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Development and characterization of a non-human primate model of disseminated synucleinopathy 非人灵长类动物散播性突触核蛋白病模型的开发和特征描述
IF 2.9 4区 医学 Q1 Medicine Pub Date : 2024-03-27 DOI: 10.3389/fnana.2024.1355940
Alberto J. Rico, Almudena Corcho, Julia Chocarro, Goiaz Ariznabarreta, Elvira Roda, Adriana Honrubia, Patricia Arnaiz, José L. Lanciego
IntroductionThe presence of a widespread cortical synucleinopathy is the main neuropathological hallmark underlying clinical entities such as Parkinson’s disease with dementia (PDD) and dementia with Lewy bodies (DLB). There currently is a pressing need for the development of non-human primate (NHPs) models of PDD and DLB to further overcome existing limitations in drug discovery.MethodsHere we took advantage of a retrogradely-spreading adeno-associated viral vector serotype 9 coding for the alpha-synuclein A53T mutated gene (AAV9-SynA53T) to induce a widespread synucleinopathy of cortical and subcortical territories innervating the putamen. Four weeks post-AAV deliveries animals were sacrificed and a comprehensive biodistribution study was conducted, comprising the quantification of neurons expressing alpha-synuclein, rostrocaudal distribution and their specific location.ResultsIntraputaminal deliveries of AAV9-SynA53T lead to a disseminated synucleinopathy throughout ipsi- and contralateral cerebral cortices, together with transduced neurons located in the ipsilateral caudal intralaminar nuclei and in the substantia nigra pars compacta (leading to thalamostriatal and nigrostriatal projections, respectively). Cortical afferent systems were found to be the main contributors to putaminal afferents (superior frontal and precentral gyri in particular).DiscussionObtained data extends current models of synucleinopathies in NHPs, providing a reproducible platform enabling the adequate implementation of end-stage preclinical screening of new drugs targeting alpha-synuclein.
导言广泛存在的皮质突触核蛋白病是帕金森病伴痴呆(PDD)和路易体痴呆(DLB)等临床实体的主要神经病理学特征。我们利用逆向传播的编码α-突触核蛋白A53T突变基因的9号血清型腺相关病毒载体(AAV9-SynA53T)诱导支配普鲁士门的皮层和皮层下区域发生广泛的突触核蛋白病。AAV投放后四周,动物被处死,并进行了全面的生物分布研究,包括α-突触核蛋白表达神经元的数量、喙尾分布及其具体位置。结果椎管内注射 AAV9-SynA53T 会导致同侧和对侧大脑皮层出现散发性突触核蛋白病,同时同侧尾椎内核和黑质(分别导致丘脑突触和黑质突触投射)也会出现转染神经元。讨论获得的数据扩展了目前的NHPs突触核蛋白病模型,提供了一个可重复的平台,使针对α-突触核蛋白的新药的临床前筛选能够充分进行到最后阶段。
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引用次数: 0
Exploring an innovative decellularization protocol for porcine nerve grafts: a translational approach to peripheral nerve repair 探索猪神经移植物的创新脱细胞方案:外周神经修复的转化方法
IF 2.9 4区 医学 Q1 Medicine Pub Date : 2024-03-19 DOI: 10.3389/fnana.2024.1380520
Luisa Muratori, Alessandro Crosio, Giulia Ronchi, Debora Molinaro, Pierluigi Tos, Arianna B. Lovati, Stefania Raimondo
IntroductionPeripheral nerves are frequently affected by lesions caused by traumatic or iatrogenic damages, resulting in loss of motor and sensory function, crucial in orthopedic outcomes and with a significant impact on patients’ quality of life. Many strategies have been proposed over years to repair nerve injuries with substance loss, to achieve musculoskeletal reinnervation and functional recovery. Allograft have been tested as an alternative to the gold standard, the autograft technique, but nerves from donors frequently cause immunogenic response. For this reason, several studies are focusing to find the best way to decellularize nerves preserving either the extracellular matrix, either the basal lamina, as the key elements used by Schwann cells and axons during the regenerative process.MethodsThis study focuses on a novel decellularization protocol for porcine nerves, aimed at reducing immunogenicity while preserving essential elements like the extracellular matrix and basal lamina, vital for nerve regeneration. To investigate the efficacy of the decellularization protocol to remove immunogenic cellular components of the nerve tissue and to preserve the basal lamina and extracellular matrix, morphological analysis was performed through Masson’s Trichrome staining, immunofluorescence, high resolution light microscopy and transmission electron microscopy. Decellularized porcine nerve graft were then employed in vivo to repair a rat median nerve lesion. Morphological analysis was also used to study the ability of the porcine decellularized graft to support the nerve regeneration.Results and DiscussionThe decellularization method was effective in preparing porcine superficial peroneal nerves for grafting as evidenced by the removal of immunogenic components and preservation of the ECM. Morphological analysis demonstrated that four weeks after injury, regenerating fibers colonized the graft suggesting a promising use to repair severe nerve lesions. The idea of using a porcine nerve graft arises from a translational perspective. This approach offers a promising direction in the orthopedic field for nerve repair, especially in severe cases where conventional methods are limited.
