Development and growth of the temporal fascia: a histological study using human fetuses.

The temporal fascia is a double lamina sandwiching a thick fat layer above the zygomatic bony arch. To characterize each lamina, their developmental processes were examined in fetuses. We observed histological sections from 22 half-heads of 10 mid-term fetuses at 14-18 weeks (crown-rump length, 95-150 mm) and 12 near-term fetuses at 26-40 weeks (crown-rump length, 215-334 mm). The superficial lamina of the temporal fascia was not evident at mid-term. Instead, a loose subcutaneous tissue was attached to the thin, deep lamina of the temporal fascia covering the temporalis muscle. At near-term, the deep lamina became thick, while the superficial lamina appeared and exhibited several variations: i) a mono-layered thick membrane (5 specimens); ii) a multi-layered membranous structure (6) and; iii) a cluster of independent thick fasciae each of which were separated by fatty tissues (1). In the second and third patterns, fatty tissue between the two laminae was likely to contain longitudinal fibrous bands in parallel with the deep lamina. Varying proportions of the multi-layered superficial lamina were not attached to the zygomatic arch, but extended below the bony arch. Whether or not lobulation or septation of fatty tissues was evident was not dependent on age. The deep lamina seemed to develop from the temporalis muscle depending on the muscle contraction. In contrast, the superficial lamina developed from subcutaneous collagenous bundles continuous to the cheek. Therein, a difference in development was clearly seen between two categories of the fasciae.

Growing stylohyoideus muscle insertion to the hyoid bone with special reference to its topographical relation to the intermediate tendon of digastricus muscle: A histological study using human fetuses.

BACKGROUND: In adults, the intermediate tendon of digastricus muscle usually runs along the medial or lateral side of the stylohyoideus muscle insertion. To provide a better understanding of the variations, we examined the topographical anatomy of the muscle and tendon in fetuses. METHODS: We examined histological sections from six early-term, 26 mid-term and six near-term fetuses (approximately 8-9, 12-18 weeks and 25-33 weeks). RESULTS: At early-term, an initial sheath of intermediate tendon of digastricus muscle received the stylohyoideus muscle at the superior aspect. The muscle and tendon was distant from the hyoid. At mid-term, near the insertion to the hyoid greater horn, the stylohyoideus muscle consistently surrounded more than 2/3 of the intermediate tendon circumference. In contrast, we found no near-term specimen in which the stylohyoideus muscle surrounded the intermediate tendon. The multilayered tendon sheath was fully developed until near-term and connected to the body of hyoid by an intermuscular septum between the thyrohyoideus muscle and one or two of suprahyoid muscles. Therefore, the hyoid insertion of the styloglossus muscle was a transient morphology at mid-term. CONCLUSION: The stylohyoideus muscle insertion was appeared to move from the tendon sheath to the hyoid greater horn and, until near-term, return to the tendon sheath. A fascia connecting the tendon sheath to the body of hyoid was strengthened by the suprahyoid and infrahyoid muscles. The latter muscles seemed to regulate fixation/relaxation of the intermediate tendon to the hyoid. The stylohyoideus muscle slips sandwiching the intermediate tendon might be a rare morphology.

Development and growth of median structures in the human tongue: A histological study using human fetuses and adult cadavers.

Glossectomy is a surgical procedure performed to remove all or part of the tongue in patients with cancer. The removal of a significant part of the tongue has a marked effect on speech and swallowing function, as patients may lose not only the tongue muscles but also the median lingual septum (MLS). Therefore, to achieve successful tongue regeneration, it is necessary to investigate the developmental processes of not only the tongue muscles but also the MLS. This study was conducted to clarify the mutual development of the tongue muscles and the MLS in human fetuses. Serial or semi-serial histological sections from 37 embryos and fetuses (aged 5-39 weeks) as well as nine adults were analyzed. The MLS appeared at Carnegie stage 15 (CS15), and until 12 weeks of gestation, abundant fibers of the intrinsic transverse muscle crossed the septum in the entire tongue. However, in near-term fetuses and adults, the contralaterally extending muscles were restricted to the deepest layer just above the genioglossus muscle. This finding indicates that the crossing transverse muscle showed the highest density at mid-term. A thorough understanding of both the MLS and the tongue muscles is necessary for successful tongue regeneration.

