Growth patterns of facial muscles at the angle of the mouth: A histological study using midterm and near-term human fetuses.

At the angle of the mouth, spoke-like muscle bundles converge at the "modiolus," which is believed to appear in utero. The aim of this study was to investigate the growth of the modiolus histologically. We studied frontal histological sections of the face from 12 midterm and six near-term fetuses. At midterm, a convergence of the levator anguli oris (LAOM) and depressor anguli oris (DAOM) was frequently present, and another convergence of the LAOM with the platysma (PM) or orbicularis oris (OOM) was also often evident. At near-term, muscle fiber merging or interdigitation was classified into nine combinations, five of which were frequently seen: LAOM-PM, LAOM-DAOM, zygomaticus major (ZMM)-orbicularis oris (OOM), buccinator (BM)-LAOM, and BM-PM. These combinations existed at slightly different depths and/or sites, thus allowing the angle of the mouth to receive multiple muscles. Notably, tissues interposed between the muscle fibers were limited to a thin epimysium at each crossing or interdigitation. Therefore, the LAOM, DAOM, OOM, BM, and PM appear to form a basic configuration at birth, but the development and growth were much delayed than the classical description. The modiolus is not a specific fibromuscular structure but simply represents a cluster of muscle convergence sites. Even at meeting between an elevator and depressor, a specific fibrous structure seems unlikely to connect the epimysium for the muscle convergence. Instead, the central nervous system appears to regulate the activity of related muscles to minimize tension or friction stress at the meeting site.

Mentalis nerve branches supplying the lower lip revisited: a study of human fetuses and donated elderly cadavers.

PURPOSE: Little information is known about the mentalis nerve course from the lower lip approximation margin (free margin) to the upper lip. Likewise, no difference in nerve distribution has been observed between the cutaneous and mucosal parts of the lip. Therefore, this study reexamined mentalis nerve morphology. METHODS: For macroscopic observations, three fresh cadavers were dissected (one male and two females; aged 78-93). We also evaluated histological sections obtained from five donated elderly cadavers (two males and three females, aged 82-96 years) and 15 human fetuses (11-40 weeks or crown-rump length 80-372 mm). Immunohistochemical analysis for S100 protein and tyrosine hydroxylase was performed. RESULTS: In both fetuses and adult cadavers, one to three nerve branches ran upward in the submucosal tissue from the mental foramen. Near the free margin of the lip, some branches passed through the orbicularis oris muscle layer toward the lip skin, whereas others followed a reversed J-shaped course along the free margin. Nerve twigs ran in parallel beneath the mucosa, whereas wavy nerve twigs attached to the basal lamina of the lip epidermis. The difference in nerve endings abruptly occurred at the skin-mucosal junction. Tyrosine hydroxylase-positive sympathetic nerve twigs surrounded arteries and formed a branch composed of S100-negative unmyelinated fibers. CONCLUSION: The lower lip skin was innervated by a perforating branch passing through the orbicularis oris muscle, that was different from the lip mucosa. A sudden change in the nerve ending configuration at the mucocutaneous junction seemed to develop postnatally.

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.

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.

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.

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.

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.

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.

Ecological, physiological, and biomechanical differences between gametophytes and sporophytes of Chondrus ocellatus (Gigartinales, Rhodophyta)1.

Although variation among habitats in the ratio of gametophytes to sporophytes has been reported in various gigartinacean species, factors controlling the phase ratio remain poorly understood. Over 18 months, we examined the phase ratio of Chondrus ocellatus at three sites: a sheltered intertidal site, Hiruga A; an exposed intertidal site, Hiruga B; and a subtidal site, Shikimi. The mean proportion of gametophytes at Hiruga A (73.1%) was significantly higher than that at Shikimi (51.2%) and Hiruga B (44.7%). Due to a significantly higher water retention ability of the gametophytes, it was expected that the gametophytes would exhibit higher desiccation tolerance. After dehydration treatments, however, neither the photosynthetic rate of vegetative blades nor the survival rate of spores was significantly different between the phases. Measurements of blade strength indicated that the sporophytic blades were less stiff and more flexible, and a culture experiment revealed that the sporophytic germlings showed a significantly higher growth rate. Flexible blades and fast-growing germlings are considered advantageous for colonizing wave-swept intertidal habitats, so these properties may have caused the different fluctuation pattern of phase ratio among the sites. The present data demonstrate that biomechanical and physiological differences between the two phases of C. ocellatus make one phase advantageous in certain environmental conditions, and that these differences likely cause an unequal ratio of isomorphic phases.

