Immunohistochemical localization of vascular factors in tooth germ of amphibian (Cynops pyrrhogaster) / Localización inmunohistoquímica de factores vasculares en germen dentario de anfibios (Cynops pyrrhogaster)

SUMMARY: Vascular endothelial growth factor (VEGF) and its receptor, VEGFR-2, are known to regulate blood vessel endothelium growth. They play important role in human and rodents teeth development. In newt jaws, there are sequential developmental teeth germs following behind the mature teeth. We examined the immunohistochemical localization of VEGF and its receptor and showed the specific expression pattern of VEGF and VEGF receptor in Cynops pyrrhogaster sequential tooth development. The intensity of immunoreactivity for VEGF in the inner enamel epithelium was weaker than that in the outer enamel epithelium in the dentine matrix formation and mineralization stages. Finally, at the enameloid maturation and enamel-like matrix formation stage, immunoreactivity for VEGF in inner enamel epithelium was stronger than in the outer enamel epithelium. The intensity of immunoreactivity for VEGFR-2 was positive for the outer enamel epithelium throughout tooth development. The crown sides of the odontoblasts were stained especially strongly for VEGF and VEGFR-2 during the dentine matrix formation and mineralization stage of the enameloid maturation and enamel- like matrix formation stage. We postulate that the expression of VEGF in the inner enamel epithelium and odontoblast widely effects tooth development in newts, as well as in human and rodents.

RESUMEN: Se sabe que el factor de crecimiento endotelial vascular (VEGF) y su receptor, VEGFR-2, regulan el crecimiento del endotelio de los vasos sanguíneos. Desempeñan un papel importante en el desarrollo de los dientes humanos y de los roedores. En las mandíbulas de tritón, hay gérmenes dentales de desarrollo secuenciales que siguen a los dientes maduros. Examinamos la localización inmunohistoquímica de VEGF y su receptor y mostramos el patrón de expresión específico de VEGF y receptor de VEGF en el desarrollo secuencial de dientes de Cynops pyrrhogaster. La intensidad de la inmunorreactividad para VEGF en el epitelio interno del esmalte era más débil que en el epitelio externo del esmalte en las etapas de formación y mineralización de la matriz de dentina. Finalmente, en la etapa de maduración del esmalte y de formación de la matriz similar al esmalte, la inmunorreactividad para VEGF en el epitelio interno del esmalte fue más fuerte que en el epitelio externo del esmalte. La intensidad de la inmunorreactividad para VEGFR- 2 fue positiva para el epitelio externo del esmalte durante el desarrollo del diente. Los márgenes de la corona de los odontoblastos se tiñeron especialmente para VEGF y VEGFR-2 durante la etapa de formación de la matriz de dentina y mineralización de la etapa de maduración del esmalte y la etapa de formación de la matriz similar al esmalte. Postulamos que la expresión de VEGF en el epitelio interno del esmalte y odontoblastos afecta ampliamente el desarrollo de los dientes en tritones, así como en humanos y roedores.

The mesopancreas and pancreatic head plexus: morphological, developmental, and clinical perspectives.

PURPOSE: Total mesopancreas excision has been found to be helpful for increasing no residual tumor resection rate and improving the prognosis of pancreatic cancer. This study analyzed the relationships among the mesopancreas and pancreatic head plexus from the morphological, developmental, and clinical perspectives. METHODS: Twenty-four cadavers were employed. The upper abdominal viscera were resected en-bloc with the hepatoduodenal ligament, abdominal aorta, and nerve plexuses, and the innervation of the pancreas was dissected. Ten additional cadavers were used for histological examination of the pancreatic head and neck, part of the duodenum, the superior mesenteric artery (SMA) and its surrounding tissues, and the related arteries and veins. RESULTS: As results, cross-sections of the SMA revealed 6-9 layers of membranous structures resembling the layers of an onion, and the nerve fibers of the superior mesenteric plexus ran between the layers. Loose areolar tissue, adipose tissue, and lymphatics existed between the SMA and the pancreatic head/uncinate process, along with abundant thin blood vessels and capillaries, but very few nerves were found approaching the pancreas. Several parallel layers of collagen fibers (so-called Treitz's fusion fascia) existed between the dorsal aspect of the pancreatic head and the aortocaval plane. CONCLUSION: The mesopancreas was continuous and connected with the para-aortic area. It may be better termed the mesopancreatoduodenum than the mesopancreas, as the duodenum-pancreas-SMA forms a complex morphological, developmental, functional, and pathological structure.

