Functions of GCN5 and P /CAF acetyltransferases in mammalian development

Histone acetyltransferases are important chromatin modifiers that function as transcriptional co-activators. The identification of the transcriptional regulator GCN5 as the first nuclear histone acetyltransferase in yeast directly linked chromatin remodeling to transcriptional regulation. Although emerging evidence suggests that acetyltransferases participate in multiple cellular processes, their roles in mammalian development remain undefined. In this study, I have cloned and characterized the mouse homolog of GCN5 and a closely related protein P/CAF that interacts with p300/CBP. In contrast to yeast GCN5, but similar to P/CAF, mouse GCN5 possesses an additional N-terminal domain that confers the ability to acetylate nucleosomal histones. GCN5 and P/CAF exhibit identical substrate specificity and both interact with p300/CBP. Interestingly, expression levels of GCN5 and P/CAF display a complementary pattern in mouse embryos and in adult tissues, suggesting that they have distinct tissue or developmental stage specific roles. To define the in vivo function of GCN5 and P/CAF, I have generated mice that are nullizygous for GCN5 or P/CAF. P/CAF null mice are viable and fertile with no gross morphological defects, indicating that P/CAF is dispensable for development and p300/CBP function in vivo. In contrast, mice lacking GCN5 die between 10.5–11 days of gestation. GCN5 null mice are severely retarded but have anterior ectopic outgrowth. Molecular marker analyses reveal that early mesoderm is formed in GCN5 null mice but further differentiation into distinct mesodermal lineages is perturbed. While presomitic mesoderm and chodamesoderm are missing in GCN5 mutant mice, extraembryonic tissues and lateral mesoderm are unaffected. This is consistent with our finding that GCN5 expression is absent in the heart and extraembryonic tissues but is uniform throughout the rest of the embryo. Remarkably, GCN5 mutant mice exhibit an unusually high incidence of apoptosis in the embryonic ectoderm and mesoderm. Finally, mice doubly null for GCN5 and P/CAF die much earlier than mice harboring the GCN5 mutation alone, suggesting that P/CAF and GCN5 share some overlapping function during embryogenesis. This work is the first study to show that specific acetyltransferase is important for cell survival as well as mesoderm differentiation or maintenance during early mammalian development. ^

Seawater carbonate chemistry and auditory behavior of Orange clownfish (Amphiprion percula) during experiments, 2011

Ocean acidification is predicted to affect marine ecosystems in many ways, including modification of fish behaviour. Previous studies have identified effects of CO2-enriched conditions on the sensory behaviour of fishes, including the loss of natural responses to odours resulting in ecologically deleterious decisions. Many fishes also rely on hearing for orientation, habitat selection, predator avoidance and communication. We used an auditory choice chamber to study the influence of CO2-enriched conditions on directional responses of juvenile clownfish (Amphiprion percula) to daytime reef noise. Rearing and test conditions were based on Intergovernmental Panel on Climate Change predictions for the twenty-first century: current-day ambient, 600, 700 and 900 µatm pCO2. Juveniles from ambient CO2-conditions significantly avoided the reef noise, as expected, but this behaviour was absent in juveniles from CO2-enriched conditions. This study provides, to our knowledge, the first evidence that ocean acidification affects the auditory response of fishes, with potentially detrimental impacts on early survival.

Inner-Hair Cells Parameterized-Hardware Implementation for Personalized Auditory Nerve Stimulation

In this paper the hardware implementation of an inner hair cell model is presented. Main features of the design are the use of Meddis’ transduction structure and the methodology for Design with Reusability. Which allows future migration to new hardware and design refinements for speech processing and custom-made hearing aids

Dyrk1A Influences Neuronal Morphogenesis Through Regulation of Cytoskeletal Dynamics in Mammalian Cortical Neurons

Down syndrome (DS) is the most frequent genetic cause of mental retardation. Cognitive dysfunction in these patients is correlated with reduced dendritic branching and complexity, along with fewer spines of abnormal shape that characterize the cortical neuronal profile of DS. DS phenotypes are caused by the disruptive effect of specific trisomic genes. Here, we report that overexpression of dual-specificity tyrosine phosphorylation-regulated kinase 1A, DYRK1A, is sufficient to produce the dendritic alterations observed in DS patients. Engineered changes in Dyrk1A gene dosage in vivo strongly alter the postnatal dendritic arborization processes with a similar progression than in humans. In cultured mammalian cortical neurons, we determined a reduction of neurite outgrowth and synaptogenesis. The mechanism underlying neurite dysgenesia involves changes in the dynamic reorganization of the cytoskeleton.

