Rat Eustachian Tube Research Articles

The rat eustachian tube: Anatomical, histological, and radiological features

PurposeThis study investigated the anatomical and histological characteristics of the rat Eustachian tube (E-tube) and the feasibility of Eustachian tubography in a rat model. Materials and methodsFifteen male Wistar rats were used in this study, and the bilateral E-tubes of each rat were examined. Ten E-tubes were used for anatomical studies, another ten for histological analysis, and the other ten for Eustachian tubography. Five rats were euthanized and decapitated, and ten E-tubes were dissected to describe the anatomy of the E-tube. Ten E-tube specimens obtained from five other rats were sectioned to investigate E-tube histology. Eustachian tubography was performed on the bilateral E-tubes of the other five rats using the trans-tympanic approach. ResultsThe rat E-tubes consisted of bony and membranous parts. Cartilage and bone tissue covered only the bony part. The E-tubes’ mean diameter and overall length were 2.97 ​mm and 4.96 ​mm, respectively. The tympanic orifices’ mean diameter was 1.21 ​mm. The epithelium of E-tubes was mainly composed of pseudostratified ciliated and goblet cells. Eustachian tubography was successfully performed on both sides of the E-tube for each rat. The technical success rate was 100%, the average running time was 4.9 ​min, and no procedure-related complications occurred. On tubography images, the E-tube, tympanic cavity, and nasopharynx could be identified because of the visualization of bony landmarks. ConclusionIn this study, we described the anatomical and histological features of rat E-tubes. With the aid of these findings, E-tube angiography was successfully performed using a transtympanic approach. These results will facilitate further investigation of E-tube dysfunction.

Bacterial adherence in otitis media: Determination of N-acetylgalactosamine (GalNAc) residues in the submucosal glands and surface epithelium of the normal and diseased Eustachian tube

Background Acute otitis media (AOM) is the most common childhood infection caused by bacteria. The pathogenesis of AOM implicates initial adherence of a pathogen to the nasopharyngeal epithelium, which is followed by bacterial colonization of the middle ear cavity through the Eustachian tube. N-acetylgalactosamine (GalNAc) is an important constituent of mucins and GalNAc containing sugar residues seem to be essential for initial adherence of respiratory bacteria to the surface of epithelial cells. Objective To explore the localization of GalNAc residues, we incubated Eustachian tube sections from Streptococcus pneumoniae infected and normal control rats with seven biotinylated, GalNAc recognizing lectins: Bauhinia purpurea lectin (BPA), Psophocarpus tetragonolobus lectin (PTA), Helix aspersa lectin (HAA), Helix pomatia lectin (HPA), Phaseolus lunatus lectin (PLA), Sophora japonica lectin (SJA) and Vicia Villosa isolectin B4 (VVA-B4). Results The mucin producing epithelium and submucosal glands of the normal Eustachian tube contained GalNAc residues, as evidenced by binding of several of the lectins. Lectin binding specificity and intensity changed following acute middle ear infection. BPA was the only lectin that exclusively stained the surface epithelium and the serous acini of the submucosal glands in the infected animals, whereas no binding was detected in the normal controls. HPA, HAA, PTA and VVA-B4 binding to surface epithelial cells increased after infection, indicating an active secretion of GalNAc containing glycans. Quantitative analysis of submucosal gland staining intensity showed significantly more GalNAc residues in the normal Eustachian tube, compared to infected animals. Conclusion We conclude that the mucous producing elements of the normal rat Eustachian tube contain GalNAc residues essential for respiratory pathogen adherence. In addition, the GalNAc residue specificity and reacting intensity change in relation to acute infection, which may be important in relation to subsequent development of secretory otitis media or formation of a bacterial biofilm in the middle ear. The results show that GalNAc residues increased in both the submucosal serous glands and in the surface epithelium of the Eustachian tube after middle ear infection with S. pneumoniae

Expression of water channel proteins (aquaporins) in the rat Eustachian tube and middle ear mucosa

Conclusion. Diverse expression of the different subtypes of aquaporins in different parts of the Eustachian tube and middle ear suggests region-specific functions of the aquaporins in the normal physiology of the tubotympanum and also suggests that they may play roles in the pathophysiology of otitis media. Objectives. The epithelial cells of the middle ear and Eustachian tube must maintain adequate water balance for normal function of the mucociliary system. Since aquaporins (AQPs) are known to play critical roles in water homeostasis, we investigated their expression in the tubotympanum of the rat. Methods. The expression of AQP subtypes 1, 2, 4, 5, and 7 were examined in the rat Eustachian tube and middle ear using RT-PCR, Western blotting, and immunohistochemistry. Results. Transcripts for AQP 1, 4, and 5 were detected in the Eustachian tube and middle ear. Expression of these molecules at the protein level was confirmed by Western blot analysis. Immunohistochemical analysis demonstrated that AQP 4 was localized to the basolateral membranes of ciliated epithelial cells while AQP 5 was localized to the apical surface of serous gland cells, but not goblet cells, in the rat Eustachian tube. AQP 1 was found to be expressed by the subepithelial fibroblasts.