Immunohistochemical study for relationship between vessel and lymphatic properties and tooth marks in human oral mucosa.

Angiogenesis is an important issue related to normal growth and differentiation, and it is a critical issue in the progression of human disease in oral mucosa. Tooth marks occur after clenching the teeth for a long period under muscle tension in the human oral cavity. However, the sites of angiogenesis, cell differentiation and microvessel density are not known for human mucosa with tooth marks. Therefore, we investigated the relationship between the markers of differentiation (Ki-67), angiogenesis (CD31, D2-40, VEGF-A), and marks from teeth in the second molar region using immunohistochemical methods. In addition, we compared these areas with the mucous membrane. Our results revealed blood and lymphoid vessels in irregular mucosa structures, and the vessels in the oral mucosa were observed in three types of samples: dentulous, denture attachment (containing partial teeth), and edentulous samples. The localization of the angiogenesis was related to the structure of the oral mucosa of connective tissue in humans, such as the mucosal fold-like of the buccal region. Using principal component analysis (PCA), we found that tooth occlusal condition, gender, anti-VEGF-A reaction levels in oral mucosa of the epithelium were positive factors in all groups, which is in contrast to the negative association of Ki-67 reaction in the epithelium and CD31 expression. In addition, Ki-67 reaction in oral mucosa had negative impacts, in contrast to the positive association of D2-40. These PCA properties provide useful information for future study of tumour progression or mechanical stress in remodelling of oral mucosa and angiogenesis. Moreover, mechanical stress of the occlusal condition may be correlated with tumour angiogenic activity and cell differentiation in human oral mucosa.

Temporal and dynamic changes in gingival blood flow during progression of ligature-induced periodontitis.

OBJECTIVES: To evaluate temporal changes in gingival blood flow (GBF) during progression of periodontitis in rats using a laser Doppler flowmeter (LDF) approach and to characterize morphological and biochemical features in the periodontium associated with GBF. MATERIALS AND METHODS: Forty-two Wistar rats were divided into a ligature-induced periodontitis group and a control group. To induce periodontitis, ligatures were tied around maxillary first molars bilaterally. GBF was measured in palatal gingiva at pretreatment and following ligature placement after 30 min, 1, 3, 7, 14, 21, and 28 days using LDF with a non-contact probe. Bone loss and gene expression in gingival tissues were assessed using micro-computed tomography (µCT) and quantitative polymerase chain reaction (PCR), respectively. Immunostaining for vascular endothelial growth factor (VEGF) in the maxilla was also histologically evaluated. RESULTS: GBF in the ligature group increased significantly compared with the control group 30 min after ligation. However, on days 3 and 7, GBF decreased in the ligature group. Also, after day 10, there was no difference in GBF between groups. The levels of alveolar bone loss, gene expression (interleukin-6, cluster of differentiation-31, VEGF-A, and lymphatic vessel endothelial hyaluronan receptor-1), and immunostained VEGF-positive vessels correlated well with changes in GBF. CONCLUSION PROGRESSION OF PERIODONTITIS: In rats was associated with a triphasic pattern of GBF, consisting of a short initial increase, followed by a rapid decrease, and then a gradual plateau phase.

Precise anatomical resection based on structures of nerve and fibrous tissue around the superior mesenteric artery for mesopancreas dissection in pancreaticoduodenectomy for pancreatic cancer.

BACKGROUND: The aim of the present study was to investigate the feasibility of resection based on the nerve and fibrous tissue (NFT) structures around the superior mesenteric artery (SMA) for resectable pancreatic adenocarcinoma (R-PDAC) patients. METHODS: NFTs around the SMA were classified into four "intensive NTFs area" with spreading the NFTs around the SMA and three SMA nerve plexus regions without branching nerves according to autopsy findings. Complete dissection of four "intensive NTFs areas" was performed by pre-exposing three SMA nerve plexus regions without branching nerves as "dissection-guiding points" with SMA nerve plexus preservation (NFT-based resection). Among 157 R-PDAC patients undergoing pancreaticoduodenectomy, surgical outcomes of 78 patients with NFT-based resection were compared with 59 patients with half-SMA nerve plexus dissection and 20 patients without NFTs dissection. RESULTS: In the NFT-based resection group, 76.5% had tumor involvement and metastasis in each intensive NTFs area. Operative time, blood loss, and postoperative diarrhea rate were significantly lower in NFT-based resection than in half-SMA nerve plexus group (321 vs 390 min; P < .01, 228 vs 550 mL; P < .01, 5.1% vs 15.3%; P = .04, respectively). R0 rate and median overall survival significantly improved in NFT-based resection than in non-NFT dissection group (93.6% vs 65.0%; P < .01, 49.6 vs 23.6 months, P = .01). CONCLUSION: NFT-based resection may become a novel method for R-PDAC patients.

