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1.
Science ; 383(6682): eadj9198, 2024 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-38300992

RESUMO

Mapping single-neuron projections is essential for understanding brain-wide connectivity and diverse functions of the hippocampus (HIP). Here, we reconstructed 10,100 single-neuron projectomes of mouse HIP and classified 43 projectome subtypes with distinct projection patterns. The number of projection targets and axon-tip distribution depended on the soma location along HIP longitudinal and transverse axes. Many projectome subtypes were enriched in specific HIP subdomains defined by spatial transcriptomic profiles. Furthermore, we delineated comprehensive wiring diagrams for HIP neurons projecting exclusively within the HIP formation (HPF) and for those projecting to both intra- and extra-HPF targets. Bihemispheric projecting neurons generally projected to one pair of homologous targets with ipsilateral preference. These organization principles of single-neuron projectomes provide a structural basis for understanding the function of HIP neurons.


Assuntos
Axônios , Mapeamento Encefálico , Hipocampo , Neurônios , Animais , Camundongos , Axônios/fisiologia , Axônios/ultraestrutura , Hipocampo/ultraestrutura , Neurônios/classificação , Neurônios/ultraestrutura , Análise de Célula Única/métodos , Rede Nervosa , Masculino , Camundongos Endogâmicos C57BL
2.
Heliyon ; 10(2): e24341, 2024 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-38293334

RESUMO

Militarine is the lead member of secondary metabolites found in multiple medicinal plants of the orchid family. It acts as not only an important inhibitor on plant growth, but also functions as the quality marker for medicinal materials. In addition, Militarine has been shown to possess remarkably medicinal value, with a definite potential for finding widespread adoption of treating various diseases, including lung injury, brain nerve injury, cognitive impairment, aging, tumors, inflammation, peptic ulcers, and more. Thus, it can serve as a material carrier for pharmacophore upon, so much so that it probes as natural source of lead compounds in the research and development of medication. The study reported herein makes an overview on the physicochemical properties and pharmacological mechanisms of Militarine compounds, summarizes the biogenic pathways of Militarine and organically integrates the biological characteristics of Militarine with multiple omics techniques. Besides, this review also constructs a regulatory system for the biological accumulation of Militarine around its precursor compounds, characteristic gene elements, key enzymes, important metabolic products, and critical steps and links. Exceptionally, emphasis on the biosynthesis of Militarine under both abiotic and biotic stress, as well as an elaboration of the signaling pathways and critical regulatory mechanisms that govern the metabolic flow of Militarine have been represented accordingly in this paper. These findings are expected to provide reference schemes and theoretical foundations for acquiring high-quality resources of Militarine and advancing its large-scale industrial production, drug development, and clinical applications to comprehensively elucidate the biosynthetic and metabolic pathways.

3.
BMC Biol ; 21(1): 135, 2023 06 06.
Artigo em Inglês | MEDLINE | ID: mdl-37280580

RESUMO

BACKGROUND: Based on their anatomical location, rostral projections of nuclei are classified as ascending circuits, while caudal projections are classified as descending circuits. Upper brainstem neurons participate in complex information processing and specific sub-populations preferentially project to participating ascending or descending circuits. Cholinergic neurons in the upper brainstem have extensive collateralizations in both ascending and descending circuits; however, their single-cell projection patterns remain unclear because of the lack of comprehensive characterization of individual neurons. RESULTS: By combining fluorescent micro-optical sectional tomography with sparse labeling, we acquired a high-resolution whole-brain dataset of pontine-tegmental cholinergic neurons (PTCNs) and reconstructed their detailed morphology using semi-automatic reconstruction methods. As the main source of acetylcholine in some subcortical areas, individual PTCNs had abundant axons with lengths up to 60 cm and 5000 terminals and innervated multiple brain regions from the spinal cord to the cortex in both hemispheres. Based on various collaterals in the ascending and descending circuits, individual PTCNs were grouped into four subtypes. The morphology of cholinergic neurons in the pedunculopontine nucleus was more divergent, whereas the laterodorsal tegmental nucleus neurons contained richer axonal branches and dendrites. In the ascending circuits, individual PTCNs innervated the thalamus in three different patterns and projected to the cortex via two separate pathways. Moreover, PTCNs targeting the ventral tegmental area and substantia nigra had abundant collaterals in the pontine reticular nuclei, and these two circuits contributed oppositely to locomotion. CONCLUSIONS: Our results suggest that individual PTCNs have abundant axons, and most project to various collaterals in the ascending and descending circuits simultaneously. They target regions with multiple patterns, such as the thalamus and cortex. These results provide a detailed organizational characterization of cholinergic neurons to understand the connexional logic of the upper brainstem.


