Anatomical Science International最新文献

ATAC-seq (assay for transposase-accessible chromatin using sequencing) enables high resolution mapping of chromatin accessibility, providing critical insights into gene regulatory mechanisms in neuroscience research. This protocol presents an optimized approach for applying ATAC-seq to neural tissues and cells, addressing the unique challenges of preserving chromatin integrity in these delicate samples. We detail the key steps of tissue processing, nuclear isolation, transposition reactions, and library preparation specifically adapted for neural applications. The method requires minimal starting material (20,000–50,000 cells) while maintaining high sensitivity for detecting regulatory elements involved in neural development, plasticity, and neurological disorders. This tailored protocol facilitates robust investigation of neuroepigenomic regulation in diverse experimental contexts.

High-speed atomic force microscopy (HS-AFM) enables real-time visualization of biological processes with nanometer-level resolution. This review highlights how HS-AFM has been instrumental in uncovering the dynamic interplay between nuclear pore complexes (NPCs)—which regulate nucleocytoplasmic transport—and genome guardians, including DNA repair proteins and chromatin regulators. Structurally, the NPCs resemble a multi-layered spider cobweb, serving as crucial molecular gatekeepers for maintaining cellular homeostasis, while genome guardians safeguard genomic integrity through DNA repair and chromatin organization. Through HS-AFM, the researchers have gained unprecedented insights into NPC dynamics, revealing their adaptability during nuclear transport, chromatin reorganization, and viral infection. It has also elucidated how genome guardians interact with NPCs, influencing chromatin organization at the nuclear periphery and regulating nucleocytoplasmic trafficking. These discoveries underscore the critical role of NPC-genome interactions in genome stability, gene expression, and nuclear transport, with broad implications for diseases such as cancer, viral infections, and neurodegenerative disorders. In conclusion, HS-AFM has transformed our ability to study the nuclear landscape at the nanoscale, bridging the gap between structural biology and functional genomics. By capturing the real-time molecular dynamics of NPCs and chromatin, HS-AFM provides an essential tool for unraveling the mechanisms that govern nuclear transport and genome regulation. Future advancements in HS-AFM technology, including higher temporal resolution, correlative imaging, and AI-driven analysis, will further expand its potential in biomedical research, paving the way for novel diagnostic and therapeutic strategies.

Tyramide signal amplification (TSA) is a highly sensitive enzymatic method for histochemical analysis. However, its application to multiplex staining is limited by quenching the catalytic activity of peroxidase (POD). Here, we provide a detailed protocol for multiplex immunofluorescence (IF) staining in mouse brain sections using a fluorescent TSA system, fluorochromized tyramide‑glucose oxidase (FT-GO). FT-GO utilizes hydrogen peroxide produced by oxidation of glucose by glucose oxidase for covalent deposition of FT onto tissue sections. For multiplex labeling with the TSA system, we inactivate antibody-conjugated POD using sodium azide. We describe tissue section preparation, triple FT-GO IF staining and confocal laser scanning microscopy. For complete details on the use and execution of this protocol, please refer to Yamauchi et al. (2022).

Mammalian visual and genital (hereafter mainly penile) organs have been extensively studied albeit separately. Both organ systems contain sensation devices necessary for visual perception and sexual intercourse. Their terminal structures are covered with eyelid/prepuce followed by the sensitive epithelia of cornea/glans facing the eyeball and glans. These structures have been closely studied in humans for appropriate visual perception and copulation and have thus been treated by numerous surgeries for long periods. Despite the vastly divergent anatomy and physiological functions, there are a few intriguing topohistological similarities for both structures, functions, and pathology. The current article focuses on such features from various viewpoints.

This article explores the historical progression of studying neuronal connections, beginning with nineteenth-century advancements in light microscopy and histological techniques. Early methods were limited in terms of their capacity to trace neuronal connections, but a breakthrough came with Camillo Golgi’s “black reaction” staining method, later refined and extensively used by Santiago Ramón y Cajal. Cajal’s observations supported the Neuron Theory, which proposed that neurons communicate via specialized points of contact, contradicting the prevailing Reticular Theory of a continuous neural network, which was supported by Golgi. This contrast is particularly intriguing because, although Golgi and Cajal used the same black reaction technique and similar microscopes, their interpretations of the microscopic world diverged significantly. An important consequence of the Neuron Theory was Cajal’s Law of Dynamic Polarization, which proposed that neurons generally consist of three functionally distinct regions: a receptor apparatus (dendrites and soma), an emission apparatus (axon), and a distribution apparatus (terminal axonal arborization). He applied this principle across various parts of the nervous system and to different neuron types, enabling the generation of the first detailed circuit diagrams of the brain. Cajal’s observations, concepts, and theories had a profound impact—not only on researchers of his time, but also on modern neuroscience. This article reflects on the early studies of neuronal connections, highlighting the scientific climate in which Golgi and Cajal initiated their groundbreaking research.

Os peroneum (OP) is one of the most common accessory bones in the human foot. The literature indicates that its prevalence varies significantly and ranges between 0.4 and 20.3%. In the clinical context, OP is the main factor responsible for the pain condition known as os peroneum syndrome, which usually manifests as pain in the lateral side of the foot. The purpose of this study is to provide a comprehensive synthesis of data regarding the prevalence of this ossicle. To find the pertinent articles, a thorough search of the major electronic bases was carried out. Data on imaging modality, sex distribution, prevalence, and geographic origin of OP were extracted. Throughout the study, the PRISMA guidelines were strictly followed. The AQUA tool was used to evaluate the reliability of the included studies. 26 studies (22,948 feet) were included in the meta-analysis. The pooled prevalence estimate (PPE) of OP was found to be 6.6% (95% CI: 5.1–8.5) (95% PI: 0.02–0.22) of the analyzed feet. In the X-ray-based subgroup, the PPE of OP was 6.7% (95% CI: 5.1–8.7) and in the cadaveric dissection-based subgroup was 11.1% (95% CI: 5.1–22.4). The highest prevalence of OP was observed in North America (8.6% (95% CI: 5.9–12.3)), followed by Europe (6.0%, (95% CI: 3.8–9.4)) and Asia (5.9% (95% CI: 3.9–9.0)). Os peroneum is a very common accessory ossicle which occurs approximately every fifteen feet. The highest prevalence of OP was found in the North American population. The occurrence of pain in the lateral part of the foot should draw physicians attention into considering a potential presence of OP during the differential diagnosis.