Osamu Mizukoshi

Substance P Nerve Fibres in the Canine Larynx by PAP Immunohistochemistry

The distribution of the Substance P (SP) immunoreactive nerve fibres in the canine larynx and laryngeal nerves was studied by PAP immunohistochemistry. Many individual SP immunoreactive nerve fibres with varicosities were observed within the epithelial layer and in the connective tissue below the epithelium of the laryngeal mucosa. Small numbers of SP immunoreactive nerve fibres were also found in the submucosal gland region and some of them appeared to terminate in glandular cells. These findings are consistent with the view that SP might be involved in the laryngeal sensory innervation system and the laryngeal glandular secretion. No SP immunoreactive nerve fibres were found in any intrinsic laryngeal muscles. The recurrent laryngeal nerve and the superior laryngeal nerve contained SP immunoreactive nerve fibres and were considered to lie in the pathway of the SP nerve fibres to the larynx.

The Site of Involvement of Hypertension within the Cochlea:A Comparative Study of Normotensive and Spontaneously Hypertensive Rats

The function and morphology of the cochlea of the spontaneously hypertensive rat (SHR) were examined and compared with the age-paired normotensive Wistar Kyoto rat (WKY). Electro- cochleographic study revealed that the function of the cochlea in the SHR declined with increasing age to a greater extent than that of WKY. Electronmicroscopic study revealed that the primary site of the cochlear deterioration of the SHR was the vascular stria, followed by the organ of Corti. Some hypotheses to explain this phenomenon are proposed for further study.

Prostaglandin synthesis by the cochlea.

The exogenous and endogenous syntheses of prostaglandins (PGs) by the cochlea of adult mongolian gerbils were studied in vitro. After incubation of the whole membraneous cochlea with [3H]-arachidonic acid (AA), syntheses of PGF2 alpha, 6-keto PGF1 alpha, PGE2, thromboxane (TX) P2 and PGD2 were evidenced in this order. The synthesis of radioactive PGs was almost completely inhibited by incubation with 10(-5) M indomethacin. No significant amounts of those PGs were detected by radioimmunoassay (RIA) in the cochlea obtained from animals killed by microwave irradiation at 5.0 kw for 0.8 sec. However, when the homogenate of the whole membraneous cochlea obtained from animals without microwave irradiation was incubated at 37 degrees C for 0-15 min, PGD2, PGE2, PGF2 alpha and 6-keto PGF1 alpha were found to be formed from endogenous AA in the cochlea by RIA. PGs were formed already at 0 time to considerable level (PGD2, PGF2 alpha and 6-keto PGF1 alpha, 90-120 pg/cochlea; PGE2, 370 pg/cochlea), reached to the maximum level (PGD2, PGF2 alpha and 6-keto PGF1 alpha, 170-200 pg/cochlea; PGE2, 500 pg/cochlea) at a 5-min incubation, and then gradually decreased. On the other hand, the amount of TXB2 was lower than the detection limit by RIA (less than 50 pg/cochlea) even after the incubation. The cochlea was dissected into three parts: organ of Corti + modiolus (OC + M), lateral wall (LW), and cochlear nerve (CN), and then PGs formed by these tissues were determined after a 5-min incubation of the homogenates. In the CN and OC + M, PGE2 was the major PG (100 and 160 pg/tissue, respectively), and the amounts of PGD2, PGF2 alpha and 6-keto PGF1 alpha were about 1/3 of those of PGE2. In the LW, the amounts of PGD2, PGE2, PGF2 alpha and 6-keto PGF1 alpha were about the same level (70-100 pg/LW).

Experimental otitis media with effusion induced by intratympanic lipid A instillation

Otitis media with effusion was induced in guinea pigs by intratympanic instillation of lipid A, the lipid moiety of gram-negative bacterial lipopolysaccharide from Salmonella minnesota Re595. Lipid A was chosen as an inducer because of its similar composition among various bacterial species. Animals were killed from the first to 14th day after instillation of various concentrations (0.2, 2, 20, 200 micrograms/ml) of lipid A in 0.5% triethylamine. By 3 days after instillation, all experimental animals developed serous middle ear effusion. The histologic findings included hemorrhage, mucosal edema, capillary engorgement, and migration of infiltrative cells including macrophages, polymorphonuclear neutrophils, and lymphocytes. These findings were most prominent 3 days after instillation, and the recovery of the middle ear epithelium was observed within 14 days. Repeated instillation of lipid A (2 micrograms/ml) at an interval of 14 days reinforced the local response accompanied by serous middle ear effusion. These findings indicate that lipid A can induce the inflammatory changes with middle ear effusion and that lipid A plays an important role in the pathogenesis of otitis media with effusion.

