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Shibata S.,Tokyo Medical and Dental University | Sakamoto Y.,Tokyo Medical and Dental University | Yokohama-Tamaki T.,Health Sciences University of Hokkaido | Murakami G.,Iwamizawa Koujin kai Hospital | Cho B.H.,Chonbuk National University
Anatomical Record | Year: 2014

Immunohistochemical localization of versican and tenascin-C were performed; the periosteum of ossifying mandible and the perichondrium of Meckel's cartilage, of vertebral cartilage, and of mandibular condylar cartilage were examined in midterm human fetuses. Versican immunoreactivity was restricted and evident only in perichondrium of Meckel's cartilage and vertebral cartilage; conversely, tenascin-C immunoreactivity was only evident in periosteum. Therefore, versican and tenascin-C can be used as molecular markers for human fetal perichondrium and fetal periosteum, respectively. Meckel's cartilage underwent endochondral ossification when it was incorporated into the ossifying mandible at the deciduous lateral incisor region. Versican immunoreactivity in the perichondrium gradually became weak toward the anterior primary bone marrow. Tenascin-C immunoreactivity in the primary bone marrow was also weak, but tenascin-C positive areas did not overlap with versican-positive areas; therefore, degradation of the perichondrium probably progressed slowly. Meanwhile, versican-positive perichondrium and tenascin-C-positive periosteum around the bone collar in vertebral cartilage were clearly discriminated. Therefore, the degradation of Meckel's cartilage perichondrium during endochondral ossification occurred at a different rate than did degradation of vertebral cartilage perichondrium. Additionally, the perichondrium of mandibular condylar cartilage showed tenascin-C immunoreactivity, but not versican immunoreactivity. That perichondrium of mandibular condylar cartilage has immunoreactivity characteristic of other periosteum tissues may indicate that this cartilage is actually distinct from primary cartilage and representative of secondary cartilage. © 2014 Wiley Periodicals, Inc.


Jin Z.W.,Chonbuk National University | Nakamura T.,Sapporo Medical University | Yu H.C.,Chonbuk National University | Kimura W.,Yamagata University | And 2 more authors.
Journal of Anatomy | Year: 2010

We demonstrated fetal peripheral lymphatic vessels (LVs) using D2-40 immunohistochemistry in a whole female fetus (18 weeks of gestation, CRL 155 mm) except for the head. There were abundant LVs in the thyroid gland, lung, stomach, small intestine, rectum and pancreas, whereas no LVs were seen in the parathyroid gland, spleen and adrenal cortex. In the liver, except for the gallbladder bed, LVs were still restricted to around hilar thick portal veins and around the hepatic vein terminals. Subcutaneous LVs were well developed throughout the body even in areas where no or few perforating LVs connected with the deep LVs. The diaphragm contained abundant, dilated LVs in the pleural half of its thickness. LVs were also seen not only along supplying arteries of muscles and cartilage but also along the epimysium and perichondrium. LVs ran in a space between the obliquus internus and transversus abdominis but not between the obliquus internus and obliquus externus. Some tight connective tissues such as the sacrotuberous ligament contained abundant LVs. The intervertebral foramen contained a lymphatic plexus. The present observations provide a better understanding of peripheral lymphatic development. The fetal lymphatic morphology seems not to represent a mini-version of the adult morphology. © 2010 The Authors. Journal compilation © 2010 Anatomical Society of Great Britain and Ireland.


Shibata S.,Tokyo Medical and Dental University | Sato R.,Tokyo Medical and Dental University | Murakamib G.,Iwamizawa Koujin kai Hospital | Fukuoka H.,Tokyo Medical and Dental University | Rodriguez-Vazquez J.F.,Complutense University of Madrid
Journal of Oral Biosciences | Year: 2013