Histology of the optic nerve head with special reference to the layer-specific distribution of composite fibers at and near the lamina cribrosa: An immunohistochemical study using specimens from elderly donated cadavers.

BACKGROUND: This study aimed to demonstrate the composite fibers of the lamina cribrosa (LC) and their layer-specific distributions. The elastic fiber-rich septa, showing a cribriform arrangement in the optic nerve, may continue into the LC. METHODS: Orbital content, including the long course of the optic nerve, was obtained from 25 elderly cadavers. Sagittal and cross-sections were prepared from each specimen. In addition to elastica Masson staining, immunohistochemistry was performed for elastin, glial fibrillary acidic protein (GFAP), S100 protein (S100), and CD68 in microglia. RESULTS: The LC beam usually had fewer elastic fibers than the septa, but an elastic fiber-rich zone was observed along the scleral flange. GFAP-positive fibers were rich in the prelaminar area, whereas S100-positive fibers were rich in all layers of the LC. Double-positive (GFAP+/S100+) fibers were present in the prelaminar area. In contrast, S100-single positive fibers were evident in the LC and retrolaminar areas and were likely to insert into a sclera-choroid border area. The density of macrophages and microglia was not different between the septa and LC. Individual variations were observed in the distribution and density of the nerve-associated fibrous tissues. CONCLUSION: The LC beam was quite different from the septa in the composite fibers and architecture. Transverse fibers, dominant in the LC beam, corresponded to fibrous processes of astrocytes and other nerve-associated fibrous tissues. Many of these nerve elements suggest low mechanical properties of the LC.

Changing the topographical anatomy among the maxilla, palatine bone, and greater palatine nerve: a histological study using human fetuses.

PURPOSE: The palatine bone (PAL) rides over the maxilla (MX) without an end-to-end suture in the bony palate of fetuses. However, changes in the topographical relationship among bones was unknown at and along the pterygopalatomaxillary suture, including the palatine canals. METHODS: Using sagittal, frontal, and horizontal histological sections of the head from 15 midterm fetuses to 12 near-term fetuses, we depicted the changes in the topographical anatomy of the MX, PAL, and greater palatine nerve (GPN). RESULTS: In the bony greater palatine canal of these fetuses, the medial and posterior walls facing the GPN were consistently made up of the PAL. At midterm, the entire course of the GPN was embedded in the PAL (six fetuses), or the MX contributed to the lateral wall of the nerve canal (nine). At near-term, the anterior and lateral walls showed individual variations: an MX in the anterior and lateral walls (three fetuses), an anterior MX and a lateral PAL (five), an anterior PAL and a lateral MX (two), and a PAL surrounding the GPN (four). CONCLUSION: These increasing variations suggested that the pterygopalatomaxillary suture was actually growing and that the PAL transiently expanded anteriorly and/or laterally to push the MX in fetuses. The "usual" morphology in which the GPN is sandwiched by the MX and PAL is likely established after birth, possibly during adolescence. The driving force of this change may not be produced by the masticatory apparatus. Rather, it might be triggered by the growing maxillary sinus.

Selective somatostatin receptor 5 inhibition improves hepatic insulin sensitivity.

Diabetes is a metabolic disorder with an increasing global prevalence. Somatostatin (SST), a peptide hormone, regulates hormone secretion via five SST receptor (SSTR) subtypes (SSTR1-5) in a tissue-specific manner. As SSTR5 is expressed in pancreatic ß-cells and intestinal L-cells, studies have suggested that SSTR5 regulates glucose tolerance through insulin and incretin secretion, thereby having a prominent role in diabetes. Moreover, SSTR5 knockout (KO) mice display enhanced insulin sensitivity; however, the underlying mechanism has not been clarified. Therefore, in this study, we investigate the effect of SSTR5 blockade on insulin resistance and the target organ using SSTR5 KO mice and a selective SSTR5 antagonist (compound-1). High-fat diet (HFD)-fed SSTR5 KO mice exhibited significantly lower homeostasis model assessment of insulin resistance (HOMA-IR) than HFD-fed wild-type mice. Two-week oral administration of compound-1 dose-dependently and significantly reduced changes in the levels of glycosylated hemoglobin (GHb), plasma glucose, plasma insulin, and HOMA-IR in male KK-Ay /Ta Jcl mice (KK-Ay mice), a model of obese type 2 diabetes with severe insulin resistance. Additionally, compound-1 significantly increased the glucose infusion rate while decreasing hepatic glucose production in male KK-Ay mice, as evidenced by hyperinsulinemic-euglycemic clamp analyses. In addition, compound-1 ameliorated the insulin-induced Akt phosphorylation suppression by octreotide in the liver of male C57BL/6J mice. Collectively, our results demonstrate that selective SSTR5 inhibition can improve insulin sensitivity by enhancing liver insulin action; thus, selective SSTR5 antagonists represent potentially novel therapeutic agents for type 2 diabetes.

