Specific localization of fibroblasts at the intercalated duct in the major salivary glands of rats.

OBJECTIVES: Immunohistochemical methods were employed to investigate the morphological heterogeneity and localization of fibroblasts associated with the function of major salivary glands in rats. METHODS: Histochemical and electron microscopic observations were made in rat parotid, submandibular, and sublingual glands and pancreas. Fibroblasts were immunostained using their specific marker, 47 kDa heat shock protein (Hsp47). RESULTS: Hsp47-immunopositive fibroblasts within the intralobular connective tissue exhibited a notably smaller size compared with the interlobular connective tissue. They were loosely distributed throughout the connective tissue. However, fibroblasts with elongated long processes were explicitly identified at the intercalated ducts in parotid, sublingual, and submandibular glands. Fibroblastic bodies and processes were tightly approximated with the basement membrane of the duct. Electron microscopy confirmed these findings, revealing a thin layer consisting of collagen fibers was found between the fibroblasts and the basement membrane. Double staining of Hsp47 and α-smooth muscle actin (αSMA) in parotid glands indicating that Hsp47-positive fibroblasts enveloped both the duct and αSMA-positive myoepithelial cells. Additionally, They projected long and thin processes longitudinally at the straight portion or circularly at the bifurcated portion of the duct. The three-dimensional reconstruction showed a frame-like structure of fibroblasts surrounding the intercalated duct with longitudinal myoepithelial cells. However, such specific localization of fibroblasts was not detected in the exocrine pancreas lacking myoepithelium. CONCLUSIONS: Small fibroblasts with long processes connecting or overwrapping each other and thin collagen layers surround the intercalated ducts in rat major salivary glands, presumably contributing to protecting the ducts from salivary flow and myoepithelial contraction.

Live imaging observation of elevation of the anterior palatal shelf in mouse embryos.

The mammalian secondary palate develops through complex processes including palatal shelf growth, elevation, and fusion. Palatal shelf elevation is a process accompanied by large-scale morphological changes over a short period. The elevation pattern changes along the anterior-posterior axis; the anterior region elevates by the "flip-up" model, and the middle and posterior regions reorient through the "flow" model. However, the mechanisms of both models are unclear because of the rapid progression of the elevation in utero. To observe palatal elevation in real time in detail, we aimed to establish a live imaging method using explants of the anterior region of the palatal shelf in mouse embryos before the beginning of elevation. Changes in the degree of shelf orientation were measured, which showed that the palatal shelf continuously changed shape toward the lingual side. The changes in the angle between the lingual and buccal bases of the palatal shelf were different; the morphological change at the lingual side resulted in a more acute angle, and the change at the buccal side resulted in a more obtuse angle. The morphological changes of the lingual and buccal sides occurred nearly simultaneously, suggesting that the anterior region of the palatal shelf in vitro elevated according to the "flip-up" model. This live imaging method enables the continuous observation of palatal shelf elevation and provides new insights into palatogenesis.

Integrin expression and extracellular matrix adhesion of septoclasts, pericytes, and endothelial cells at the chondro-osseous junction and the metaphysis of the proximal tibia in young mice.

We previously reported that septoclasts, which are uncalcified growth plate (GP) cartilage matrix-resorbing cells, are derived from pericytes surrounding capillary endothelial cells. Resorption of the GP is assumed to be regulated synchronously by septoclasts, pericytes, and endothelial cells. To reveal the contribution of the extracellular matrix (ECM) to the regulatory mechanisms of septoclastic cartilage resorption, we investigated the spatial correlation between the cells and the ECM in the GP matrix and basement membrane (BM) and investigated the expression of integrins-ECM receptors-in the cells. Septoclasts attached to the transverse septa containing collagen-II/-X at the tip of their processes and to the longitudinal septa containing collagen-II/-X at the spine-like processes extending from their bodies and processes. Collagen-IV and laminin α4 in the BM were sparsely detected between septoclasts and capillary endothelial cells at the chondro-osseous junction (COJ) and were absent in the outer surface of pericytes at the metaphysis. Integrin α1/α2, integrin α1, and integrin α2/α6 were detected in the cell membranes of septoclasts, pericytes, and endothelial cells, respectively. These results suggest that the adhesion between septoclasts and the cartilage ECM forming the scaffolds for cartilage resorption and migration is provided by integrin α2-collagen-II/-X interaction and that the adhesions between the BM and pericytes or endothelial cells are mediated by integrin α1-collagen-IV and integrin α2/α6-laminin interaction, respectively.

