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Misexpression of acetylcholinesterases in the C. elegans pha-2 mutant accompanies ultrastructural defects in pharyngeal muscle cells.

pha-2 is the Caenorhabditis elegans homolog of the vertebrate homeobox gene Hex. Embryonic expression of pha-2 is mostly pharyngeal and the only described mutant allele of pha-2 results in a severe pharyngeal defect in which certain muscle cells (pm5 cells) and neurons are grossly deformed. Here, we performed a detailed characterization of the pha-2 phenotype using cell-type-specific reporters, physical manipulation of the nuclei in pharyngeal muscle cells using "optical tweezers", electron microscopy, staining of the actin cytoskeleton as well as phenotypic rescue and ectopic expression experiments. The main findings of the present study are (i) the pha-2 (ad472) mutation specifically impairs the pharyngeal expression of pha-2; (ii) in the pha-2 mutant, the cytoskeleton of the pm5 cells is measurably weaker than in normal cells and is severely disrupted by large tubular structures and organelles; (iii) the pm5 cells of the pha-2 mutant fail to express the acetylcholinesterase genes ace-1 and ace-2; (iv) ectopic expression of pha-2 can induce ectopic expression of ace-1 and ace-2; and (v) the anc-1 mutant with mislocalized pm5 cell nuclei occasionally shows an isthmus phenotype similar to that of pha-2 worms.

Acetylcholinesterase↗

Direct parabrachial nuclear projections to the pharyngeal motoneurons in the rat: an anterograde and retrograde double-labeling study.

The parabrachial nucleus consists of several subnuclei which contains autonomic, gustatory, visceral sensory, nociceptive, and respiratory related neurons. We have investigated the direct projections from the rat parabrachial region, including the K olliker-Fuse nucleus, to the pharyngeal motoneurons with an anterograde and retrograde double-tracing technique. The cholera toxin subunit-B was injected into the lower pharynx or the esophagus after injection of biotinylated dextran amine into the ventrolateral parabrachial nuclear region, including the external medial, the external lateral, and the crescent area of the central lateral parabrachial nuclei and into the Kölliker-Fuse nucleus. The anterogradely dextran amine-labeled fibers from these nuclei projected to the semicompact, loose and external formations besides the compact formation of the nucleus ambiguus. Many anterogradely labeled fibers and terminals were found to contact retrogradely cholera toxin-labeled pharyngeal neuronal soma and dendrites in the semicompact formation of the nucleus ambiguus. The medial half of the parabrachial nucleus, including the medial and the medial part of the central lateral parabrachial nuclei, sent a few fibers to the reticular formation just dorsal to the esophageal motoneurons but no fibers to either the pharyngeal or to the esophageal motoneurons. These results suggested that the visceral sensory, gustatory, nociceptive or respiratory related neurons in the parabrachial nucleus project directly to the pharyngeal motoneurons, but there are no parabrachial projections to the esophageal motoneurons in the nucleus ambiguus.

Animals↗

Galphaq and Galpha11 proteins mediate endothelin-1 signaling in neural crest-derived pharyngeal arch mesenchyme.

Endothelin-A (ET(A)) is a G-protein-coupled receptor expressed in the neural crest-derived mesenchyme of the pharyngeal arches during craniofacial development. Targeted deletion of the ET(A) receptor or its ligand endothelin-1 (ET-1) causes cleft palate and hypoplasia of the mandible, otic cup, and tympanic ring. Previously we showed that Galpha(q)/Galpha(11)-null mice die around E11.0, whereas Galpha(q)((-/-))Galpha(11)((+/-)) mice survive to birth with hypomorphic phenotypes similar to, but less severe than, ET(A) or ET-1-null mice. To determine whether ET-1 signaling is transduced by Galpha(q)/Galpha(11) proteins, we examined the expression patterns of several ET-1 dependent and independent transcription factors in Galpha(q)/Galpha(11)-deficient embryos. Expression of genes encoding the ET-1-dependent transcription factors Dlx3, Dlx6, dHAND, and eHAND was specifically downregulated in the pharyngeal arches of Galpha(q)/Galpha(11)-deficient mice. In contrast, pharyngeal arch expression of the homeobox gene Msx1, which is not regulated by ET-1 signaling, was maintained in these embryos. We conclude that the Galpha(q) and Galpha(11) proteins serve as the intracellular mediators of ET-1 signaling in the pharyngeal arch mesenchyme.

Animals↗

Damage of the pharyngeal mucosa and hyperresponsiveness of airway in sinusitis.

