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Solvent Dehydration and Low Temperature Vacuum Drying for SEM Imaging of Pre-Hatching Frog Embryos
IF 1.5 4区 医学 Q2 ANATOMY & MORPHOLOGY Pub Date : 2025-02-08 DOI: 10.1002/jmor.70030
Daniela Zurita-Paredes, Daniela Flores-Bolaños, Karla Vizuete, Alexis Debut, Andrés Romero-Carvajal

Scanning electron microscopy (SEM) is a fundamental technique to study the morphology of anuran embryos and tadpoles. Here, we present a drying method for SEM imaging of late frog embryos using commonly available dehydration solvents such as ethanol or methanol, xylene, and applying low temperature vacuum freeze drying. Briefly, embryos from early embryonic gills development to hatching were fixed with a paraformaldehyde—glutaraldehyde mix, then dehydrated to ethanol or methanol, and then slowly dried using low temperature and constant vacuum pressure. An extra step of clearing using xylene after ethanol dehydration improved results considerably. Our protocol successfully preserved embryo shape and the morphology of fragile and delicate superficial structures (e.g., external embryonic gills, apical ectodermal microridges and surface ciliation), while avoiding the use of some SEM toxic reagents.

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引用次数: 0
Anatomy of the Critically-Endangered Anji Salamander (Hynobius amjiensis) Provides New Insights Into Morphological Evolution of Salamanders
IF 1.5 4区 医学 Q2 ANATOMY & MORPHOLOGY Pub Date : 2025-01-29 DOI: 10.1002/jmor.70028
Cang-Song Chen, Jia Jia, Xian-Ting Wang, Jia Yang, Ke-Qin Gao

The Anji Salamander (Hynobius amjiensis) is a critically-endangered amphibian endemic to the Tianmushan Mountain area in southeastern China. As most of its congeneric species in the ancestral salamander family Hynobiidae, the osteology of H. amjiensis has remained essentially unknown and has hampered efforts in understanding morphological evolutionary patterns of early salamanders. Here, we investigate the skeletal anatomy of H. amjiensis based on microcomputed tomography scans of post-metamorphosed juvenile and adult specimens. Our results reveal Hynobiidae has more early-tetrapod-like plesiomorphic characters than expected, as H. amjiensis has a stapedial foramen in the middle ear and two centralia and a centrale-radius contact in the limb. We demonstrate that Hynobius amjiensis is the first known living salamander species with a stapedial foramen whose absence was believed to unite salamanders and anurans, and hence opens major questions on the evolution of the middle ear in modern amphibians: if some salamanders and caecilians had a stapedial foramen inherited from their common ancestor, when and how many times was the foramen lost independently in modern amphibians, and how did this structural loss impact the phylogenetic evolution of salamander clades? Our findings of hyper-ossified pectoral and pelvic girdles and loss of postminimus in the pes in H. amjiensis demonstrate that functional morphological features in hynobiids are potentially informative in phylogeny and ontogeny of early salamanders.

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引用次数: 0
Morphological Criteria for Staging Near-Hatching Embryos of the Domesticated Mallard (Anas platyrhynchos) and Swan Goose (Anser cygnoides)
IF 1.5 4区 医学 Q2 ANATOMY & MORPHOLOGY Pub Date : 2025-01-29 DOI: 10.1002/jmor.70027
Bassel Arnaout, Kaylen Brzezinski, Benjamin Steventon, Daniel J. Field