引言 周围神经经常受到外伤或先天性损伤引起的病变的影响,从而导致运动和感觉功能的丧失,这对骨科治疗效果至关重要,并对患者的生活质量产生重大影响。多年来,人们提出了许多策略来修复物质缺失的神经损伤,以实现肌肉骨骼神经再支配和功能恢复。异体移植已作为自体移植技术这一黄金标准的替代方法进行了测试,但来自供体的神经经常会引起免疫原性反应。因此,多项研究都在寻找神经脱细胞的最佳方法,以保留细胞外基质或基底层,因为它们是许旺细胞和轴突在再生过程中使用的关键元素。本研究重点关注猪神经的新型脱细胞方案,旨在降低免疫原性,同时保留细胞外基质和基底层等对神经再生至关重要的元素。为了研究脱细胞方案在去除神经组织中的免疫原性细胞成分以及保留基底层和细胞外基质方面的功效,我们通过马森三色染色、免疫荧光、高分辨率光学显微镜和透射电子显微镜进行了形态学分析。然后采用脱细胞猪神经移植物在体内修复大鼠正中神经损伤。结果与讨论脱细胞方法能有效地制备猪腓浅神经用于移植,这体现在免疫原性成分的去除和 ECM 的保留上。形态学分析表明,损伤四周后,再生纤维在移植物上定植,这表明该方法有望用于修复严重的神经损伤。使用猪神经移植物的想法源于转化的角度。这种方法为骨科领域的神经修复提供了一个前景广阔的方向,尤其是在传统方法受限的严重病例中。
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引用次数: 0
Phylogenetic reduction of the magnocellular red nucleus in primates and inter-subject variability in humans 灵长类动物大细胞红核的系统发育减少和人类受试者间的变异性
IF 2.9 4区 医学 Q1 Medicine Pub Date : 2024-03-13 DOI: 10.3389/fnana.2024.1331305
Martin Stacho, A. Niklas Häusler, Andrea Brandstetter, Francesca Iannilli, Hartmut Mohlberg, Christian Schiffer, Jeroen B. Smaers, Katrin Amunts
IntroductionThe red nucleus is part of the motor system controlling limb movements. While this seems to be a function common in many vertebrates, its organization and circuitry have undergone massive changes during evolution. In primates, it is sub-divided into the magnocellular and parvocellular parts that give rise to rubrospinal and rubro-olivary connection, respectively. These two subdivisions are subject to striking variation within the primates and the size of the magnocellular part is markedly reduced in bipedal primates including humans. The parvocellular part is part of the olivo-cerebellar circuitry that is prominent in humans. Despite the well-described differences between species in the literature, systematic comparative studies of the red nucleus remain rare.MethodsWe therefore mapped the red nucleus in cytoarchitectonic sections of 20 primate species belonging to 5 primate groups including prosimians, new world monkeys, old world monkeys, non-human apes and humans. We used Ornstein-Uhlenbeck modelling, ancestral state estimation and phylogenetic analysis of covariance to scrutinize the phylogenetic relations of the red nucleus volume.ResultsWe created openly available high-resolution cytoarchitectonic delineations of the human red nucleus in the microscopic BigBrain model and human probabilistic maps that capture inter-subject variations in quantitative terms. Further, we compared the volume of the nucleus across primates and showed that the parvocellular subdivision scaled proportionally to the brain volume across the groups while the magnocellular part deviated significantly from the scaling in humans and non-human apes. These two groups showed the lowest size of the magnocellular red nucleus relative to the whole brain volume and the largest relative difference between the parvocellular and magnocellular subdivision.DiscussionThat is, the red nucleus has transformed from a magnocellular-dominated to a parvocellular-dominated station. It is reasonable to assume that these changes are intertwined with evolutionary developments in other brain regions, in particular the motor system. We speculate that the interspecies variations might partly reflect the differences in hand dexterity but also the tentative involvement of the red nucleus in sensory and cognitive functions.
