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Single cell labeling combined with electrophysiological identification 单细胞标记与电生理鉴定相结合。
IF 1.7 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2025-08-12 DOI: 10.1007/s12565-025-00882-x
Takahiro Furuta

Gaining insight into the circuit architecture of neural tissue significantly aids in the investigation of nervous system mechanisms. Since neural networks are constituted by connections between axons and dendrites derived from neurons, morphological analysis of each neuron is useful to reconstruct circuit diagrams of neural circuits in detail. Here, I introduce a method where we can combine single unit recordings and visualization of neuronal morphology at a single neuron level. This technique reveals morphological characteristics of neurons which are electrophysiologically identified.

深入了解神经组织的电路结构有助于神经系统机制的研究。由于神经网络是由来自神经元的轴突和树突之间的连接构成的,因此对每个神经元的形态学分析有助于详细地重建神经回路的电路图。在这里,我介绍了一种方法,我们可以在单个神经元水平上结合单个单元记录和神经元形态的可视化。这项技术揭示了电生理学鉴定的神经元的形态特征。
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引用次数: 0
A detailed protocol for sparse and bright labeling of neurons via dual adeno-associated virus vectors with Cre recombination 通过双腺相关病毒载体与Cre重组对神经元进行稀疏和明亮标记的详细方案。
IF 1.7 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2025-08-08 DOI: 10.1007/s12565-025-00867-w
Megumu Takahashi, Hidenobu Mizuno, Masato Koike, Takuji Iwasato, Hiroyuki Hioki

Adeno-associated virus (AAV) vectors are widely used to label individual neurons by expressing fluorescent proteins. To analyze neuronal morphology in detail, it is important to achieve sparse and strong expression of fluorescent proteins for clearly visualizing cell bodies, dendrites, and axons. However, injections of high-titer AAV vectors often result in excessive cell labeling, while injecting low-titer AAV vectors reduces signal intensity. Here, we provide a detailed, step-by-step protocol for achieving sparse and bright neuronal labeling using dual AAV vectors with Cre recombination, based on the Supernova system. We prepared two types of AAV vectors: one is a driver vector (AAV2/1-SYN-iCre-BGHpA) that expresses Cre recombinase under a neuron-specific promoter, and the other is a reporter vector (AAV2/1-SynTetOff FLEX-GFP) that encodes GFP with a flip-excision switch (FLEX) and the Tet-Off system. When a mixture of a serially diluted reporter vector and a high-titer driver vector was stereotaxically injected into the mouse brain, the number of labeled neurons and the fluorescence intensity of GFP decreased in a dose-dependent manner. Conversely, diluting the driver vector decreased the number of GFP-expressing cells while the fluorescence intensity of GFP remained high. Furthermore, this approach proved effective for systemic delivery via retro-orbital injection of AAV2/B10 vectors. This protocol includes comprehensive reagent lists, stepwise injection procedures, and troubleshooting tips, allowing researchers to reproducibly achieve sparse and bright labeling through both stereotaxic and retro-orbital injections.