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.

Morphology of the Upper Esophageal Sphincter or Cricopharyngeus Muscle Revisited: A Study Using Adult and Fetal Specimens.

Histological examination of specimens from 22 donated elderly cadavers and 15 human fetuses revealed that the cricopharyngeus muscle (CPM) provided (1) posterior circular muscle fibers adjacent to the external aspect of the uppermost esophageal circular muscle and (2) a thin anterior sling connecting to that same muscle. Another thick lateral bundle of longitudinal muscle originated independently from a fascia covering the posterior cricoarytenoideus muscle, extended laterally and posteriorly, and occupied a space after the CPM had disappeared at the anterolateral angle of the esophagus below the cricoid. The thick fascia contained abundant elastic fibers along the internal surface of the pharyngeal constrictors (posteromedial elastic lamina), but was interrupted or discontinued near the cricoid origin of the CPM. As no submucosal smooth muscles or elastic fibers were connected to it, the CPM did not accompany a specific elastic structure at the interface between the pharyngeal and esophageal muscles. In fetuses, the medial half of the CPM was inserted into the cricoid while the lateral half continued to the sternothyroideus muscle or ended at a fascia covering the cricothyroideus. These anterolateral ends provided a mechanical load for longitudinal growth of the pharyngeal constrictors. Consequently, the CPM was unlikely to develop and grow to form the upper esophageal sphincter, and the muscle bundle crossing the lateral aspect of the pharyngo-esophageal junction appeared to have a secondary passive role as a sphincter. This situation contrasts with that of another sphincter in the human body formed from striated muscle. Clin. Anat., 33:782-794, 2020. © 2019 Wiley Periodicals, Inc.

SCO-267, a GPR40 Full Agonist, Improves Glycemic and Body Weight Control in Rat Models of Diabetes and Obesity.

The GPR40/FFA1 receptor is a G-protein-coupled receptor expressed in the pancreatic islets and enteroendocrine cells. Here, we report the pharmacological profiles of (3S)-3-cyclopropyl-3-{2-[(1-{2-[(2,2-dimethylpropyl)(6-methylpyridin-2-yl)carbamoyl]-5-methoxyphenyl}piperidin-4-yl)methoxy]pyridin-4-yl}propanoic acid (SCO-267), a novel full agonist of GPR40. Ca2+ signaling and insulin and glucagon-like peptide-1 (GLP-1) secretion were evaluated in GPR40-expressing CHO, MIN6, and GLUTag cells. Hormone secretions and effects on fasting glucose were tested in rats. Single or repeated dosing effects were evaluated in neonatally streptozotocin-induced diabetic rats (N-STZ-1.5 rats), diet-induced obese (DIO) rats, and GPR40-knockout (Ffar1-/- ) mice. Treatment with SCO-267 activated Gq signaling in both high- and low-FFAR1-expressing CHO cells, stimulated insulin secretion in MIN6 cells, and induced GLP-1 release in GLUTag cells. When administered to normal rats, SCO-267 increased insulin, glucagon, GLP-1, glucose-dependent insulinotropic peptide, and peptide YY (PYY) secretions under nonfasting conditions. These results show the full agonistic property of SCO-267 against GPR40. Hypoglycemia was not induced in SCO-267-treated rats during the fasting condition. In diabetic N-STZ-1.5 rats, SCO-267 was highly effective in improving glucose tolerance in single and 2-week dosing studies. DIO rats treated with SCO-267 for 2 weeks showed elevated plasma GLP-1 and PYY levels, reduced food intake, and decreased body weight. In wild-type mice, SCO-267 induced GLP-1 secretion, food intake inhibition, and body weight reduction; however, these effects were abolished in Ffar1-/- mice, indicating a GPR40-dependent mechanism. In conclusion, SCO-267 stimulated islet and gut hormone secretion, improved glycemic control in diabetic rats, and decreased body weight in obese rats. These data suggest the therapeutic potential of SCO-267 for the treatment of diabetes and obesity.

