74. KanahashiT, ImaiH, Otani H, YamadaS, Männer J, Takakuwa T. Boundary Formation of the Human Caudal Foregut During the Early Fetal Period: Three-Dimensional Analysis Using T1-Weighted and Diffusion Tensor Images. Cells Tissues Organs, 2025, in press. doi: 10.1159/000546997
Introduction: While caudal foregut development in human fetuses has been outlined in previous research, the formation of its border region remains unclear. This study aimed to visualize the precise timeline of caudal foregut boundary formation. Methods: Three-dimensional images of the foregut from T1-weighted scans of 24 fetuses (crown–rump length [CRL]: 34–103 mm) were analyzed to measure the wall thickness and lumen diameter at nine specific sites. The internal structure in the border region was verified using histological sections and diffusion tensor imaging (DTI) tractography. Results: The lower esophageal and pyloric canal walls in samples with CRL ≥50 mm were relatively thicker. The esophageal wall at the esophageal hiatus, where the lower esophageal sphincter is located, was particularly thick in samples with CRL ≥88 mm. Increased wall thickness at the esophageal hiatus and pyloric canal resulted in a narrower lumen. The pyloric canal lumen narrowed from its distal to proximal sections. The lumen diameter-to-wall thickness ratio at the esophageal hiatus and proximal pyloric was negatively correlated with CRL. The thickened esophageal wall at the esophageal hiatus had a thick submucosa layer, and all layers in the pyloric canal thickened with growth. DTI tractography revealed that the lower esophageal wall mainly comprised longitudinal fibers, whereas the pyloric canal wall consisted solely of circular fibers, with fractional anisotropy increasing with growth. Conclusion: This study provides a comprehensive timeline of normal caudal foregut boundary formation during the early human fetal period, thereby improving the understanding of congenital foregut obstruction pathogenesis.
Ishida, N, Kanahashi T, Matsubayashi J, Imai, H, Männer J, Yamada S, Takakuwa,T. Change in diameters of the small intestine according to embryonic and early fetal growth. 2025. J Anatomy in press. doi: 10.1111/joa.14285.
Abstract No previous studies have examined the diameter of the small intestine successively from the oral to the anal side of the small intestine. Therefore, the objectives of this study were to determine the successive intestinal diameters from the oral to the anal side (proximal to the distal) of the intestine, evaluate changes in diameter associated with growth, examine the effects of positional variation along the intestinal tract, investigate dynamic positional change from the extraembryonic coelom to the abdominal cavity, and assess the impact of complex tertiary intestinal loop formation. To this end, 14 human embryonic and fetal specimens with crown-rump lengths (CRLs) ranging from 25.6 to 69.0 mm were selected for high-resolution magnetic resonance imaging acquisition. The small intestines of the specimens were located in the extraembryonic coelom (herniation phase), transitioning phase, or abdominal cavity (return phase). The small intestine and mesentery were reconstructed in three dimensions, and the resulting morphological changes were observed and analyzed. Successive intestinal diameters from the oral to anal side of the small intestine were determined. Specifically, we observed the following: 1) gradual changes in the diameter of the position from the oral to the anal side in the jejunum-ileum, 2) the difference between the duodenum and jejunum-ileum, and 3) the difference between the superior part of the duodenum derived from the foregut and the remaining parts derived from the midgut. 4) Notably, the dynamic positional change from the extraembryonic coelom to the abdominal cavity, along with the rapid elongation and complex intestinal loop formation—a conspicuous phenomenon in the embryonic and early fetal periods—had little effect on the changes in diameter. This study indicates that increased diameter may serve as a useful indicator of intestinal development and differentiation, independent of tertiary intestinal loop formation and positional changes into and out of the abdominal cavity.
Introduction: Features of the superior mesenteric artery (SMA) and its intestinal branches during the embryonic and early fetal periods have not been fully described. We aimed to comprehensively elucidate the characteristics of intestinal branch artery formation in the SMA. Methods: Serial tissue sections of seven early fetal specimens belonging to the Blechschmidt Collection were digitalized and used for segmentation and reconstruction of the intestinal loop, SMA trunk, intestinal branch arteries, and mesentery for further analysis. Results: The intestinal branch arteries fed the intestinal tract from the oral side to the anal side, according to the order of their origin from the root to the periphery of the SMA trunk. SMA and intestinal branches were not as strongly conserved in their morphology as indicated in previous research but varied between specimens. Most intestinal branch arteries exhibited frequent branching with small intervals at the periphery, whereas the proximal branch exhibited few branches. Only a few peripheral branches made contact with the neighboring intestinal branch arteries. The fetal intestinal branch artery architecture differed greatly from that of adults. There were considerable inter- and intra-specimen variations in the intestinal tract length per feeding intestinal branch artery. The SMA branching arteries did not always supply each tertiary loop individually, and not every loop is connected to one branching artery. Conclusion: This study elucidates the characteristics of forming the SMA intestinal branch arteries. Specifically, the findings suggest that the SMA is similar to other arteries in that its branches show a level of variability in feeding tissues.
