Denise Bonente, Tiziana Tamborrino, Niccolò Fagni, Sandra Bracco, Sara Leonini, Sara Ottolenghi, Virginia Barone, Eugenio Bertelli
{"title":"Radiological anatomy of the trochlear spine and associated bony structures around the superior oblique tendon: a CT-based study.","authors":"Denise Bonente, Tiziana Tamborrino, Niccolò Fagni, Sandra Bracco, Sara Leonini, Sara Ottolenghi, Virginia Barone, Eugenio Bertelli","doi":"10.1007/s12565-025-00871-0","DOIUrl":"https://doi.org/10.1007/s12565-025-00871-0","url":null,"abstract":"<p><p>The superior oblique muscle tendon is known to bend in the anterior orbit around a cartilaginous trochlea. The site where the tendon bends is frequently interested by the presence of a small depression, the trochlear fovea, and/or the trochlear spine. Exact topography, size and frequency of these items are still undetermined. For this purpose, we studied 120 orbits of individuals that underwent computed tomography for pathologies not involving the anterior orbit. We detected the presence of the trochlear spine in 10% of orbits and we determined its location and size. We also observed the presence of two tubercles (TT<sub>1</sub> and TT<sub>2</sub>), with distinct positions relative to the tendon. TT<sub>1</sub> was present in 5% of orbits and was located on the same spot of the spine differing from the latter only for its morphology. TT<sub>2</sub> lied in a more advanced position and it was rarer (1,67% of orbits). The spine and the first type of tubercle were located above and behind the tendon reflection; the second tubercle lied below and ahead of the tendon reflection. A distinct trochlear fovea was detected in 25.83% of orbits and lied 3.42 ± 0.97 mm behind the orbital rim. Fovea diameters were 4.16 ± 1.08 mm × 3.84 ± 0.97 mm. In conclusion we demonstrate that in the anterior orbit a bony process is present in at least 15% or orbits. It is a note of interest for strabismus surgery when it is necessary to intervene on the superior oblique muscle or when it is needed access to the medial orbital wall.</p>","PeriodicalId":7816,"journal":{"name":"Anatomical Science International","volume":" ","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144641579","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"A condylar imprint of Farabeuf's knocker and associated cervical variants.","authors":"Mugurel Constantin Rusu, Răzvan Costin Tudose","doi":"10.1007/s12565-025-00870-1","DOIUrl":"https://doi.org/10.1007/s12565-025-00870-1","url":null,"abstract":"<p><p>Different head and neck anatomical structures, such as the temporomandibular joint, carotid arteries and their branches, styloid process, hyoid and thyroid cartilage, are subjected to anatomical variations. A range of novel and rare anatomical variants were found during the anatomical exploration of a DICOM file. The archived angioCT file of a 63-year-old male was examined anatomically. There were no pathological processes or other artefacts to distort the anatomical patterns. The imprint of Farabeuf's knocker (posterior zygomatic tubercle) on the posterior side of the right mandibular condyle was found. This is an anatomical novelty. Rare neck variations were found: a right-tilted and collapsed hyoid bone over the thyroid cartilage and the agenesis of the left superior horn of this cartilage. Variations of the external carotid arteries' branches were observed. A right thyrolingual trunk gave off a superior thyroid artery that supplied both thyroid lobes with distinctive right and left branches, the opposite superior thyroid artery being absent. On both sides, the superficial temporal artery coursed over the temporomandibular joints. The right internal carotid artery coiled immediately beneath the skull base. It is recommended that CT scans be carefully explored, as various concomitant anatomical variations, including rare or previously unreported findings, may be encountered.</p>","PeriodicalId":7816,"journal":{"name":"Anatomical Science International","volume":" ","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144635935","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Single-cell endogenous protein labeling via CRISPR-Cas9-mediated genome editing in the mouse brain","authors":"Motokazu Uchigashima, Takayasu Mikuni","doi":"10.1007/s12565-025-00866-x","DOIUrl":"10.1007/s12565-025-00866-x","url":null,"abstract":"<div><p>High-precision mapping of endogenous proteins is essential for understanding the molecular mechanism underlying neuronal functions in the brain. The SLENDR (single-cell labeling of endogenous proteins by clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9-mediated homology-directed repair) technique provides single-cell endogenous protein labeling with genetically encoded tags within the mammalian brain through precise genome editing via homology-directed repair (HDR). This technique is based on the introduction of HDR-mediated genome editing into neuronal progenitors in embryonic brains by in utero electroporation. Subsequent histological analyses enable high-resolution interrogation of the subcellular distribution of endogenous proteins within a single neuron using conventional fluorescent microscopy. Here, we describe a step-by-step protocol for the SLENDR technique to label endogenous proteins with genetically encoded tags in single pyramidal cells of the mouse primary somatosensory cortex. This protocol would be helpful to visualize the molecular organization underlying biological processes at single-neuron levels in the brain, such as signal processing from synaptic inputs to neuronal outputs across different scales.</p></div>","PeriodicalId":7816,"journal":{"name":"Anatomical Science International","volume":"100 :","pages":"579 - 590"},"PeriodicalIF":1.7,"publicationDate":"2025-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144625288","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Kazunori Toida, Keita Satoh, Haruyo Yamanishi, Yukari Minami-Ogawa, Simone N. T. Kurial, Emi Kiyokage