腺相关病毒(AAV)载体通过表达荧光蛋白被广泛用于标记单个神经元。为了详细分析神经元形态,实现荧光蛋白的稀疏和强表达对于清晰地观察细胞体、树突和轴突是很重要的。然而,注射高滴度的AAV载体往往会导致过度的细胞标记,而注射低滴度的AAV载体会降低信号强度。在这里,我们提供了一个详细的,一步一步的协议,实现稀疏和明亮的神经元标记使用双AAV载体与Cre重组,基于超新星系统。我们制备了两种类型的AAV载体:一种是驱动载体(AAV2/1-SYN-iCre-BGHpA),在神经元特异性启动子下表达Cre重组酶;另一种是报告载体(AAV2/1-SynTetOff FLEX-GFP),通过翻转切除开关(FLEX)和Tet-Off系统编码GFP。将连续稀释的报告载体和高滴度驱动载体的混合物立体定向注射到小鼠大脑中,标记神经元的数量和GFP的荧光强度呈剂量依赖性下降。相反,稀释驱动载体减少了表达GFP的细胞数量,而GFP的荧光强度仍然很高。此外,该方法被证明可以通过眶后注射AAV2/B10载体进行全身递送。该方案包括全面的试剂清单,逐步注射程序和故障排除提示,允许研究人员通过立体定向和后眼眶注射可重复地实现稀疏和明亮的标记。
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引用次数: 0
Correction: Introduction to the CLEM technique developed in the field of neuroanatomy 更正:介绍在神经解剖学领域发展起来的CLEM技术。
IF 1.7 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2025-08-07 DOI: 10.1007/s12565-025-00893-8
Takaichi Fukuda
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引用次数: 0
Internal carotid artery agenesis: a systematic review of prevalence, basal brain circulation variations, and associated pathologies. 颈内动脉发育不全:患病率、基础脑循环变化和相关病理的系统综述。
IF 1.7 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2025-08-06 DOI: 10.1007/s12565-025-00873-y
Mathias Orellana-Donoso, Juan Sanchis-Gimeno, Sary Afandi-Rebolledo, Pablo Nova-Baeza, Paloma García-Mena, Alejandra Suazo-Santibañez, Rocío Peñailillo-Ibarra, Iván Valdés-Orrego, Juan José Valenzuela-Fuenzalida

The internal carotid artery (ICA) is a crucial vessel, and its congenital absence, known as ICA agenesis, is a rare vascular anomaly. This systematic review aimed to provide a comprehensive overview of the available evidence on this condition. We systematically searched multiple databases from inception until January 2024. Two authors independently screened titles and abstracts, and a third reviewer was involved if consensus could not be reached. Data extraction and quality assessment were performed using validated the AQUA tool and the JBI critical appraisal checklist tool. A total of 138 (7.8%) articles out of 1,778 met the inclusion criteria, reporting 2040 subjects from which 176 presented (8.6%) ICA agenesis. Most of the studies were case reports (96%), and the included studies originated from various countries, with the United States (17.39%), Japan (14.49%), Turkey (10.87%), and India (10.14%) contributing the most. Left-sided agenesis was the most common (67.39%), followed by right-sided (42.03%) and bilateral (22.46%) agenesis. The most reported clinical manifestations were Horner's syndrome, increased risk of intracranial aneurysms, and pituitary gland abnormalities. ICA agenesis is a rare vascular anomaly with a left-sided predominance. While some patients may develop associated clinical conditions, many remain asymptomatic due to the development of robust collateral circulation. Comprehensive preoperative evaluation and awareness of these anatomical variations are crucial to guide surgical planning and minimize the risk of complications. PROSPERO registration number: CRD42024592673.

颈内动脉(ICA)是至关重要的血管,其先天性缺失,称为ICA发育不全,是一种罕见的血管异常。本系统综述的目的是对这一疾病的现有证据提供一个全面的概述。从成立到2024年1月,我们系统地检索了多个数据库。两位作者独立筛选标题和摘要,如果无法达成共识,则会有第三位审稿人参与。使用经过验证的AQUA工具和JBI关键评估清单工具进行数据提取和质量评估。1778篇文章中有138篇(7.8%)符合纳入标准,报告了2040名受试者,其中176篇(8.6%)出现ICA发病。大多数研究为病例报告(96%),纳入的研究来自不同国家,其中美国(17.39%)、日本(14.49%)、土耳其(10.87%)和印度(10.14%)贡献最多。左侧发育不全最常见(67.39%),其次为右侧(42.03%)和双侧(22.46%)。报告最多的临床表现为霍纳综合征,颅内动脉瘤的风险增加,脑垂体异常。ICA发育不全是一种罕见的血管异常,以左侧为主。虽然一些患者可能会出现相关的临床症状,但由于侧支循环的健全,许多患者仍无症状。全面的术前评估和了解这些解剖变异对指导手术计划和减少并发症的风险至关重要。普洛斯彼罗注册号:CRD42024592673。
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引用次数: 0
Direct imaging of the three-dimensional ultrastructure of neuronal organelles 神经元细胞器三维超微结构的直接成像。
IF 1.7 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2025-08-05 DOI: 10.1007/s12565-025-00888-5
Daisuke Koga, Ryosuke Morinaga, Satoshi Kusumi

