The rat neostriatum has a mosaic organization composed of striosome/patch compartments embedded in more extensive matrix compartment, which are distinguished from each other by the input-output as well expression many molecular markers. compartment gives rise to dual γ-aminobutyric acid (GABA)ergic striatofugal systems, i.e. direct and indirect pathway neurons, whereas striosome is considered involve neurons alone. Although whole axonal arborization been examined vivo intracellular staining,...

Parvalbumin (PV)-producing fast-spiking neurons are well known to generate gamma oscillation by mutual chemical and electrical connections in the neocortex. Although it was clearly demonstrated that PV form a dense gap junction network with each other not only at proximal sites but also distal dendrites, comprehensive quantitative data on still lacking. To elucidate connectivity, we investigated inhibitory inputs somatosensory cortex, using transgenic mice which dendrites cell bodies of were...

Automated tape-collecting ultramicrotomy in conjunction with scanning electron microscopy (SEM) is a powerful approach for volume and three-dimensional neuronal circuit analysis. Current tapes are limited by section wrinkle formation, surface scratches sample charging during imaging. Here we show that plasma-hydrophilized carbon nanotube (CNT)-coated polyethylene terephthalate (PET) tape effectively resolves these issues produces SEM images of comparable quality to those from transmission...

Odor information is regulated by olfactory inputs, bulbar interneurons, and centrifugal inputs in the bulb (OB). Cholinergic neurons projecting from nucleus of horizontal limb diagonal band Broca magnocellular preoptic are one primary to OB. In this study, we focused on cholinergic regulation OB analyzed neural morphology with a particular emphasis projection pathways neurons. Single-cell imaging specific neuron within dense fibers critical evaluate structure function circuits. We labeled...

Abstract To examine inputs to parvalbumin (PV)‐producing interneurons, we generated transgenic mice expressing somatodendritic membrane‐targeted green fluorescent protein specifically in the and completely visualized their dendrites somata. Using immunolabeling for vesicular glutamate transporter (VGluT)1, VGluT2, GABA transporter, found that VGluT1‐positive terminals made contacts 4‐ 3.1‐fold more frequently with PV‐producing interneurons than VGluT2‐positive GABAergic terminals,...

Olfactory processing is well known to be regulated by centrifugal afferents from other brain regions, such as noradrenergic, acetylcholinergic, and serotonergic neurons. Serotonergic neurons widely innervate regulate the functions of various regions. In present study, we focused on regulation olfactory bulb (OB), one most structurally functionally well-defined Visualization a single neuron among abundant dense fibers essential characterize understand neuronal circuits. We accomplished this...

Visualization of neurons is indispensable for the investigation neuronal circuits in central nervous system. Virus vectors have been widely used labeling particular subsets neurons, and adeno-associated virus (AAV) vector has gained popularity as a tool gene transfer. Here, we developed single AAV Tet-Off platform, AAV-SynTetOff, to improve gene-transduction efficiency, specifically neurons. The platform composed regulator response elements genome. After infection Neuro-2a cells with...

ABSTRACT Dynorphins, leumorphin, and neoendorphins are preprodynorphin (PPD)‐derived peptides ligands for κ‐opioid receptors. Using an antibody to PPD C‐terminal, we investigated the chemical molecular characteristics of PPD‐expressing neurons in mouse neocortex. PPD‐immunopositive neuronal somata were distributed most frequently layer 5 less layers 2–4 6 throughout neocortical regions. Combined labeling immunofluorescence fluorescent mRNA signals revealed that almost all expressed glutamic...

The recurrent network composed of excitatory and inhibitory neurons is fundamental to neocortical function. Inhibitory in the mammalian neocortex are molecularly diverse, individual cell types play unique functional roles microcircuit. Recently, vasoactive intestinal polypeptide-positive (VIP+) neurons, comprising a subclass have attracted particular attention because they can disinhibit pyramidal cells through inhibition other such as parvalbumin- (PV+) somatostatin-positive (SOM+)...