Associations among species traits, distribution, and demographic performance after typhoon disturbance for 22 co-occurring woody species in a mesic forest on a subtropical oceanic island.

To support conservation and restoration, it is important to understand how differences in species functional traits relate to the distribution and demographic performance (i.e., changes in rates of growth, survival and recruitment) of co-occurring endemic, indigenous, and alien species on oceanic islands, where species are especially vulnerable to invasion. We examined interspecific differences in leaf and wood traits, and their associations with species origin (endemic, indigenous, and alien), distribution patterns, and demographic performance after typhoon disturbance for 22 co-occurring woody species at Sekimon on Hahajima Island in the Ogasawara Islands. Principal components analysis showed that the first and second principal components were associated with trait variations along spectra of leaf and wood economics (LES and WES). Species origin was not significantly associated with these components. Conservative species with low resource acquisition along the LES were abundant before the typhoon but acquisitive species had higher recruitment after the typhoon. After the typhoon, acquisitive species along the WES showed higher recruitment, relative growth rates but low survival rates on soil substrates. Endemic and indigenous species had lower recruitment and relative growth rates and endemic species had lower survival rates than alien species. Alien, endemic and indigenous species have similar functional space along the LES and WES, but these functional differences do not simply explain high demographic performance of alien species after the typhoon under the conditions of the species composition shifting from endemic species to alien species after repeated typhoon disturbances.

Biphenyl-induced cytotoxicity is mediated by an increase in intracellular Zn2.

Biphenyl is found both in natural and anthropogenic sources and is used as a fungistat in the packaging of citrus fruits. Acute exposure to high levels of biphenyl has been observed to cause skin irritation and toxic effects on the liver and kidneys. However, the mechanisms of cytotoxicity induced by biphenyl are not yet well understood. In the present study, the cytotoxicity of biphenyl was studied by flow cytometry with fluorescent probes. Biphenyl at 100 µM significantly increased cell lethality after 3 h in rat thymocytes. In addition, biphenyl at 100 µM or more elevated intracellular Zn2+ levels. N,N,N',N'-Tetrakis(2-pyridylmethyl)ethylenediamine (TPEN), an intracellular and extracellular Zn2+ chelator, but not diethylenetriamine-N,N,N',N″,N″-pentaacetic acid (DTPA), a membrane-impermeable Zn2+ chelator, attenuated the biphenyl-induced increase in intracellular Zn2+ levels and cell death. These results suggested that biphenyl-induced cytotoxicity caused an increase in intracellular Zn2+ levels, which was dependent on internal Zn2+. Moreover, biphenyl led to an increase in sensitivity to oxidative stress, while TPEN inhibited this biphenyl-induced increase. Our findings revealed that biphenyl caused an increase in the intracellular free Zn2+ concentration, inducing cytotoxicity, cell death, and an increase in sensitivity to oxidative stress.

Suboccipital myodural bridges revisited: Application to cervicogenic headaches.