导言红核是控制肢体运动的运动系统的一部分。虽然这似乎是许多脊椎动物的共同功能,但其组织和电路在进化过程中发生了巨大变化。在灵长类动物中,它被细分为大细胞和副细胞部分,分别产生红脊髓和红髓鞘连接。在灵长类动物中,这两个细分部分存在显著差异,在包括人类在内的两足灵长类动物中,大细胞部分的大小明显缩小。副小脑部分是橄榄-小脑回路的一部分,在人类中非常突出。因此,我们绘制了属于 5 个灵长类的 20 种灵长类动物(包括拟猴、新世界猴、旧世界猴、非人类猿和人类)的红核细胞结构切片。我们利用奥恩斯坦-乌伦贝克模型、祖先状态估计和系统发育协方差分析,仔细研究了红核体积的系统发育关系。结果我们在显微 BigBrain 模型中创建了公开可用的人类红核高分辨率细胞结构图,并创建了人类概率图,以定量方式捕捉受试者之间的变化。此外,我们还比较了不同灵长类动物的细胞核体积,结果表明,在不同组别中,细胞旁部分与脑体积成比例,而在人类和非人猿中,细胞大部明显偏离了这一比例。讨论也就是说,红核已从以镁细胞为主转变为以副镁细胞为主。我们有理由认为,这些变化与其他脑区(尤其是运动系统)的进化发展交织在一起。我们推测,种间变化可能部分反映了手部灵活性的差异,但也初步反映了红核参与感觉和认知功能的情况。
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引用次数: 0
Localization of hyperpolarization-activated cyclic nucleotide-gated channels in the vertebrate retinas across species and their physiological roles 超极化激活的环核苷酸门控通道在脊椎动物视网膜中的跨物种定位及其生理作用
IF 2.9 4区 医学 Q1 Medicine Pub Date : 2024-03-06 DOI: 10.3389/fnana.2024.1385932
Daniel Kim, Hyeonhee Roh, Hyung-Min Lee, Sang Jeong Kim, Maesoon Im

Transmembrane proteins known as hyperpolarization-activated cyclic nucleotide-gated (HCN) channels control the movement of Na+ and K+ ions across cellular membranes. HCN channels are known to be involved in crucial physiological functions in regulating neuronal excitability and rhythmicity, and pacemaker activity in the heart. Although HCN channels have been relatively well investigated in the brain, their distribution and function in the retina have received less attention, remaining their physiological roles to be comprehensively understood. Also, because recent studies reported HCN channels have been somewhat linked with the dysfunction of photoreceptors which are affected by retinal diseases, investigating HCN channels in the retina may offer valuable insights into disease mechanisms and potentially contribute to identifying novel therapeutic targets for retinal degenerative disorders. This paper endeavors to summarize the existing literature on the distribution and function of HCN channels reported in the vertebrate retinas of various species and discuss the potential implications for the treatment of retinal diseases. Then, we recapitulate current knowledge regarding the function and regulation of HCN channels, as well as their relevance to various neurological disorders.