In mammalian neocortex, learning triggers the formation and turnover of new postsynaptic spines on pyramidal cell dendrites. However, biological principles spine reorganization during remain elusive because identity their presynaptic neuronal partners is unknown. Here, we show that two neural circuits supervise distinct programs dynamics to execute learning. We imaged in motor cortex performed post hoc identification afferent neurons. New appeared formed small transient contacts with...

Indirect pathway medium-sized spiny neurons (iMSNs) in the neostriatum are well known to project external segment of globus pallidus (GPe). Although direct MSNs (dMSNs) also send axon collaterals GPe, it remains unclear how dMSNs and iMSNs converge within GPe. Here, we selectively labeled neighboring with green red fluorescent proteins using an adeno-associated virus vector examined axonal projections GPe mice. Both formed two arborizations displaying topographical dorsoventral mediolateral...

Large GABAergic (LG) neurons form a distinct cell type in the inferior colliculus (IC), identified by presence of dense VGLUT2-containing axosomatic terminals. Although some terminals originate from local and commissural IC neurons, it has been unclear whether LG also receive inputs lower auditory brainstem nuclei, i.e., cochlear nuclei (CN), superior olivary complex (SOC), lateral lemniscus (NLL). In this study we injected recombinant viral tracers that force infected cells to express GFP...

Recent progress in psychiatric research has accumulated many mouse models relevant to developmental neuropsychiatric disorders using numerous genetic and environmental manipulations. Since the prefrontal cortex (PFC) is essential for cognitive functions whose impairments are central symptoms associated with humans, it become crucial clarify altered processes of PFC circuits these mice. To that end, we aimed understand a sequence molecular events during normal development. Expression profiles...

We developed an adeno-associated virus (AAV) vector-based technique to label mouse neostriatal neurons comprising direct and indirect pathways with different fluorescent proteins analyze their axonal projections. The AAV vector expresses GFP or RFP in the presence absence of Cre recombinase should be useful for labeling two cell populations exclusively dependent on its expression. Here, we describe design, stereotaxic injection vector, a highly sensitive immunoperoxidase method axon...

In the neocortex, both layer 2/3 and 5 contain corticocortical pyramidal cells projecting to other cortices. We previously found that among L5 of secondary motor cortex (M2), not only intratelencephalic projection but also tract innervate ipsilateral cortices two subtypes are different in diversity axonal laminar distributions. Layer houses intratelencephalically multiple contralateral However, it remained unclear whether can be divided into each with distinct innervation specific targets....

Olfactory input is processed in the glomerulus of main olfactory bulb (OB) and relayed to higher centers brain by projection neurons. Conversely, centrifugal inputs from other regions project OB. We have previously analyzed into OB several using single-neuron labeling. In this study, we noradrenergic (NA) fibers derived locus coeruleus (LC), because their pathways synaptic connections not been clarified detail. NA projections labeling immunoelectron microscopy. Individual neurons labeled...

Abstract Proprioception from muscle spindles is necessary for motor function executed by the cerebellum. In particular, cerebellar nuclear neurons that receive proprioceptive signals and send projections to lower brainstem or spinal cord play key roles in control. However, little known about which regions orofacial proprioception. Here, we investigated nuclei supratrigeminal nucleus (Su5), conveys proprioception arising jaw-closing (JCMSs). Injections of an anterograde tracer into Su5...

脳が，認知・思考・記憶・感情といった高次機能を実現する仕組みを解き明かすには，その構造的基盤である神経回路網の理解が必要不可欠である．「構造無き機能は無い」からである．国内外で続々と開発されている脳透明化技術は，高速かつ大規模な三次元構造解析を可能にする革新的技術であり，神経回路解析に新たなブレイクスルーをもたらすと期待される．透明化技術を比較する上で重要なポイントが「clearing-preservation spectrum」である．透明化能力（clearing capability）の向上は散乱光の抑制につながり，深部まで安定して高精細な画像を取得するために大事である．一方で，透明化処理の影響によって組織の構築や標識シグナルの保持が悪くなるというトレードオフが厳存する．標識された構造物を再現よく定量的に観察するためには，構造や各種シグナルを適切に保存・維持する能力（preservation...