Ryo Aoki

The Habenulo-Raphe Serotonergic Circuit Encodes an Aversive Expectation Value Essential for Adaptive Active Avoidance of Danger

Anticipation of danger at first elicits panic in animals, but later it helps them to avoid the real threat adaptively. In zebrafish, as fish experience more and more danger, neurons in the ventral habenula (vHb) showed tonic increase in the activity to the presented cue and activated serotonergic neurons in the median raphe (MR). This neuronal activity could represent the expectation of a dangerous outcome and be used for comparison with a real outcome when the fish is learning how to escape from a dangerous to a safer environment. Indeed, inhibiting synaptic transmission from vHb to MR impaired adaptive avoidance learning, while panic behavior induced by classical fear conditioning remained intact. Furthermore, artificially triggering this negative outcome expectation signal by optogenetic stimulation of vHb neurons evoked place avoidance behavior. Thus, vHb-MR circuit is essential for representing the level of expected danger and behavioral programming to adaptively avoid potential hazard.

Dual Roles of Notch in Regulation of Apically Restricted Mitosis and Apicobasal Polarity of Neuroepithelial Cells

How the mitosis of neuroepithelial stem cells is restricted to the apical ventricular area remains unclear. In zebrafish, the mosaic eyes(rw306) (moe/epb41l5(rw306)) mutation disrupts the interaction between the putative adaptor protein Moe and the apicobasal polarity regulator Crumbs (Crb), and impairs the maintenance of neuroepithelial apicobasal polarity. While Crb interacts directly with Notch and inhibits its activity, Moe reverses this inhibition. In the moe(rw306) hindbrain, Notch activity is significantly reduced, and the number of cells that proliferate basally away from the apical area is increased. Surprisingly, activation of Notch in the moe(rw306) mutant rescues not only the basally localized proliferation but also the aberrant neuroepithelial apicobasal polarity. We present evidence that the Crb⋅Moe complex and Notch play key roles in a positive feedback loop to maintain the apicobasal polarity and the apical-high basal-low gradient of Notch activity in neuroepithelial cells, both of which are essential for their apically restricted mitosis.

Imaging of Neural Ensemble for the Retrieval of a Learned Behavioral Program

The encoding of long-term associative memories for learned behaviors is a fundamental brain function. Yet, how behavior is stably consolidated and retrieved in the vertebrate cortex is poorly understood. We trained zebrafish in aversive reinforcement learning and measured calcium signals across their entire brain during retrieval of the learned response. A discrete area of dorsal telencephalon that was inactive immediately after training became active the next day. Analysis of the identified area indicated that it was specific and essential for long-term memory retrieval and contained electrophysiological responses entrained to the learning stimulus. When the behavioral rule changed, a rapid spatial shift in the functional map across the telencephalon was observed. These results demonstrate that the retrieval of long-term memories for learned behaviors can be studied at the whole-brain scale in behaving zebrafish in vivo. Moreover, the findings indicate that consolidated memory traces can be rapidly modified during reinforcement learning.

Neuroepithelial cells require fucosylated glycans to guide the migration of vagus motor neuron progenitors in the developing zebrafish hindbrain.

The molecular mechanisms by which neurons migrate and accumulate to form the neural layers and nuclei remain unclear. The formation of vagus motor nuclei in zebrafish embryos is an ideal model system in which to address this issue because of the transparency of the embryos and the availability of established genetic and molecular biological techniques. To determine the genes required for the formation of the vagus motor nuclei, we performed N-ethyl-N-nitrosourea-based mutant screening using a zebrafish line that expresses green fluorescent protein in the motor neurons. In wild-type embryos, the vagus motor neuron progenitors are born in the ventral ventricular zone, then migrate tangentially in the dorsolateral direction, forming the nuclei. However, in towhead (twdrw685) mutant embryos, the vagus motor neuron progenitors stray medially away from the normal migratory pathway and fail to stop in the right location. The twdrw685 mutant has a defect in the GDP-mannose 4,6 dehydratase (gmds) gene, which encodes a key enzyme in the fucosylation pathway. Levels of fucosylated glycans were markedly and specifically reduced in twdrw685 mutant embryos. Cell transplantation analysis revealed that GMDS is not essential in the vagus motor neuron progenitors for correct formation of the vagus motor nuclei, but is required in the neuroepithelial cells that surround the progenitors. Together, these findings suggest that fucosylated glycans expressed in neuroepithelial cells are required to guide the migration of vagus motor neuron progenitors.