In the context of cell morphological analyses observing organelles embedded within the cell matrix is difficult. The osmium maceration method is a unique technique used to directly observe the three-dimensional structure of organelles through scanning electron microscopy, without requiring time-consuming and labor-intensive reconstruction. In this method, tissues are immersed in a diluted osmium solution for several days to remove cytosolic soluble proteins and filamentous structures, including microfilaments, intermediate filaments, and microtubules, from the freeze-cracked surfaces of cells, leaving the subcellular structures, Golgi apparatus, mitochondria, and smooth and rough endoplasmic reticulum intact. Specimen preparation involves several key steps, specifically pre-fixation with aldehyde fixatives, tissue excision, trimming, post-fixation with osmium tetroxide solution, dimethyl sulfoxide cracking (i.e., freeze-cracking), the thawing of cracked tissues, osmium maceration, osmium fixation, conductive staining (tannin–osmium method), dehydration, drying, mounting, metal coating, and scanning electron microscopy observations. Here, we present a step-by-step protocol based on the maceration method using neural cells as an example, ensuring reproducibility and consistent results for neurons and various other cell types. Moreover, the results presented indicate that the osmium maceration method is effective for elucidating the three-dimensional intracellular ultrastructure of neurons.

在细胞形态学分析的背景下,观察嵌入细胞基质中的细胞器是困难的。锇浸渍法是一种独特的技术,可以通过扫描电子显微镜直接观察细胞器的三维结构,不需要耗时费力的重建。在这种方法中,将组织浸泡在稀释的锇溶液中数天,从细胞冻裂的表面去除胞质可溶性蛋白和丝状结构,包括微丝、中间丝和微管,留下亚细胞结构、高尔基体、线粒体和光滑和粗糙的内质网。标本制备包括几个关键步骤,特别是乙醛固定剂预固定、组织切除、修剪、四氧化锇溶液后固定、二甲亚砜裂解(即冷冻裂解)、裂解组织解冻、锇浸渍、锇固定、导电染色(单宁-锇法)、脱水、干燥、安装、金属涂层和扫描电镜观察。在这里,我们提出了一个基于浸渍法的一步一步的协议,以神经细胞为例,确保神经元和各种其他细胞类型的可重复性和一致的结果。此外,实验结果表明锇浸渍法可以有效地阐明神经元细胞内三维超微结构。
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引用次数: 0
Hippocampal architecture viewed through the eyes of methodological development 从方法论发展的角度看海马体结构。
IF 1.7 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2025-08-05 DOI: 10.1007/s12565-025-00878-7
Ling Zhao, Nicola Palomero-Gallagher

This review focuses on the structural organization of the hippocampus and how our understanding of its cellular architecture and functional circuits has been enabled over the last 400 years by the development of methods as varied as the Golgi impregnation, immunohistochemical staining procedures, and polarized light imaging. We provide an outlook on how cutting-edge techniques such as high-resolution imaging and artificial intelligence may continue to shed light on the structural organization of the hippocampus and emphasize the importance of collaborative multidisciplinary efforts including classical neuroanatomists in this endeavor.

这篇综述的重点是海马的结构组织,以及我们对其细胞结构和功能电路的理解是如何在过去的400年里通过高尔基浸渍、免疫组织化学染色和偏振光成像等多种方法的发展而得以实现的。我们展望了高分辨率成像和人工智能等尖端技术如何继续揭示海马体的结构组织,并强调了包括经典神经解剖学家在内的多学科合作的重要性。
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引用次数: 0
Correction: A systematic review of the use of human body donor models for postgraduate laparoscopic surgical training. 更正:对研究生腹腔镜手术训练中使用人体供体模型的系统回顾。
IF 1.7 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2025-08-04 DOI: 10.1007/s12565-025-00889-4
Sedat Alp Pinar, Joseph J Morrow, Chia Yew Kong
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引用次数: 0
Seven (quadruple right and triple left) renal arteries and their topological relationship with other branches of the abdominal aorta. 七根肾动脉(四根右动脉和三根左动脉)及其与腹主动脉其他分支的拓扑关系。
IF 1.7 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2025-08-02 DOI: 10.1007/s12565-025-00890-x
Shigeyuki Esumi, Yoshikazu Koba, Yoshihiro Kumagai, Yoko Nishitani, Takaichi Fukuda