Assuntos
Axônios , Tronco Encefálico , Tronco Encefálico/fisiologia , Axônios/fisiologia , Ponte/anatomia & histologia , Ponte/fisiologia , Encéfalo , Neurônios Colinérgicos
4.
Front Pharmacol ; 14: 1091530, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37361204

RESUMO

Introduction: The gastric mucosa (GM) is the first barrier and vital interface in the stomach that protects the host from hydrochloric acid in gastric juice and defends against exogenous insults to gastric tissues. The use of traditional Chinese medications (TCMs) for the treatment of gastric mucosal injury (GMI) has long-standing history and a good curative effect. Whereas there are poor overall reports on the intrinsic mechanisms of these TCM preparations that pharmacology uses to protect body from GMI, which is crucial to treating this disease. These existing reviews have deficiencies that limit the clinical application and development of both customary prescriptions and new drugs. Methods: Further basic and translational studies must be done to elucidate the intrinsic mechanisms of influence of these TCM preparations. Moreover, well-designed and well-conducted experiences and clinical trials are necessary to ascertain the efficacy and mechanisms of these agents. Therefore, this paper presents a focused overview of currently published literature to assess how TCMs action that facilitates the cures for GMI. It offers a whole train of current state of pharmacological evidence, identifies the pharmacological mechanisms of TCMs on GM, and highlights that remarkable capacity of TCMs to restore GM after damage. Results: These TCMs preparations promote the repair of multicomponent targets such as the gastric mucus, epithelial layer, blood flow (GMBF) and lamina propria barrier. Summary: Overall, this study has summarized the essential regulatory mechanisms and pharmacological efficacy of TCMs on new and productive therapeutic targets. Discussion: This review provides an avenue for studying various drugs with potentially promising effects on mucosal integrity, as well as subsequent pharmacological studies, clinical applications, and new drug development.

5.
iScience ; 26(5): 106705, 2023 May 19.
Artigo em Inglês | MEDLINE | ID: mdl-37216109

RESUMO

Resin embedding combined with ultra-thin sectioning has been widely used in microscopic and electron imaging to acquire precise structural information of biological tissues. However, the existing embedding method was detrimental to quenchable fluorescent signals of precise structures and pH-insensitive fluorescent dyes. Here, we developed a low-temperature chemical polymerization method named HM20-T to maintain weak signals of various precise structures and to decrease background fluorescence. The fluorescence preservation ratio of green fluorescent protein (GFP) tagged presynaptic elements and tdTomato labeled axons doubled. The HM20-T method was suitable for a variety of fluorescent dyes, such as DyLight 488 conjugated Lycopersicon esculentum lectin. Moreover, the brains also retained immunoreactivity after embedding. In summary, the HM20-T method was suitable for the characterization of multi-color labeled precise structures, which would contribute to the acquisition of complete morphology of various biological tissues and to the investigation of composition and circuit connection in the whole brain.