Role of the polybasic sequence in the Doc2α C2B domain in dense-core vesicle exocytosis in PC12 cells

J. Neurochem. (2010) 114, 171–181.

A proliferative probiotic Bifidobacterium strain in the gut ameliorates progression of metabolic disorders via microbiota modulation and acetate elevation

The gut microbiota is an important contributor to the worldwide prevalence of metabolic syndrome (MS), which includes obesity and diabetes. The anti-MS effects exerted by Bifidobacterium animalis ssp. lactis GCL2505 (BlaG), a highly proliferative Bifidobacterium strain in the gut, and B. longum ssp. longum JCM1217T (BloJ) were comparatively examined. BlaG treatment reduced visceral fat accumulation and improved glucose tolerance, whereas BloJ had no effect on these parameters. Gut microbial analysis revealed that BlaG exerted stronger effects on the overall bacterial structure of the gut microbiota than BloJ, including enrichment of the genus Bifidobacterium. The levels of acetate and glucagon-like peptide-1 were increased by BlaG treatment in both the gut and plasma, but not by BloJ treatment. Correlation analysis suggested that the elevation of gut acetate levels by BlaG treatment plays a pivotal role in the BlaG-induced anti-MS effects. These findings indicated that BlaG, a highly viable and proliferative probiotic, improves metabolic disorders by modulating gut microbiota, which results in the elevation of SCFAs, especially acetate.

Y-maze avoidance: an automated and rapid associative learning paradigm in zebrafish.

Adult zebrafish are gaining attention in behavioral neuroscience. By combining video tracking and computer-controlled visual cue presentations on a liquid crystal display screen under the tank, we have developed a new method by which zebrafish can be trained to avoid one arm of a Y-shaped tank by presenting a specific color on the floor paired with an electric shock. Within two hours, zebrafish learn to choose the correct arm at an efficiency of 89.0%. In addition, we also developed a graphical user interface to modify the paradigm assessment parameters such as shape of the tank and time schedules.

Measurement of mechanomyogram

We have proposed an approach for measuring mechanomyogram (MMG) by taking advantage of the acoustic impedance matching between liquid and human skin. Using liquid, we could convey the pressure signal of MMG to a piezo-resistive cantilever efficiently. In experiments, the sensor was placed on the skin surface over biceps brachii (a large muscle that is located in the upperarm). The MMG signal, of which the frequency was in range of 10-15 Hz, was able to be detected using silicone oil as the propagating medium. On the other hand, there was no response in the case of using air as the medium. Experimental results also indicated that the proposed sensor was able to detect the vascular oscillations.

An automated platform for high-throughput mouse behavior and physiology with voluntary head-fixation

Recording neural activity during animal behavior is a cornerstone of modern brain research. However, integration of cutting-edge technologies for neural circuit analysis with complex behavioral measurements poses a severe experimental bottleneck for researchers. Critical problems include a lack of standardization for psychometric and neurometric integration, and lack of tools that can generate large, sharable data sets for the research community in a time and cost effective way. Here, we introduce a novel mouse behavioral learning platform featuring voluntary head fixation and automated high-throughput data collection for integrating complex behavioral assays with virtually any physiological device. We provide experimental validation by demonstrating behavioral training of mice in visual discrimination and auditory detection tasks. To examine facile integration with physiology systems, we coupled the platform to a two-photon microscope for imaging of cortical networks at single-cell resolution. Our behavioral learning and recording platform is a prototype for the next generation of mouse cognitive studies.Transgenic approaches and improvements in functional imaging have necessitated an advance in the behavioral toolkit. Here the authors describe an automated high-throughput voluntary head fixation system for training mice on complex psychophysical decision tasks compatible with concurrent two-photon microscopy.

Detection of high-frequency component of mechanomyogram

This paper reports a method to detect high-frequency component of mechanomyogram (a record of muscle sound, MMG). We propose an MMG sensor with sector-shaped piezoresistive cantilevers and measure MMG of humans biceps brachii. Experimental results show that the frequency component of the MMG of biceps brachii is present in 10 Hz-10 kHz whereas the previous studies have not been able to detect above 100 Hz.