{"title":"Correlative light and electron microscopy analysis for neurobiological applications","authors":"Kazunori Toida, Keita Satoh, Haruyo Yamanishi, Yukari Minami-Ogawa, Simone N. T. Kurial, Emi Kiyokage","doi":"10.1007/s12565-025-00868-9","DOIUrl":"10.1007/s12565-025-00868-9","url":null,"abstract":"<div><p>In this article, we will introduce the significance of correlative light and electron microscopy analyses, an integrated morphological analysis method that directly combines laser and electron microscopes. This protocol comprehensively instructs the appropriate methods for the visualization of genetically labeled neurons in mice using digitalized electron microscopes. We wish to consider with the readers the importance and functional potential of electron microscopic analysis for medical and biological applications, especially for neurobiological applications.</p></div>","PeriodicalId":7816,"journal":{"name":"Anatomical Science International","volume":"100 :","pages":"455 - 464"},"PeriodicalIF":1.7,"publicationDate":"2025-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144590290","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Acceleration of axonal regeneration by GABA/Gly excitation.","authors":"Chitoshi Takayama, Tsukasa Yafuso","doi":"10.1007/s12565-025-00869-8","DOIUrl":"https://doi.org/10.1007/s12565-025-00869-8","url":null,"abstract":"<p><p>In the mature central nervous system (CNS), γ-aminobutyric acid (GABA) and glycine (Gly) are predominant inhibitory neurotransmitters that negatively regulate neural activities. In contrast, GABA mediates membrane potential depolarization during development, and GABA/Gly become excitatory after nerve injury because of the high intracellular Cl<sup>-</sup> concentration induced by low expression of K<sup>+</sup>, Cl<sup>-</sup> cotransporter 2 (KCC2), which transports Cl<sup>-</sup> out of neurons. Many studies have reported that during CNS development, GABAergic excitatory action might play crucial roles in neurogenesis through Ca<sup>2+</sup> influx. Nevertheless, its involvement in neurogenesis has not been proven because the CNS can develop normally without GABAergic signals. Recently, two research groups demonstrated that low level of KCC2 (i.e., GABA/Gly excitation) after nerve injury is involved in axonal regeneration and in enhancement of functional recovery. In this manuscript, we review GABA/Gly excitation and introduce recent findings describing its involvement in axonal regeneration.</p>","PeriodicalId":7816,"journal":{"name":"Anatomical Science International","volume":" ","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144590289","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sven Hildebrand, Johannes Franz, Shubharthi Sengupta, Anna Schueth, Andreas Herrler, Alard Roebroeck
{"title":"ht-MASH: a high-throughput, cost-effective, and robust protocol for microscopic 3D imaging of human angio- and cytoarchitecture in large human brain samples","authors":"Sven Hildebrand, Johannes Franz, Shubharthi Sengupta, Anna Schueth, Andreas Herrler, Alard Roebroeck","doi":"10.1007/s12565-025-00859-w","DOIUrl":"10.1007/s12565-025-00859-w","url":null,"abstract":"<div><p>The performance of many optical tissue clearing protocols has considerably improved in the last few years, so that now even notoriously difficult specimen such as highly myelinated human brain tissue can be rendered highly transparent. However, optical tissue clearing is still routinely performed on relatively small samples, especially in the case of the human brain. Recent advances in histological tissue processing now allow scaling up the clearing process considerably towards much larger samples. Yet so far, these methods can have considerable drawbacks in their feasibility to be implemented routinely, especially in smaller laboratories. Here, we present an updated version of our MASH protocol, which allows optical tissue clearing of very large human brain tissue samples and labelling of angio- and cytoarchitecture therein. This pipeline is cost-efficient and easy to implement, so that even smaller labs can apply it at scale. At the same time, the use of rapid prototyping using 3D printing to create custom clearing equipment is versatile enough to be adjusted to other optical tissue clearing methods than the one used in this study (e.g., aqueous methods such as CUBIC or other solvent-based methods of the DISCO family), sample sizes or tissue types. Our pipeline has, therefore, the potential to advance optical tissue clearing and labelling of large human tissue samples towards a more robust and routine implementation in the blooming field of 3D histology.</p><h3>Graphical abstract</h3>\u0000<div><figure><div><div><picture><source><img></source></picture></div></div></figure></div></div>","PeriodicalId":7816,"journal":{"name":"Anatomical Science International","volume":"100 :","pages":"498 - 513"},"PeriodicalIF":1.7,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s12565-025-00859-w.