CD34 is Expressed in Endothelial Cells in Embryonic Testes and is Additionally Expressed in Non-Endothelial Cells in Postnatal Mouse Testes.

CD34 is expressed in various cell types in various tissues/organs, and has been regarded as being expressed in progenitors in various differentiation pathways. On the other hand, morphological studies have reported the presence of a special type of interstitial cells, telocytes, which generally express CD34, and have extremely long moniliform prolongations in various tissues/organs in vertebrates. We have recently reported the successful reconstruction of testicular structures by 3-D re-aggregation culture of dissociated prepubertal mouse testicular cells, and the roles of CD34 + cells in the reconstruction. However, it was unknown whether CD34 is expressed in embryonic through adult testes, and if so, in what cell type it is expressed. In order to clarify the expression of CD34 and behavior of CD34 + cells during development of mouse testes, we performed immunohistochemical studies. The results show that CD34 is expressed in two cell types in testes; one is endothelial cells which co-express CD31, VE-cadherin, and integrin ß1, but barely express PDGFRα and integrin α4 and α9, throughout development, while the other one is non-endothelial cells in which CD34 expression is initiated after birth, and which co-express PDGFRα and integrin α4, α9, and ß1. The latter corresponds to telocytes. The present findings will lead to clarifying the roles of these two types of CD34 + cells in spermatogenesis.

Topographical relationships of the yolk sac remnant and vitelline vessels with the midgut loop in the extra-embryonic coelom of human embryos.

The yolk sac is supplied by the vitelline artery and vein (VA, VV), which run through the yolk stalk in combination with the omphaloenteric duct. Moreover, the VV takes a free posterior course outside the midgut mesentery containing the secondarily-developed superior mesenteric vein (SMV). However, the regression process of these structures has not been demonstrated photographically. The present study evaluated serial histological sections from 20 embryos of stages 15-19 or crown-rump length (CRL) 7.5-20 mm. All specimens carried the SMV as sequential tissue slits. However, an omphaloenteric duct with epithelia continuous with the midgut loop was not observed. In smaller embryos (CRL <13 mm) the VA extended distally or anteriorly from the midgut apex in the extra-embryonic coelom, whereas in larger embryos (CRL 16-20 mm) the artery was absent from the distal side of the apex. The entire course or part of the VV outside the mesentery was always seen, but four larger embryos lacked the venous terminal near the duodenum. A vacuole-like remnant of the yolk sac was present in all smaller embryos (CRL <10 mm), but was absent from 7 of the 11 larger embryos. The size of the remnant was equal to the thickness of the VA or VV, with the remnant being sandwiched between the VA and VV. Moreover, the regressing yolk sac often communicated with or opened to the VV. Consequently, the yolk sac regressed first, followed by the regression of the VA until 6 weeks. The yolk stalk was clearly observed until 5 weeks.

Behavior and Functional Roles of CD34+ Mesenchymal Cells in Mammalian Testes.

Mammalian testes consist of seminiferous tubules within which Sertoli cells line up at the periphery and nurse germ cells, and of interstitia that harbor various cells such as peritubular myoid cells (PMCs), Leydig cells (LCs), vascular endothelial cells, immune cells such as macrophages, and mesenchymal (stromal) cells. Morphological studies have recently reported the presence of telocytes with telopodes in the interstitium of adult mouse, rat, and human testes. CD34+PDGFRα+ telocytes with long and moniliform telopodes form reticular networks with various cell types such as LCs, PMCs, and vessels, indicating their potential functions in cell-cell communications and tissue homeostasis. Functional studies have recently been performed on testicular interstitial cells and CD34+ cells, using 3D re-aggregate cultures of dissociated testicular cells, and cell cultures. Direct observation of CD34+ cells and adult LCs (ALCs) revealed that CD34+ cells extend thin cytoplasmic processes (telopodes), move toward the LC-CD34+ cell-re-aggregates, and finally enter into the re-aggregates, indicating the chemotactic behavior of CD34+ telocytes toward ALCs. In mammalian testes, important roles of mesenchymal interstitial cells as stem/progenitors in the differentiation and regeneration of LCs have been reported. Here, reports on testicular telocytes so far obtained are reviewed, and future perspectives on the studies of testicular telocytes are noted.