Spatiotemporal Gene Expression Regions along the Anterior-Posterior Axis in Mouse Embryos before and after Palatal Elevation.

The mammalian secondary palate is formed through complex developmental processes: growth, elevation, and fusion. Although it is known that the palatal elevation pattern changes along the anterior-posterior axis, it is unclear what molecules are expressed and whether their locations change before and after elevation. We examined the expression regions of molecules associated with palatal shelf elevation (Pax9, Osr2, and Tgfß3) and tissue deformation (F-actin, E-cadherin, and Ki67) using immunohistochemistry and RT-PCR in mouse embryos at E13.5 (before elevation) and E14.5 (after elevation). Pax9 was expressed at significantly higher levels in the lingual/nasal region in the anterior and middle parts, as well as in the buccal/oral region in the posterior part at E13.5. At E14.5, Pax9 was expressed at significantly higher levels in both the lingual/nasal and buccal/oral regions in the anterior and middle parts and the buccal/oral regions in the posterior part. Osr2 was expressed at significantly higher levels in the buccal/oral region in all parts at E13.5 and was more strongly expressed at E13.5 than at E14.5 in all regions. No spatiotemporal changes were found in the other molecules. These results suggested that Pax9 and Osr2 are critical molecules leading to differences in the elevation pattern in palatogenesis.

Development and Regeneration of Muscle, Tendon, and Myotendinous Junctions in Striated Skeletal Muscle.

Owing to a rapid increase in aging population in recent years, the deterioration of motor function in older adults has become an important social problem, and several studies have aimed to investigate the mechanisms underlying muscle function decline. Furthermore, structural maintenance of the muscle-tendon-bone complexes in the muscle attachment sites is important for motor function, particularly for joints; however, the development and regeneration of these complexes have not been studied thoroughly and require further elucidation. Recent studies have provided insights into the roles of mesenchymal progenitors in the development and regeneration of muscles and myotendinous junctions. In particular, studies on muscles and myotendinous junctions have-through the use of the recently developed scRNA-seq-reported the presence of syncytia, thereby suggesting that fibroblasts may be transformed into myoblasts in a BMP-dependent manner. In addition, the high mobility group box 1-a DNA-binding protein found in nuclei-is reportedly involved in muscle regeneration. Furthermore, studies have identified several factors required for the formation of locomotor apparatuses, e.g., tenomodulin (Tnmd) and mohawk (Mkx), which are essential for tendon maturation.

Spatial and chronological localization of septoclasts in the mouse Meckel's cartilage.

Meckel's cartilage (MC) in the first branchial arch of mammals is a transient structure that disappears before birth, except for the most anterior and posterior portions. Recent studies reported that some congenital abnormalities in craniofacial regions are linked with the persistence or dysplasia of MC. However, the mechanisms underlying the resorption of MC have not been elucidated. Cartilage resorption in endochondral ossification is performed by multinuclear osteoclasts/chondroclasts as well as mononuclear septoclasts, which were newly added to the list of cartilage phagocytes. Septoclasts located exclusively at the chondro-osseous junction of the growth plate resorb the uncalcified cartilage matrix. We hypothesized that septoclasts participate in the resorption of MC and attempted to clarify the localization and roles of septoclasts in MC of mouse using a specific immunohistochemistry marker, epidermal type-fatty acid-binding protein (E-FABP/FABP5). E-FABP-immunopositive septoclasts were detected for the first time at the beginning of MC resorption and localized along the resorption surface. Septoclasts of MC in embryonic mice possessed several processes that elongated toward the uncalcified cartilage matrix, expressed cathepsin B, and exhibited characteristic pericapillary localization. Additionally, they localized between hypertrophied cartilage and osteoclasts/chondroclasts in the resorption surface. Confocal laser-scanning microscopy revealed a decrease in the numbers of septoclasts and their processes with the progression of MC disappearance before birth. The present study showed that E-FABP-immunopositive septoclasts participated in the disappearance of MC through the resorption of the uncalcified cartilage matrix and that they have different roles from osteoclasts/chondroclasts.