Morphogenesis of the ventral pancreas anlagen is influenced by the SMA branching pattern.

Developmentally, the uncinate process of the pancreas is derived from the ventral pancreatic anlagen, supplied by the superior mesenteric artery (SMA), and contains pancreatic polypeptide (PP)-rich islets of Langerhans. In contrast, the other parts of the pancreas originate from the dorsal anlagen supplied by the celiac system and contain PP-poor islets. This study was performed to investigate whether morphogenesis of the ventral pancreas anlagen is associated with the pattern of SMA branching. SMA branches to the pancreatic body were dissected in 44 cadavers. The cadavers were divided into two groups: the SMA group in which the SMA gave off branches to the pancreatic body and the General group in which it did not. In the SMA group, the ratio of the diameter of the SMA branch supplying the pancreatic body (SMA branch) to that of the SMA itself was calculated. After dissection was completed, tissues were collected from all pancreatic specimens for HE staining and for immunohistochemistry with PP and insulin antibodies. There were 25 cadavers in the General group and 19 in the SMA group. In 10/19 cadavers from the SMA group, PP-rich islets were confirmed in the pancreatic body. The SMA branch diameter ratio was significantly smaller in the SMA group cadavers with PP-poor islets (n = 9) than in cadavers with PP-rich islets (n = 10) (P < 0.001). These findings suggest a relation between the SMA branching pattern and the distribution of PP cells.

Distribution of glutamate receptor, ionotropic, kainate 1 and neuropeptide calcitonin gene-related peptide mRNAs during formation of the embryonic and postnatal mouse molar in the maxilla.

The neuropeptide calcitonin gene-related peptide (CGRP) is a well-characterized neurotransmitter. Glutamate receptor, ionotropic, kainate 1 (Grik1) has also been demonstrated to generate high-affinity kainate receptors. However, little is known about the roles of CGRP and Grik1 during the developmental formation of teeth. In this study, we endeavoured to analyse the expression and localization of CGRP and Grik1 mRNAs using in situ hybridization on the mouse maxilla during development from the embryonic stage (E18.5) to after birth (P10, P15 and P20). We found that hybridization with an anti-sense probe for CGRP clearly localized in the maxilla at E18.5 in contrast to that of P15 and P20. Hybridization with an anti-sense probe for CGRP was not detected in the dental pulp of molars in the maxilla at P10, which is in contrast to Grik1 mRNA at the same developmental stage. Hybridization with an anti-sense probe for Grik1 mRNA was detected in the basal region of the dental pulp of molars at P10 and P15. Finally, these markers were not detected in molars in the mouse maxilla at P20. The ratio of positive cells for the hybridization signals of Grik1and CGRP in the dental pulp decreased from E18.5 (p<0.001). These features in CGRP and Grik1r mRNAs may indicate roles of function during tooth development between embryonic and postnatal stages with root formation and erupted movements.

Clinical anatomy of the anterior and posterior hepatic plexuses, including relations with the pancreatic plexus: A cadaver study.

The poor prognosis after surgery for pancreatic cancer or extrahepatic bile duct cancer has mainly been attributed to early lymph node metastasis, as well as a high frequency of perineural invasion along the peripancreatic neural plexuses or extrahepatic bile duct plexus. However, there has been no detailed morphological description of the anterior and posterior hepatic plexuses (AHP and PHP). In addition, the concepts of the pancreatic plexus and PHP are confused by surgeons. To assess the relations of the pancreatic plexus and hepatic plexuses from the morphological, developmental, and clinical perspectives, these plexuses were dissected in 24 cadavers. The PHP was found to be completely independent of the AHP. The PHP ran behind the portal vein, with most nerve fibers ascending along the bile duct to the gallbladder and the liver or descending to the distal common bile duct and duodenal papilla. Some branches of the PHP contributed to the pancreatic plexus, corresponding to pancreatic head plexus I as defined by the Japan Pancreas Society. The differences between the PHP and pancreatic head plexus I should be understood, even though liver function is not obviously affected after PHP excision for pancreatic head cancer. Further study is needed to determine whether there are functional differences between the AHP and PHP. Clin. Anat., 33:630-636, 2020. © 2019 Wiley Periodicals, Inc.