Liquid chromatography-mass spectrometry measurements of blood diphosphoinositol pentakisphosphate levels.

Diphosphoinositol pentakisphosphate (InsP7) is an inositol pyrophosphate generated by inositol hexakisphosphate kinases (IP6Ks) that regulate diverse biological functions in cells. To date, we have a limited understanding of the InsP7 biology owing to limited data on InsP7 levels in blood or other tissues. Given the significant role of InsP7 in maintaining biological homeostasis, further advancement in InsP7 measurement is essential. In this study, we report a highly sensitive liquid chromatography with tandem mass spectrometry method for determining InsP7 levels in whole blood, which is an easily accessible tissue and for which knowledge on InsP7 levels is limited. We applied a perchloric acid-based method to increase the extraction efficiency of InsP7 from the cells. Subsequently, we combined a YMC-Triart C18 metal-free column, ion-pair reagents, EDTA, methylenediphosphonic acid, and N,N,N',N'-Ethylenediaminetetrakis(methylenephosphonic acid) to minimize InsP7 adsorption to the detection system. We prepared blood quality control samples that were highly exposed to IP6K inhibitor, showing minimum InsP7 levels, to decrease the lower quantification limit of InsP7. Furthermore, we applied rat plasma, which was found to show no InsP7 levels, as a surrogate matrix. This setting resulted in the highly sensitive detection of InsP7 levels in blood obtained from rats, with a quantification sensitivity of 1 ng of InsP7 per mL of blood. The current method demonstrated acceptable accuracy (100 ± 15%) and precision (coefficient of variation ≤ 15%) in rat blood. Using this method, we revealed endogenous basal levels of InsP7 (37.4 ng/mL) in blood obtained from normal rats, and decreased levels of InsP7 (4.1-21.7 ng/mL) in blood obtained from IP6K inhibitor-administered rats. In summary, we established a highly sensitive method for measuring InsP7 and revealed its levels in rat blood. The current findings may help in understanding InsP7 biology.

Anatomy of the vitelline vein remnant in human embryos and fetuses.

PURPOSE: To demonstrate the entire course of the human vitelline vein (VV) in specimens after degeneration of the yolk sac. METHODS: Sagittal and horizontal histological sections from 8 embryos and 19 fetuses (gestational age approximately 6-12 weeks; crown-rump length 11-61 mm) were examined. RESULTS: Two types of VV remnants were observed: a long VV on the right superior side of the mesentery of the jejunum (VV1) and a short VV on the left inferior side of the mesentery (VV2). The VV1, observed in 12 specimens, was 20-30 microns in diameter and ran dorsally between the right liver lobe and the jejunum, subsequently merging with an initial superior mesenteric vein on the pancreatic head immediately below the superior portion of the duodenum. The VV2, observed in four specimens, passed dorsally between loops of the ileum on the left side of the mesentery of the ileum and connected to the mesentery. Many of the VVs did not originate from the umbilical cord but suddenly started in the sack of physiological herniation. At 10-12 weeks, after herniation, the VVs originated from the umbilicus and were involved by the expanding greater omentum. CONCLUSIONS: The right-sided and left-sided VVs seemed to correspond to right and left VV remnants, respectively, and both took an upstream course outside the mesentery of the jejunum and ileum. The right VV upstream portion was likely to disappear later than the left one, but the timing of degeneration varied greatly among individuals, depending on the topographical relationship between the right liver lobe and the jejunum.

Optic nerve-associated connective tissue structures revisited: A histological study using human fetuses and adult cadavers.