6.
Bioinformatics ; 39(4)2023 04 03.
Artigo em Inglês | MEDLINE | ID: mdl-36946294

RESUMO

MOTIVATION: Reconstructing and analyzing all blood vessels throughout the brain is significant for understanding brain function, revealing the mechanisms of brain disease, and mapping the whole-brain vascular atlas. Vessel segmentation is a fundamental step in reconstruction and analysis. The whole-brain optical microscopic imaging method enables the acquisition of whole-brain vessel images at the capillary resolution. Due to the massive amount of data and the complex vascular features generated by high-resolution whole-brain imaging, achieving rapid and accurate segmentation of whole-brain vasculature becomes a challenge. RESULTS: We introduce HP-VSP, a high-performance vessel segmentation pipeline based on deep learning. The pipeline consists of three processes: data blocking, block prediction, and block fusion. We used parallel computing to parallelize this pipeline to improve the efficiency of whole-brain vessel segmentation. We also designed a lightweight deep neural network based on multi-resolution vessel feature extraction to segment vessels at different scales throughout the brain accurately. We validated our approach on whole-brain vascular data from three transgenic mice collected by HD-fMOST. The results show that our proposed segmentation network achieves the state-of-the-art level under various evaluation metrics. In contrast, the parameters of the network are only 1% of those of similar networks. The established segmentation pipeline could be used on various computing platforms and complete the whole-brain vessel segmentation in 3 h. We also demonstrated that our pipeline could be applied to the vascular analysis. AVAILABILITY AND IMPLEMENTATION: The dataset is available at http://atlas.brainsmatics.org/a/li2301. The source code is freely available at https://github.com/visionlyx/HP-VSP.


Assuntos
Aprendizado Profundo , Camundongos , Animais , Encéfalo/diagnóstico por imagem , Imageamento Tridimensional/métodos , Redes Neurais de Computação , Software , Processamento de Imagem Assistida por Computador/métodos
7.
Front Neurosci ; 16: 1033880, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36278018

RESUMO

Visualizing the relationships and interactions among different biological components in the whole brain is crucial to our understanding of brain structures and functions. However, an automatic multicolor whole-brain imaging technique is still lacking. Here, we developed a multicolor wide-field large-volume tomography (multicolor WVT) to simultaneously acquire fluorescent signals in blue, green, and red channels in the whole brain. To facilitate the segmentation of brain regions and anatomical annotation, we used 4', 6-diamidino-2-phenylindole (DAPI) to provide cytoarchitecture through real-time counterstaining. We optimized the imaging planes and modes of three channels to overcome the axial chromatic aberration of the illumination path and avoid the crosstalk from DAPI to the green channel without the modification of system configuration. We also developed an automatic contour recognition algorithm based on DAPI-staining cytoarchitecture to shorten data acquisition time and reduce data redundancy. To demonstrate the potential of our system in deciphering the relationship of the multiple components of neural circuits, we acquired and quantified the brain-wide distributions of cholinergic neurons and input of ventral Caudoputamen (CP) with the anatomical annotation in the same brain. We further identified the cholinergic type of upstream neurons projecting to CP through the triple-color collocated analysis and quantified its proportions in the two brain-wide distributions. Both accounted for 0.22%, implying CP might be modulated by non-cholinergic neurons. Our method provides a new research tool for studying the different biological components in the same organ and potentially facilitates the understanding of the processing mechanism of neural circuits and other biological activities.

8.
Adv Sci (Weinh) ; 9(33): e2202553, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-36228099

RESUMO

Axonal projection conveys neural information. The divergent and diverse projections of individual neurons imply the complexity of information flow. It is necessary to investigate the relationship between the projection and functional information at the single neuron level for understanding the rules of neural circuit assembly, but a gap remains due to a lack of methods to map the function to whole-brain projection. Here an approach is developed to bridge two-photon calcium imaging in vivo with high-resolution whole-brain imaging based on sparse labeling with the genetically encoded calcium indicator GCaMP6. Reliable whole-brain projections are captured by the high-definition fluorescent micro-optical sectioning tomography (HD-fMOST). A cross-modality cell matching is performed and the functional annotation of whole-brain projection at the single-neuron level (FAWPS) is obtained. Applying it to the layer 2/3 (L2/3) neurons in mouse visual cortex, the relationship is investigated between functional preferences and axonal projection features. The functional preference of projection motifs and the correlation between axonal length in MOs and neuronal orientation selectivity, suggest that projection motif-defined neurons form a functionally specific information flow, and the projection strength in specific targets relates to the information clarity. This pipeline provides a new way to understand the principle of neuronal information transmission.