Studying avian embryology necessitates reliable and precise staging tables—descriptions of embryonic features appearing during development that are used to approximate the extent of embryonic development from fertilisation to hatching. Staging tables for waterfowl (Anseriformes) have previously been established based on morphological features from fertilisation to approximately 10 days before hatching. Embryonic changes over the final 10 days of pre-hatching development have also been documented and proposed as useful staging criteria. However, the reliability of these changes—which focus on the size of the bill and middle toes—as useful staging criteria across different waterfowl breeds has not been fully examined. To evaluate the reliability of these criteria for staging near-hatching embryos, we examined 27 embryos of Mallard (Anas platyrhynchos) and Swan Goose (Anser cygnoides). Comparisons with previously published data revealed that size variation within the same developmental stage across breeds is equivalent to within-breed variation across different stages, suggesting limited reliability of bill and middle toe size for staging waterfowl embryos. Consequently, we devised novel staging criteria for waterfowl based on four easily measurable morphological traits and show that these criteria allow correct stage identification with over 70% accuracy. Our results highlight the importance of quantifying staging accuracy for improving the reliability of embryonic staging tables.

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引用次数: 0
Functional Morphology of Hatching: Ontogeny and Distribution of Hatching Gland Cells in Red-Eyed Treefrogs and a New Marker for Anuran Hatching Enzyme
IF 1.5 4区 医学 Q2 ANATOMY & MORPHOLOGY Pub Date : 2025-01-28 DOI: 10.1002/jmor.70029
María José Salazar-Nicholls, Henry Macías Bazante, Karen M. Warkentin

Environmentally cued hatching (ECH) is widespread in animals and requires regulation of hatching mechanisms. Enzymatic digestion of the egg membrane is a common hatching mechanism in vertebrates and invertebrates. In amphibians and fishes, hatching enzymes (HE) are synthesized and released by hatching gland cells (HGC), whose functional ontogeny determines when hatching can occur. Ontogenetic studies of HGC development or HE expression are limited, based largely on external cell morphology; few markers for HGC or HE are available, and those appear specific for Xenopus. Moreover, mechanisms regulating HE release are unknown in anurans. To investigate variation in the hatching process, we need tools to identify and analyze its components. Agalychnis callidryas (Hylidae) is a well-established model of ECH, showing plastically timed, acute HE release, unlike the gradual release described for some aquatic anurans. We developed a new antibody marker for A. callidryas HE that also labels HGC/HE in glassfrogs (Centrolenidae). As glassfrogs and treefrogs diverged 62 mya, the antibody may be broadly useful in anurans. We used the AcHE antibody to examine the development and distribution of HGC and accumulation of HE, two key elements of hatching mechanisms, in A. callidryas. We found a much larger number (ca. 4200) and broader distribution of HGC than has been documented in any amphibian, with HGC densely but non-contiguously distributed over the front of the head and eyes and scattered along the dorsal midline. HE expression begins before hatching competence and is strong throughout the plastic hatching period, unlike HE gene expression which diminishes after competence. The distribution and expression ontogeny of A. callidryas' HE/HGC appear related to their hatching performance, plasticity, and embryo morphology. The AcHE antibody will enable comparative research to elucidate co-variation in the functional morphology, performance, and ecological context of hatching.

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引用次数: 0
Ecomorphology of South American Penguins 南美洲企鹅的生态形态学。
IF 1.5 4区 医学 Q2 ANATOMY & MORPHOLOGY Pub Date : 2025-01-19 DOI: 10.1002/jmor.70025
Elliott Bloom, Juan Carlos Torres-Mura, Marcelo Bertellotti, Fritz Hertel

A major goal of evolutionary ecology is to understand the interaction between ecological differences and the functional morphology of organisms. Studies of this type are common among flying birds but less so in penguins. Penguins (Spheniscidae) are the most derived extant underwater flying birds using their wings for swimming and beak when foraging. The Humboldt Penguin (Spheniscus humboldti) and Magellanic Penguin (S. magellanicus) occur along the coast of South America and their morphology was compared in allopatry and sympatry throughout their ranges. Measurements included: mass, tarsus length, four beak/head dimensions, bite force, wing loading, and aspect ratio. A thin-plate spline/relative warp analysis was also used to detect subtle differences in wing shape. Both species generally overlapped in trait morphology, but Magellanic Penguins showed greater trait diversity. Wing morphology was more homogenous between species than beak morphology indicating a similar mode of locomotion but potential differences in prey procurement. Morphological character displacement in sympatry was only evident in beak length. Local adaptation was common in other traits, and Punta Norte (Argentina) was often distinct in having high variation, notably in beak depth, wing loading, and wing shape (relative warp 1). This may be attributed to the fact that penguins here dive deep and forage farther from their colony; they also have a greater colony size that may contribute to greater intraspecific competition for resources. These results support a potentially optimal wing design for aquatic movement, which likely applies to other penguin species. Differences in morphology may also be related to differences between Atlantic and Pacific ecosystems.