Ultrastructural and Fluorescence Histochemical Studies on the Sympathetic Innervation of the Canine Laryngeal Glands

The existence of the adrenergic terminals of the canine laryngeal glands has been revealed by electron microscopy and fluorescence histochemistry. Adrenergic fibres with fluorescent varicosities were observed around the base of th acini in the submucosa. In dogs treated with 5-OHDA, varicosities containing both small and large, dense-cored vesicles containing both small and large, dense-cored vesicles, believed to be adrenergic terminals, were found near blood vessels, gland cells and myoepithelial cells in the submucosal gland region. The role of these adrenergic terminals is briefly discussed.

Effect of anti-interferon serum of influenza virus infection in mice

Mice were infected by an aerosol of influenza virus Type A (0.5 LD50) and subsequently treated with 4 intranasal instillations of anti-interferon antiserum over a period of 72 h. All the mice treated with antiserum died within 7 days post-infection, whilst the mice in the control groups survived. In mice that did not receive the antibody, virus titers in the lung peaked on day 3 and then decreased again. Also, interferon was detectable both in lung homogenates and serum. In mice treated with antiserum, no interferon was detectable and the virus concentrations in the lung increased until death. These results suggest that interferon produced in the respiratory tract plays an important role in the early stages of influenza virus infection.

Wheat germ agglutinin binding sites in the organ of Corti as revealed by lectin-gold labeling

The distribution of wheat germ agglutinin(WGA)-binding sites in the organ of Corti of the guinea pig and mongolian gerbil was studied. WGA was conjugated with gold particles and applied on thin sections of the cochlea embedded in Spurrs resin and in Lowicryl K4M. WGA-binding sites were found on the plasma membrane, lysosomes and cytoskeletons of hair and supporting cells as well as on the tectorial and basilar membranes. No distinct difference was discovered between hair cells and supporting cells in terms of WGA-binding activities.

Cytochemical localization of specific carbohydrates in the cochlea using wheat germ agglutinin-gold

Localization of specific carbohydrates in the cochlea was examined by postembedding labelling by wheat germ agglutinin (WGA)-gold at the electron microscopic level. Gold labelling was observed in the tectorial membrane, basilar membrane, basal membrane of the capillary and other connective tissues. In these areas, the labelling was observed over either fibrous structures or the ground substance. WGA is known to bind specifically with N-acetylneuraminic acid, N-acetyl-D-galactosamine, N-acetyl-D-glucosamine and its beta-(1-4)-linked oligosaccharides. The labelled sites, therefore, are considered to indicate the presence of glycoconjugates that contain these carbohydrates as their constituent.

Cupulogenesis and glycoconjugates in the labyrinthine ampulla as revealed by WGA-gold labeling

SummaryWe studied the distribution of wheat germ agglutinin (WGA)-bindable glycoconjugates in the vestibular ampulla of mongolian gerbils. WGA was conjugated with gold particles and applied to Lowicryl K4M sections of the ampulla. WGA-binding sites were found on the cupula and some of the secretory granules and Golgi apparatuses in the supporting cells of the sensory epithelia. The granules were seen to secrete into the endolymphatic space through reticular membrane. It is likely, therefore, that glycoconjugates are glycosylated at the Golgi apparatus in the supporting cells, stored in the granules, and secreted through the reticular membrane into the endolymphatic space to be used as a component of the cupula. The cell membranes of various cells, connective tissue filaments in the perilymphatic space and the cytoplasm of melanocytes were also labeled with WGA-gold.

Quantitative Analysis of Kanamycin Ototoxicosis

The morphological changes after kanamycin intoxication of the inner ear, including both the cochlea and the vestibule, were quantitatively analysed by the surface preparation technique after succinic dehydrogenase staining. 75 guinea pigs were used. The outer hair cells in the basal coil and the inner hair cells in the upper coils of the cochlea were the most severely damaged, but many unusual modes of damage were also revealed. For example, the initial hair cell damage in the cochlea appeared in the upper hair cells. The clearly observed vestibular damage contradicts the general belief that kanamycin is not so toxic to the vestibular hair cells. The utricular macula and the lateral crista were most severely damaged. The delayed ototoxicity of kanamycin was observed for the first time in the vestibular hair cells.