Assuntos
Neurônios , Córtex Visual , Animais , Camundongos , Neurônios/fisiologia , Encéfalo , Córtex Visual/fisiologia , Axônios/fisiologia , Mapeamento Encefálico/métodos
9.
Neurosci Bull ; 38(11): 1315-1329, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-35984621

RESUMO

The zona incerta (ZI) is involved in various functions and may serve as an integrative node of the circuits for global behavioral modulation. However, the long-range connectivity of different sectors in the mouse ZI has not been comprehensively mapped. Here, we obtained whole-brain images of the input and output connections via fluorescence micro-optical sectioning tomography and viral tracing. The principal regions in the input-output circuits of ZI GABAergic neurons were topologically organized. The 3D distribution of cortical inputs showed rostro-caudal correspondence with different ZI sectors, while the projection fibers from ZI sectors were longitudinally organized in the superior colliculus. Clustering results show that the medial and lateral ZI are two different major functional compartments, and they can be further divided into more subdomains based on projection and input connectivity. This study provides a comprehensive anatomical foundation for understanding how the ZI is involved in integrating different information, conveying motivational states, and modulating global behaviors.


Assuntos
Conectoma , Zona Incerta , Animais , Camundongos , Neurônios GABAérgicos
10.
Transl Cancer Res ; 11(3): 548-558, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35402172

RESUMO

Background: Muscle relaxation training is a method of gradually relaxing the whole body by consciously controlling the process of muscle contraction and relaxation, which is mostly used to improve the physical and mental health of breast cancer patients and improve the quality of life of patients. We conducted a systematic review to compare the effects of muscle relaxation training and conventional nursing on the psychological health and quality of life (QoL) of breast cancer patients. The results of this study provide a basis for nursing program selection of breast cancer patients. Methods: The PubMed, EMbase, Web of Science, The Cochrane Library, China national knowledge infrastructure (CNKI), WanFang Data, (China Biology Medicine disc) CBM, and WWW.CQVIP.COM (VIP) databases were searched to retrieve articles on randomized controlled trials (RCTs) or quasi-RCTs on the effects of muscle relaxation training on the mental health and QoL of breast cancer patients. The search period ran from the establishment of the databases to August 31st, 2021. Two researchers independently screened the literature, extracted the data, and The Cochrane Handbook for Systematic Reviews of Interventions assessed the risk of bias in the included studies. Stata 15.0 software was then used for the meta-analysis. Results: Funnel plots were analyzed by E Egger's test and Begg's test. The results of the test (P>0.05) showed that the possibility of publication bias was small. A total of 13 RCTs and quasi-RCTs, comprising 1,355 patients, were included in the meta-analysis. The results for the outcome measures were as follows: level of depression [weighted mean difference (WMD) =-9.31, 95% confidence interval (CI): -11.96 to -6.65, level of anxiety (WMD =-8.96, 95% CI: -10.06 to -7.86)], and QoL (WMD =13.13, 95% CI: 7.24, 19.02). The results showed that muscle relaxation training can significantly reduce depression and anxiety in breast cancer patients, improve their quality of life, and can be used as the first choice for breast cancer patients to improve negative emotions. Discussion: Muscle relaxation training significantly reduced the depression and anxiety of breast cancer patients, improved their QoL, and brought about both psychological and QoL improvements.

11.
Neurosci Bull ; 38(6): 576-590, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35312957

RESUMO

Neurons in the primary auditory area (AUDp) innervate multiple brain regions with long-range projections while receiving informative inputs for diverse functions. However, the brain-wide connections of these neurons have not been comprehensively investigated. Here, we simultaneously applied virus-based anterograde and retrograde tracing, labeled the connections of excitatory and inhibitory neurons in the mouse AUDp, and acquired whole-brain information using a dual-channel fluorescence micro-optical sectioning tomography system. Quantified results showed that the two types of neurons received inputs with similar patterns but sent heterogeneous projections to downstream regions. In the isocortex, functionally different areas consistently sent feedback-dominated projections to these neurons, with concomitant laterally-dominated projections from the sensory and limbic cortices to inhibitory neurons. In subcortical regions, the dorsal and medial parts of the non-lemniscal auditory thalamus (AT) were reciprocally connected to the AUDp, while the ventral part contained the most fibers of passage from the excitatory neurons and barely sent projections back, indicating the regional heterogeneity of the AUDp-AT circuit. Our results reveal details of the whole-brain network and provide new insights for further physiological and functional studies of the AUDp.