被称为超极化激活环核苷酸门控(HCN)通道的跨膜蛋白控制着 Na+ 和 K+ 离子在细胞膜上的移动。众所周知,HCN 通道参与了调节神经元兴奋性和节律性以及心脏起搏器活动等重要生理功能。虽然 HCN 通道在大脑中的研究相对较多,但其在视网膜中的分布和功能却较少受到关注,其生理作用仍有待全面了解。此外,由于最近的研究报告称 HCN 通道与受视网膜疾病影响的光感受器的功能障碍有一定联系,因此研究视网膜中的 HCN 通道可为了解疾病机制提供有价值的见解,并可能有助于确定视网膜退行性疾病的新型治疗靶点。本文试图总结现有文献中关于不同物种脊椎动物视网膜中 HCN 通道分布和功能的报道,并讨论其对视网膜疾病治疗的潜在影响。然后,我们概述了目前有关 HCN 通道功能和调控的知识,以及它们与各种神经系统疾病的相关性。
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引用次数: 0
Anterograde trans-neuronal labeling of striatal interneurons in relation to dopamine neurons in the substantia nigra pars compacta 纹状体中间神经元与黑质紧密团多巴胺神经元的前向跨神经元标记
IF 2.9 4区 医学 Q1 Medicine Pub Date : 2024-02-28 DOI: 10.3389/fnana.2024.1325368
Fuyuki Karube, Yang Yang, Kenta Kobayashi, Fumino Fujiyama
Recent advances in neural tracing have unveiled numerous neural circuits characterized by brain region and cell type specificity, illuminating the underpinnings of specific functions and behaviors. Dopaminergic (DA) neurons in the midbrain are highly heterogeneous in terms of gene and protein expression and axonal projections. Different cell types within the substantia nigra pars compacta (SNc) tend to project to the striatum in a cell-type-dependent manner characterized by specific topography. Given the wide and dense distribution of DA axons, coupled with a combination of synaptic and volume transmission, it remains unclear how DA release is spatially and temporally regulated, to appropriately achieve specific behaviors and functions. Our hypothesis posits that hidden rules governing synapse formation between pre-synaptic DA neuron types and striatal neuron types may modulate the effect of DA at a single-cell level. To address this conjecture, we employed adeno-associated virus serotype 1 (AAV1) to visualize the neural circuitry of DA neurons. AAV1 has emerged as a potent anatomical instrument capable of labeling and visualizing pre- and post-synaptic neurons simultaneously through anterograde trans-synaptic labeling. First, AAV1-Cre was injected into the SNc, resulting in Cre expression in both medium spiny neurons and interneurons in the striatum. Due to the potential occurrence of the retrograde transfer of AAV1, only striatal interneurons were considered for trans-synaptic or trans-neuronal labeling. Interneuron types expressing parvalbumin, choline acetyltransferase, somatostatin, or nitrogen oxide synthase exhibited Cre expression. Using a combination of AAV1-Cre and Cre-driven fluorophore expressing AAVs, striatal interneurons and the axons originating from the SNc were visualized in distinct colors. Using immunofluorescence against neurotransmitter transporters, almost all axons in the striatum visualized using this approach were confirmed to be dopaminergic. Moreover, individual DA axons established multiple appositions on the somata and proximal dendrites of interneurons. This finding suggests that irrespective of the extensive and widespread axonal arborization of DA neurons, a particular DA neuron may exert a significant influence on specific interneurons. Thus, AAV1-based labeling of the DA system can be a valuable tool to uncover the concealed rules governing these intricate relationships.