Analysis of the development of human foetal nasal turbinates using CBCT imaging.

PURPOSE: The morphological structure of the nasal cavity (NC) is important for endoscopic surgical treatment. The location of nasal turbinates, including the superior turbinate (ST), middle turbinate (MT) and inferior turbinate (IT), are well presented during the formation of the human NC in cone beam CT (CBCT) images. There is a complex relationship between the nasal sinuses, the maxillary sinus (MS), ethmoidal sinus and sphenoid sinus, during formation of the NC structure at the morphological level. There is a need to clearly define the relationships of these nasal elements at the ossification level, during development. METHODS: We investigated the three-dimensional construction of human foetal NC elements, including ST, MT, IT and vomer, using CBCT images from 16 weeks gestation (E16) to E31 (25 foetuses) and compared it to histochemical observations (E25). RESULTS: At the stage of ossification, the studied elements are elongated in the posterior region near the sphenoidal bone, showing that the locations of the ST, MT, and IT are important during formation of the NC. CBCT analysis revealed that the horizontal and vertical directions of nasal turbinates affect the formation of the human NC. CONCLUSION: The location and elongated development of the MT is one of the most important elements for NC formation. The relationship between the nasal sinus and nasal turbine at the level of ossification may provide useful information in clinical treatment of children.

Distribution of the neuropeptide calcitonin gene-related peptide-α of tooth germ during formation of the mouse mandible.

Calcitonin gene-related peptide-α (CGRPα) is a neurotransmitter that is related to bone formation during development. However, CGRP expression is not well known to affect the formation of teeth during development. During tooth germ development, the relationships among CGRPα, calcitonin receptor-like receptor (CRLR), amelogenin (AMELX), dentin sialophosphoprotein (DSPP), osteopontin (OPN) and osteocalcin (OCN) are unclear despite various tooth and osteogenesis markers. Our real-time RT-PCR results showed that the expression levels of CGRPα mRNA gradually decreased, in contrast to the mRNA abundances of CRLR, AMELX, DSPP, OPN, and OCN, which rapidly increased from E14.5 to P1 in the mandible. In situ hybridization using an antisense probe for CGRPα mRNA showed significant localized expression levels around the tooth bud at E14.5 and epithelial cells near the dental ledge and outer and inner enamel epithelium at E17.5 compared to those at P1. The localization of the anti-CGRPα antibody reaction revealed a strong positive reaction at the surface layer of oral epithelial cells at E14.5 and oral epithelial cells of the dental lamina around the dental ledge depression in the mandible of E17.5 mice using immunohistochemical methods The different anti-CGRPα reaction revealed its important roles during tooth formation at the postnatal stage. CGRPα mRNA was also detected in the interactions of tooth germ with the formation of odontoblast and amelobast layers from dental papilla and inner enamel epithelium. CGRPα may also be related to tooth germ development. Furthermore, CGRPα is an important tooth and bone formation marker, and bone cells provide further evidence of a role in mandibular development in contrast to inflammatory systems.

The distribution of nerves supplying the testis, epididymis and accessory sex glands of Suncus murinus.

Chronic testicular pain remains an important challenge for urologists. Investigation of the innervation of male gonads thus becomes essential for deepening our understanding of their regulatory roles in male reproductive physiology and pathophysiology. Studies of testicular innervation are mainly limited to the intratesticular peptidergic nerves of the testis by immunohistochemical and acetylcholinesterase histochemical investigations in some animals. Little is known about the detailed, overall distribution in general experimental animal testis. In this study, the distribution of nerves supplying the testis, epididymis and accessory sex glands of Suncus murinus was investigated by whole mount immunohistochemistry staining using a neurofilament protein antibody. Testicular nerves arose through three routes: nerves deriving from the mesenteric and renal plexuses accompanied the testicular artery, entering into the testicular hilum through the superior ligament of the testis. The nerves originating from the hypogastric plexus then ran along the internal iliac artery, deferential artery, and passed through the mesoductus deferens or mesoepididymis, innervating the cauda and corpus of the epididymis, the vas deferens and the inferior pole of the testis. The third route arose from the pelvic plexus, distributed in the seminal vesicle and the prostate. The density of nerve fibers was higher in the cauda epididymidis than in the testis, and more abundant in the vas deferens. The different origins and distribution densities of testicular nerves in S. murinus may serve different neuronal regulatory functions, and, therefore, S. murinus may be an important model animal for understanding the different characteristics of testicular pain.