Assuntos
Córtex Auditivo , Animais , Córtex Auditivo/fisiologia , Axônios , Mapeamento Encefálico , Camundongos , Neurônios/fisiologia , Tálamo/fisiologia
13.
Biomed Opt Express ; 12(11): 6730-6745, 2021 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-34858677

RESUMO

Resin embedding of multi-color labeled whole organs is the primary step to preserve structural information for visualization of fine structures in three dimensions. It is essential to study the morphological characteristics, spatial and positional relationships of the millions of neurons, and the intricate network of blood vessels with fluorescent labels in the brain. However, the current resin embedding method is inadequate because of incompatibilities with fluorescent dyes, making it difficult to reconstruct a variety of structures for the interpretation of their complex spatial relationships. We modified the resin embedding method for large biological tissues labeled with multiple fluorescent dyes and proteins through different labeling strategies. With TrueBlack as the background fluorescence inhibitor in the glycol methacrylate (GMA) embedding, we referred to the method as GMA-T (Glycol methacrylate with TB). In the GMA-T embedded mouse brains, structures labeled with fluorescent proteins and dyes were visualized in millimeter-scale networks with sub-cellular resolution, allowing quantitative analysis of different anatomical structures in the same brain, including neurons and blood vessels. In combination with high-resolution whole-brain imaging, it is possible to obtain a variety of fluorescence labeled structures in just a few days. We quantified the distribution and morphology of the tdTomato-labeled vasoactive intestinal polypeptide (VIP) neurons and the BSA-FITC labeled blood vessels in the same brain. These results demonstrated that VIP neurons and blood vessels have their own unique distribution patterns and morphological characteristics among cortical regions and different layers in cerebral cortex, and there was no significant correlation between VIP neurons and vessels. This approach provides a novel approach to study the interaction among different anatomical structures within large-volume biological samples labeled with multiple fluorescent dyes and proteins, which helps elucidating the complex anatomical characteristics of biological organs.

14.
Elife ; 102021 11 18.
Artigo em Inglês | MEDLINE | ID: mdl-34792021

RESUMO

The dorsal raphe nucleus (DR) and median raphe nucleus (MR) contain populations of glutamatergic and GABAergic neurons that regulate diverse behavioral functions. However, their whole-brain input-output circuits remain incompletely elucidated. We used viral tracing combined with fluorescence micro-optical sectioning tomography to generate a comprehensive whole-brain atlas of inputs and outputs of glutamatergic and GABAergic neurons in the DR and MR. We found that these neurons received inputs from similar upstream brain regions. The glutamatergic and GABAergic neurons in the same raphe nucleus had divergent projection patterns with differences in critical brain regions. Specifically, MR glutamatergic neurons projected to the lateral habenula through multiple pathways. Correlation and cluster analysis revealed that glutamatergic and GABAergic neurons in the same raphe nucleus received heterogeneous inputs and sent different collateral projections. This connectivity atlas further elucidates the anatomical architecture of the raphe nuclei, which could facilitate better understanding of their behavioral functions.


Assuntos
Conectoma , Núcleo Dorsal da Rafe/fisiologia , Núcleos da Rafe do Mesencéfalo/fisiologia , Neurônios/fisiologia , Animais , Neurônios GABAérgicos/fisiologia , Camundongos
15.
Nature ; 598(7879): 188-194, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34616074

RESUMO

The cortico-basal ganglia-thalamo-cortical loop is one of the fundamental network motifs in the brain. Revealing its structural and functional organization is critical to understanding cognition, sensorimotor behaviour, and the natural history of many neurological and neuropsychiatric disorders. Classically, this network is conceptualized to contain three information channels: motor, limbic and associative1-4. Yet this three-channel view cannot explain the myriad functions of the basal ganglia. We previously subdivided the dorsal striatum into 29 functional domains on the basis of the topography of inputs from the entire cortex5. Here we map the multi-synaptic output pathways of these striatal domains through the globus pallidus external part (GPe), substantia nigra reticular part (SNr), thalamic nuclei and cortex. Accordingly, we identify 14 SNr and 36 GPe domains and a direct cortico-SNr projection. The striatonigral direct pathway displays a greater convergence of striatal inputs than the more parallel striatopallidal indirect pathway, although direct and indirect pathways originating from the same striatal domain ultimately converge onto the same postsynaptic SNr neurons. Following the SNr outputs, we delineate six domains in the parafascicular and ventromedial thalamic nuclei. Subsequently, we identify six parallel cortico-basal ganglia-thalamic subnetworks that sequentially transduce specific subsets of cortical information through every elemental node of the cortico-basal ganglia-thalamic loop. Thalamic domains relay this output back to the originating corticostriatal neurons of each subnetwork in a bona fide closed loop.