The effect of the sympathetic and sensory nervous system on active eustachian tube function in the rat

Conclusion. We propose that simultaneous activation of the sensory and sympathetic nervous system may adversely affect eustachian tube function, which may have a role in the genesis of Ménière's disease. Objective. We have determined the distribution of sympathetic and nociceptive sensory axons in the mucosa of the rat eustachian tube and investigated whether sympathetic or nociceptive neurons influence the function of the eustachian tube. Materials and methods. We tested whether the ability of a rat to equalize air pressure in the middle ear during evoked swallowing was altered by activation or blockade of the local sympathetic nervous system, or by stimulation of nociceptive neurons with capsaicin. Results. Sympathetic axons were sparse, but CGRP-immunoreactive, nociceptive axons formed a dense subepithelial plexus beneath the eustachian tube epithelium. Neither the adrenergic blocking drug, bretylium, nor electrical stimulation of the superior cervical ganglion significantly altered eustachian tube function. Capsaicin alone did not affect eustachian tube function but capsaicin applied with an alpha adrenoceptor agonist impaired the function of the eustachian tube. Capsaicin applied to the bulla also increased spontaneous swallowing in anaesthetized rats and this effect was enhanced by addition of an alpha adrenoceptor agonist and by stimulation of the superior cervical ganglion.

Effect of exogenous surfactant on ventilatory and clearance function of the rat's eustachian tube.

The Eustachian tube has three important functions with respect to the middle ear: ventilation, clearance, and protection. Surfactants are assumed to be important to maintain these functions. The administration of exogenous surfactant may therefore be effective to improve the function of the Eustachian tube. This randomized, double-blind, placebo-controlled study was designed to investigate the effect of exogenous surfactant on the function of the Eustachian tube in rats. Exogenous surfactant was administered into the middle ear of 10 otologically healthy rats, and 10 other rats received placebo. The effect on the opening and closing pressure (passive ventilatory function) and the dye clearance time (clearance function) of the rat's Eustachian tube was measured. A significant decrease in the opening pressure was seen after the administration of surfactant. Both surfactant and placebo caused an increase in the closing pressure. A serious disturbance of the dye clearance time was induced in 13 rats, and the test failed in 1 rat. In the remaining 6 rats, no significant differences in the dye clearance time were observed between the two groups. Exogenous surfactant decreased the closing forces of the Eustachian tube even in otologically healthy rats. No significant effect on the mucociliary clearance was observed, but this may have resulted from the small number of rats. Additional randomized, double-blind, placebo-controlled trials should be conducted to determine the clinical relevance of these changes and to further assess the effect of surfactant on the function of the Eustachian tube.

Rat eustachian tube and its musculature.

The objective of this study was to evaluate the anatomy of the eustachian tube (ET) of the rat and the paratubal musculature. Microdissection and serial sections were used. The ET consists of collapsible membranous and membranocartilaginous segments and a noncollapsible bony segment. Tubal muscles are attached to the collapsible part; the salpingopharyngeus muscle (SPM) is well developed and consists of 3 distinct groups of muscle fibers; the tensor veli palatini muscle (TVPM) consists of 2 functionally different groups of fibers, but only 1 group assists in opening the ET. Attachment of the fibers of the SPM and TVPM that are involved in tubal opening is confined to the dorsal portion of the ET. This finding, together with the earlier observation that this part is mainly lined by squamous epithelium, strongly suggests that the dorsal part has a ventilatory function. The ventral portion of the ET, which is lined by ciliated-secretory epithelium and lacks the attachment of muscle fibers that can dilate the lumen, is assumed to serve clearance. The anatomic position of the levator veli palatini muscle suggests that this muscle contributes to the protective function of the ET. These findings are discussed with regard to the ET in humans.

Sympathetic innervation of the eustachian tube in rats.