Unlike the usual peripheral nerve, the optic nerve accompanies a thick "dural sheath," a thin "sheath of pia mater" (SPM), and multiple "septa," which divides the nerve fibers into fascicles. We collected specimens from 25 adult cadavers and 15 fetuses and revisited the histological architecture of the optic and oculomotor nerves. In the optic chiasma, the meningeal layer of the dura joins the pia to form a thick SPM, and the periosteum of the sphenoid is continuous with the dural sheath at the orbital exit of the bony optic canal. The septa appeared as a cluster of irregularly arrayed fibrous plates in the intracranial course near the chiasma. Thus, the septa were not derived from either the SPM or the dural sheath. In the orbit, the central artery of the retina accompanies collagenous fibers from the dural sheath and the SPM to provide the vascular sheath in the optic nerve. These connective tissue configurations were the same between adult and fetal specimens. At the optic disk, the dural sheath and SPM merged with the sclera, whereas the septa appeared to end at the lamina cribrosa. However, in fetuses without lamina cribrosa, the septa extend into the nerve fiber layer of the retina. The SPM and septa showed strong elastin immunoreactivity, in contrast to the absence of reactivity in the sheaths of the oculomotor nerve. Each S100 protein-positive Schwann sheath of the oculomotor nerve was surrounded by collagenous endoneurium. Glial fibrillary acidic protein-positive astrocytes showed a linear arrangement along the septa.

Lost or fragmented bony septum of the optic canal facing the sphenoid sinus: a histological study using elderly donated cadavers.

PURPOSE: To histologically describe a direct contact (the so-called dehiscence) of the optic nerve (ON) and/or internal carotid artery (ICA) to the mucosa of posterior paranasal sinuses represented by the sphenoid sinus (SS). METHODS: Observations of histological sections of unilateral or bilateral skull bases (parasellar area and orbital apex) from 22 elderly cadavers were made. RESULTS: A bony septum was less than 300 µm between the SS and ICA and 200 µm between the SS and optic nerve. Parts of the septa were sometimes absent due to fragmentation and holes of the bony lamella (2/22 facing the ICA; 4 facing the ICA in combination with an absent bony septum facing the nerve). In these dehiscence sites, the SS submucosal tissue attached to a thick sheath (50-100 µm in thickness) enclosing the optic nerve and ophthalmic artery and/or the ICA adventitia (50-200 µm in thickness). The ICA sometimes contained a sclerotic plaque that attached to or even protruded into the SS. With or without dehiscence, the SS mucosa was always thin (50-100 µm in thickness) and accompanied no mononuclear cellular infiltration or tumor. CONCLUSIONS: A thin bony septum of the optic nerve or ICA had been notable as a danger point during surgery, but even a 0.05-mm-thick bone lamella might be an effective barrier against cellular infiltration or bacterial invasion from the SS. Fragmentation and holes of the bony lamella in 4 cadavers might allow cellular invasion to the optic nerve. Accordingly, unknown immunological cross talks might occur to cause demyelination.

Growth in fetuses of the constrictor pharyngis superior with special reference to its meeting with the buccinator: an embryological basis of adult variations in palatopharyngeal anatomy.

PURPOSE: The constrictor pharyngis superior (CPS) initially develops along the posterior wall of the pharyngeal mucosal tube, whereas, during the early phase, the buccinators (BC) are far anterolateral to the CPS. The process and timing of their meeting during fetal growth have not been determined. METHODS: The topographical relationship between the growing BC and CPS was assessed in histological sections from 22 early- and mid-term fetuses of approximate gestational age (GA) 8-16 weeks, and eight late-term fetuses of approximate GA 31-39 weeks. RESULTS: At 8-9 weeks, the palatopharyngeus appeared to pull the CPS up and forward. Until 11 weeks, the CPS was attached to the hamulus of the pterygoid (pterygopharyngeal part). Until 13 weeks, the CPS extended anterolaterally beyond the hamulus to meet the BC. Some BC muscle fibers originated from the oral mucosa. Notably, by 30 weeks, the CPS-BC interface had become covered by or attached to the palatopharyngeus. Muscle fibers of the palatopharyngeus, however, were thinner than those of the CPS and BC. At and near the interface, BC muscle fibers tended to run along the left-right axis, whereas those of the CPS ran anteroposteriorly. A definite fascia (i.e., a future pterygomandibular raphe) was usually absent between these muscles in fetuses. CONCLUSIONS: The excess anterior growth of the CPS with its subsequent degeneration might cause individual anatomical variations in composite muscle bundles of the palatopharyngeus-CPS complex or palatopharyngeal sphincter. A tensile transduction from the BC to the CPS through the raphe seemed unnecessary for cooperative suckling and swallowing after birth.