To investigate if mandibular condylar cartilage is derived from the periosteum of the ossifying mandible or from a separate, programmed blastema. Materials and methods: Fetal mice at E14.0-16.0, fetal rats at E16.0-18.0, and human embryos at 9 and 10 wks of gestation were used. The initial formation of rat condylar cartilage was investigated by using serial sections and enzyme-histochemistry to detect alkaline phosphatase activity. Histological observations of serial sections of human fetuses as well as 3D-reconstruction models were also analyzed. The expression of collagen type mRNA in developing rat condylar cartilage was directly compared with that in mice by performing in situ hybridization. Results: An anlage of the rat condylar process (condylar anlage) was clearly identified in the posterior position of the ossifying mandible and was continuous with it at E16.0. Newly formed rat condylar cartilage was observed at E16.5 and was continuous with the ossifying mandible. Mesenchymal cells in the condylar anlage at E16.0 showed alkaline phosphatase activity and chondrocytes in the newly formed condylar cartilage also showed enzymatic activity. Thus, rat mandibular condylar cartilage that derives from alkaline phosphatase-positive periosteum-like cells is continuous with the ossifying mandible, as previously demonstrated in mice, but rapid differentiation into hypertrophic chondrocytes in rats is not remarkable compared to that in mice. The condylar anlage and the newly formed cartilage were also continuous with the ossifying mandible in human embryos. Conclusions: Mammalian mandibular condylar cartilage derives from the periosteum of the ossifying mandible in mice, rats, and humans.© 2013 Japanese Association for Oral Biology. Published by Elsevier B.V. All rights reserved.


Katori Y.,Tohoku University | Katori Y.,Sendai Municipal Hospital | Cho B.H.,Chonbuk National University | Song C.H.,Chonbuk National University | And 3 more authors.
Annals of Anatomy | Year: 2010

Background: A craniocaudal transition from smooth to striated muscle occurs in the fetal mouse esophagus muscularis propria, until finally the entire muscle component becomes striated. Although no such investigation has been conducted using human fetuses, the transition appears to be incomplete. Methods: In horizontal sections of 10 human fetuses between 9 and 16 weeks of gestation, we identified immunoreactivity for smooth muscle actin (SMA), striated muscle myosin heavy chain (MyH), desmin, PGP9.5, S100 protein, c-kit, and CD68 in the thoracic esophagus. The TUNEL method was used to identify apoptosis. For comparison, the same immunohistochemistry was conducted using 10 adult esophaguses. Results: In fetuses at all stages examined, a transition zone was found in the upper thoracic esophagus that was attached to the middle one-third of the trachea. In the transition zone, the MyH-positive longitudinal muscle fibers were surrounded by flat, SMA-positive cells, whereas the MyH-positive circular fibers were sometimes located adjacent to the SMA-positive fibers. However, in adults, smooth muscle tended to be clearly separated from striated muscle. The distribution of cells showing immunoreactivity for PGP9.5, S100 or c-kit did not differ between the oral and anal sides of the transition zone. Desmin was positive in the muscularis propria, but negative in the muscularis mucosae. Neither CD68-positive macrophages nor TUNEL-positive cells were present in the esophagus. Conclusions: In the human esophagus, the smooth-to-striated muscle transition appears to stop at the mid-thoracic level. Cell death or transdifferentiation of smooth muscle appears unlikely, but phenotypic transformation into desmin-positive myofibroblasts is a possibility. © 2009 Elsevier GmbH. All rights reserved.


Kim J.-H.,Chonbuk National University | Han E.-H.,Chonbuk National University | Jin Z.-W.,Chonbuk National University | Lee H.-K.,Chonbuk National University | And 3 more authors.
Anatomical Record | Year: 2012

Using semiserial sections from 19 human fetuses of 8-30 weeks gestation, we examined the topohistology of the upper abdominal lymphatics and compared it with that of the lower abdominal and pelvic lymphatics. The upper abdominal lymphatics were characterized by an intimate relationship with the peritoneal lining, a common mesentery for the celiac trunk and superior mesenteric artery (SMA). Lymphatic connections from the upper abdominal viscera to the paraaortic and paracaval areas followed two routes: (1) from the intestinal mesentery, along the peritoneum on the left aspect of the proximal SMA, via the chain of lymph follicles (LFs) lying along the retropancreatic fusion fascia, to drain into the LFs around the left renal vein; (2) from sites along the peritoneum on the posterior wall of the omental bursa, via the root of the hepatoduodenal ligament, to drain into LFs around the vena cava. The development of these two posterior drainage routes seemed to be promoted by the peritoneum or a peritoneal remnant (i.e., fusion fascia) attaching to the great vessels, and inhibited or impeded by the developing nerves and diaphragm. No paraaortic, paracaval, or pelvic LFs lay along the peritoneum. The pelvic LFs were usually located along the bundle of lymphatic vessels originating from the femoral canal. © 2011 Wiley Periodicals, Inc.