Supernumerary renal arteries are frequently observed, but their developmental origin remains controversial. One hypothesis, the ladder theory, ascribed origins of multiple renal arteries to a segmental array of mesonephric arteries. However, recent studies in both rodents and humans demonstrated evidence against this theory. We describe a rare variation in a Japanese female cadaver exhibiting seven renal arteries encountered during student dissection practice in 2023. On the right side, the main renal artery branched off from the aorta 29 mm below the origin of the superior mesenteric artery. Three additional renal arteries branched off at the level of or below the inferior mesenteric artery. On the left side, two renal arteries branched off at usual high positions, but another branch below the inferior mesenteric artery. In 43 cadavers dissected in 2023, all renal arteries including duplicate type, but except for the present case, arose at high positions near the superior mesenteric artery. Incidence of connection between multiple renal arteries and adrenal and/or gonadal arteries was 78.6% (11 out of 14 kidneys). Importantly, all these connections were observed at high-positioned branches. Combining present results with previous data in literatures revealed the close relationship between high-positional branching and connections with adrenal/gonadal arteries (p = 0.0056), suggesting two factors influencing renal artery development: (1) prenatal, interconnected arteries at high positions, and (2) low-positioned individual branches arising from the aorta during kidney ascent with poor interconnectivity. Arteries involved in these two events might regress with various degrees, leading to diversity in number and connectivity of renal arteries.

肾多余动脉经常被观察到,但其发育起源仍有争议。一种假说,阶梯理论,将多肾动脉的起源归因于肾中动脉的节段性排列。然而,最近对啮齿动物和人类的研究证明了反对这一理论的证据。我们描述了一个罕见的变异在日本女性尸体表现出七个肾动脉在学生解剖实践中遇到的2023。在右侧,肾主动脉在肠系膜上动脉起源下方29毫米处从主动脉分叉。另外三条肾动脉在肠系膜下动脉的水平或以下分支。在左侧,两条肾动脉在通常的高位分支,但另一条分支位于肠系膜下动脉下方。在2023年解剖的43具尸体中,除本病例外,所有肾动脉(包括重复型)均出现在靠近肠系膜上动脉的高位。多肾动脉与肾上腺和/或性腺动脉连接的发生率为78.6%(14个肾脏中的11个)。重要的是,所有这些连接都是在高位分支上观察到的。将本研究结果与以往文献数据相结合,发现高位置分支与肾上腺/性腺动脉连接密切相关(p = 0.0056),提示影响肾动脉发育的因素有两个:(1)产前高位置动脉相互连接,(2)肾上升时主动脉产生的低位置单个分支相互连接差。参与这两种事件的动脉可能有不同程度的退化,导致肾动脉数量和连通性的多样性。
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引用次数: 0
Magnetic resonance angiography-based assessment of the renal artery and its branching pattern. 基于磁共振血管造影的肾动脉及其分支形态评估。
IF 1.7 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2025-08-01 DOI: 10.1007/s12565-025-00891-w
Necmi Keskin, Belgin Bamac, Ozgur Cakir, Tuncay Colak, Cagatay Barut