Localization of CGRP and VEGF mRNAs in the mouse superior cervical ganglion during pre- and postnatal development.

The neuropeptide calcitonin gene-related peptide (CGRP) mediates inflammation and head pain by influencing the functional vascular blood supply. CGRP is a well-characterized mediator of receptor-regulated neurotransmitter release. However, knowledge regarding the role of CGRP during the development of the superior cervical ganglion (SCG) is limited. In the present study, we observed the localization of CGRP and vascular endothelial growth factor (VEGF-A) mRNAs during prenatal development at embryonic day 14.5 (E14.5), E17.5 and postnatal day 1 (P1) using in situ hybridization. The antisense probe for CGRP was detected by in situ hybridization at E14.5, E17.5, and P1, and the highest levels were detected at E17.5. In contrast, the antisense probe for VEGF-A was detected by in situ hybridization in gradually increasing intensity from E14.5 to P1. The differences in the expression of these two markers revealed specific characteristics related to CGRP concentration and release compared to those of VEGF-A during development. The correlation between CGRP and VEGF-A may influence functional stress and the vascular blood supply during prenatal and postnatal development.

CBCT imaging of the alveolar bone structure in maxilla of elderly donor cadavers and PCA analysis.

There is an important bone matrix with remodelling between dentate and edentulous samples of the human maxilla for bone metabolism. Cone beam computed tomography (CBCT) is useful for structural analysis of bone. The objective of this study was to investigate morphological data of donor cadavers in detail using CBCT imaging and principal component analysis (PCA). We analysed 38 donor cadavers using a CBCT apparatus. The analytical results defined differences in skull measurement parameters and dentate and edentulous levels using PCA. We observed cortical bone, trabecular bone, and the distance from the bottom of the maxillary sinus to the oral mucosa at a right angle to the palatal plane of the first molar region between dentate and edentulous samples of the human maxilla using CBCT imaging. In the dentate sample of the maxilla, component 1 was defined by negative contributions from gender (-0.84) and age (-0.54) to positive contributions such as cortical bone structure (CBS, 0.68) and trabecular bone structure (TBS, 0.50). There was a difference in CBS between dentate and edentulous human maxilla samples. This study of CBCT data provides useful basal information for planning dental implant surgery using PCA.

Expression of CGRP neurotransmitter and vascular genesis marker mRNA is age-dependent in superior cervical ganglia of senescence-accelerated prone mice.

Calcitonin gene-related peptide (CGRP) is a neurotransmitter that is released from the superior cervical ganglion (SCG) and causes head and neck pain. The morphological properties of human SCG neurons, including neurotransmitter content, are altered during aging. However, morphological changes in CGRP in the SCG during aging are not known. Therefore, we investigated CGRP and other markers in the SCG during aging in an aging model of senescence-accelerated prone mouse (SAMP8) and senescence-accelerated resistant mice (SAMR1) using real-time RT-PCR mRNA analyses and in situ hybridization. The abundance of neurotransmitter (CGRP, NPY, TRPV1), vascular genesis marker (CD31, LYVE-1), and cytochrome C mRNA differed between 12-week-old and 24-week-old SAMP8 and SAMR1. Abundance of TRPV1, CD31 and cytochrome C mRNAs of SAMP8 decreased between 12- and 24-week-old. The ratio of CGRP mRNA positive cells and CGRP mRNA abundance levels of the SCG of aging mouse such as SAMP8 have already been also higher than that of SAMR1 at 12-week-old. The CGRP positive shrunken ganglion cells was increased from 12- to 24-weeks-old mouse in SAMR1 and SAMP8. The SCG primarily affected the internal and external carotid arteries, larynx thyroid gland, and pharyngeal muscle during aging.

Morphological observation and CBCT of the bony canal structure of the groove and the location of blood vessels and nerves in the palatine of elderly human cadavers.