Tendinous annulus of Zinn for a common origin of the extraocular rectus muscles: a histological study of the orbital apex from donated elderly cadavers.

The medial, inferior, lateral, and superior rectus muscles (MR, IR, LR, SR), levator palpebrae superioris (LPS), and superior oblique muscle (SO) seem to originate from the tendinous annulus of Zinn, ring-like fibrous tissue crossing the bony orbital fissure. We observed the histological annulus structure using semi-serial histological sections of the orbital apex from 30 elderly donated cadavers. Nearly frontal sections demonstrated a ring-like fibrous structure (a candidate annulus) connecting or embedding four rectus muscles. The candidate annulus did not contain the LPS and SO, and, in the anterior side, the latter muscles originated from the optic canal opening. Far posterior to the annulus, there was a common tendon of the MR, IR, and LR attached to the infero-medial wall of the bony orbital fissure. At the superior part, the annulus is tightly attached to the optic nerve sheath and the periosteum. Sagittal (or Horizontal) sections clearly exhibited parts of the annulus at the MR (SR) origin. Both sagittal and horizontal sections displayed (1) the common origin of the three rectus muscles near the oculomotor nerve in the bony fissure and (2) an accessory, independent muscle bundle of the MR originating from the superomedial margin of the optic canal near the origins of the LPS or SO. Consequently, the so-called tendinous annulus appeared not to provide origins of all six muscles.

Superior labial artery and vein anastomosis configuration to be considered in lip augmentation.

The treatment of cleft lip and palate is performed over a long period, starting immediately after birth. However, esthetic problems remain after lip augmentation. Endothelial cells of new capillaries are important for wound healing. Thus, the reconstruction of vascular networks is key to postoperative wound healing during lip augmentation. However, studies describing the superior labial artery (SLA) and superior labial vein (SLV) are rare, and their mutual positional relationship thus remains unclear. We procured 29 adult cadavers and ten fetuses. Macroscopic and histological examinations were performed on adult cadavers. We extracted soft tissues and blood vessels after micro-computed tomography (CT) and 3D tissue reconstruction. We performed histological investigations of vascular networks within the cleft lip in fetal samples. In adults, the SLV was distributed throughout the cutaneous side of the orbicularis oris muscle and the SLA, throughout the mucosal side. The SLV and SLA were separated by this muscle. Micro-CT images revealed that the SLA on the mucosal side transversed the orbicularis oris muscle to the SLV (55％). Histological analysis of fetuses revealed that the SLA was on the mucosal side, similar to that in adults, and traversed the orbicularis oris muscle in continuity with the SLV of the cutaneous side (100%). In lip augmentation, the reconstruction of the vascular structure, which involves the anastomosis of SLA and SLV passing through the orbicularis oris muscle, is an important factor when considering esthetic repair.

Septoclasts expressing epidermal fatty acid-binding protein (E-FABP, FABP5) in endochondral ossification.

BACKGROUND: Long-chain fatty acids (LCFAs) and retinoic acid (RA) are abundant in the growth plates (GPs) of long bones; however, their roles have not been elucidated. We observed that epidermal fatty acid-binding protein (E-FABP/FABP5) with a high affinity for both LCFAs and RA is exclusively expressed in the septoclasts located at the chondro-osseous junction (COJ) of the GP. HIGHLIGHTS: E-FABP expressed in septoclasts is involved in both LCFA metabolism and RA signaling as an intracellular transporter of both LCFAs and RA. Septoclasts with shortened cytoplasmic processes are associated with cartilage resorptive activity downregulation because of E-FABP deficiency or excess or deficiency of RA. In ontogeny, the septoclasts are differentiated from the pericytes and involved in the resorption of the uncalcified matrix of the cartilage templates in endochondral ossification. CONCLUSION: Septoclasts originate from pericytes and express E-FABP to play crucial roles in uncalcified matrix resorption by LCFA metabolism and RA signaling during endochondral ossification.