进化生态学的一个主要目标是了解生态差异和生物功能形态之间的相互作用。这种类型的研究在鸟类中很常见,但在企鹅中就不那么常见了。企鹅(企鹅科)是现存的最衍生的水下飞禽,它们用翅膀游泳,用喙觅食。洪堡企鹅(Spheniscus humboldti)和麦哲伦企鹅(S. magellanicus)分布在南美洲海岸,并在其分布范围内对其异种和同属形态进行了比较。测量包括:质量,跗骨长度,四个喙/头尺寸,咬合力,机翼载荷和长径比。薄板样条/相对翘曲分析也用于检测机翼形状的细微差异。麦哲伦企鹅和麦哲伦企鹅在性状形态上基本重叠,但麦哲伦企鹅表现出更大的性状多样性。翅的形态在物种之间比喙的形态更为均匀,这表明它们的运动模式相似,但在捕食方面存在潜在的差异。同感区形态性状位移仅在喙长上表现明显。当地适应在其他特征上是常见的,Punta Norte(阿根廷)在高变异上通常是独特的,特别是在喙的深度,翅膀的载荷和翅膀的形状(相对翘曲1)。这可能是由于企鹅在这里潜水深,觅食离他们的殖民地更远;它们也有更大的群体规模,这可能有助于更大的种内资源竞争。这些结果支持了一种潜在的最佳水生运动翅膀设计,这可能适用于其他企鹅物种。形态上的差异也可能与大西洋和太平洋生态系统之间的差异有关。
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引用次数: 0
Heads and Tails: Comparative Osteology of Nearctic Dipsadid Snakes 头与尾:新北极双翅蛇的比较骨学。
IF 1.5 4区 医学 Q2 ANATOMY & MORPHOLOGY Pub Date : 2025-01-15 DOI: 10.1002/jmor.70019
Roberta Azeredo Murta-Fonseca, Daniel Silva Fernandes, Angele Martins

Although numerous studies have addressed some aspects of the cranial osteology of Nearctic dipsadid species, only the species within the genera Heterodon and Carphophis have a formal published description of their skull. Similarly, vertebral data on such species are extremely scarce, and most of the available literature is focused on fossils. Such group has a complex phylogenetic history, being recovered as monophyletic or nonmonophyletic depending on the approach. In this paper, we provide detailed and comparative descriptions of the osteology of dipsadid species distributed in the Nearctic region based on 69 specimens of dry material and high-resolution computed tomography (CT) scans. Additionally, we explore the morphological variation of the skull and cervical vertebrae within the context of distinct phylogenetic hypotheses previously proposed. Only two suprageneric groups previously proposed shared exclusive morphological traits: (Carphophis amoenus + Contia tenuis), proposed by three studies, and (Diadophis punctatus (Ca. amoenus + Co. tenuis)), proposed by one study. Large and detailed studies on the skull, mandible, and vertebrae represent an important step toward the understanding of the evolution of species, especially when they also show intraspecific variation.