Takakuwa T,Kakeya M, Ishida N, Kanahashi T, Fujii S, Männer J, Yamada S. Superior mesenteric artery during intestinal loop formation and its positional changes from the extracoelom to the abdominal cavity. Cells Tissues Organs 2025, in press, DOI: 10.1159/000545751
71. Ishida N, Ueda Y, Kanahashi T, Matsubayashi J, Imai H, Yamada S, Takakuwa T.Hierarchical loop formation in human midgut during physiological umbilical herniation, J Anatomy 2024, in press DOI:10.1111/joa.14228
CS16からCS18までのすべての標本で一次ループの形成が観察された。
腸間膜狭窄により中腸がを 4 つのセグメントに分割可能。
中腸の二次ループは、最初にCS19のセグメント2と4（S2とS4）で識別。
三次ループの形成は、CS21 で最初に確認
CS23 までに、三次ループはほとんどの標本のS2,3,4のセグメントで観察
S1は1つの二次ループのまま。
三次ループの数は頭殿長に応じて増加し、中腸の S2 から S4 で最大 19 個
ループ形成における遺伝的要因と生体力学的要因の役割を包括的に理解する上で極めて重要
Abstract
The primary loop, a single hairpin-shaped loop, becomes recognizable at the Carnegie stage (CS) 16. This loop projects toward the umbilical cord and subsequently gives rise to four secondary loops in the midgut of human embryos. As development advances, the segments corresponding to each secondary loop further develop into an increasing number of loops, referred to as tertiary loops. The mesenteric leaves and the narrowing parts, which serve as the borders of the secondary loops, remain identifiable throughout the subsequent stages of development. This study aimed to describe the morphological alterations that occur in the midgut and mesentery over time during the herniated phase of the midgut. A total of 47 human embryos between CS16 and CS23 and two fetuses in the physiological umbilical herniated stage were selected for high-resolution magnetic resonance imaging acquisition. Specimens were obtained from the Congenital Anomaly Research Center of Kyoto University. Serial tissue sections obtained from four embryos were subjected to histological observation. The midgut and mesentery were reconstructed in three dimensions, and the resulting morphological changes were observed and analyzed. Formation of the primary loop was observed in all specimens between CS16 and CS18. Secondary loops in the midgut were initially discerned at CS19 in segments 2 and 4 (S2 and S4). The border between S3 and S4 was identified at the apex of the midgut hernia, where traces of the vitelline artery and duct enter the mesentery. At CS21 and later stages of development, the presence of three borders at the exact location delineated by mesenteric narrowing was consistently observed, which resulted in the midgut being divided into four segments in all specimens. The formation of tertiary loops was initially identified at Carnegie stage (CS) 21, occurring in either segment S2 or S3. By CS23, tertiary loops were observed in three segments in most specimens. Notably, the initial formation of tertiary loops in S4 occurred one Carnegie stage later than in S2 or S3. Additionally, the increase in the number of folds and the length per fold in S4 was delayed compared with the number and length of folds observed in both S2 and S3. The number of loops in S1 remained constant (one secondary loop) across all specimens. Upon reaching a critical threshold length, the number of loops exhibited a marked increase, accompanied by rapid elongation in S2, S3, and S4. The number of tertiary loops increased in accordance with the crown-rump length, which exhibited a maximum of 19 tertiary loops in S2 to S4 of the midgut. These findings support the hypothesis that tertiary loops develop biomechanically through the rapid elongation of the midgut and slow growth of the mesentery. This study describes the morphological alterations occurring in the midgut and mesentery over time during the herniated phase of the midgut and provides a comprehensive understanding of the roles of genetic and biomechanical factors in loop formation.
70. Kumagai M, Kanahashi M, Matsubayashi J, Imai H, Otani H, Takakuwa T. Primary sulci formation in human cerebral cortex development. Anat Rec (Hoboken) 2025, doi: 10.1002/ar.25637
Abstract
We aimed to determine the timing of appearance and the morphologic and morphometric features of the initial human cerebral sulcal formation. Using high-resolution magnetic resonance images obtained from 33 samples between 11 and 16 weeks (w) of gestation (crown-rump length <130 mm), the cerebral surface and internal structures on serial two-dimensional planes and all possible sulci on three-dimensional reconstructions were marked, allowing comparison of the positions of the sulci in the samples and inter-samples. Our method provided accurate conclusions regarding the timing of sulcal formation. Detection timing was as early as and earlier than those in previous studies using anatomical dissection and magnetic resonance imaging (MRI), respectively: <12 w for the callosum, <13 w for the hippocampal, calcarine, and parieto-occipital sulci, and <15 w for the lateral sulcus. Occasionally, an olfactory sulcus was detected. However, the cingulate sulcus could not be definitely identified. The lateral sulcus gradually appeared and changed shape. The lengths of the left and right sides of the olfactory sulci and the left side of the hippocampal sulcus increased linearly with the CRL. The length of the right side of the hippocampal sulcus and the left and right sides of the calcarine, parieto-occipital, and not determined_a sulci did not increase with the CRL The depth of the all sulci, except for the parieto-occipital sulci, increased linearly with the CRL. The sulci might not arise as if they elongate gradually but arise simultaneously over some distance. We determined the timing of the initial sulcal formation using high-resolution MRI. Our findings may significantly impact prenatal diagnosis and research on neurodevelopmental disorders.
