Positive and negative regulator Junb : impact on chromatin remodeling and stress response

The AP-1 transcription factor is a central player in a multitude of biological processes from normal development to neoplastic transformation causing cancer. Junb, a subunit of AP-1, is special by the fact that it has at the same time activator and repressor functions. While positively regulated Junb target genes are principally required for proper vascular development, negative regulation of cytokines is of crucial importance to suppress pro-inflammatory and tumorigenic phenotypes. In this work, I approached this double-edge role of Junb by addressing two scientific questions: the mode of operation of Junb as negative transcription regulator and its impact in the ER stress response and apoptosis. First, I could show that, in addition to the general view of being an inhibitor of AP-1 by absorbing Jun activity, Junb also represses genes by epigenetic mechanisms. Although Junb did regulate neither the levels of histone acetylation nor the expression of HDACs, DNMTs and co-repressor complexes, few genes showed differential induction by HDAC inhibitors in wild-type and Junb-deficient fibroblasts. Presumably, these genes may be regulated through a yet to be identified Junb-dependent mechanism involving HDACs. Moreover, Junb regulated the DNA methylation of the imprinting control region of the gene H19. The molecular mechanisms involved in Junb-dependent epigenetic regulation appear to be novel and very unusual for an AP-1 member and remains to be fully solved. Secondly, I investigated the role of Junb in ER stress, a condition that has been described to contribute to hypoxia tolerance and tumor progression. Although Junb deficiency resulted in minor changes in the ER stress-triggered unfolded protein response (UPR), Junb-ablated MEFs were resistant towards apoptosis. Very high levels of activated pro-survival kinases resulted in aberrant post-translational modification of BH3-only proteins Bim and Bad and subsequent failure in mitochondria permeabilization and caspases activation. A soluble factor, most likely Pdgfb, elicited a pro-survival autocrine loop causative for the apoptosis resistance in absence of Junb. In summary, the negative regulation of cytokines and growth factors by Junb accounts for most of the deleterious effects observed in absence of Junb, except for the angiogenesis phenotype. Thus, the understanding of how Junb represses genes and the targeting of this specific mechanism would represent a promising therapeutic approach to treat in the future inflammatory disease and cancer

Computational Modeling of Motile Cilia-Driven Cerebrospinal Flow in the Brain Ventricles of Zebrafish Embryo.

Motile cilia are hair-like microscopic structures which generate directional flow to provide fluid transport in various biological processes. Ciliary beating is one of the sources of cerebrospinal flow (CSF) in brain ventricles. In this study, we investigated how the tilt angle, quantity, and phase relationship of cilia affect CSF flow patterns in the brain ventricles of zebrafish embryos. For this purpose, two-dimensional computational fluid dynamics (CFD) simulations are performed to determine the flow fields generated by the motile cilia. The cilia are modeled as thin membranes with prescribed motions. The cilia motions were obtained from a two-day post-fertilization zebrafish embryo previously imaged via light sheet fluorescence microscopy. We observed that the cilium angle significantly alters the generated flow velocity and mass flow rates. As the cilium angle gets closer to the wall, higher flow velocities are observed. Phase difference between two adjacent beating cilia also affects the flow field as the cilia with no phase difference produce significantly lower mass flow rates. In conclusion, our simulations revealed that the most efficient method for cilia-driven fluid transport relies on the alignment of multiple cilia beating with a phase difference, which is also observed in vivo in the developing zebrafish brain.Part of this work was supported by a FRIPRO grant from the Research Council of Norway (N.J.-Y. grant number 314189). The publication of this article was funded by the Qatar National Library

Elevated photic response is followed by a rapid decay and depressed state in ictogenic networks

Objective: The switch between nonseizure and seizure states involves profound alterations in network excitability and synchrony. In this study, we aimed to identify and compare features of neural excitability and dynamics across multiple zebrafish seizure and epilepsy models. Methods: Inspired by video-electroencephalographic recordings in patients, we developed a framework to study spontaneous and photically evoked neural and locomotor activity in zebrafish larvae, by combining high-throughput behavioral tracking and whole-brain in vivo two-photon calcium imaging. Results: Our setup allowed us to dissect behavioral and physiological features that are divergent or convergent across multiple models. We observed that spontaneous locomotor and neural activity exhibit great diversity across models. Nonetheless, during photic stimulation, hyperexcitability and rapid response dynamics were well conserved across multiple models, highlighting the reliability of photically evoked activity for high-throughput assays. Intriguingly, in several models, we observed that the initial elevated photic response is often followed by rapid decay of neural activity and a prominent depressed state. Elevated photic response and following depressed state in seizure-prone networks are significantly reduced by the antiseizure medication valproic acid. Finally, rapid decay and depression of neural activity following photic stimulation temporally overlap with slow recruitment of astroglial calcium signals that are enhanced in seizure-prone networks. Significance: We argue that fast decay of neural activity and depressed states following photic response are likely due to homeostatic mechanisms triggered by excessive neural activity. An improved understanding of the interplay between elevated and depressed excitability states might suggest tailored epilepsy therapies. Keywords: astroglia; calcium imaging; depressed state; elevated state; epilepsy; high-throughput behavior; hyperexcitability; photic stimulation; seizure; zebrafis