Localization of T-cell factor 4 positive fibroblasts and CD206-positive macrophages during skeletal muscle regeneration in mice.

Skeletal muscle regeneration is initiated by the activation of the transcription factor paired box 7 (Pax7), which is expressed in the satellite cells. The nuclear transcription factor T-cell factor 4 (Tcf4) is expressed in the fibroblasts and is involved in muscle tissue repair, while M2-like macrophages play an important role in skeletal muscle regeneration. However, the localization of M2-like macrophages and the expression of Tcf4 over a period of time during skeletal muscle regeneration remain unknown. Therefore, the murine masseter muscle was immunofluorescence investigated for the surface protein CD206 of M2-like macrophages and Tcf4 of fibroblasts during skeletal muscle regeneration to understand the changes in the CD206 and Tcf4 expression over time. We observed that CD206 entered the cytoplasm of some regenerating muscle fibers 5-7 days after the experimental muscle damage, that is, in the early stage of maturation of the regenerating muscle fibers with central nuclei. In addition, Tcf4 was expressed in the nuclei of the fibroblasts around the regenerating muscle fibers and in the central nuclei of the regenerating muscle fibers. Furthermore, the expression of laminin adjacent to Tcf4-positive cells was observed to partially disappear, and the shape of this missing part was observed to be identical to that of the nuclei of Tcf4-positive cells adjacent to the laminin. Clathrin was also expressed in these sites, demonstrating endocytosis. Thus, these results suggest that in the early stage of maturation of the regenerating muscle fibers, M2-like macrophages and Tcf4-positive fibroblasts enter the cytoplasm of the regenerating muscle fibers, thereby regulating the expression of various maturation factors.

Expression and enhancement of FABP4 in septoclasts of the growth plate in FABP5-deficient mouse tibiae.

In our previous study, fatty acid-binding protein 5 (FABP5) was expressed in septoclasts with long processes which are considered to resorb uncalcified matrix of the growth plate (GP) cartilage, and no apparent abnormalities were detected in the histo-architecture of the GP of FABP5-deficient (FABP5-/-) mice. Those finding lead us to hypothesize that another FABP can compensate the deletion of FABP5 in septoclasts of its gene-mutant mice. Based on the hypothesis, the present study examined the expression levels of several other FABPs in septoclasts and their morphology in FABP5-/- mouse tibiae. Processes of FABP5-/- septoclasts tend to be shorter than wild septoclasts. FABP4-positive septoclasts in FABP5-/- mice were more numerous than those cells in wild mice.Peroxisome proliferator-activated receptor (PPAR) γ was expressed in FABP4-positive septoclasts of FABP5-/- mice as well as mice administered with GW1929, a PPARγ agonist, suggesting that the occurrence of PPARγ induces an increase of FABP4-positive septoclasts. The present finding suggests that the functional exertion of FABP5 in septoclasts is supplemented by FABP4 in normal and FABP5-/- mice, and that the expression of FABP4 is up-regulated in accompany with PPARγ in FABP5-/- for maintenance of resorptive activity in the GP.

Developmental characteristics of secondary cartilage in the mandibular condyle and sphenoid bone in mice.

OBJECTIVE: Secondary cartilage develops from osteochondral progenitor cells. Hypertrophic chondrocytes in secondary cartilage increase within a very short time and then ossify rapidly. In the present study, we investigated the sequential development process of osteochondral progenitor cells, and the morphology and size of hypertrophic chondrocytes in secondary cartilage. DESIGN: ICR mice at embryonic days (E) 14.5-17.5 were used. The mandibular condyle and the medial pterygoid process of the sphenoid bone were observed as secondary cartilage, and the cranial base and the lateral pterygoid process of the sphenoid bone, which is primary cartilage, were observed as a control. Thin sections were subjected to immunostaining and alkaline phosphatase (ALP) staining. Using a confocal laser microscope, 3D stereoscopic reconstruction of hypertrophic cells was performed. To evaluate the size of hypertrophic chondrocytes objectively, the cell size was measured in each cartilage. RESULTS: Hypertrophic chondrocytes of secondary cartilage first expressed type X collagen (Col X) at E15.5. SRY-box 9 (Sox 9) and ALP were co-expressed in the fibroblastic/polymorphic tissue layer of secondary cartilage. This layer was very thick at E15.5, and then rapidly became thin. Hypertrophic cells in secondary cartilage were markedly smaller than those in primary cartilage. CONCLUSIONS: The small hypertrophic cells present in secondary cartilage may have been a characteristic acquired in order for the cartilage to smoothly promote a marked increase in hypertrophic cells and rapid calcification.