BACKGROUND: In sinusitis bronchoconstriction is supposed to originate from pharyngobronchial reflexes triggered by seeding of the inflammatory process into the pharynx. OBJECTIVE: Our aim was to evaluate whether in sinusitis bronchial and extrathoracic airway (EA) dysfunction correlate with morphologic abnormalities of the pharyngeal mucosa. METHODS: We performed histamine inhalation challenge, nasal lavage, and nasopharyngeal biopsies in 24 nonasthmatic patients with exacerbation of chronic sinusitis. The histamine PC20 was the threshold of bronchial responsiveness, and that causing 25% fall in maximal midinspiratory flow was the threshold of EA responsiveness (PC25MIF50). Thresholds of 8 mg/ml or less were assumed to indicate bronchial hyperresponsiveness (BHR) or EA hyperresponsiveness (EAHR). PC20 and PC25MIF50 values were related to clinical data, nasal lavage fluid eosinophils, pharyngeal epithelium and basement membrane thickness, and density of submucosal vessels and nervous fibers. RESULTS: The PC20 was closely related to PC25MIF50 (p = 0.0004). Ten patients had EAHR, 9 had combined EAHR and BHR, and 5 had neither EAHR nor BHR. EAHR was strongly associated with epithelial thinning, and BHR with long-standing sinusitis, a lower PC25MIF50, increased submucosal nerve density and increased nasal lavage fluid eosinophils. CONCLUSIONS: Our findings suggest that in nonasthmatic patients with sinusitis, pharyngeal damage may contribute to airway dysfunction by favoring the access of irritants to submucosal nerve endings, with activation of constrictive reflexes to the EA. Proliferation of sensory neurons, consequent to long-lasting pharyngeal inflammation, may cause more severe EA narrowing and activate pharyngobronchial reflexes.

Adult↗

Pharyngeal sensation and gag reflex in healthy subjects.

The gag reflex is often used in the assessment of swallowing, yet its absence does not predict aspiration in acute stroke. Disordered pharyngeal sensation has been found to be a sensitive predictor. The occurrence of gag reflex and pharyngeal sensation in healthy people is unknown. We studied these tests in 140 healthy subjects (half elderly and half young). Gag reflex was absent in 37% of subjects whereas pharyngeal sensation was absent in only 1. The results largely explain the low predictive value of gag reflex in the assessment of aspiration in acute stroke. Testing pharyngeal sensation would be more likely to be useful in these circumstances.

Adult↗

Role of pharyngeal length in patients with a lack of overbite.

Anterior open bites may be localized dental manifestations, which are caused by habits or skeletal disharmony, with or without functional aberration. Previous studies suggest various associations between open bites and underlying etiologic factors. We hypothesize that respiratory efficiency may be associated with anterior open bites. Under the assumption that breathing efficiency of the oropharynx and hypopharynx may be related to pharyngeal airway length, a cephalometric variable, vertical airway length (VAL), was measured on lateral cephalograms obtained from a total of 58 subjects with, so called, "open bite tendencies" (hereafter, open bite tendency). By means of the variable VAL, the association between pharyngeal length and open bite was investigated. In addition, the difference between actual open bite and open bite tendencies was also examined. The samples were randomly collected under stringent selection criteria from an existing database. A series of statistical analyses, such as unpaired t test, multiple regression, and discriminant analysis, was used to test the proposed hypothesis. The study found that none of the open bite tendency indicators used can segregate open bite subjects from nonopen bite subjects. The obtained discriminant function clearly divides the samples into two groups, i.e., open bite group and nonopen bite group, which were based on VAL and lower facial height. The study concludes that, first, an open bite tendency may be a different entity from an actual open bite or may be a misconceptualized term. Second, pharyngeal length may be a convenient indicator to diagnose open bite. We speculate that open bite may be different from an open bite tendency in pharyngeal length.

Adolescent↗

[The dynamics of pharyngeal structures in obstructive sleep apnea (during spontaneous, continuous positive pressure and BiPAP ventilation)].