神经追踪技术的最新进展揭示了以脑区和细胞类型特异性为特征的众多神经回路,阐明了特定功能和行为的基础。中脑的多巴胺能(DA)神经元在基因和蛋白质表达以及轴突投射方面具有高度异质性。黑质紧实旁(SNc)中的不同细胞类型倾向于以细胞类型依赖的方式向纹状体投射,其特征是特定的地形。鉴于 DA 轴突分布广泛而密集,再加上突触和体积传输的结合,目前仍不清楚 DA 释放是如何在空间和时间上进行调节,以适当实现特定行为和功能的。我们的假设认为,支配突触前 DA 神经元类型和纹状体神经元类型之间突触形成的隐藏规则可能会在单细胞水平上调节 DA 的效应。为了解决这一猜想,我们采用了血清型 1(AAV1)腺相关病毒来观察 DA 神经元的神经回路。AAV1 已成为一种有效的解剖学工具,能够通过前向跨突触标记同时标记和观察突触前后的神经元。首先,将AAV1-Cre注射到SNc,使Cre在纹状体的中刺神经元和中间神经元中表达。由于AAV1可能发生逆向转移,因此只考虑对纹状体中间神经元进行跨突触或跨神经元标记。表达副神经元、胆碱乙酰转移酶、体生长抑素或氧化氮合酶的中间神经元类型表现出 Cre 表达。通过结合使用 AAV1-Cre 和 Cre 驱动的荧光团表达 AAV,纹状体中间神经元和源自 SNc 的轴突以不同的颜色显现出来。利用针对神经递质转运体的免疫荧光技术,用这种方法观察到的纹状体中几乎所有轴突都被证实是多巴胺能轴突。此外,单个 DA 轴突在中间神经元的体节和近端树突上建立了多个附着点。这一发现表明,尽管DA神经元的轴突分布广泛,但特定的DA神经元可能会对特定的中间神经元产生重大影响。因此,基于 AAV1 对 DA 系统进行标记可以成为揭示这些错综复杂关系的隐藏规则的重要工具。
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引用次数: 0
Arterial supply and morphological characteristics of sympathetic neurons in the human superior cervical ganglion 人体颈上神经节交感神经元的动脉供应和形态特征
IF 2.9 4区 医学 Q1 Medicine Pub Date : 2024-02-22 DOI: 10.3389/fnana.2024.1372180
Jelena Boljanović, Milan Milisavljević, Milan Latas, Laslo Puškaš, Nikola Bogosavljević, Marko Vujačić, Dejan Aleksandrić, Dejan Ćetković, Nemanja Branković, Aleksandra Dožić, Mila Ćetković

The aim of this study was the micromorphological analysis of the distribution of microvessels, mast cells and ganglionic neurons in two parts, proximal and distal of the human superior cervical sympathetic ganglions (SCSGs). Statistical analyses were applied to detect the possible metric regional differences in their densities. Five injected human SCSGs with colored India ink and gelatin were microdissected and examined. Second group of five human SCSGs was prepared and serially sliced for CD34 and mast cell tryptase immunostaining. The microscopic fields of two parts of the SCSGs were analyzed for the following quantifications: microvessel density (MVD), mast cell density (MCD), and ganglionic cell count and measurements. The mean number of CD34-positive microvessels in microscopic fields, the MVD, had a value of 83 for the upper parts, and 82.7 for the lower parts of SCSGs. The mean number of tryptase-positive mast cells in microscopic fields, the MCD, was 4.5 in the proximal parts, and 4.7 in the distal parts of SCSGs. The mean number of ganglionic neurons in microscopic fields was 19.5 in the proximal parts, and 19.8 in the distal parts of SCSGs. The density of CD34-positive microvessels, the density of tryptase-positive mast cells, and the density, mean diameters and mean areas of ganglionic neurons were not significantly different in two observed parts, upper and lower of the SCSGs. In conclusion, the distributions of microvessels, mast cells, and neurons in two parts of the SCSGs were uniform with no specific micromorphological variations, there is a homogenous vascular and cellular pattern within the SCSGs.