Origin and development of septoclasts in endochondral ossification of mice.

Septoclasts are mononuclear spindle-shaped phagocytes with their long processes in uncalcified cartilage matrices and locate adjacent to the capillary endothelium at the chondro-osseous junction of the growth plate. We have previously revealed a selective expression of epidermal-type fatty acid-binding protein (E-FABP/FABP5) in septoclasts. Although, pericytes are known to distribute along capillaries and directly surround their endothelial cells in a situation similar to septoclasts, no clear evidence is available on the relationship between septoclasts and pericytes. We investigated the chronological localization and morphological change of septoclasts during development of the tibia of mice to clarify the development of septoclasts and the immune-localization of pericyte markers in septoclasts to clarify the origin of septoclasts. E-FABP-immunoreactive septoclasts emerged at the perichondrium in the middle of the cartilaginous templates of the tibia in prenatal development. Septoclasts migrated to the surface of the cartilage adjacent to invading blood vessels. Processes of septoclasts became longer and their apexes attached to Von Kossa-negative uncalcified matrices during the formation process of the primary ossification center. Not only platelet-derived growth factor receptor beta, but also neuron-glial antigen 2 was localized in septoclasts of mice from E15 (embryonic day 15) to P6w (postnatal 6 week). Our results suggest that septoclasts are originated from pericytes and involved in the blood vessel invasion during formation of the primary ossification center.

Morphology and relationships of the biceps brachii and brachialis with the musculocutaneous nerve.

INTRODUCTION: Major anatomical textbooks generally state that the biceps brachii muscle (BB) is composed of long and short heads, whereas the brachialis muscle (BR) consists of a single head. However, the numbers of heads comprising the BB and the BR are very variable. The purpose of this study was to investigate how the branching patterns of the musculocutaneous nerve (MC) influence the number of heads of the BB and the BR. MATERIALS AND METHODS: Morphological examinations of the BB and MC were conducted using cadavers of 22 Japanese individuals, and morphological examinations of the BR and the MC were conducted in 9 of those 22 individuals. RESULTS: A three-headed BB was observed in 7 of the 22 specimens (31.8%). Most of these specimens showed a Type III branch pattern (after penetrating the long head or the short head, the MC innervated the supernumerary head or communicated with the main root again). The number of BR heads was categorized into three types: Type A, two heads (superficial and deep heads, 22.2%); Type B, three or four heads (two or three superficial heads and one deep head, 44.4%); and Type C, multiple heads (33.3%). Among these categories, branches of the MC in Type A specimens were most simple. CONCLUSION: A supernumerary head of the BB seemed to be present if the MC penetrates it. The BR basically consists of superficial and deep heads, and the number of superficial heads is affected by branches of the MC.

Retinoic acid regulates cell-shape and -death of E-FABP (FABP5)-immunoreactive septoclasts in the growth plate cartilage of mice.