There seems to be no complete demonstration of the suboccipital fascial configuration. In 30 human fetuses near term, we found two types of candidate myodural bridge: (1) a thick connective tissue band running between the rectus capitis posterior major and minor muscles (rectus capitis posterior major [Rma], rectus capitis posterior minori [Rmi]; Type 1 bridge; 27 fetuses); and (2) a thin fascia extending from the upper margin of the Rmi (Type 2 bridge; 20 fetuses). Neither of these bridge candidates contained elastic fibers. The Type 1 bridge originated from: (1) fatty tissue located beneath the semispinalis capitis (four fetuses); (2) a fascia covering the multifidus (nine); (3) a fascia bordering between the Rma and Rmi or lining the Rma (13); (4) a fascia covering the inferior aspect of the Rmi (three); and (5) a common fascia covering the Rma and obliquus capitis inferior muscle (nine). Multiple origins usually coexisted in the 27 fetuses. In the minor Type 2 bridge, composite fibers were aligned in the same direction as striated muscle fibers. Thus, force transmission via the thin fascia seemed to be effective along a straight line. However, in the major Type 1 bridges, striated muscle fibers almost always did not insert into or originate from the covering fascia. Moreover, at and near the dural attachment, most composite fibers of Type 1 bridges were interrupted by subdural veins and dispersed around the veins. In newborns, force transmission via myodural bridges was likely to be limited or ineffective. The postnatal growth might determine a likely connection between the bridge and headache. Clin. Anat. 32:914-928, 2019. © 2019 Wiley Periodicals, Inc.

Site-dependent differences in the composite fibers of male pelvic plexus branches: an immunohistochemical analysis of donated elderly cadavers.

BACKGROUND: Although the pelvic autonomic plexus branches are considered to be a mixture of sympathetic and parasympathetic nerves, little is known regarding the composite fibers of the pelvic plexus branches. This study aimed to investigate the immunohistochemical features of sympathetic and parasympathetic nerves in the pelvic autonomic plexus branches. METHODS: Using 10 donated elderly male cadavers, the detailed topohistology of nerve fibers at and around the bladder, seminal vesicle, prostate, and rectum was examined. Neuronal nitric oxide synthase (nNOS) and vasoactive intestinal polypeptide (VIP) were used as parasympathetic nerve markers; tyrosine hydroxylase (TH) was used as a sympathetic nerve marker. The myenteric plexus of the colon was utilized as a positive control. RESULTS: Most nerve fibers in the bladder, seminal vesicle, prostate, and rectum were both nNOS- and TH-positive. Thus, pelvic plexus branches were classified into two types: 1) triple-positive mixed nerves (nNOS+, VIP+, TH+, thick myelinated fibers + or -) and 2) double-positive mixed nerves (nNOS+, VIP-, TH+, thick myelinated fibers + or -). Notably, triple-positive nerves were localized within the posterosuperior part of the plexus (near the rectum) and travelled anteroinferiorly toward the posterolateral corner of the prostate. The posteriorly and inferiorly located nerves were predominantly composed of parasympathetic, rather than sympathetic, fibers. In contrast, nerve fibers within and along the bladder and seminal vesicle contained either no or few VIP-positive nerves. These superiorly located nerves were characterized by clear sympathetic nerve dominance. CONCLUSIONS: The nerves of the pelvic plexus branches were clearly classified into nerves around the bladder and seminal vesicle (VIP-negative) and nerves around the prostate (VIP-positive). Although nNOS- and VIP-positive nerve fibers are candidate cavernous nerves, cavernous nerve identity cannot be definitively concluded for these nerves in the periprostatic region.

A RET-ER81-NRG1 Signaling Pathway Drives the Development of Pacinian Corpuscles.

Axon-Schwann cell interactions are crucial for the development, function, and repair of the peripheral nervous system, but mechanisms underlying communication between axons and nonmyelinating Schwann cells are unclear. Here, we show that ER81 is functionally required in a subset of mouse RET+ mechanosensory neurons for formation of Pacinian corpuscles, which are composed of a single myelinated axon and multiple layers of nonmyelinating Schwann cells, and Ret is required for the maintenance of Er81 expression. Interestingly, Er81 mutants have normal myelination but exhibit deficient interactions between axons and corpuscle-forming nonmyelinating Schwann cells. Finally, ablating Neuregulin-1 (Nrg1) in mechanosensory neurons results in no Pacinian corpuscles, and an Nrg1 isoform not required for communication with myelinating Schwann cells is specifically decreased in Er81-null somatosensory neurons. Collectively, our results suggest that a RET-ER81-NRG1 signaling pathway promotes axon communication with nonmyelinating Schwann cells, and that neurons use distinct mechanisms to interact with different types of Schwann cells. SIGNIFICANCE STATEMENT: Communication between neurons and Schwann cells is critical for development, normal function, and regeneration of the peripheral nervous system. Despite many studies about axonal communication with myelinating Schwann cells, mostly via a specific isoform of Neuregulin1, the molecular nature of axonal communication with nonmyelinating Schwann cells is poorly understood. Here, we described a RET-ER81-Neuregulin1 signaling pathway in neurons innervating Pacinian corpuscle somatosensory end organs, which is essential for communication between the innervating axon and the end organ nonmyelinating Schwann cells. We also showed that this signaling pathway uses isoforms of Neuregulin1 that are not involved in myelination, providing evidence that neurons use different isoforms of Neuregulin1 to interact with different types of Schwann cells.

