP1 neuropathy mice model thus revealed that some of the pathophysiological changes present in axonal recessive form of the GDAP1-related CMT might be the consequence of changes in the mitochondrial network biology and mitochondria-endoplasmic reticulum interaction leading to abnormalities in calcium homeostasis.

Review of quantitative phase-digital holographic microscopy: promising novel imaging technique to resolve neuronal network activity and identify cellular biomarkers of psychiatric disorders.

Quantitative phase microscopy (QPM) has recently emerged as a new powerful quantitative imaging technique well suited to noninvasively explore a transparent specimen with a nanometric axial sensitivity. In this review, we expose the recent developments of quantitative phase-digital holographic microscopy (QP-DHM). Quantitative phase-digital holographic microscopy (QP-DHM) represents an important and efficient quantitative phase method to explore cell structure and dynamics. In a second part, the most relevant QPM applications in the field of cell biology are summarized. A particular emphasis is placed on the original biological information, which can be derived from the quantitative phase signal. In a third part, recent applications obtained, with QP-DHM in the field of cellular neuroscience, namely the possibility to optically resolve neuronal network activity and spine dynamics, are presented. Furthermore, potential applications of QPM related to psychiatry through the identification of new and original cell biomarkers that, when combined with a range of other biomarkers, could significantly contribute to the determination of high risk developmental trajectories for psychiatric disorders, are discussed.

Identification of neuronal network properties from the spectral analysis of calcium imaging signals in neuronal cultures

Neuronal networks in vitro are prominent systems to study the development of connections in living neuronal networks and the interplay between connectivity, activity and function. These cultured networks show a rich spontaneous activity that evolves concurrently with the connectivity of the underlying network. In this work we monitor the development of neuronal cultures, and record their activity using calcium fluorescence imaging. We use spectral analysis to characterize global dynamical and structural traits of the neuronal cultures. We first observe that the power spectrum can be used as a signature of the state of the network, for instance when inhibition is active or silent, as well as a measure of the network's connectivity strength. Second, the power spectrum identifies prominent developmental changes in the network such as GABAA switch. And third, the analysis of the spatial distribution of the spectral density, in experiments with a controlled disintegration of the network through CNQX, an AMPA-glutamate receptor antagonist in excitatory neurons, reveals the existence of communities of strongly connected, highly active neurons that display synchronous oscillations. Our work illustrates the interest of spectral analysis for the study of in vitro networks, and its potential use as a network-state indicator, for instance to compare healthy and diseased neuronal networks.

Methylglyoxal Produced by Amyloid- Peptide-Induced Nitrotyrosination of Triosephosphate Isomerase Triggers Neuronal Death in Alzheimer’s Disease

Amyloid-β peptide (Aβ) aggregates induce nitro-oxidative stress, contributing to the characteristic neurodegeneration found in Alzheimer's disease (AD). One of the most strongly nitrotyrosinated proteins in AD is the triosephosphate isomerase (TPI) enzyme which regulates glycolytic flow, and its efficiency decreased when it is nitrotyrosinated. The main aims of this study were to analyze the impact of TPI nitrotyrosination on cell viability and to identify the mechanism behind this effect. In human neuroblastoma cells (SH-SY5Y), we evaluated the effects of Aβ42 oligomers on TPI nitrotyrosination. We found an increased production of methylglyoxal (MG), a toxic byproduct of the inefficient nitro-TPI function. The proapoptotic effects of Aβ42 oligomers, such as decreasing the protective Bcl2 and increasing the proapoptotic caspase-3 and Bax, were prevented with a MG chelator. Moreover, we used a double mutant TPI (Y165F and Y209F) to mimic nitrosative modifications due to Aβ action. Neuroblastoma cells transfected with the double mutant TPI consistently triggered MG production and a decrease in cell viability due to apoptotic mechanisms. Our data show for the first time that MG is playing a key role in the neuronal death induced by Aβ oligomers. This occurs because of TPI nitrotyrosination, which affects both tyrosines associated with the catalytic center.