本研究的目的是对人体颈上交感神经节(SCSGs)近端和远端两部分的微血管、肥大细胞和神经节神经元的分布进行微形态学分析。统计分析用于检测其密度可能存在的度量区域差异。用有色印度墨水和明胶注射 5 个人体 SCSG,并对其进行显微解剖和检查。制备第二组 5 个人类 SCSG,并将其连续切片用于 CD34 和肥大细胞胰蛋白酶免疫染色。对 SCSG 两个部分的显微镜视野进行以下量化分析:微血管密度(MVD)、肥大细胞密度(MCD)、神经节细胞计数和测量。显微视野中 CD34 阳性微血管的平均数量,即 MVD,SCSG 上部的值为 83,下部为 82.7。显微视野中胰蛋白酶阳性肥大细胞的平均数量(MCD)在 SCSG 的近端部分为 4.5 个,远端部分为 4.7 个。在显微视野中,神经节神经元的平均数量在SCSG近端为19.5个,远端为19.8个。CD34 阳性微血管的密度、胰蛋白酶阳性肥大细胞的密度、神经节神经元的密度、平均直径和平均面积在 SCSG 上部和下部的两个观察部位没有显著差异。总之,SCSG 两个部分的微血管、肥大细胞和神经元分布均匀,没有特殊的微观形态变化,SCSG 内部存在同质的血管和细胞模式。
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引用次数: 0
Editorial: Subcortical and spinal control of motor networks across vertebrates. 社论:脊椎动物运动网络的皮层下和脊髓控制
IF 2.9 4区 医学 Q1 Medicine Pub Date : 2024-02-21 eCollection Date: 2024-01-01 DOI: 10.3389/fnana.2024.1378811
Jean-Luc Boulland, Marie-Claude Perreault
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引用次数: 0
Developmental patterns of extracellular matrix molecules in the embryonic and postnatal mouse hindbrain 胚胎和出生后小鼠后脑细胞外基质分子的发育模式
IF 2.9 4区 医学 Q1 Medicine Pub Date : 2024-02-16 DOI: 10.3389/fnana.2024.1369103
Ildikó Wéber, Adél Dakos, Zoltán Mészár, Clara Matesz, András Birinyi

Normal brain development requires continuous communication between developing neurons and their environment filled by a complex network referred to as extracellular matrix (ECM). The ECM is divided into distinct families of molecules including hyaluronic acid, proteoglycans, glycoproteins such as tenascins, and link proteins. In this study, we characterize the temporal and spatial distribution of the extracellular matrix molecules in the embryonic and postnatal mouse hindbrain by using antibodies and lectin histochemistry. In the embryo, hyaluronan and neurocan were found in high amounts until the time of birth whereas versican and tenascin-R were detected in lower intensities during the whole embryonic period. After birth, both hyaluronic acid and neurocan still produced intense staining in almost all areas of the hindbrain, while tenascin-R labeling showed a continuous increase during postnatal development. The reaction with WFA and aggrecan was revealed first 4th postnatal day (P4) with low staining intensities, while HAPLN was detected two weeks after birth (P14). The perineuronal net appeared first around the facial and vestibular neurons at P4 with hyaluronic acid cytochemistry. One week after birth aggrecan, neurocan, tenascin-R, and WFA were also accumulated around the neurons located in several hindbrain nuclei, but HAPLN1 was detected on the second postnatal week. Our results provide further evidence that many extracellular macromolecules that will be incorporated into the perineuronal net are already expressed at embryonic and early postnatal stages of development to control differentiation, migration, and synaptogenesis of neurons. In late postnatal period, the experience-driven neuronal activity induces formation of perineuronal net to stabilize synaptic connections.