Septoclasts, which are mononuclear and spindle-shaped cells with many processes, have been considered to resorb the transverse septa of the growth plate (GP) cartilage at the chondro-osseous junction (COJ). We previously reported the expression of epidermal-type fatty acid-binding protein (E-FABP, FABP5) and localization of peroxisome proliferator-activated receptor (PPAR)ß/δ, which mediates the cell survival or proliferation, in septoclasts. On the other hand, retinoic acid (RA) can bind to E-FABP and is stored abundantly in the GP cartilage. From these information, it is possible to hypothesize that RA in the GP is incorporated into septoclasts during the cartilage resorption and regulates the growth and/or death of septoclasts. To clarify the mechanism of the cartilage resorption induced by RA, we administered an overdose of RA or its precursor vitamin A (VA)-deficient diet to young mice. In mice of both RA excess and VA deficiency, septoclasts decreased in the number and cell size in association with shorter and lesser processes than those in normal mice, suggesting a substantial suppression of resorption by septoclasts in the GP cartilage. Lack of PPARß/δ-expression, TUNEL reaction, RA receptor (RAR)ß, and cellular retinoic acid-binding protein (CRABP)-II were induced in E-FABP-positive septoclasts under RA excess, suggesting the growth arrest/cell-death of septoclasts, whereas cartilage-derived retinoic acid-sensitive protein (CD-RAP) inducing the cell growth arrest or morphological changes was induced in septoclasts under VA deficiency. These results support and do not conflict with our hypothesis, suggesting that endogenous RA in the GP is possibly incorporated in septoclasts and utilized to regulate the activity of septoclasts resorbing the GP cartilage.

Fetal Tendinous Connection Between the Tensor Tympani and Tensor Veli Palatini Muscles: A Single Digastric Muscle Acting for Morphogenesis of the Cranial Base.

Some researchers contend that in adults the tensor tympani muscle (TT) connects with the tensor veli palatini muscle (TVP) by an intermediate tendon, in disagreement with the other researchers. To resolve this controversy, we examined serial sections of 50 human embryos and fetuses at 6-17 weeks of development. At 6 weeks, in the first pharyngeal arch, a mesenchymal connection was found first to divide a single anlage into the TT and TVP. At and after 7 weeks, the TT was connected continuously with the TVP by a definite tendinous tissue mediolaterally crossing the pharyngotympanic tube. At 11 weeks another fascia was visible covering the cranial and lateral sides of the tube. This "gonial fascia" had two thickened borders: the superior one corresponded to a part of the connecting tendon between the TT and TVP; the inferior one was a fibrous band ending at the os goniale near the lateral end of the TVP. In association with the gonial fascia, the fetal TT and TVP seemed to provide a functional complex. The TT-TVP complex might first help elevate the palatal shelves in association with the developing tongue. Next, the tubal passage, maintained by contraction of the muscle complex, seems to facilitate the removal of loose mesenchymal tissues from the tympanic cavity. Third, the muscle complex most likely determined the final morphology of the pterygoid process. Consequently, despite the controversial morphologies in adults, the TT and TVP seemed to make a single digastric muscle acting for the morphogenesis of the cranial base.

Morphological Changes of Myoepithelial Cells in the Rat Submandibular Gland Following the Application of Surgical Stimuli.

Myoepithelial cells (MECs) exist on the basal surface of acini in major exocrine glands, include myofilaments and various constructive proteins, and share characteristics with smooth muscle and epithelial cells. MECs project several ramified processes to invest acini, and possibly contract to compress acini to support the secretion by the glandular cells. However, the functional roles of MECs in salivary secretion are still unclear. We investigated morphological changes in immunostained MECs using the anti-α-smooth muscle actin (αSMA) antibody in operated or non-operated contralateral (NC) submandibular glands after partial or total resection. Furthermore, we investigated and discuss other salivary glands of rats. MECs in the parotid, sublingual and submandibular gland of adult rats exhibited different shapes and localizations. After surgery, in both operated and NC glands, the number of MECs and αSMA-immunopositive areas increased significantly. Three-dimensional analysis using a confocal laser-scanning microscope revealed that substantial and significant enhancement became evident in the number, length, and thickness of MEC-processes covering acini of the operated and NC submandibular glands. The preset findings indicate that MECs alter the morphology of their processes in operated and NC glands after surgery of the partial or total resection. It is suggested that MECs promote salivary secretion using elongated, thickened, and more ramified processes.

Development and regression of the thyroglossal duct in mice.