Obstructive sleep apnea results from a pharyngeal collapse. Upper airway can be investigated using either static or dynamic methods during wakefulness or when the patient is sleeping. Somnofluoroscopy is one of the dynamic methods allowing a visualization of the upper airway during sleep. A lateral projection of the pharynx is obtained during fluoroscopic examination which allows visualization of the upper airway dimensions and the bone structures (hyoid bone, cervical spine, mandible). Standard polygraphic parameters (EEG, EOG, flow rate, thoracic and abdominal movements) and fluoroscopic image are simultaneously acquired on the same videotape. Using this technique, we have described the typical pattern of events occurring during an episode of apnea: 1-beginning of airway occlusion in the oropharynx with anterior or posterior hooking of the soft palate, 2-suction on the uvula downwards and complete occlusion of the oropharynx with further extension to the hypopharynx, 3-active movements of the cervical spine and hyoid bone as if the patient is choking, 4-overcoming of the occlusion usually accompanied by opening of the jaw and occurring either as a sudden event throughout the length of the pharyngeal airway or as a progressive reopening from the hypopharynx. In a recent study, we have investigated upper airway dynamics when a continuous positive pressure with one level (CPAP) or two levels of pressure (BiPAP) was applied. When using CPAP with pressure below the optimal pressure, uvula movements were the first changes we observed, preceding the pharyngeal collapse. Lowering the expiratory pressure alone lead to a significant reduction in pharyngeal dimensions starting at expiration and extending also to inspiration when the expiratory pressure is further reduced. Using BiPAP may lead to upper airway instability. The frequency and the variability of this phenomenon need further studies to be established.

Fluoroscopy↗

Combined videomanometric identification of abnormalities related to pharyngeal retention.

RATIONALE AND OBJECTIVES: The authors determined the usefulness of performing videoradiography and pharyngeal solid-state manometry during barium swallow in dysphagic patients with pharyngeal retention. METHODS: Results were retrospectively analyzed of simultaneous videoradiography and manometry examinations in 14 patients with retention of barium in the pharynx. Twelve age-matched patients without retention served as a control group. RESULTS: Patients with retention regularly had less opening of the upper esophageal sphincter than patients without retention (7.6 vs 10.3 mm, respectively; P = .003). In patients with retention, the laryngeal elevation was lower (17.1 vs 23.8 mm, respectively; P = .001), and the resting pressure of the upper esophageal sphincter was significantly lower (42.4 vs 54.0 mm Hg, respectively; P = .04). The duration of upper esophageal sphincter relaxation was also shorter in patients with retention (374 vs 603 msec, respectively; P = .003). The peak pharyngeal contraction pressure was not significantly different. CONCLUSION: The constrictors play a minor role in the conveyance of the bolus through the pharynx. Pharyngeal shortening could be the most important mechanism in bolus transport.

Adult↗

Pharyngeal trauma in children--accidental and otherwise.

Pharyngeal perforation is an uncommon injury in children. Most reported cases to date have been secondary to instrumentation or penetrating wounds. Laceration to the pharyngeal wall may introduce air, secretions and bacteria into the parapharyngeal space and mediastinum and consequently has potentially life-threatening sequelae. The management of these injuries is controversial. We present a series of four children who suffered pharyngeal trauma, accidentally and otherwise, and discuss their management. We recommend a high index of suspicion of pharyngeal injury in all cases of oropharyngeal trauma and overnight admission to hospital for observation until an accurate diagnosis has been established. Non-accidental injury of the child must be seriously considered in all cases.

Accidents↗

Acquisition of esophageal speech subsequent to learning pharyngeal speech: an unusual case study.

Laryngectomees who are delayed in obtaining alaryngeal voice therapy may develop pharyngeal voice, an undesirable type of alaryngeal voice. Pharyngeal voice is usually self-developed; however, it may also be acquired in the clinical setting when the patient is being taught to inject air for esophageal sound production. This report describes the principal findings involving a laryngectomee who had developed and used pharyngeal speech as his primary method of communication for two years prior to enrollment in alaryngeal voice therapy. A clinical technique that was found to be successful in altering the pharyngeal voice to esophageal voice is reported. Findings pertaining to the overall intelligibility as well as vocal fundamental frequency of the two different types of alaryngeal speech are included.

Aged↗

Simultaneous assessment of pharyngeal wall and velar displacement for selected speech sounds.

Velar and pharyngeal wall displacements were studied simultaneously by using conventional lateral-view radiography and frontal-view tomography. Twenty-five normal adult male and female subjects were utilized. Major results pertaining to lateral pharyngeal wall displacements during speech include the following. 1) At more superior levels of the pharynx, greater mesial displacements occurred for nonnasal compared to nasal speech sounds and no significant differences were noted among the nonnasal speech sounds studied. 2) At an inferior level of the pharynx, mesial displacements were greater for low vowels compared to the other speech sounds studied but some lateral (outward) movements were observed during high vowel production for some subjects. 3) Greatest mesial displacements for all subjects ad speech sounds studied occurred at the level of and just below the plane of the hard palate. Low correlations between velar and lateral pharyngeal wall displacements were obtained for most subjects. The results do not support the hypothesis that the levator veli palatini muscle is solely involved in both velar and lateral pharyngeal wall movements.

Adult↗

Biomechanical analysis of the pharyngeal swallow in postsurgical patients with anterior tongue and floor of mouth resection and distal flap reconstruction.