正常的大脑发育需要发育中的神经元与周围环境之间通过被称为细胞外基质(ECM)的复杂网络进行持续沟通。细胞外基质分为不同的分子家族,包括透明质酸、蛋白多糖、糖蛋白(如腱鞘蛋白)和连接蛋白。在这项研究中,我们利用抗体和凝集素组织化学方法描述了细胞外基质分子在胚胎和出生后小鼠后脑中的时空分布特征。在胚胎中,透明质酸和神经蛋白在出生前含量较高,而在整个胚胎期,透明质酸和神经蛋白的含量较低。出生后,透明质酸和神经钙在后脑几乎所有区域仍能产生强烈的染色,而tenascin-R标记在出生后的发育过程中显示出持续的增加。与 WFA 和 aggrecan 的反应最早出现在出生后第 4 天(P4),染色强度较低,而 HAPLN 则在出生后两周(P14)才被检测到。通过透明质酸细胞化学作用,神经元周围网在出生后第 4 天首先出现在面部神经元和前庭神经元周围。出生后一周,位于几个后脑核的神经元周围也积累了凝集素、神经凝集素、tenascin-R 和 WFA,但 HAPLN1 在出生后第二周才被检测到。我们的研究结果进一步证明,许多将被纳入神经元周围网的细胞外大分子已在胚胎期和出生后早期表达,以控制神经元的分化、迁移和突触发生。在出生后晚期,经验驱动的神经元活动会诱导神经元周围网的形成,以稳定突触连接。
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引用次数: 0
Glycine is a transmitter in the human and chimpanzee cochlear nuclei 甘氨酸是人类和黑猩猩耳蜗核的一种递质
IF 2.9 4区 医学 Q1 Medicine Pub Date : 2024-02-15 DOI: 10.3389/fnana.2024.1331230
Joan S. Baizer, Chet C. Sherwood, Patrick R. Hof, James F. Baker, Sandra F. Witelson
IntroductionAuditory information is relayed from the cochlea via the eighth cranial nerve to the dorsal and ventral cochlear nuclei (DCN, VCN). The organization, neurochemistry and circuitry of the cochlear nuclei (CN) have been studied in many species. It is well-established that glycine is an inhibitory transmitter in the CN of rodents and cats, with glycinergic cells in the DCN and VCN. There are, however, major differences in the laminar and cellular organization of the DCN between humans (and other primates) and rodents and cats. We therefore asked whether there might also be differences in glycinergic neurotransmission in the CN.MethodsWe studied brainstem sections from humans, chimpanzees, and cats. We used antibodies to glycine receptors (GLYR) to identify neurons receiving glycinergic input, and antibodies to the neuronal glycine transporter (GLYT2) to immunolabel glycinergic axons and terminals. We also examined archival sections immunostained for calretinin (CR) and nonphosphorylated neurofilament protein (NPNFP) to try to locate the octopus cell area (OCA), a region in the VCN that rodents has minimal glycinergic input.ResultsIn humans and chimpanzees we found widespread immunolabel for glycine receptors in DCN and in the posterior (PVCN) and anterior (AVCN) divisions of the VCN. We found a parallel distribution of GLYT2-immunolabeled fibers and puncta. The data also suggest that, as in rodents, a region containing octopus cells in cats, humans and chimpanzees has little glycinergic input.DiscussionOur results show that glycine is a major transmitter in the human and chimpanzee CN, despite the species differences in DCN organization. The sources of the glycinergic input to the CN in humans and chimpanzees are not known.
引言 听觉信息从耳蜗经由第八颅神经传递到背侧和腹侧耳蜗核(DCN、VCN)。许多物种都对耳蜗核(CN)的组织、神经化学和电路进行过研究。在啮齿动物和猫的耳蜗核中,甘氨酸是一种抑制性递质,在 DCN 和 VCN 中都有甘氨酸能细胞。然而,人类(及其他灵长类动物)与啮齿动物和猫在直流神经网的层状和细胞组织上存在很大差异。因此,我们提出了一个问题:CN 中的甘氨酸能神经递质是否也存在差异?方法我们研究了人类、黑猩猩和猫的脑干切片。我们使用甘氨酸受体(GLYR)抗体识别接受甘氨酸能输入的神经元,并使用神经元甘氨酸转运体(GLYT2)抗体免疫标记甘氨酸能轴突和末梢。我们还检查了免疫染色钙黄绿素(CR)和非磷酸化神经丝蛋白(NPNFP)的档案切片,试图确定章鱼细胞区(OCA)的位置,章鱼细胞区是啮齿类动物VCN中甘氨酸能输入最小的区域。结果在人类和黑猩猩中,我们发现DCN以及VCN的后部(PVCN)和前部(AVCN)中广泛存在甘氨酸受体免疫标记。我们发现 GLYT2 免疫标记的纤维和点状物呈平行分布。这些数据还表明,与啮齿类动物一样,猫、人和黑猩猩中含有章鱼细胞的区域几乎没有甘氨酸能输入。讨论我们的研究结果表明,尽管DCN的组织结构存在物种差异,但甘氨酸是人和黑猩猩CN的主要递质。人类和黑猩猩神经中枢的甘氨酸能输入来源尚不清楚。
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引用次数: 0