尽管许多研究已经解决了新北极双足动物的颅骨骨学方面的一些问题,但只有异齿龙属和carphophhis属的物种对其头骨有正式发表的描述。同样,关于这类物种的椎体数据也非常少,而且大多数可用的文献都集中在化石上。这一群体具有复杂的系统发育历史,根据不同的方法可以被恢复为单系或非单系。在本文中,我们基于69个干燥材料标本和高分辨率计算机断层扫描(CT)提供了分布在新北极地区的dipsadid物种的骨学详细和比较描述。此外,我们在先前提出的不同系统发育假说的背景下探讨了头骨和颈椎的形态变异。只有两个先前提出的超属类群具有独特的形态特征:(carphophhis amoenus + conttia tenuis),由三个研究提出,(Diadophis punctatus (Ca. amoenus + Co.)。Tenuis)),由一项研究提出。对头骨、下颌骨和椎骨进行大规模而详细的研究是了解物种进化的重要一步,特别是当它们也显示出种内变异时。
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引用次数: 0
Dental Anomalies and Cranio-Dental Ontogeny in a Captive Wild Boar Population From France 法国圈养野猪种群的牙齿畸形和颅牙个体发育。
IF 1.5 4区 医学 Q2 ANATOMY & MORPHOLOGY Pub Date : 2025-01-09 DOI: 10.1002/jmor.70024
Helder Gomes Rodrigues, Clémence Le Gouellec, Katia Ortiz, Yann Locatelli, Dimitri Neaux, Thomas Cucchi

Dental anomalies are frequent in boars and pigs, and they generally affect the first premolar loci. The prevalence of these dental anomalies was investigated in a large number of populations around the world. These studies mainly focused on the influence of domestication, size, sexual dimorphism or food hardness on these anomalies. However, they rarely considered ontogenetic aspects, while these are crucial for understanding their aetiology during animal growth and how the dental row-jaw complex is affected. Here, we studied the incidence of missing first upper and lower premolars in a French population of captive wild boars to discuss the functional and developmental reasons for missing teeth and to assess the impact of missing teeth on the growth of the dental row-jaw complex. Using the CT-scan data of the cranium and mandible of 24 wild boars investigated six times each during their growth, and presenting a balanced sex ratio, we recorded the number of missing teeth. We then quantified the shape of the upper and lower jaws using 3D geometric morphometrics. We found a similar prevalence of missing first premolar (37.5%) between the upper and the lower jaws, which is higher than the frequencies observed in most continental populations of wild boars. The increasing number of anomalies during ontogeny suggests a relaxed constraint on the dentition associated with a different feeding behaviour in captivity. The absence of first premolars does not appear to be associated with size variation or sexual dimorphism, nor does it affect the place of the dentition within the jaw, the latter being more influenced by the dimorphic shape of the canines and the timing of dental eruption.

牙齿异常在公猪和猪中很常见,它们通常影响第一前臼齿的位置。在世界各地的大量人口中调查了这些牙齿异常的患病率。这些研究主要集中在驯化、大小、性别二态性或食物硬度对这些异常的影响上。然而,他们很少考虑个体发生方面,而这些对于理解动物生长过程中的病因学以及牙齿排颌复合体如何受到影响至关重要。在这里,我们研究了法国圈养野猪第一上颌和下前臼齿缺失的发生率,以讨论缺失牙齿的功能和发育原因,并评估缺失牙齿对牙齿排颌复体生长的影响。我们利用24头野猪在其生长过程中对其头盖骨和下颌骨各进行了6次ct扫描,并呈现出平衡的性别比例,记录了缺失牙齿的数量。然后,我们使用三维几何形态计量学量化了上颚和下颚的形状。我们发现,在上颚和下颚之间缺失第一前臼齿的发生率相似(37.5%),这比在大多数大陆野猪种群中观察到的频率要高。个体发育过程中越来越多的异常现象表明,在圈养环境中,与不同摄食行为相关的齿列受到了宽松的约束。第一前臼齿的缺失似乎与大小变化或两性二态性无关,也不影响齿列在颌骨内的位置,后者更多地受到犬科动物二态形状和牙齿出牙时间的影响。
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引用次数: 0
Vascular and Osteological Morphology of Expanded Digit Tips Suggests Specialization in the Wandering Salamander (Aneides vagrans) 扩张趾尖的血管和骨形态学表明流浪蝾螈(Aneides vagrans)的特化。
IF 1.5 4区 医学 Q2 ANATOMY & MORPHOLOGY Pub Date : 2025-01-08 DOI: 10.1002/jmor.70026
Christian E. Brown, William P. Goldenberg, Olivia M. Hinds, Mary Kate O'Donnell, Nancy L. Staub