PURPOSE: The greater and lesser palatine nerves and vessels supply the hard and soft palates, and the roots of these vessels and nerves run through a bony structure. However, the arrangement of blood vessels in the maxilla requires attention during clinical treatments, but detailed morphological information about changes in the greater and lesser palatine arteries and nerves during aging is unavailable. We therefore need detailed investigations of the morphology of the donor cadaver palatine using cone-beam computed tomography (CBCT) and macroscopic observations. METHODS: We investigated 72 donor cadavers using macroscopic segmentation and CBCT. The results' analysis examined differences in skull measurement parameters and differences between dentate and edentulous cases. RESULTS: The greater palatine artery and nerve showed different macroscopic arrangements in dentate and edentulous cadavers. We also classified three types of bony structures of the nerve and vessel roots in the molar regions of the palatine using CBCT images: the shallow groove, deep groove, and flat groove. The deep groove is the deepest of the three and is remarkable in edentulous elderly cadavers. CONCLUSION: This study of macroscopic and CBCT data provides information useful for planning dental implant surgeries and autogenous bone harvesting.

A morphological study of the foramina of the mandible in the Japanese macaque by cone-beam computed tomography.

The mandibular canal (MC) contains vessels and nerves in the mandible of the Japanese macaque (JM). The inferior alveolar nerves and vessels of the mandible insert from the mandibular foramen and then run through the MC, the mental foramen and spinal foramen to the outside of the mandible. However, the detailed morphological properties of multiple canals, such as the accessory canal (AC) of the mandible, are unknown in JMs. The purpose of this study was to describe the multiple canals of JMs and to determine the location and analyse the measurements of the JM mandible. In this study, we also showed the course of the lingual foramen in 17 JMs (male: n = 8; female: n = 9) using cone-beam computed tomography (CBCT). In our results, we classified multiple mental foramina and multiple lingual foramina found on the mandibular body at the premolar or molar region. However, there was no significance between the formation of mandibular properties and the lingual foramen. These multiple foramina contain nerves and blood vessels have a few branched canals; these branches run downward and connect with the inferior mandibular nerve and artery. These morphological features may provide useful information about surgical treatment of the alveolus in a human model.

Expression of CGRP, vasculogenesis and osteogenesis associated mRNAs in the developing mouse mandible and tibia.

The neuropeptide Calcitonin Gene-Related Peptide (CGRP) is a well-characterized neurotransmitter. However, little is known about the role of CGRP in osteogenesis and vascular genesis during the developmental formation of bone. In the present study, we assessed the abundance of CGRP mRNA and the mRNA of osteogenesis and vascular genesis markers in the foetal mouse mandible and leg bone (tibia). We also analysed the expression and localization of CGRP, osteopontin (OPN) and vascular endothelial growth factor (VEGF-A) using in situ hybridization and immunohistochemical localization in the mouse mandible and tibia at embryonic days 12.5 (E12.5), E14.5, E17.5, and postnatal day 1 (P1). CGRP was clearly detected in the mandible relative to the tibia at E14.5. Hybridization using an anti-sense probe for CGRP was not detected in the mandible at P1. Hybridization with an anti-sense probe for OPN was detected at E14.5, later in the mandible and at P1 in Meckel's cartilage. However, OPN was only detected in the tibia at E17.5 and later. The abundance of CGRP mRNA differed between the mandible and tibia. The level of vasculogenesis markers, such as VEGF-A, was similar to that of CGRP in the mandible. The levels of VEGF-A, cluster of differentiation 31 (CD31) and lymphatic vessel endothelial hyaluronan receptor 1 (LIVE-1) differed from that of OPN in the mandible. In contrast, the levels of VEGF-A, CD31, matrix metalloproteinase-2 (MMP-2), collagen I (Col I), collagen II (Col II) and OPN mRNA differed from E12.5 to P1 (P<0.001) in the tibia. The abundance of mRNA of CGRP and bone matrix markers (Col I, Col II, and OPN) was low at P5 in the tibia. These differences in CGRP and other mRNAs may induce a different manner of ossification between the mandible and tibia. Therefore, a time lag of ossification occurs between the mandible and tibia during foetal development.

A morphological study of the multi-posterior superior alveolar canals of maxilla in the Japanese macaque by cone-beam computed tomography.