69. Iwasa Y, Kanahashi T, Imai H, Otani H, Yamada S, Takakuwa T. Human trapezius muscle development during early fetal period. J Anatomy 2024, 245, 663-673, doi: 10.1111/joa.14116
Abstract
J Anatomy 2024, 245巻, 11号(僧帽筋のDTI）
This study aimed to observe human trapezius muscle (TpzM) development during the early fetal period and apply diffusion tensor imaging (DTI) analysis to describe the muscle architecture that leads to physiological functions. Human embryonic and early fetal specimens were selected for this study. TpzM was first detected at Carnegie stage 20. The position of the TpzM changed with the formation of the scapula, clavicle, and vertebrae, which are its insertions and origins. DTI revealed the fiber orientation from each vertebral level to dissect each muscle. Fiber orientation in the ventral view gradually changed from the cervical to thoracic vertebrae, except for the middle part at which the insertions changed, which was almost similar in all early fetal specimens. The TpzM volume increased from C1 to C7 in the upper part, reached local maxima at C6 and C7 in the middle, and then decreased. These muscles can be categorized into three parts according to their insertions and presented with the features of each part. The fiber orientation and distribution of the three parts at the vertebral level were almost constant during the early fetal period. The border between the upper and middle parts was mainly located around the C6 and C7 vertebral levels, whereas the middle and lower parts were between the Th1 and Th2 vertebral levels. A three-dimensional change in the fiber orientation in the upper part of the TpzM according to the vertebral level was noticeable. Our data will help to elucidate the developmental processes of TpzM.
Tanima M, Ishida K, Ishikawa A, Yamada S, Takakuwa T, Aoyama T, Three-dimensional imaging analysis of the developmental process of posterior meniscofemoral ligaments in rat embryos. Cells Tissues Organs 2024, in press, , DOI: 10.1159/000536108
The posterior meniscofemoral ligament (pMFL) of knee joint is a ligament that runs posterior to the posterior cruciate ligament (PCL) and it is known that the height of the pMFL attachment site causes meniscus avulsion. Therefore, understanding the three-dimensional (3D) structure of the pMFL attachment site is essential to better understand the pathogenesis of meniscus disorders. However, the developmental process of pMFL has not been well investigated. The purpose of this study was to analyze pMFL development in rat knee jointsusing 3D reconstructed images produced from episcopic fluorescence image capture (EFIC) images and examine its relationship with other knee joint components. Knee joints of Wistar rat embryos between embryonic day (E) 16 and E21 were observed with HE stained tissues. Serial EFIC images of the hindlimbs of E17-E21 were respectively captured, from which 3Dimages were reconstructed and the features of pMFL structure: length and angle, were measured. Besides, the chronological volume changes and the volume ratio of the knee joint components compared to E17 were calculated to identify the differences in growth by components. pMFL was observed from E17 and was attached to the medial femoral condyle and lateral meniscus at all developmental stages, as in mature rats. The lack of marked variation in the attachment site and angle of the pMFL with the developmental stage indicates that the pMFL and surrounding knee joint components developed while maintaining their positional relationship from the onset of development. Current results may support to congenital etiology of meniscus disorder.
67. Isotani N, Kanahashi T, Imai H, Yoneyama A, Yamada S, Takakuwa T. Regional differences in the umbilical vein and ductus venosus at different stages of normal human development. Anat Rec (Hoboken), 2024, 307, 3306-3326.DOI:10.1002/ar.25421
During the fetal period, oxygenated blood from the placenta flows through the umbilical vein (UV), portal sinus, ductus venosus (DV), and inferior vena cava (IVC) to the heart. This venous route varies regionally in many aspects. Herein, we sought to characterize the venous route’s morphological features and regional differences during embryonic and early-fetal periods. Twenty-nine specimens were selected for high-resolution digitized imaging; 18 embryos were chosen for histological analysis. The venous route showed a primitive, large, S-shaped curved morphology with regional narrowing and dilation at Carnegie stage (CS) 15. Regional differences in vessel-wall differentiation became apparent from approximately CS20. The vessel wall was poorly developed in most DV parts; local vessel-wall thickness at the inlet was first detected at CS20. The lumen of the venous route changed from a non-uniform shape to a relatively round and uniform morphology after CS21. During the early-fetal period, two large bends were observed around the passage of the umbilical ring and at the inlet of the liver. The length ratio of the extrahepatic UV to the total venous route increased. The sectional area gradually increased during embryonic development, whereas differences in sectional area between the DV, UV, and IVC became more pronounced in the early-fetal period. Furthermore, differences in the sectional area between the narrowest part of the DV and other hepatic veins and the transverse sinus became more pronounced. In summary, the present study described morphological, morphometric, and histological changes in the venous route throughout embryonic and early-fetal development, clarifying regional characteristics.