VCAM1-α4ß1 integrin interaction mediates interstitial tissue reconstruction in 3-D re-aggregate culture of dissociated prepubertal mouse testicular cells.

Roles of interstitial tissue in morphogenesis of testicular structures remain less well understood. To analyze the roles of CD34+ cells in the reconstruction of interstitial tissue containing Leydig cells (LCs), and testicular structures, we used 3D-reaggregate culture of dissociated testicular cells from prepubertal mouse. After a week of culture, adult Leydig cells (ALCs) were preferentially incorporated within CD34+ cell-aggregates, but fetal LCs (FLCs) were not. Immunofluorescence studies showed that integrins α4, α9 and ß1, and VCAM1, one of the ligands for integrins α4ß1 and α9ß1, are expressed mainly in CD34+ cells and ALCs, but not in FLCs. Addition of function-blocking antibodies against each integrin and VCAM1 to the culture disturbed the reconstruction of testicular structures. Antibodies against α4 and ß1 integrins and VCAM1 robustly inhibited cell-to-cell adhesion between testicular cells and between CD34+ cells. Cell-adhesion assays indicated that CD34+ cells adhere to VCAM1 through the interaction with α4ß1 integrin. Live cell imaging showed that CD34+ cells adhered around ALC-aggregates. CD34+ cells on the dish moved toward the aggregates, extending filopodia, and entered into them, which was disturbed by VCAM1 antibody. These results indicate that VCAM1-α4ß1 integrin interaction plays pivotal roles in formation of testicular interstitial tissues in vitro and also in vivo.

Human orbital muscle in adult cadavers and near-term fetuses: its bony attachments and individual variation identified by immunohistochemistry.

PURPOSE: To compare fetal and adult morphologies of the orbital muscle (OM) and to describe the detailed topographical anatomy in adults. METHODS: Using unilateral orbits from 15 near-term fetuses and 21 elderly cadavers, semiserial horizontal or sagittal paraffin sections were prepared at intervals of 20-100 µm. In addition to routine histology, we performed immunohistochemistry for smooth muscle actin. RESULTS: At near term, the OM consistently extended widely from the zygomatic bone or the greater wing of the sphenoid to the maxilla or ethmoid. Thus, it was a large sheet covering the future inferior orbital fissure. In contrast, the adult OM was a thin and small muscle bundle connecting (1) the greater wing of the sphenoid to the maxilla (11/19 cadavers), (2) the lesser wing of the sphenoid to the maxilla (5/19) or the greater wing (3/19). The small OM was likely to be restricted within the greater wing (5/19 cadavers) or the maxilla (3/19). Two of these five types of OM coexisted in eight orbits. OM attachment to the lesser wing was not seen in fetuses, whereas ethmoid attachment was absent in adults. CONCLUSIONS: The lesser wing attachment of the OM seemed to establish after birth. A growing common origin of the three recti was likely involved in "stealing" the near-term OM attachment from the ethmoid. The strong immunoreactivity of remnant-like OM in the elderly suggests that OM contraction is still likely to occur against the increased flow through a thin vein. However, the contraction might have no clinical significance.

Characteristic Distribution of Hematopoietic Cells in Bone Marrow of Xenopus Laevis.

Bone marrow is the principal site of hematopoiesis in mammals. Amphibians were the first phylogenetic group in vertebrates to acquire bone marrow, but the distribution of hematopoietic cells in the bone marrow of the primitive frog, Xenopus laevis (X. laevis) has not been well documented. The purpose of this study was to perform a histological investigation of the distribution of hematopoietic cells in femoral bone marrow at various stages of development in X. laevis. Hematopoietic cells showed preferential distribution on the endosteal surface of cortical bone throughout all stages of development, from tadpole to aged frog. In mature frogs, hematopoietic cells appeared at the boundary between the epiphysis and the bone marrow. The distribution of hematopoietic cells around the blood vessels was limited to a small number of vessels in the bone marrow. Abundant adipose tissue was observed in the bone marrow cavity from the tadpole stage to the mature frog stage. Hematopoietic cells showed preferential distribution in a belt-like fashion on the surface of newly-formed bones in a bone regeneration model in the diaphysis of X. laevis. These results indicate that the distribution of hematopoietic cells in bone marrow in X. laevis differs from that in mammals, and that the bone marrow of X. laevis constitutes a useful model for exploring the mechanism underlying the phylogenetic differentiation of bone marrow hematopoiesis.