The purpose of this study was to examine changes in the biomechanics of pharyngeal swallow after surgery in eight patients (six men and two women) with anterior tongue and floor of mouth resections with distal flap reconstruction. Eight normal age-matched subjects were also studied. Swallowing performance was assessed following a standardized protocol with videofluoroscopy preoperatively and at 1 and 3 months postoperatively for the oral cancer patients. The normal subjects received a single videofluoroscopic study. Computer-assisted biomechanical analysis was used to mark the movements of specific oropharyngeal structures over time throughout the swallow of calibrated boluses. Statistical analyses revealed that tongue base, pharyngeal wall, hyoid, laryngeal, and cricopharyngeal movements during the swallow were altered significantly after surgery for the cancer patients. Some oropharyngeal structural movements differed from those of normal control subjects before surgery. In this study, biomechanical measures indicated that there was recovery in some aspects of the pharyngeal swallow in this patient group. The duration of tongue base to pharyngeal wall contact, which was significantly reduced preoperatively and at 1 month after surgery, increased significantly to within normal levels by the 3-month postoperative evaluation. Duration of laryngeal closure and the onset of laryngeal closure relative to cricopharyngeal opening also improved significantly to within normal levels by the 3-month postoperative evaluation.

Adult↗

SER-7b, a constitutively active Galphas coupled 5-HT7-like receptor expressed in the Caenorhabditis elegans M4 pharyngeal motorneuron.

Serotonin plays a key role in the regulation of pharyngeal pumping in nematodes. We have isolated a Caenorhabditis elegans cDNA (C09B7.1b, ser-7b) with greatest identity to the 5-HT7 receptor family. Membranes from COS-7 cells expressing SER-7b exhibit saturable [3H]-LSD binding (Kd = 45 nm) that is inhibited by serotonin (5-HT) and tryptamine, but not by other physiological biogenic amines. Expression of SER-7b in COS-7 cells results in dramatic increase in basal cAMP levels over untransfected cells that is dependent on expression level. 5-HT further elevates cAMP levels in a dose-dependent manner (pEC50 = 7.5 +/- 0.5). Mammalian 5-HT7 receptor inverse agonists reduce constitutive activity, with methiothepin the most potent (pIC50 = 7.8 +/- 0.1). Ser-7::GFP transcriptional fusions reveal that SER-7b appears to be expressed solely in the M4 pharyngeal motorneuron after hatching. This is the first report of a Galphas coupled biogenic amine receptor in nematodes and the localization of SER-7b in the M4 pharyngeal motorneuron suggests that SER-7b may play a role in the regulation of pharyngeal pumping.

Amino Acid Sequence↗

Characteristics of lower esophageal sphincter relaxation induced by pharyngeal stimulation with minute amounts of water.

BACKGROUND & AIMS: Injection of minute amounts of water in the pharynx leads to a long-duration lower esophageal sphincter (LES) relaxation that is an important feature of a transient LES relaxation. The aims of this study were to determine if a pharyngeal stimulus-induced LES relaxation is associated with crural diaphragm inhibition, esophageal common cavity, and acid reflux in normal subjects. METHODS: Esophageal and LES pressure, crural diaphragm electromyographs, and pH were recorded in the fasting and postprandial states in normal subjects. The pharynx was stimulated by injection of water at bolus volumes of 0.05 and 0.1 mL and then in 0.1-mL increments. RESULTS: Pharyngeal stimulation induced a long-duration LES relaxation in the absence of swallow and esophageal peristalsis. In the fasting state, none of the LES relaxations induced by pharyngeal stimulation were associated with inhibition of crural diaphragm, esophageal common cavity, and acid reflux. In the postprandial periods, only 8% of the pharyngeal stimuli resulted in crural diaphragm inhibition and esophageal common cavity. Esophageal reflux was found only at the times of simultaneous LES and crural diaphragm relaxation. CONCLUSIONS: Stimulation of the pharynx has different effects on the LES and crural diaphragm. Relaxation of crural diaphragm along with LES relaxation is essential for the occurrence of gastroesophageal reflux in normal subjects.

Adult↗

The effect of head posture on the pharyngeal airway space (PAS).