This study aimed to investigate the detailed anatomy of the renal arteries through magnetic resonance angiography (MRA), offering an alternative to conventional imaging techniques. By identifying the variations, spatial orientation, and vertebral origin of the renal arteries, the study seeks to provide comprehensive anatomical data that may support surgeons and interventional radiologists in clinical planning and decision-making. Magnetic resonance angiography images of 196 individuals who underwent abdominal imaging were retrospectively retrieved from the Picture Archiving and Communication System (PACS). The analysis focused on the vertebral level of renal artery origin, angle of emergence, distances from major abdominal aortic branches, and renal artery branching patterns. These parameters were statistically assessed based on age and sex using SPSS 25. In male individuals, the distances between the renal arteries and both the inferior mesenteric artery and the aortic bifurcation were significantly greater than in females (p < 0.05). A statistically significant positive correlation was observed between age and the coronal angle of the left renal artery in both sexes and in the overall group (p = 0.001). The most frequent origin point of the right renal artery was identified at the mid-level between the L1 and L2 vertebrae, whereas the left renal artery commonly originated at the upper level of L2 in both sexes. The anatomical parameters defined by age and sex in this study may improve diagnostic accuracy for clinicians. These findings can aid interventional radiologists in catheter selection and procedure planning, supporting more personalized and effective interventions.

本研究旨在通过磁共振血管造影(MRA)研究肾动脉的详细解剖结构,提供一种替代传统成像技术的方法。通过确定肾动脉的变异、空间方向和椎体起源,本研究旨在提供全面的解剖学数据,以支持外科医生和介入放射科医生进行临床计划和决策。从图片存档和通信系统(PACS)中回顾性检索196例接受腹部成像的患者的磁共振血管成像图像。分析主要集中在肾动脉起源的椎体水平、出现角度、与腹主动脉主要分支的距离和肾动脉分支模式。这些参数根据年龄和性别使用SPSS 25进行统计评估。在男性个体中,肾动脉与肠系膜下动脉和主动脉分叉之间的距离明显大于女性(p
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引用次数: 0
Morphometric analysis of mental foramen in retained cadaveric specimens of mandibles of Sri Lankan population. 斯里兰卡人下颌骨尸体标本颏孔形态计量学分析。
IF 1.7 4区 医学 Q3 ANATOMY & MORPHOLOGY Pub Date : 2025-07-31 DOI: 10.1007/s12565-025-00883-w
Dadallage Tharanga De Silva, Usliyanage Clifford Priyantha Perera

Understanding the variation of the location of the mental foramen in the mandible will aid in the prevention of injury to associated structures during invasive procedures in the mental area. This study was carried out using 228 mental foramina in 114 dry mandibles belonging to the adult Sri Lankan population of the southern region of Sri Lanka to determine the shape, size, direction, existence of accessory foramina, and location with respect to anatomical landmarks. A single mental foramen was the most common in Sri Lankan adults, and 7% of them had accessory mental foramen on either side. The most common characteristic of the mental foramen was an oval shape, located below the 2nd premolar tooth in the posterior-superior direction. The mean distance from the mandibular midline to the mental foramen was 25.98 ± 1.9 mm and 25.64 ± 2.02 mm on the right and left sides, respectively. This study offers substantial data about possible variations in physical characteristics and the location of the mental foramen in a population in the southern region of Sri Lanka. Hence, it is possible to use the relative position of teeth or distances from various borders of the mandible to precisely locate the mental foramen. Therefore, this information is crucial for clinicians, who must be aware of these variances and the necessity of determining the precise location of the mental foramen before doing surgery in this region.

了解下颌骨颏孔位置的变化将有助于预防在颏区侵入性手术中对相关结构的损伤。本研究利用斯里兰卡南部地区斯里兰卡成年人口的114个干下颌骨的228个颏孔来确定其形状、大小、方向、附属孔的存在以及与解剖标志相关的位置。斯里兰卡成年人最常见的是单一的精神孔,其中7%的人在两侧都有副精神孔。颏孔最常见的特征为椭圆形,位于第2前磨牙下方后上方向。下颌中线至颏孔的平均左右距离分别为25.98±1.9 mm和25.64±2.02 mm。这项研究为斯里兰卡南部地区人群的身体特征和心理孔位置的可能变化提供了大量数据。因此,可以使用牙齿的相对位置或与下颌骨各边界的距离来精确定位颏孔。因此,这一信息对临床医生来说是至关重要的,他们必须意识到这些差异,并在该区域进行手术前确定精神孔的精确位置。
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引用次数: 0
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Anatomical Science International
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