Past, present and future of zebrafish in epilepsy research

Animal models contribute greatly to our understanding of brain development and function as well as its dysfunction in neurological diseases. Epilepsy research is a very good example of how animal models can provide us with a mechanistic understanding of the genes, molecules, and pathophysiological processes involved in disease. Over the course of the last two decades, zebrafish came in as a new player in epilepsy research, with an expanding number of laboratories using this animal to understand epilepsy and to discover new strategies for preventing seizures. Yet, zebrafish as a model offers a lot more for epilepsy research. In this viewpoint, we aim to highlight some key contributions of zebrafish to epilepsy research, and we want to emphasize the great untapped potential of this animal model for expanding these contributions. We hope that our suggestions will trigger further discussions between clinicians and researchers with a common goal to understand and cure epilepsy

The role of motile cilia in the development and physiology of the nervous system

Motile cilia are miniature, whip-like organelles whose beating generates a directional fluid flow. The flow generated by ciliated epithelia is a subject of great interest, as defective ciliary motility results in severe human diseases called motile ciliopathies. Despite the abundance of motile cilia in diverse organs including the nervous system, their role in organ development and homeostasis remains poorly understood. Recently, much progress has been made regarding the identity of motile ciliated cells and the role of motile-cilia-mediated flow in the development and physiology of the nervous system. In this review, we will discuss these recent advances from sensory organs, specifically the nose and the ear, to the spinal cord and brain ventricles

Loss of glutamate transporter eaat2a leads to aberrant neuronal excitability, recurrent epileptic seizures, and basal hypoactivity

Astroglial excitatory amino acid transporter 2 (EAAT2, GLT-1, and SLC1A2) regulates the duration and extent of neuronal excitation by removing glutamate from the synaptic cleft. Hence, an impairment in EAAT2 function could lead to an imbalanced brain network excitability. Here, we investigated the functional alterations of neuronal and astroglial networks associated with the loss of function in the astroglia predominant eaat2a gene in zebrafish. We observed that eaat2a-/- mutant zebrafish larvae display recurrent spontaneous and light-induced seizures in neurons and astroglia, which coincide with an abrupt increase in extracellular glutamate levels. In stark contrast to this hyperexcitability, basal neuronal and astroglial activity was surprisingly reduced in eaat2a-/- mutant animals, which manifested in decreased overall locomotion. Our results reveal an essential and mechanistic contribution of EAAT2a in balancing brain excitability, and its direct link to epileptic seizures. Keywords: astroglia; brain excitability; calcium imaging; eaat2; epilepsy; glutamate; zebrafish

Loss of glutamate transporter eaat2a leads to aberrant neuronal excitability, recurrent epileptic seizures, and basal hypoactivity

Astroglial excitatory amino acid transporter 2 (EAAT2, GLT-1, and SLC1A2) regulates the duration and extent of neuronal excitation by removing glutamate from the synaptic cleft. Hence, an impairment in EAAT2 function could lead to an imbalanced brain network excitability. Here, we investigated the functional alterations of neuronal and astroglial networks associated with the loss of function in the astroglia predominant eaat2a gene in zebrafish. We observed that eaat2a−/− mutant zebrafish larvae display recurrent spontaneous and light-induced seizures in neurons and astroglia, which coincide with an abrupt increase in extracellular glutamate levels. In stark contrast to this hyperexcitability, basal neuronal and astroglial activity was surprisingly reduced in eaat2a−/− mutant animals, which manifested in decreased overall locomotion. Our results reveal an essential and mechanistic contribution of EAAT2a in balancing brain excitability, and its direct link to epileptic seizures

Motile-Cilia-Mediated Flow Improves Sensitivity and Temporal Resolution of Olfactory Computations

Motile cilia are actively beating hair-like structures that cover the surface of multiple epithelia. The flow that ciliary beating generates is utilized for diverse functions and depends on the spatial location and biophysical properties of cilia. Here we show that the motile cilia in the nose of aquatic vertebrates are spatially organized and stably beat with an asymmetric pattern, resulting in a robust and stereotypical flow around the nose. Our results demonstrate that these flow fields attract odors to the nose pit and facilitate detection of odors by the olfactory system in stagnant environments. Moreover, we show that ciliary beating quickly exchanges the content of the nose, thereby improving the temporal resolution of the olfactory system for detecting dynamic changes of odor plumes in turbulent environments. Altogether, our work unravels a central function of ciliary beating for generating flow fields that increase the sensitivity and the temporal resolution of olfactory computations in the vertebrate brain