The posterior superior alveolar canal (PSAC) composed of several canals which contains vessels and nerve in molar region of the maxilla of Japanese macaque. The PSAC of maxilla run to the maxillary sinus. However, the PSAC and accessory canal (AC) of the maxilla in the Japanese macaque (JM) is unknown in morphological features in the maxilla. The purpose of this study was to describe the PSAC of the primates and to determine whether this structure could be used as a model for the human clinical condition. In this study, we showed the course of PSAC structure of the 23 JMs (male: n = 15; female: n = 8) using a cone-beam computed tomography apparatus. In the results, we classified a type to have one AC toward, a type to have two ACs toward, and three ACs in a type to have in PSAC. The main canal have some bony branch canals (BBCs) composed of 3 types (no BBC, one BBC, two BBCs). These canals and they run downward and supply to MS, these roots of maxillary molar region of the craniofacial skeleton in contrast to numerous small accessory canals with no nerve and vessels observed in the posterior regions in maxilla. These morphology features may give useful information about MS in dental treatment human model.

Expression of CGRP in embryonic mouse masseter muscle.

Neuropeptide calcitonin gene-related peptide (CGRP) is a mediator of inflammation and head pain that influences the functional vascular blood supply. The CGRP also regulate myoblast and acetylcholine receptors on neuromuscular junctions in development. However, little is known about its appearance and location during mouse masseter muscle (MM) development. We detected the mRNA abundance of CGRP, vascular genesis markers (Vascular endothelial growth factor A (VEGF-A), PECAM (CD31), lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1)) and embryonic and adult myosin heavy chain (MyHCs) (embryonic, IIa, IIb, and IIx) using real-time RT-PCR during development from the embryonic stage to after birth (E12.5, E14.5, E17.5, E18.5, P0, P1 and P5). We also endeavored to analyze the expression and localization of CGRP in situ hybridization in the developing mouse MM during development from the embryonic stage to after birth (E12.5, E14.5, E17.5, and P1). The antisense probe for CGRP was detected by in situ hybridization at E12.5, E14.5 E17.5 and then no longer detected after birth. The CGRP, CD31, embryonic MyHC abundance levels are highest at E17.5 (p<0.001) and they show a pattern similar to that of the other markers from E12.5 to P5. PCA analysis indicates a specific relation between CGRP and embryonic MyHC, CD31, and LYVE-1 in MM development. Cluster analyses identified the following distinct clusters for mRNA abundance in the MM: cluster 1, P5; cluster 2, E12.5, E14.5, E17.5, E18.5, P0, and P1. The positive correlation between CGRP and embryonic MyHC (Pearson's r>0.65; p<0.01) was analyzed. These data suggested that CGRP may have an influence on embryonic MyHC during mouse MM development. CGRP also affects the angiogenesis markers at embryonic stages.

Expression of myostatin in early postnatal mouse masseter and rectus femoris muscles.

AIMS: Myostatin (Mstn) is a member of the transforming growth factor-ß (TGF-ß) family that inhibits muscle differentiation. In this study, we aimed to identify the relationships between Mstn, thyroid hormone receptor alpha (TRα), and myosin heavy chain (MyHC) isoform expression during early postnatal development. METHODS: We investigated the expression of Mstn, TRα, and MyHCs (embryonic, slow, IIa, IIb, and IIx) using quantitative real-time RT-PCR and ELISA (Mstn) in postnatal mouse muscles between day 0 and day 10. We also examined the correlations between Mstn, TR and MyHCs during the early development of mouse masseter muscle (MM) and rectus femoris muscle (RFM). RESULTS: Distinct Mstn mRNA expression patterns were observed in the two muscles despite nearly non-significant changes in the Mstn protein abundance in MM. The expression pattern of the TRα mRNA in the MM differed from that observed in the RFM. The expression of MyHC IIa, IIb and IIx mRNAs increased in the MM and decreased in the RFM from day 0 to day 10, whereas embryonic fiber MyHC mRNA expression was similar in both muscle types. Principal component analysis showed the existence of a correlation between: (1) TRα and MyHC, (2) Mstn and MyHC, and (3) TRα and Mstn in MM. The correlations were different in RFM and MM. Cluster analyses identified the distinct clusters: cluster 1, days 0-4 for the MM and day 0 for the RFM; cluster 2, day 6 for the MM and day 2 for the RFM; and cluster 3, days 8-10 for the MM and days 4-10 for the RFM. CONCLUSIONS: These data suggest that TRα influences MyHC expression in both muscle types. In addition, Mstn has a limited effect in the MM related to the expression of individual MyHCs, as opposed to its role in the RFM, at early postnatal developmental stages. TRα could be involved in regulating both the temporal expression of MyHCs and Mstn at the early postnatal stages in the MM and RFM.