The thyroid anlage develops in the foramen caecum area of the tongue, and migrates through the anterior neck towards its final position in front of the laryngeal cartilages. During migration, the thyroglossal duct, a temporary structure connecting the thyroid anlage and the foramen caecum, is recognized. In the present study, chronological changes and apoptosis in the thyroglossal duct of mice were investigated histochemically using an antibody against Nkx2-1, initially identified as a thyroid transcription factor 1 (TTF1), and the TUNEL reaction in consecutive serial sagittal sections. At embryonic day 10.00 (E10.00), the thyroid anlage was Nkx2-1-immunoreactive and located just below the foramen caecum. As the thyroid anlage descended, the thyroglossal duct was formed at E10.25, being less than 10µm in diameter. By E10.75, the Nkx2-1-positive thyroglossal duct had progressively elongated up to 100µm. At E11.00 the thyroglossal duct began to disappear, beginning in its mid-portion, and finally became invisible at E11.50. At E11.00-12.00, apoptotic cells were found in an area where the thyroglossal duct was partially discontinuous. After E12.00, cartilaginous tissue of the hyoid bone anlage developed in the mid-portion of the area where the thyroglossal duct had regressed. Immunoreactivity for thyroglobulin, a marker of differentiated thyroid endocrine cells, was detected at E13.00. These results strongly suggest that the mouse thyroglossal duct disappears as a result of apoptosis before differentiation of the endocrine thyroid.

Expression of epidermal fatty acid binding protein (E-FABP) in septoclasts in the growth plate cartilage of mice.

n-3 Polyunsaturated fatty acids play a role in regulating the growth of the long bones. Fatty acid-binding proteins (FABPs) bind and transport hydrophobic long-chain fatty acids intracellularly, and epidermal-type FABP (E-FABP) has an affinity for n-3 fatty acids. This study aimed to clarify the localization of E-FABP in the growth plate of the mouse tibia. At the chondro-osseous junction (COJ) of the growth plate, E-FABP-immunoreactivity was exclusively localized in mononuclear, spindle-shaped cells with several long processes. These E-FABP-immunoreactive cells were identified as being septoclasts, i.e., cells that resorb uncalcified transverse septa. The processes of these immunoreactive septoclasts terminated between the longitudinal and transverse septa. E-FABP-immunoreactivity was found in the entire cytoplasm and on the mitochondrial outer membrane. In ontogeny, immunoreactive septoclasts were observed immediately after emergence of the primary ossifying center and were distributed not only at the COJ but also in the metaphysis near the COJ. The number of septoclasts increased at the postnatal age of 1 week (P1w)-P2w, and thereafter gradually decreased; and the cells became concentrated at the COJ after P3w-P4w. The immunoreactivity for peroxisome proliferator-activated receptor (PPAR)ß/δ was detected in these E-FABP-immunoreactive septoclasts. The present results suggest that fatty acids, preferably n-3 ones, are intracellularly transported by E-FABP to various targets, including mitochondria and nucleus, in which PPARß/δ may play functional roles in the transcriptional regulation of genes involved in the endochondral ossification.

Localization of hsp27 in the rat submandibular gland following the application of various surgical treatments.

Salivary glands repair and regenerate following various types of injuries and surgical procedures. However, the tissue responses induced in the contralateral glands have yet to be elucidated in detail. Hsp27, a member of the heat-shock protein (Hsp) family, is strongly expressed in physiological environments, particularly during development. Hsp27 was previously shown to play a role in the regulation of acinar cell proliferation and differentiation in the rat submandibular gland. The present study performed the following surgical treatments on the right submandibular glands of adult rats: 1) duct ligation followed by unligation after one week; 2) partial sialoadenectomy; and 3) total sialoadenectomy. Immunohistochemistry for Hsp27 and Ki67 was performed in the experimental and normal contralateral glands, and localization was histologically and morphometrically analyzed. The results obtained revealed the localization of Hsp27 to the intercalated duct in the submandibular glands of non-treated rats. The expression of Hsp27 was strongly induced in both the uninjured contralateral control glands as well as treated glands of experimental rats regardless of the surgical procedure performed. The number of Hsp27-immunopositive cells increased rapidly following surgery, and subsequently returned to the same level as that in non-treated rats after 4 weeks. However, no marked changes were observed in the number of Ki67-immunopositive proliferating cells. Therefore, the change in the number of Hsp27-immunopositive cells may have contributed to compensatory hypertrophy. The results of the present study indicate that the expression of Hsp27 in the intercalated duct in the submandibular gland may play a role in the differentiation of acinar cells.