NeuroEditor: a tool to edit and visualize neuronal morphologies 神经编辑器:编辑和可视化神经元形态的工具
IF 2.9 4区 医学 Q1 Medicine Pub Date : 2024-02-14 DOI: 10.3389/fnana.2024.1342762
Ivan Velasco, Juan J. Garcia-Cantero, Juan P. Brito, Sofia Bayona, Luis Pastor, Susana Mata
The digital extraction of detailed neuronal morphologies from microscopy data is an essential step in the study of neurons. Ever since Cajal’s work, the acquisition and analysis of neuron anatomy has yielded invaluable insight into the nervous system, which has led to our present understanding of many structural and functional aspects of the brain and the nervous system, well beyond the anatomical perspective. Obtaining detailed anatomical data, though, is not a simple task. Despite recent progress, acquiring neuron details still involves using labor-intensive, error prone methods that facilitate the introduction of inaccuracies and mistakes. In consequence, getting reliable morphological tracings usually needs the completion of post-processing steps that require user intervention to ensure the extracted data accuracy. Within this framework, this paper presents NeuroEditor, a new software tool for visualization, editing and correction of previously reconstructed neuronal tracings. This tool has been developed specifically for alleviating the burden associated with the acquisition of detailed morphologies. NeuroEditor offers a set of algorithms that can automatically detect the presence of potential errors in tracings. The tool facilitates users to explore an error with a simple mouse click so that it can be corrected manually or, where applicable, automatically. In some cases, this tool can also propose a set of actions to automatically correct a particular type of error. Additionally, this tool allows users to visualize and compare the original and modified tracings, also providing a 3D mesh that approximates the neuronal membrane. The approximation of this mesh is computed and recomputed on-the-fly, reflecting any instantaneous changes during the tracing process. Moreover, NeuroEditor can be easily extended by users, who can program their own algorithms in Python and run them within the tool. Last, this paper includes an example showing how users can easily define a customized workflow by applying a sequence of editing operations. The edited morphology can then be stored, together with the corresponding 3D mesh that approximates the neuronal membrane.
从显微镜数据中以数字方式提取详细的神经元形态是研究神经元的重要步骤。自卡哈尔的研究工作以来,神经元解剖学的获取和分析为我们深入了解神经系统提供了宝贵的资料,使我们现在对大脑和神经系统的许多结构和功能方面有了超越解剖学视角的理解。不过,获取详细的解剖数据并非易事。尽管最近取得了一些进展,但获取神经元细节仍然需要使用劳动密集型、易出错的方法,这些方法容易造成误差和错误。因此,获取可靠的形态描记通常需要完成后处理步骤,这些步骤需要用户干预,以确保提取数据的准确性。在这一框架内,本文介绍了 NeuroEditor,这是一种用于可视化、编辑和修正先前重建的神经元描记的新软件工具。该工具专为减轻与获取详细形态相关的负担而开发。NeuroEditor 提供一套算法,可自动检测描记中是否存在潜在错误。用户只需轻点鼠标,该工具就能帮助用户发现错误,从而手动或自动纠正错误。在某些情况下,该工具还能提出一套自动纠正特定类型错误的操作。此外,该工具还允许用户直观地比较原始描记和修改后的描记,并提供近似神经元膜的三维网格。该网格的近似值可即时计算和重新计算,以反映描记过程中的任何瞬时变化。此外,用户还可以轻松扩展 NeuroEditor,用 Python 编写自己的算法并在工具中运行。最后,本文举例说明了用户如何通过应用一系列编辑操作来轻松定义自定义工作流程。编辑后的形态可以连同近似神经元膜的相应三维网格一起存储。
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引用次数: 0
期刊
Frontiers in Neuroanatomy
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