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Biology subjects

Takeuchi, R.

Publications and source records attributed to Takeuchi, R..

6 recordsLinked to original sources

Causal Discovery of Synchronous Neural Oscillations based on Jacobian-informed VAR-LiNGAM

The primary objective of system neuroscience is to understand the functional mapping and its causation in the dynamics of the brain network. Some experimental and methodological studies suggest that functional modularity and its hierarchical information processing in the brain network are crucial to understanding the functional role of task-specific or state-specific information flow in the brain. However, because most of the established techniques for detecting effective network structures in the neuroscience research field are strongly based on the "Granger causality" perspective, existing causal discovery methods specified for brain network analysis cannot identify the causal hierarchy in the modular network in the brain due to spurious correlation issues and indistinguishability of causal direction under the Gaussianity of observational noise in a linear system. To address the issues, we developed a causal discovery method for synchronous neural dynamics, called the Jacobian-informed linear non-Gaussian acyclic model, "j-VAR-LiNGAM", by incorporating the information of the Jacobian matrix determined from a phase-coupled oscillator model estimated from observed neural data into the VAR-LiNGAM algorithms. The method was validated by showing that it could extract causal ordering in both synthetic data and empirical neural observed data. Moreover, by analyzing the observed neural oscillatory signals obtained from mice and humans, we confirmed that our method identified causally hierarchical structures in the brain, which aligned with the neurophysiological interpretations. These findings suggested that our proposed method can reveal the neural basis of hierarchical information processing in the brain network.

neuroscience↗

Insulin-like androgenic gland factor regulates the development of internal and external secondary sexual characteristics in juvenile male kuruma prawn Marsupenaeus japonicus

In decapod crustaceans, insulin-like androgenic gland factor (IAG) is a well-known peptide hormone produced by the androgenic gland, a male-specific endocrine organ. In certain species classified within Pleocyemata, experimental manipulation of IAG signaling has successfully altered both internal and external sexual characteristics, leading to sex reversal. Therefore, IAG is considered a key regulator of male sexual differentiation and maturation and is thought to be a crustacean androgen. In the kuruma prawn Marsupenaeus japonicus (Dendrobranchiata), an important fishery species in Japan, a homologous molecule known as Maj-IAG has been identified. However, its physiological function in males remains poorly understood. In this context, we conducted Maj-IAG knockdown (KD) via RNA interference in juvenile male M. japonicus. Over a 9-week period, Maj-IAG KD significantly inhibited the development of the petasma, male-specific external copulatory structure, and parts of the internal reproductive organs, such as the vas deferens and seminal vesicles. In contrast, somatic growth and testicular development were unaffected by Maj-IAG KD. Histological analysis of the testes revealed no evident abnormalities such as degeneration of testicular structures or abnormal arrest of germ cell differentiation. These findings indicate that Maj-IAG partially governs the development of male secondary sexual characteristics in juveniles and further suggest that once testicular differentiation is initiated, the maintenance and development of the testis proceeds without dependence on Maj-IAG. Our study provides fundamental insights that may contribute to the advancement of functional sex reversal technologies in penaeid shrimps. HighlightsO_LILong-term gene knockdown of the insulin-like androgenic gland factor gene (Maj-IAG) was successfully achieved in Marsupenaeus japonicus. C_LIO_LISomatic growth and spermatogenesis were not affected by Maj-IAG knockdown. C_LIO_LIMaj-IAG is involved in the development of male secondary sexual characteristics in juvenile prawns, including the external male copulatory structures and certain parts of the internal male reproductive organs. C_LI

zoology↗

Multiplex Gene Editing Suppresses Random Integration of Hepatitis B Virus DNA in Chronically Infected Liver