A new approach to a model of the mammalian retina with optically programmable logic cells

This paper reports a model of the mammalian retina as well as an interpretation of some functions of the visual cortex. Its main objective is to simulate some of the behaviors observed at the different retina cells depending on the characteristics of the light impinging onto the photoreceptors. This simulation is carried out with a simple structure employed previously as basic building block of some optical computer architectures. Its possibility to perform any type of Boolean function allows a wide range of behaviors.

A model of the mammalian retina and the visual cortex. Disorders of vision

A model of the mammalian retina and the behavior of the first layers in the visual cortex is reported. The building blocks are optically programmable logic cells. A model of the retina, similar to the one reported by Dowling (1987) is presented. From the model of the visual cortex obtained, some types of symmetries and asymmetries are possible to be detected

Image processing based on a model of the mammalian retina

A first study in order to construct a simple model of the mammalian retina is reported. The basic elements for this model are Optical Programmable Logic Cells, OPLCs, previously employed as a functional element for Optical Computing. The same type of circuit simulates the five types of neurons present in the retina. Different responses are obtained by modifying either internal or external connections. Two types of behaviors are reported: symmetrical and non-symmetrical with respect to light position. Some other higher functions, as the possibility to differentiate between symmetric and non-symmetric light images, are performed by another simulation of the first layers of the visual cortex. The possibility to apply these models to image processing is reported.

Unattended motion detection system based on the mammalian retina

Sensing systems in living bodies offer a large variety of possible different configurations and philosophies able to be emulated in artificial sensing systems. Motion detection is one of the areas where different animals adopt different solutions and, in most of the cases, these solutions reflect a very sophisticated form. One of them, the mammalian visual system, presents several advantages with respect to the artificial ones. The main objective of this paper is to present a system, based on this biological structure, able to detect motion, its sense and its characteristics. The configuration adopted responds to the internal structure of the mammalian retina, where just five types of cells arranged in five layers are able to differentiate a large number of characteristics of the image impinging onto it. Its main advantage is that the detection of these properties is based purely on its hardware. A simple unit, based in a previous optical logic cell employed in optical computing, is the basis for emulating the different behaviors of the biological neurons. No software is present and, in this way, no possible interference from outside affects to the final behavior. This type of structure is able to work, once the internal configuration is implemented, without any further attention. Different possibilities are present in the architecture to be presented: detection of motion, of its direction and intensity. Moreover, some other characteristics, as symmetry may be obtained.

Image characterisation based on the mammalian visual cortex

A proposal for a model of the primary visual cortex is reported. It is structured with the basis of a simple unit cell able to perform fourteen pairs of different boolean functions corresponding to the two possible inputs. As a first step, a model of the retina is presented. Different types of responses, according to the different possibilities of interconnecting the building blocks, have been obtained. These responses constitute the basis for an initial configuration of the mammalian primary visual cortex. Some qualitative functions, as symmetry or size of an optical input, have been obtained. A proposal to extend this model to some higher functions, concludes the paper.

A study on the lateralization of the effect of musical imagery on spontaneous otoacoustic emissions

It has been suggested that different pathways through the brain are followed depending on the type of information that is being processed. Although it is now known that there is a continuous exchange of information through both hemispheres, language is considered to be processed by the left hemisphere, where Broca?s and Wernicke?s areas are located. On the other hand, music is thought to be processed mainly by the right hemisphere. According to Sininger Y.S. & Cone- Wesson, B. (2004), there is a similar but contralateral specialization of the human ears; due to the fact that auditory pathways cross-over at the brainstem. A previous study showed an effect of musical imagery on spontaneous otoacoustic emissions (SOAEs) (Perez-Acosta and Ramos-Amezquita, 2006), providing evidence of an efferent influence from the auditory cortex on the basilar membrane. Based on these results, the present work is a comparative study between left and right ears of a population of eight musicians that presented SOAEs. A familiar musical tune was chosen, and the subjects were trained in the task of evoking it after having heard it. Samples of ear-canal signals were obtained and processed in order to extract frequency and amplitude data on the SOAEs. This procedure was carried out before, during and after the musical image creation task. Results were then analyzed to compare the difference between SOAE responses of left and right ears. A clear asymmetrical SOAEs response to musical imagery tasks between left and right ears was obtained. Significant changes of SOAE amplitude related to musical imagery tasks were only observed on the right ear of the subjects. These results may suggest a predominant left hemisphere activity related to a melodic image creation task.