Enteric neurons of the esophagus: an immunohistochemical study using donated elderly cadavers.

PURPOSE: To describe and discuss the normal anatomy and function of enteric neurons in the esophagus of aged individuals. METHOD: We examined ganglion cells in esophagus specimens obtained from 15 elderly cadavers without any macroscopic pathology in the mediastinum and abdomen. Neuronal nitric oxide synthase and vasoactive intestinal polypeptide were used as parasympathetic nerve markers, and tyrosine hydroxylase as a sympathetic nerve marker. RESULTS: The thoracic and abdominal esophagus contained a well-developed myenteric nerve plexus (S100 protein-positive area) in the intermuscular layer: 0.02-0.03 mm2 per 1-mm length of the circular esophageal wall. The cervical esophagus usually contained no ganglion cells. The number of parasympathetic ganglion cells was maximal in the upper or middle thoracic esophagus (mean 18-23 cells per section), whereas sympathetic cells were considerably less numerous at any sites (mean 1-3 cells). CONCLUSION: In comparison with previous data from elderly cadavers, the esophagus carried much fewer ganglion cells than the intestine and colon; sympathetic cells were particular less numerous. Esophageal smooth muscle exhibits a unique mode of peristalsis characterized by a rebound contraction with a long latency after stimulation. This type of peristalsis appears to be regulated by inhibitory, nNOS-positive nerves with a sparse distribution, which seems to account for the long-span peristalsis unique to the esophagus. The extreme sparsity of ganglion cells in the cervical esophagus suggests that enteric neuron-integrated peristalsis, like that in the intestine and colon, is unlikely. Surgical treatment of the esophagus is likely to change or impair these unique features.

Distance between intramuscular nerve and artery in the extraocular muscles: a preliminary immunohistochemical study using elderly human cadavers.

PURPOSE: Extraocular muscles are quite different from skeletal muscles in muscle fiber type and nerve supply; the small motor unit may be the most well known. As the first step to understanding the nerve-artery relationship, in this study we measured the distance from the arteriole (25-50 µm in thickness) to the nerve terminal twigs in extraocular muscles. MATERIALS AND METHODS: With the aid of immunohistochemistry for nerves and arteries, we examined the arteriole-nerve distance at 10-15 sites in each of 68 extraocular muscles obtained from ten elderly cadavers. The oblique sections were nearly tangential to the muscle plate and included both global and orbital aspects of the muscle. RESULTS: In all muscles, the nerve twigs usually took a course parallel to muscle fibers, in contrast to most arterioles that crossed muscles. Possibly due to polyinnervation, an intramuscular nerve plexus was evident in four rectus and two oblique muscles. The arteriole-nerve distance usually ranged from 300 to 400 µm. However, individual differences were more than two times greater in each of seven muscles. Moreover, in each muscle the difference between sites sometimes reached 1 mm or more. The distance was generally shorter in the rectus and oblique muscles than in the levator palpebrae muscle, which reached statistical significance (p < 0.05). CONCLUSIONS: The differences in arteriole-nerve distances between sites within each muscle, between muscles, and between individuals might lead to an individual biological rhythm of fatigue in oculomotor performance.