The enzymatic activity of inositol hexakisphosphate kinase controls circulating phosphate in mammals.

Circulating phosphate levels are tightly controlled within a narrow range in mammals. By using a novel small-molecule inhibitor, we show that the enzymatic activity of inositol hexakisphosphate kinases (IP6K) is essential for phosphate regulation in vivo. IP6K inhibition suppressed XPR1, a phosphate exporter, thereby decreasing cellular phosphate export, which resulted in increased intracellular ATP levels. The in vivo inhibition of IP6K decreased plasma phosphate levels without inhibiting gut intake or kidney reuptake of phosphate, demonstrating a pivotal role of IP6K-regulated cellular phosphate export on circulating phosphate levels. IP6K inhibition-induced decrease in intracellular inositol pyrophosphate, an enzymatic product of IP6K, was correlated with phosphate changes. Chronic IP6K inhibition alleviated hyperphosphataemia, increased kidney ATP, and improved kidney functions in chronic kidney disease rats. Our results demonstrate that the enzymatic activity of IP6K regulates circulating phosphate and intracellular ATP and suggest that IP6K inhibition is a potential novel treatment strategy against hyperphosphataemia.

Fetal development of the carotid canal with special reference to a contribution of the sphenoid bone and pharyngotympanic tube.

The bony carotid canal is a tube-like bone with a rough surface in contrast to smooth surfaces of the other parts of the temporal bone petrosal portion (petrosa): it takes an impression of the additional, out-sourcing product. No study had been conducted to evaluate a contribution of the adjacent sphenoid and pharyngotympanic tube (PTT) to the carotid canal. We examined sagittal and horizontal histological sections of hemi-heads from 37 human fetuses at 10 to 37 weeks. At 10 to 18 weeks, the future carotid canal was identified as a wide loose space between the cartilaginous cochlea and the ossified or cartilaginous sphenoid elements (ala temporalis and pterygoid). A linear mesenchymal condensation extending between the cochlear wall and ala temporalis suggested the future antero-inferior margin of the carotid canal. This delineation was more clearly identified in later stages. After 25 weeks, 1) the growing pterygoid pushed the PTT upward and, in turn, the PTT pushed the internal carotid artery (ICA) upward toward the petrosa: 2) a membranous ossification occurs in the dense mesenchymal tissue, the latter of which took an appearance of an anterior process of the petrosa; 3) the bony process of the petrosa involved the ICA inside or posteriorly. The bony carotid canal was made with membranous ossification in the dense mesenchymal tissue between the petrosa and sphenoid. The mother tissue was detached from the sphenoid by the PTT. The ossification of the septum between the ICA and tympanic cavity seemed to continue after birth.

Development and growth of the craniocervical junction with special reference to topographical relationship between the occipital basion, the anterior arch of atlas, and the odontoid process of axis: A study using human fetuses.

The embryonic occipital bone and odontoid process of the axis are attached and connected by the notochord, but become separated in later development and growth. With special attention to the process of separation, we examined sagittal sections of the craniocervical junction in 18 human fetuses at 8-16 weeks and 22 fetuses at 31-37 weeks. At 8-9 weeks, the anterior arch of atlas was always seen overriding the occipital basal part. The odontoid process was close to the occipital with or without a transient joint cavity until 16 weeks. Near term, the top of the odontoid process was usually higher than the anterior arch, but the former was sometimes (7 of 22) at a level almost equal to or lower than the latter. The apical ligament was evident in a few specimens (5 of 22). A distance between the occipital basion and odontoid process was sometimes less than 1.5 mm (8 of 22) or less than half the thickness of the arch (10 of 22). A transient joint cavity between the basion and odontoid process was often (10 of 22). In three fetuses near term, the atlanto-occipital joint cavity was continuous with the median atlanto-axial joint cavity, and the anterior arch was overriding the occipital basal part. Therefore, rather than stage or age, individual differences were evident in the topographical relationship between the three bony elements at the craniocervical junction. An understanding of the embryology and normal development will aid in the correct interpretation of radiologic images of the pediatric cervical spine.