The glyoxylic catecholaminergic histofluorescence method was employed on the mucosa of the rat's eustachian tube (ET) in order to study the sympathetic innervation present. One percent neutral red was used as counterstain. Many noradrenergic fibers were demonstrable around blood vessels, glands and submucosa of the ET, but not in the epithelium. In a group of rats following neurectomy, the superior cervical ganglia (SCG) were removed unilaterally or bilaterally. Changes in sympathetic innervation of the ET were examined 14 days after SCG ganglionectomy. In those animals after unilateral SCG ganglionectomy, no noradrenergic histofluorescence was found in the ipsilateral ET, although some scant fluorescence could be detected in the tube's nasopharyngeal (NP) orifice. However, no noradrenergic histofluorescence could be observed in animals bilateral SCG ganglionectomies. Our results indicate that sympathetic innervation of the ET in the rat originates in the SCG, with some cross-innervation of sympathetic fibers occurring in the tube's NP orifice.

Surfactant and isoprenaline effect on eustachian tube opening in rats with acute otitis media

Purulent otitis media was induced in 12 middle ears of Sprague-Dawley rats by inoculation of pneumococci type 3. The pressure required to open the eustachian tube was recorded 4 days later and was found to be 3.05 ± 0.10 kPa (mean ± SEM), which was 1.08 ± 0.15 kPa lower than the mean value obtained in a control group of healthy rats ( P ⩽ .001). We also measured the effects of pulmonary surfactant instilled into the middle ear as well as isoprenaline injected intravenously. The presence of surfactant further reduced the pressure opening level by 0.72 ± 0.05 kPa; this reduction did not differ from that found in the control group ( P > .8). Isoprenaline reduced the pressure opening level in the control group by 0.20 ± 0.03 kPa ( P ⩽ .001), and no reduction was found in the rats with otitis media. Although the part played by surface tension-lowering factors in eustachian tube function is still uncertain, such factors apparently also exert an effect in the inflamed tube. However, isoprenaline, which is considered to act by the secretion of surface tension-lowering substances, did not facilitate opening of the eustachian tube in rats with acute otitis media.

Rat eustachian tube synthesizes disaturated phosphatidylcholine

Otitis media results when the eustachian tube fails to adequately ventilate the middle ear. A surface tension-lowering substance may be required for normal tube opening, especially in young children with poorly developed naso-pharyngeal musculature. We report here that rat eustachian tube epithelium synthesizes disaturated phosphatidylcholine, which is recognized as the surface tension-lowering substance of pulmonary surfactant.

The anatomy of the eustachian tube in the rat: a macro- and microscopical study.

The rat eustachian tube (ET), from the nasopharyngeal orifice to the tympanal orifice, is about 4.5 mm long, of which the naso-medial membranous part (the nasopharyngeal orifice) measures about 1.5 mm and the occipito-lateral bony portion about 3 mm. The nasopharyngeal orifice is surrounded by two soft, lip-like, mucosal swellings--one ventral and one cranial--both easily mobile. The muscles related to the tubal opening mechanisms are the salpingopharyngeus, the tensor veli palatini and the levator veli palatini muscles. The salpingopharyngeus muscle originates partly from the cranial lip, whereas the palatal muscles originate partly from the ventral lip. The tympanal two thirds of the mucosal lining of the ET is cranially guided by a cartilage and incompletely framed by bony structures. The tympanal orifice is situated in the nasal part of the medial wall, well above the floor of the bulla. The tensor tympani muscle does not seem to take part in the opening and closing mechanisms of the ET. The mucosal lining of the ET consists of a respiratory epithelium with numerous glands in the lamina propria. It is suggested that the tubal muscles control the passage through the ET by moving the lip-like folds of the nasopharyngeal orifice. The anatomy of the rat ET is comparable to that described in Homo and it can be concluded that the rat ET might be a good model for studying the function of the human ET.

Effect of tubal occlusion on bone formation in the middle ear of the rat.

The effect of experimental occlusion of the Eustachian tube in rats have been studied. Only in germ-free animals it has been possible to eliminate infection as a complicating factor. The formation of new bone in the wall of middle ear and bulla was apparent soon after occlusion, even in the absence of infection. These findings suggest that tubal occlusion causing serous otitis media can lead to extensive changes in the bone lining the middle ear cavity and this could explain the condensation of bone found in ears without any apparent sign of infection.

Structure and Healing Capacity of the Rat Tympanic Membrane After Eustachian Tube Occlusion

Obstruction of the Eustachian tube in rats results in an accumulation of serous fluid in the middle ear cavity. the fluid was first detected in the attic, and subsequently the whole cavity became filled. However, when a perforation of the tympanic membrane was performed in connection with experimental tube obstruction, the mesotympanon was found free from fluid, but not the attic, as long as the perforation remained open. After 9-11 days of obstruction the pars flaccida was found to be maximally retracted, forming a non-adhesive clean retraction pocket. in addition, marked ultrastructural changes were found to have taken place in the pars flaccida following tube obstruction. Finally, in spite of the occluded Eustachian tube, traumatic tensa perforation was found to heal rapidly.