For over a century researchers have marveled at the square-shaped toe tips of several species of climbing salamanders (genus Aneides), speculating about the function of large blood sinuses therein. Wandering salamanders (Aneides vagrans) have been reported to exhibit exquisite locomotor control while climbing, jumping, and gliding high (88 m) within the redwood canopy; however, a detailed investigation of their digital vascular system has yet to be conducted. Here, we describe the vascular and osteological structure of, and blood circulation through, the distal regions of the toes of A. vagrans using histology in tandem with live-animal videos. Specifically, we sectioned a toe of A. vagrans at 0.90 μm, embedded it in Spurrs resin, and stained the tissue with toluidine blue. An additional three toes were sectioned at 10 μm, embedded in paraffin, and after sectioning and mounting, treated with Verhoeff and Quad stains. For living salamanders, we recorded real-time videos of blood flowing within individual toes upon a translucent surface oriented both horizontally (0°) and vertically (90°) to simulate both prostrate and vertical clinging scenarios, then analyzed the image sequences using ImageJ. We found that the vascularized toe tips have one large sinus cavity that is divided more proximally into two chambers via a septum, and there are mucous and granular glands in the dorsal and dorsolateral integument of the digit tips. Live-animal trials revealed variable sinus-filling both within and between toes, seemingly associated with variable pressure applied to the substrate when standing, stepping, clinging, and climbing. We conclude that A. vagrans, and likely other climbing salamanders, can functionally fill, trap, and drain the blood in their vascularized toe tips to optimize attachment, detachment, and complex arboreal locomotion (e.g., landing after gliding flight). Such an adaptation could provide insights for bioinspired designs.

一个多世纪以来,研究人员一直对几种爬行蝾螈(蝾螈属)的方形脚趾尖感到惊讶,并推测其中的大血窦的功能。据报道,流浪蝾螈(Aneides vagrans)在红木树冠内攀爬、跳跃和滑翔(88米)时表现出精细的运动控制能力;然而,他们的数字血管系统的详细调查尚未进行。在这里,我们描述了血管和骨结构,并通过血液循环,利用组织学串联与活体动物视频的迷走牛腿脚趾远端区域。具体来说,我们在0.90 μm处对a . vagrans的脚趾进行切片,将其嵌入Spurrs树脂中,并用甲苯胺蓝对组织进行染色。另外取3只脚趾,在10 μm处切片,石蜡包埋,切片和贴装后进行Verhoeff和Quad染色处理。对于活蝾螈,我们在一个水平(0°)和垂直(90°)方向的半透明表面上记录了单个脚趾内血液流动的实时视频,以模拟俯卧和垂直粘附的场景,然后使用ImageJ分析图像序列。我们发现血管化的趾尖有一个大的窦腔,通过隔膜更近地分为两个腔室,在趾尖的背侧和背外侧被膜中有粘液和颗粒腺体。活体动物实验显示,脚趾内部和脚趾之间的窦填充变化,似乎与站立、行走、攀爬时施加在基底上的压力变化有关。我们得出的结论是,流浪蜥蜴和其他爬行的蝾螈一样,可以在它们血管化的脚趾尖上填充、捕获和排出血液,以优化附着、分离和复杂的树栖运动(例如,滑翔飞行后着陆)。这种适应可以为生物灵感设计提供见解。
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引用次数: 0
Middle Ear Mechanics in the Barn Owl 谷仓猫头鹰的中耳机械师。
IF 1.5 4区 医学 Q2 ANATOMY & MORPHOLOGY Pub Date : 2025-01-03 DOI: 10.1002/jmor.70020
John Peacock, Monica A. Benson, Daniel J. Field, Garth M. Spellman