Gene editing technologies have opened the possibility of directly targeting viral DNA in therapeutic applications. In chronically infected hepatocytes with hepatitis B virus (HBV), covalently closed circular DNA (cccDNA) serves as the master template for viral transcripts and gene products. In the present study, we evaluated the outcomes of anti-HBV multiplex gene editing with the CRISPR-Cas9 endonuclease from Staphylococcus aureus (SaCas9) using primary human hepatocytes (PHHs) and HBV mouse models. Nonviral delivery of SaCas9-encoding mRNA and a pair of HBV-targeting guide RNAs (gRNAs) substantially reduced viral biomarkers and intrahepatic HBV DNA copies in vitro and in vivo, suggesting that fragmentation of HBV DNA primarily leads to its degradation. Hybridization capture sequencing analyses indicated that small insertions and deletions (indels) and structural variants including excisions and inversions of the viral sequences were accumulated in the residual HBV DNA. These assays also demonstrated that transient expression of the HBV-targeting SaCas9 significantly suppressed random integration of HBV DNA, while this therapeutic approach was unlikely to affect chromosomal translocations involving viral copies. Taken together, our results suggest that anti-HBV multiplex gene editing eliminates viral DNA from chronically infected hepatocytes, potentially reducing the risk of hepatocarcinogenesis associated with HBV DNA integration.

bioengineering↗

Weak and tunable adhesion-clutch drives rapid cell migration and glioblastoma motility

To move forward, migrating cells must exert backward forces against the extracellular environment. Recent studies have highlighted the importance of integrin-independent forces for cell migration; but the molecular machinery that exerts forces remains unclear. Here, we show that the clutch-linker molecule shootin1 and the cell adhesion molecule L1 transmit the backward force of treadmilling actin filaments to the adhesive environment for rapid dendritic cell migration. Notably, shootin1 and L1 transmit weak traction forces, [~]100 times weaker than integrin-based forces, by constituting an integrin-independent slippery adhesion-clutch. This adhesion-clutch system is tunable in response to the chemoattractant CCL19 and the adhesive ligand laminin and mediates chemotaxis through its polarized activation within cells. Furthermore, its aberrant activity enhances glioblastoma cell motility. Our results show that the weak adhesion-clutch is well-suited for rapid cell migration, without forming strong adhesions that impede cell motility, and provides a potential target for inhibiting abnormal cell motility.

cell biology↗

A mosaic adeno-associated virus vector as a versatile tool that exhibits high levels of transgene expression and neuron specificity in primate brain

Recent emphasis has been placed on gene transduction mediated through recombinant adeno-associated virus (AAV) vector to manipulate activity of neurons and their circuitry in the primate brain. In the present study, we created a novel AAV vector of which capsid was composed of capsid proteins derived from the serotypes 1 and 2 (AAV1 and AAV2). Following the injection into the frontal cortex of macaque monkeys, this mosaic vector, termed AAV2.1 vector, was found to exhibit the excellence in transgene expression (for the AAV1 vector) and neuron specificity (for the AAV2 vector) simultaneously. To explore its applicability to chemogenetic manipulation and in vivo calcium imaging, the AAV2.1 vector expressing excitatory DREADDs or GCaMP was injected into the striatum or the visual cortex of macaque monkeys, respectively. Our results have defined that such vectors secure intense and stable expression of the target proteins and yield conspicuous modulation and imaging of neuronal activity.

neuroscience↗

Expression and Manufacturing of Protein Therapeutics in Spirulina

Arthrospira platensis (commonly known as spirulina) is a photosynthetic cyanobacterium1. It is a highly nutritious food that has been consumed for decades in the US, and even longer by indigenous cultures2. Its widespread use as a safe food source and proven scalability have driven frequent attempts to convert it into a biomanufacturing platform. But these were repeatedly frustrated by spirulinas genetic intractability. We report here efficient and versatile genetic engineering methodology for spirulina that allows stable expression of bioactive protein therapeutics at high levels. We further describe large-scale, indoor cultivation and downstream processing methods appropriate for the manufacturing of biopharmaceuticals in spirulina. The potential of the platform is illustrated by pre-clinical development and human testing of an orally delivered antibody therapeutic against campylobacter, a major cause of infant mortality in the developing world and a growing antibiotic resistance threat3,4. This integrated development and manufacturing platform blends the safety of food-based biotechnology with the ease of genetic manipulation, rapid growth rates and high productivity characteristic of microbial platforms. These features combine for exceptionally low-cost production of biopharmaceuticals to address medical needs that are unfeasible with current biotechnology platforms.

molecular biology↗