Evaluation of the bioaccumulation kinetics of gold nanorods in vital mammalian organs by mean of TXRF spectrometry

This work presents the first application of total-reflection X-ray fluorescence (TXRF) spectrometry, a new and powerful alternative analytical method, to evaluation of the bioaccumulation kinetics of gold nanorods (GNRs) in various tissues upon intravenous administration in mice. The analytical parameters for developed methodology by TXRF were evaluated by means of the parallel analysis of bovine liver certified reference material samples (BCR-185R) doped with 10 μg/g gold. The average values (n = 5) achieved for gold measurements in lyophilized tissue weight were as follows: recovery 99.7%, expanded uncertainty (k = 2) 7%, repeatability 1.7%, detection limit 112 ng/g, and quantification limit 370 ng/g. The GNR bioaccumulation kinetics was analyzed in several vital mammalian organs such as liver, spleen, brain, and lung at different times. Additionally, urine samples were analyzed to study the kinetics of elimination of the GNRs by this excretion route. The main achievement was clearly differentiating two kinds of behaviors. GNRs were quickly bioaccumulated by highly vascular filtration organs such as liver and spleen, while GNRs do not show a bioaccumulation rates in brain and lung for the period of time investigated. In parallel, urine also shows a lack of GNR accumulation. TXRF has proven to be a powerful, versatile, and precise analytical technique for the evaluation of GNRs content in biological systems and, in a more general way, for any kind of metallic nanoparticles.

Modeling of GRBAS perceptual evaluation using spectral features obtained from an auditory-based filterbank.

Perceptual voice evaluation according to the GRBAS scale is modelled using a linear combination of acoustic parameters calculated after a filter-bank analysis of the recorded voice signals. Modelling results indicate that for breathiness and asthenia more than 55% of the variance of perceptual rates can be explained by such a model, with only 4 latent variables. Moreover, the greatest part of the explained variance can be attributed to only one or two latent variables similarly weighted by all 5 listeners involved in the experiment. Correlation factors between actual rates and model predictions around 0.6 are obtained.

Comparing in vitro, in situ, and in vivo experimental data in a three-dimensional model of mammalian cochlear mechanics

Normal mammalian hearing is refined by amplification of the motion of the cochlear partition. This partition, comprising the organ of Corti sandwiched between the basilar and tectorial membranes, contains the outer hair cells that are thought to drive this amplification process. Force generation by outer hair cells has been studied extensively in vitro and in situ, but, to understand cochlear amplification fully, it is necessary to characterize the role played by each of the components of the cochlear partition in vivo. Observations of cochlear partition motion in vivo are severely restricted by its inaccessibility and sensitivity to surgical trauma, so, for the present study, a computer model has been used to simulate the operation of the cochlea under different experimental conditions. In this model, which uniquely retains much of the three-dimensional complexity of the real cochlea, the motions of the basilar and tectorial membranes are fundamentally different during in situ- and in vivo-like conditions. Furthermore, enhanced outer hair cell force generation in vitro leads paradoxically to a decrease in the gain of the cochlear amplifier during sound stimulation to the model in vivo. These results suggest that it is not possible to extrapolate directly from experimental observations made in vitro and in situ to the normal operation of the intact organ in vivo.

Import of DNA into mammalian nuclei by proteins originating from a plant pathogenic bacterium

Import of DNA into mammalian nuclei is generally inefficient. Therefore, one of the current challenges in human gene therapy is the development of efficient DNA delivery systems. Here we tested whether bacterial proteins could be used to target DNA to mammalian cells. Agrobacterium tumefaciens, a plant pathogen, efficiently transfers DNA as a nucleoprotein complex to plant cells. Agrobacterium-mediated T-DNA transfer to plant cells is the only known example for interkingdom DNA transfer and is widely used for plant transformation. Agrobacterium virulence proteins VirD2 and VirE2 perform important functions in this process. We reconstituted complexes consisting of the bacterial virulence proteins VirD2, VirE2, and single-stranded DNA (ssDNA) in vitro. These complexes were tested for import into HeLa cell nuclei. Import of ssDNA required both VirD2 and VirE2 proteins. A VirD2 mutant lacking its C-terminal nuclear localization signal was deficient in import of the ssDNA–protein complexes into nuclei. Import of VirD2–ssDNA–VirE2 complexes was fast and efficient, and was shown to depended on importin α, Ran, and an energy source. We report here that the bacterium-derived and plant-adapted protein–DNA complex, made in vitro, can be efficiently imported into mammalian nuclei following the classical importin-dependent nuclear import pathway. This demonstrates the potential of our approach to enhance gene transfer to animal cells.