The barn owl is a common research subject in auditory science due to its exceptional capacity for high frequency hearing and superb sound source localization capabilities. Despite longstanding interest in the auditory performance of barn owls, the function of its middle ear has attracted remarkably little attention. Here, we report the middle ear transfer function measured by laser Doppler vibrometry and direct measurements of inner ear pressures. Our results illustrate that the barn owl middle ear produces a pressure gain between the ear canal and the inner ear vestibule of up to 35 dB, which is comparable to that seen in mammals. The footplate velocity transfer function magnitudes overlap with those measured in other bird species, however the differences in phase between the footplate velocity and the sound pressure stimulus indicate a middle ear group delay that is notably shorter than other birds. This work brings us closer to a more complete understanding of the physiology of hearing in a model organism in auditory science.

仓鸮具有超常的高频听觉能力和卓越的声源定位能力,是听觉科学研究的热点。尽管长期以来人们对仓鸮的听觉表现很感兴趣,但其中耳的功能却很少引起人们的注意。在这里,我们报告了用激光多普勒振动仪测量中耳传递函数和直接测量内耳压力。我们的研究结果表明,仓鸮中耳在耳道和内耳前庭之间产生高达35分贝的压力增益,这与哺乳动物的压力增益相当。足底速度传递函数的大小与其他鸟类相同,但足底速度与声压刺激的相位差异表明中耳群延迟明显短于其他鸟类。这项工作使我们对听觉科学中模式生物的听觉生理学有了更全面的了解。
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引用次数: 0
An Atlas of Anatomical Variants of the Human Talus 人类距骨解剖变异图谱。
IF 1.5 4区 医学 Q2 ANATOMY & MORPHOLOGY Pub Date : 2024-12-28 DOI: 10.1002/jmor.70023
Samuel James Cockerill, Matilde Arnay-de-la-Rosa, Emilio González-Reimers

Anatomical variants can be used effectively to identify relationships between individuals in kinship analysis and they may be useful during surgical procedures. These procedures can be better implemented when the cause, appearance and location are understood. Clear representations and definitions of anatomical traits are necessary. A similar morphology of pathologies and variations of normal morphology can lead to confusion and unnecessary intervention. Therefore, there is a considerable need for an anatomical atlas of the particular skeletal elements, such as the talus bone. We have analyzed a total of 925 tali, 591 belonging to different modern and pre-Hispanic samples. Furthermore, 334 tali were analyzed from a reference collection from Athens. We have identified and defined the anatomical variants of the talus bone, of which only some have been mentioned by other researchers. We propose standardized methods that may potentially improve future research in human skeleton anatomy and its applications. We provide information on anatomical variants with the aim of improving their identification, classification and use in medical professions and bioanthropology.

在亲属关系分析中,解剖变异可以有效地用于识别个体之间的关系,它们可能在外科手术过程中有用。当了解了原因、外观和位置后,这些程序才能更好地执行。解剖特征的清晰表述和定义是必要的。病理形态的相似和正常形态的变化可导致混淆和不必要的干预。因此,有相当大的需要对特定的骨骼元素,如距骨解剖图谱。我们总共分析了925个tali,其中591个属于不同的现代和前西班牙样本。此外,从雅典的参考收藏中分析了334个塔利。我们已经确定并定义了距骨的解剖变异,其中只有一些被其他研究人员提到。我们提出了标准化的方法,可能会潜在地改善人类骨骼解剖及其应用的未来研究。我们提供有关解剖变异的信息,目的是改进其在医学专业和生物人类学中的识别、分类和使用。
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引用次数: 0
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Journal of Morphology
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