The purpose of this study was to investigate the relationship between cranio-cervical inclination and pharyngeal airway space (PAS) by measuring these parameters at different head postures in the same subjects and to obtain a regression equation to correct the values measured. Fifty lateral cephalometric radiographs taken at five different head postures per individual were obtained from ten adults (seven males and three females) aged from 25 to 30 years with nose breathers and Class I occlusion. The changes in cranio-cervical inclination produced by head extension were correlated with changes in the variables describing the PAS. The OPT/NSL (cranio-cervical inclination in the second vertebrae) and C3-Me (distance between the third vertebrae and the Menton) correlated strongly with PAS-TP (the most proximal distance measured between the posterior pharyngeal wall and the tongue base) in the pharyngeal airway space (r = 0.807 and 0.854 respectively). The regression equations were Y = - 27.177+0.39X (Y = PAS-TP, X = OPT/NSL), and Y = -21.105+0.402X (Y = PAS-TP, X = C3-Me), respectively. From these equations we could conclude that an increase of 10 degrees in OPT/NSL or 10 mm in C3-Me increased the pharyngeal airway space (PAS-TP) by about 4 mm.

Adult↗

Refinement of velopharyngoplasty in patients with cleft palate by covering the pharyngeal flap with nasal mucosa from the velum.

The velopharyngoplasty performed using the popular Sanvenero-Rosselli method improves the speech quality of patients with cleft palate suffering from persistent velopharyngeal insufficiency despite successful closure of the hard and soft palates. However, often a relatively narrow pharyngeal bridge results due to healing not only by granulation and scar contraction, but also due to the insertion of the inferior tip of the flap into a narrow bed. Elevation of two velar flaps with nasal mucosa to cover the exposed muscular undersurface of the pharyngeal flap produces a broader recipient bed into which the pharyngeal flap can be spread. The speech quality of 27 patients treated with this modified method (group B) was compared with that of 27 patients without this modification (group A). Eight weeks after velopharyngoplasty (followed by uneventful wound healing) the speech quality of group B was significantly better than that of group A (p<0.03 MannWhitney U-test). This difference was even more evident when comparing the preoperative with the postoperative speech quality (p<0.005). Thus, covering the exposed muscular layer of the pharyngeal flap with nasal mucosa of the velum is an important modification of velopharyngoplasty for the improvement of speech quality.

Adolescent↗

Rescue of Caenorhabditis elegans pharyngeal development by a vertebrate heart specification gene.

Development of pharyngeal muscle in nematodes and cardiac muscle in vertebrates and insects involves the related homeobox genes ceh-22, nkx2.5, and tinman, respectively. To determine whether the nematode and vertebrate genes perform similar functions, we examined activity of the zebrafish nkx2.5 gene in transgenic Caenorhabditis elegans. Here, we report that ectopic expression of nkx2.5 in C. elegans body wall muscle can directly activate expression of both the endogenous myo-2 gene, a ceh-22 target normally expressed only in pharyngeal muscle, and a synthetic reporter construct controlled by a multimerized CEH-22 binding site. nkx2.5 also efficiently rescues a ceh-22 mutant when expressed in pharyngeal muscle. Together, these results indicate that nkx2.5 and ceh-22 provide a single conserved molecular function. Further, they suggest that an evolutionarily conserved mechanism underlies heart development in vertebrates and insects and pharyngeal development in nematodes.

Amino Acid Sequence↗

Insulin-like growth factor-binding protein-3 plays an important role in regulating pharyngeal skeleton and inner ear formation and differentiation.

Insulin-like growth factor-binding protein (IGFBP)-3 is the major insulin-like growth factor (IGF) carrier protein in the bloodstream. IGFBP-3 prolongs the half-life of circulating IGFs and prevents their potential hypoglycemic effect. IGFBP-3 is also expressed in many peripheral tissues in fetal and adult stages. In vitro, IGFBP-3 can inhibit or potentiate IGF actions and even possesses IGF-independent activities, suggesting that local IGFBP-3 may also have paracrine/autocrine function(s). The in vivo function of IGFBP-3, however, is unclear. In this study, we elucidate the developmental role of IGFBP-3 using the zebrafish model. IGFBP-3 mRNA expression is first detected in the migrating cranial neural crest cells and subsequently in pharyngeal arches in zebrafish embryos. IGFBP-3 mRNA is also persistently expressed in the developing inner ears. To determine the role of IGFBP-3 in these tissues, we ablated the IGFBP-3 gene product using morpholino-modified antisense oligonucleotides (MOs). The IGFBP-3 knocked down embryos had delayed pharyngeal skeleton morphogenesis and greatly reduced pharyngeal cartilage differentiation. Knockdown of IGFBP-3 also significantly decreased inner ear size and disrupted hair cell differentiation and semicircular canal formation. Furthermore, reintroduction of a MO-resistant form of IGFBP-3 "rescued" the MO-induced defects. These findings suggest that IGFBP-3 plays an important role in regulating pharyngeal cartilage and inner ear development and growth in zebrafish.

Animals↗