Jin Z.W.,Chonbuk National University | Cho B.H.,Chonbuk National University | Murakami G.,Iwamizawa Koujin kai Hospital | Fujimiya M.,Sapporo Medical University | And 2 more authors.
Clinical Anatomy | Year: 2010

The retrohepatic inferior vena cava (IVC) is commonly considered to originate from the right vitelline or omphalomesenteric vein. In contrast, Alexander Barry hypothesized that one of the hepatic veins grows to merge with the subcardinal vein and develops into the retrohepatic IVC. We re-examined fetal development of the retrohepatic IVC and other related veins using serial histological sections of 20 human fetuses between 6 and 16 weeks of gestation. At 6-7 weeks, when a basic configuration of the portal-hepatic vein systems had just been established, one of hepatic veins (i.e., the posterocaudal vein in the present study) had grown caudally to reach the posterocaudal surface of the liver, and notably, extended into the primitive right adrenal gland (five of the eight early-staged fetuses). Because the inferior right hepatic vein (IRHV) and retrohepatic IVC appeared at the same developmental stage, it is likely that any peripheral remnants of the posterocaudal vein would continue to function as primary drainage territory for the IRHV. The caudate vein developed rapidly in accordance with marked caudal and leftward extension of Spiegel's lobe at 12-16 weeks. Thin accessory hepatic veins developed later than the caudate vein and IRHV. The present results supported Barry's hypothesis. © 2010 Wiley-Liss, Inc.


Katori Y.,Sendai Medical Hospital | Shibata S.,Health Sciences University of Hokkaido | Kawase T.,Tohoku University | Cho B.H.,Chonbuk National University | Murakami G.,Iwamizawa Koujin kai Hospital
Cleft Palate-Craniofacial Journal | Year: 2012

Objective: Transient immunoreactivity for tyrosine hydroxylase, which mediates the conversion of the amino acid L-tyrosine to dihydroxyphenylalanine, in the midline epithelial seam between the bilateral palatal shelves was investigated in human fetuses. Materials and Methods: Horizontal or frontal paraffin sections of two human fetuses at 9 and 15 weeks of gestation were used to examine the distribution of tyrosine hydroxylase-immunoreactive cells in regions of the entire head other than the brain. Immunohistochemical staining for S100 protein, calretinin, cytokeratin 14, and vimentin was examined using adjacent or near sections. Results: Tyrosine hydroxylase-immunoreactive cells were large and densely distributed in the midline epithelial seam at the site of palatal fusion in fetuses at 9 weeks but not in fetuses at 15 weeks, in which the midline epithelial seam had already disappeared. No expression of S100 protein, calretinin, or vimentin was detected, but the midline epithelial seam was positive for cytokeratin 14. Tyrosine hydroxylase immunoreactivity was not detected in epithelia during the process of palatal fusion in mice from E 14.0 to 15.0. Conclusions: These findings indicate that tyrosine hydroxylase- immunoreactive cells in the midline epithelial seams are nonneural epithelial cells and suggest that the tyrosine hydroxylase is a novel factor involved in normal palatal formation, especially the fate of the midline epithelial seam in humans. © Copyright 2012 American Cleft Palate-Craniofacial Association.


Shibata S.,Tokyo Medical and Dental University | Sakamoto Y.,Tokyo Medical and Dental University | Baba O.,Ohu University | Qin C.,Texas A&M University | And 2 more authors.
European Journal of Histochemistry | Year: 2013