Assuntos
Gânglios da Base/citologia , Córtex Cerebral/citologia , Vias Neurais , Neurônios/citologia , Tálamo/citologia , Animais , Gânglios da Base/anatomia & histologia , Córtex Cerebral/anatomia & histologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Tálamo/anatomia & histologia
16.
Nat Methods ; 18(3): 309-315, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33649587

RESUMO

The microscopic visualization of large-scale three-dimensional (3D) samples by optical microscopy requires overcoming challenges in imaging quality and speed and in big data acquisition and management. We report a line-illumination modulation (LiMo) technique for imaging thick tissues with high throughput and low background. Combining LiMo with thin tissue sectioning, we further develop a high-definition fluorescent micro-optical sectioning tomography (HD-fMOST) method that features an average signal-to-noise ratio of 110, leading to substantial improvement in neuronal morphology reconstruction. We achieve a >30-fold lossless data compression at a voxel resolution of 0.32 × 0.32 × 1.00 µm3, enabling online data storage to a USB drive or in the cloud, and high-precision (95% accuracy) brain-wide 3D cell counting in real time. These results highlight the potential of HD-fMOST to facilitate large-scale acquisition and analysis of whole-brain high-resolution datasets.


Assuntos
Encéfalo/diagnóstico por imagem , Imageamento Tridimensional/métodos , Microscopia/métodos , Microtomia/métodos , Razão Sinal-Ruído , Tomografia/métodos
17.
Neuroinformatics ; 19(2): 267-284, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-32754778

RESUMO

The extreme complexity of mammalian brains requires a comprehensive deconstruction of neuroanatomical structures. Scientists normally use a brain stereotactic atlas to determine the locations of neurons and neuronal circuits. However, different brain images are normally not naturally aligned even when they are imaged with the same setup, let alone under the differing resolutions and dataset sizes used in mesoscopic imaging. As a result, it is difficult to achieve high-throughput automatic registration without manual intervention. Here, we propose a deep learning-based registration method called DeepMapi to predict a deformation field used to register mesoscopic optical images to an atlas. We use a self-feedback strategy to address the problem of imbalanced training sets (sampling at a fixed step size in nonuniform brains of structures and deformations) and use a dual-hierarchical network to capture the large and small deformations. By comparing DeepMapi with other registration methods, we demonstrate its superiority over a set of ground truth images, including both optical and MRI images. DeepMapi achieves fully automatic registration of mesoscopic micro-optical images, even macroscopic MRI datasets, in minutes, with an accuracy comparable to those of manual annotations by anatomists.


Assuntos
Mapeamento Encefálico/métodos , Encéfalo/diagnóstico por imagem , Processamento de Imagem Assistida por Computador/métodos , Imageamento por Ressonância Magnética/métodos , Rede Nervosa/diagnóstico por imagem , Redes Neurais de Computação , Animais , Mapeamento Encefálico/normas , Bases de Dados Factuais/normas , Humanos , Processamento de Imagem Assistida por Computador/normas , Imageamento Tridimensional/métodos , Imageamento Tridimensional/normas , Imageamento por Ressonância Magnética/normas , Camundongos , Camundongos Endogâmicos C57BL , Neuroimagem/métodos , Neuroimagem/normas , Projetos de Pesquisa/normas
18.
Sci Rep ; 10(1): 12209, 2020 07 22.
Artigo em Inglês | MEDLINE | ID: mdl-32699360