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144582846","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Tracing of specific neural pathways in the rat brain using lentiviral vectors for retrograde gene transfer","authors":"Saaya Akama, Yoshio Iguchi, Shigeki Kato, Kazuto Kobayashi","doi":"10.1007/s12565-025-00864-z","DOIUrl":"10.1007/s12565-025-00864-z","url":null,"abstract":"<div><p>Understanding the structural and the functional organization of neural circuits in the brain is a fundamental goal of neuroscience. Lentiviral vectors for retrograde gene transfer transduce neurons through the entry from synaptic terminals and enable tracing and manipulation of neuronal populations of interests based on the synaptic connections. The highly efficient and neuron-specific retrograde gene transfer (NeuRet) vectors are derived from the pseudotyping of human immunodeficiency virus type 1-based vectors with fusion envelope glycoproteins. Viral RNA genome is reverse-transcribed and synthesized DNA is integrated into the host genomes, allowing stable and long-term expression of transgenes. Dorsal striatum, the input layer of the basal ganglia, integrates signals from various brain regions, including cerebral cortex, thalamus, and ventral midbrain, and plays a critical role in motor control, learning, and decision-making. Consequently, understanding the input and the output connectivity of the dorsal striatum is fundamental to revealing how circuits within the basal ganglia contribute to physiological and behavioral functions, and its impairments are related to neurological disorders. This paper outlines the procedures for injecting a NeuRet vector carrying a green fluorescent protein gene into the sub-regions of dorsal striatum in rats, followed by immunohistochemistry to detect the transgene expression in the brain.</p></div>","PeriodicalId":7816,"journal":{"name":"Anatomical Science International","volume":"100 :","pages":"413 - 419"},"PeriodicalIF":1.7,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144537798","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Visualization of nanostructures in neuronal growth cones and axons using super-resolution structured illumination microscopy","authors":"Motohiro Nozumi, Michihiro Igarashi","doi":"10.1007/s12565-025-00863-0","DOIUrl":"10.1007/s12565-025-00863-0","url":null,"abstract":"<div><p>In neurons, which possess highly organized cellular structures, such as growth cones, axons, synapses, and dendritic spines, precise determination of molecular localization is a powerful approach for elucidating cellular functions. Super-resolution microscopy has revealed higher-order structures and molecular distributions that were previously undetectable using conventional confocal microscopy. Among super-resolution techniques, structured illumination microscopy (SIM) is particularly well-suited for live-cell imaging. In this study, we present a detailed methodology for observing growth cones and axons using SIM. Our SIM imaging of primary mouse neurons revealed that the phosphorylated GAP-43 localizes within axons in a pattern consistent with the membrane-associated periodic skeleton.</p></div>","PeriodicalId":7816,"journal":{"name":"Anatomical Science International","volume":"100 :","pages":"591 - 597"},"PeriodicalIF":1.7,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144526109","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Combination method of macroscopic imaging with MRI and microscopic imaging with super resolution microscopy for brain microstructure analysis","authors":"Yuka Nakano, Kenji F. Tanaka, Yoshifumi Abe","doi":"10.1007/s12565-025-00862-1","DOIUrl":"10.1007/s12565-025-00862-1","url":null,"abstract":"<div><p>Identifying disease-relevant and therapy-related brain regions remains a major challenge in studies using animal models of psychiatric disorders. Conventional hypothesis-driven approaches often result in limited or subjective identification of brain regions. In this study, we propose an integrative method combining unbiased whole-brain structural magnetic resonance imaging (MRI) screening and detailed histological analysis. Our methodology uses structural MRI to systematically detect volumetric changes across the entire brain, enabling the identification of target regions without relying on predefined hypotheses. Once brain regions are identified, super-resolution microscopy (SRM) is employed to determine microstructural alterations at the cellular level, focusing on neurons and glial cells within those regions. To exemplify the utility of this method, we applied it to a mouse model treated with electroconvulsive therapy (ECT), an intervention which is known to increase hippocampal volume. Our demonstration highlights the potential of this approach to systematically search for brain regions of interest, providing valuable insights and guiding future studies toward a more focused exploration of key aspects of psychiatric disorder research, both in terms of pathophysiology and therapeutic action.</p></div>","PeriodicalId":7816,"journal":{"name":"Anatomical Science International","volume":"100 :","pages":"465 - 479"},"PeriodicalIF":1.7,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144526106","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}