Immunohistochemical localization of collagen types I, II, and X, aggrecan, versican, dentin matrix protein (DMP)-1, martix extracellular phosphoprotein (MEPE) were performed for Meckel's cartilage, cranial base cartilage, and mandibular condylar cartilage in human midterm fetuses; staining patterns within the condylar cartilage were compared to those within other cartilaginous structures. Mandibular condylar cartilage contained aggrecan; it also had more type I collagen and a thicker hypertrophic cell layer than the other two types of cartilage; these three characteristics are similar to those of the secondary cartilage of rodents. MEPE immunoreactivity was first evident in the cartilage matrix of all types of cartilage in the human fetuses and in Meckel's cartilage of mice and rats. MEPE immunoreactivity was enhanced in the deep layer of the hypertrophic cell layer and in the cartilaginous core of the bone trabeculae in the primary spongiosa. These results indicated that MEPE is a component of cartilage matrix and may be involved in cartilage mineralization. DMP-1 immunoreactivity first became evident in human bone lacunae walls and canaliculi; this pattern of expression was comparable to the pattern seen in rodents. In addition, chondroid bone was evident in the mandibular (glenoid) fossa of the temporal bone, and it had aggrecan, collagen types I and X, MEPE, and DMP-1 immunoreactivity; these findings indicated that chondroid bone in this region has phenotypic expression indicative of both hypertrophic chondrocytes and osteocytes. © S. Shibata et al., 2013 Licensee PAGEPress, Italy.


Cho K.H.,Wonkwang University | Cheong J.S.,Wonkwang University | Ha Y.S.,Wonkwang University | Cho B.H.,Chonbuk National University | And 2 more authors.
Journal of Anatomy | Year: 2012

Using D2-40 immunohistochemistry, we assessed the distribution of peripheral lymphatic vessels (LVs) in the head-and-neck region of four midterm fetuses without nuchal edema, two of 10weeks and two of 15weeks' gestation. We observed abundant LVs in the subcutaneous layer, especially in and along the facial muscles. In the occipital region, only a few LVs were identified perforating the back muscles. The parotid and thyroid glands were surrounded by LVs, but the sublingual and submandibular glands were not. The numbers of submucosal LVs increased from 10 to 15weeks' gestation in all of the nasal, oral, pharyngeal, and laryngeal cavities, but not in the palate. The laryngeal submucosa had an extremely high density of LVs. In contrast, we found few LVs along bone and cartilage except for those of the mandible as well as along the pharyngotympanic tube, middle ear, tooth germ, and the cranial nerves and ganglia. Some of these results suggested that cerebrospinal fluid outflow to the head LVs commences after 15weeks' gestation. The subcutaneous LVs of the head appear to grow from the neck side, whereas initial submucosal LVs likely develop in situ because no communication was evident with other sites during early developmental stages. In addition, CD68-positive macrophages did not accompany the developing LVs. © 2011 The Authors. Journal of Anatomy © 2011 Anatomical Society of Great Britain and Ireland.


Hwang S.E.,Chonbuk National University | Cho B.H.,Chonbuk National University | Hirai I.,Yamagata University | Kim H.T.,Chonbuk National University | And 4 more authors.
Clinical Anatomy | Year: 2010

At 8-16 weeks of gestation, Spiegel's lobe of the caudate lobe appears as a sac-like herniation of the liver parenchyma between the inferior vena cava and ductus venosus or Arantius' duct. In 5 of 11 fetuses at 20-30 weeks of gestation, we found that an external notch was formed into the posterior aspect of the caudate lobe by a peritoneal fold containing the left gastric artery. This notch appeared to correspond to that observed in adults, which is usually seen at the antero-inferior margin of the lobe after rotation of the lobe along the horizontal or transverse axis. However, the notch did not accompany two of the three fetuses in which the left hepatic artery originated from the left gastric artery. Notably, until 9-10 weeks of gestation, the inferior and left part of Spiegel's lobe rode over the hepatoduodenal ligament and protruded medially into the lesser sac (bursa omentalis) behind the stomach. Thus, the fetal Winslow's foramen was located at the "superior" side of the ligament. However, as seen in adults, the protruding Spiegel's lobe was located at the posterior side of the lesser omentum. Therefore, a hypothetical rotation along the transverse axis in the later stages of development seems necessary to explain this repositioning. Considering that Spiegel's lobe develops faster than surrounding structures, it is likely that the lesser sac resulting from the rotation of the gastrointestinal tract, which actively contributes to facilitate the growth of the Spiegel lobe. © 2010 Wiley-Liss, Inc.

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