RESUMO

Dissection of the neural circuits of the cerebral cortex is essential for studying mechanisms underlying brain function. Herein, combining a retrograde rabies tracing system with fluorescent micro-optical sectional tomography, we investigated long-range input neurons of corticotropin-releasing hormone containing neurons in the six main cortical areas, including the prefrontal, somatosensory, motor, auditory, and visual cortices. The whole brain distribution of input neurons showed similar patterns to input neurons distributed mainly in the adjacent cortical areas, thalamus, and basal forebrain. Reconstruction of continuous three-dimensional datasets showed the anterior and middle thalamus projected mainly to the rostral cortex whereas the posterior and lateral projected to the caudal cortex. In the basal forebrain, immunohistochemical staining showed these cortical areas received afferent information from cholinergic neurons in the substantia innominata and lateral globus pallidus, whereas cholinergic neurons in the diagonal band nucleus projected strongly to the prefrontal and visual cortex. Additionally, dense neurons in the zona incerta and ventral hippocampus were found to project to the prefrontal cortex. These results showed general patterns of cortical input circuits and unique connection patterns of each individual area, allowing for valuable comparisons among the organisation of different cortical areas and new insight into cortical functions.


Assuntos
Encéfalo/metabolismo , Hormônio Liberador da Corticotropina/metabolismo , Neurônios/metabolismo , Animais , Mapeamento Encefálico/métodos , Hormônio Liberador da Corticotropina/genética , Vetores Genéticos/genética , Vetores Genéticos/metabolismo , Globo Pálido/metabolismo , Processamento de Imagem Assistida por Computador , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Microscopia Confocal , Tálamo/metabolismo
19.
Sci Rep ; 10(1): 7916, 2020 05 13.
Artigo em Inglês | MEDLINE | ID: mdl-32405029

RESUMO

Neuronal cell types are essential to the comprehensive understanding of the neuronal function and neuron can be categorized by their anatomical property. However, complete morphology data for neurons with a whole brain projection, for example the pyramidal neurons in the cortex, are sparse because it is difficult to trace the neuronal fibers across the whole brain and acquire the neuron morphology at the single axon resolution. Thus the cell types of pyramidal neurons have yet to be studied at the single axon resolution thoroughly. In this work, we acquire images for a Thy1 H-line mouse brain using a fluorescence micro-optical sectioning tomography system. Then we sample 42 pyramidal neurons whose somata are in the layer 5 of the motor cortex and reconstruct their morphology across the whole brain. Based on the reconstructed neuronal anatomy, we analyze the axonal and dendritic fibers of the neurons in addition to the soma spatial distributions, and identify two axonal projection pattern of pyramidal tract neurons and two dendritic spreading patterns of intratelencephalic neurons. The raw image data are available upon request as an additional asset to the community. The morphological patterns identified in this work can be a typical representation of neuron subtypes and reveal the possible input-output function of a single pyramidal neuron.


Assuntos
Córtex Motor/citologia , Neurônios Motores/citologia , Células Piramidais/citologia , Animais , Axônios , Dendritos , Imunofluorescência , Camundongos , Córtex Motor/metabolismo , Neurônios Motores/metabolismo , Células Piramidais/metabolismo
20.
Front Neurosci ; 14: 322, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32351352

RESUMO

Mapping the cytoarchitecture of the whole brain can reveal the organizational logic of neural systems. However, this remains a significant challenge, especially for gyrencephalic brains with a large volume. Here we propose an integrated pipeline for generating a cytoarchitectonic atlas with single-cell resolution of the whole brain. To analyze a large-volume brain, we used a modified en-bloc Nissl staining protocol to achieve uniform staining of large-scale brain specimens from ferret (Mustela putorius furo). By combining whole-brain imaging and big data processing, we established strategies for parsing cytoarchitectural information at a voxel resolution of 0.33 µm × 0.33 µm × 1 µm and terabyte-scale data analysis. Using the cytoarchitectonic datasets for adult ferret brain, we identified giant pyramidal neurons in ferret brains and provide the first report of their morphological diversity, neurochemical phenotype, and distribution patterns in the whole brain in three dimensions. This pipeline will facilitate studies on the organization and development of the mammalian brains, from that of rodents to the gyrencephalic brains of ferret and even primates.

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