Biology subjects

Hosoi, N.

Publications and source records attributed to Hosoi, N..

4 recordsLinked to original sources

bioRxiv2025

A compact glutamic acid decarboxylase 67 promoter enables inhibitory neuron-targeted AAV gene therapy for treatment-resistant epilepsy

Epilepsy often becomes treatment-resistant, partly due to impaired inhibitory neurotransmission and reduced {gamma}-aminobutyric acid (GABA) function. Enhancing inhibitory neuron activity via gene therapy may restore excitation-inhibition (E/I) balance. We developed a compact 410-bp glutamic acid decarboxylase 67 promoter (cmGAD67) that enables strong, selective transgene expression in inhibitory neurons while preserving AAV packaging capacity. When delivered systemically, AAV vectors carrying cmGAD67 preferentially targeted parvalbumin interneurons and supported effective circuit manipulation. To evaluate therapeutic potential, we expressed glutamic acid decarboxylase 65 (GAD65) under cmGAD67 (AAV-GAD65) in pentylenetetrazole (PTZ) epilepsy models. Systemic AAV-GAD65 suppressed abnormal delta oscillations, reduced seizure-like events, normalized anxiety-like behavior, and improved survival in a severe PTZ paradigm. Biochemical analyses confirmed increased cortical and hippocampal GABA levels, linking behavioral and electrophysiological improvements to enhanced inhibitory neurotransmitter synthesis. Prior clinical evidence indicates that AAV-GAD65 delivery to the subthalamic nucleus is safe and effective in Parkinsons disease. Building on this foundation, our findings establish the cmGAD67 promoter as a powerful platform for inhibitory neuron-targeted AAV gene therapy and highlight AAV-cmGAD67-GAD65 as a promising approach for treatment-resistant epilepsy and other disorders involving disrupted E/I balance.

neuroscience↗

bioRxiv2025

Axis-specific neuro-musculoskeletal contributions to postural stability and vibration avoidance in mice

Animals must appropriately regulate their responses to external mechanical oscillations such as whole-body (WB) vibration to ensure postural stability. However, knowledge of mouse responses to WB vibration remains limited. Here, we used high-speed video-based quantitative analyses to characterize body-part movements in awake and anesthetized mice under vertical, longitudinal, and lateral WB vibration. Awake mice exhibited lower resonance frequencies and smaller displacement amplitudes than anesthetized mice during vertical and lateral WB vibration, but not longitudinal vibration, indicating that the neuro-musculoskeletal (NMS) system contributes to postural stability in a vibration axis-dependent manner. Vibration modeling suggests that the NMS system acts as an active vibration absorber by reducing effective stiffness and dynamically adjusting effective damping, analogous to active vehicle suspensions. Axis-specific vibration control may reflect an evolutionary balance between postural stability and locomotor efficiency in quadrupeds, paralleling the design principles of rear-wheel-drive vehicles. Behavioral preference tests revealed that mice selectively avoid vertical WB vibration at specific frequencies, but neither lateral nor longitudinal WB vibration. These findings reveal an axis-specific NMS control principle analogous to vehicle design, linking postural regulation, locomotor efficiency and avoidance behavior, and advancing our understanding of the biomechanics and behavior of quadrupeds exposed to WB vibration.

biophysics↗

bioRxiv2024

Optimization of microRNA Target Sequence Arrangement in AAV Vectors Dramatically Enhances Specificity and Efficiency of Transgene Expression in Cortical Microglia

Microglia play a critical role in diseases such as Alzheimers and stroke, making them a significant target for therapeutic intervention. However, due to their immune functions in detecting and combating viral invasion, efficient gene delivery to microglia remains challenging. We achieved specific and efficient gene delivery to microglia using an adeno- associated virus (AAV) vector designed for this purpose. This microglia-targeting AAV vector includes the mouse microglia/macrophage-specific ionized calcium-binding adaptor molecule 1 (mIba1) promoter, green fluorescent protein (GFP), microRNA target sequences (miR.Ts), woodchuck hepatitis virus posttranscriptional regulatory element (WPRE), and polyadenylation (polyA) signal, positioned between inverted terminal repeats. When miR.Ts were placed downstream of WPRE (between WPRE and polyA), gene expression occurred not only in microglia but also in a substantial number of neurons. However, when miR.Ts were positioned upstream of WPRE (between GFP and WPRE) or on both sides of WPRE, neuronal expression was significantly suppressed, resulting in selective GFP expression in microglia. Notably, positioning miR.Ts on both sides of WPRE achieved over 90% specificity and more than 60% efficiency in transgene expression in microglia three weeks after viral administration. This vector also enabled GCaMP expression in microglia, facilitating real-time monitoring of calcium dynamics and microglial process activity in the cortex. Additionally, intravenous administration of this vector with the blood-brain barrier-penetrant AAV-9P31 capsid variant resulted in extensive GFP expression selectively in microglia throughout the brain. These findings establish this AAV vector system as a robust tool for long-term, specific, and efficient gene expression in microglia.

neuroscience↗

bioRxiv2018

Deletion of class II ARF causes essential tremors through Nav1.6 traffic impairment

ADP-ribosylation factors (ARFs) are a family of small monomeric GTPases consisting of three classes. In the present study, we generated class II ARF-deficient mice (ARF4+/-/ARF5-/-) and found that they exhibited severe movement-associated tremors. Treatment of the mice with propranolol and gabapentin, which alleviate symptoms in patients with essential tremors, similarly reduced the amplitude of the pathologic tremors. In vivo electrophysiological recordings of the ARF4+/-/ARF5-/- mice revealed that they exhibited reduced excitability of their cerebellar Purkinje cells. Immunohistochemical studies revealed that ARF4+/-/ARF5-/- mice exhibit a severe, selective reduction of Nav1.6 proteins that are important for maintaining repetitive action potential firing in the axon initial segments (AISs) of the Purkinje cells. This decrease in Nav1.6 protein expression and the consequent tremors were alleviated by Purkinje cell-specific expression of ARF5. These results indicate that class II ARF mediates the selective trafficking of Nav1.6 to the AISs in cerebellar Purkinje cells, and suggest that the essential tremors can be ascribed to the reduced intrinsic excitability of Purkinje cells, caused by the selective decrease of Nav1.6 proteins in the AISs.

neuroscience↗