Fine structural localization of peroxidase activity in the epithelium and the gland of the rat larynx

Summary Endogenous peroxidase activity was demonstrated in ciliated cells and secretory cells of the laryngeal epithelium and gland of rats, using the diaminobenzidine method for cytochemical demonstration of peroxidase activity. The intensity of peroxidase activity was greatly varied from cell to cell, but the fine structural localization of the activity was similar in various cell types. The activity was localized in cisternae of rough-surfaced endoplasmic reticulum including nuclear envelope, some vesicles and saccules of the Golgi complex, large membrane-limited granules, multivesicular bodies and probable lysosomes. In secretory cells, the activity was also found in secretory granules.The significance of peroxidase activity is not unclear, while the activity, at least a part of it, seems to be secreted into the cavity of the larynx. The possibility that peroxidase participates bactericidal mechanism, deserves further investigation.     

Time-course of apoptosis in the olfactory epithelium of rainbow trout exposed to a low copper level

Copper at low doses is known to specifically induce olfactory neuron death in fish olfactory epithelium. Using light and electron transmission microscopy, we have investigated the features and the time-course of receptor cell death in rainbow trout exposed for 15 days to 20 mug Cu(2+)/l. Ultrastructural observations demonstrate that degenerating cells, which included both mature and immature neurons, exhibited morphological changes characteristic of a cell death by apoptosis. Quantitative analysis shows that the number of apoptotic cells increased significantly already after 1 day of exposure, reaching a peak at day 5. From this timepoint of exposure, no more mature neuron was noted in the olfactory epithelium. Following a significant decrease in the number of apoptotic cells at day 10, a second wave of neuron death was noted at day 15. These findings argue for the occurrence of a neurogenesis process to balance the receptor cell death, despite continued copper exposure, and for a higher vulnerability to the metal of olfactory neurons presenting more advanced stages of cell differentiation. The molecular mechanisms by which copper may induce olfactory neuron apoptosis are discussed.     

Olfaction and neurogenesis in the olfactory epithelium

Anosmia was experimentally produced in strain C57BL/6 laboratory mice by treatment with 1% zinc sulfate solution. Structural and functional changes taking place in the olfactory epithelium were investigated during this process and during reinstatement of olfaction. Isoamyl acetate, butyl acetate, and substances present in murine urine were used as olfactory stimuli. Response to these odorants was found to recover from zinc sulfate action at different rates. The highest (both relative and absolute) daily rise in amplitude response was that induced by isoamyl acetate and butyl acetate and lowest in the case of odors of biological origin. Response to olfactory stimuli recovered most rapidly in the areas of the epithelium where maximum response to the same stimuli had been seen in intact animals."Biopharmautomatica" Combined Research and Production Unit, Gor'kii. Translated from Neirofiziologiya, Vol. 22, No. 4, pp. 500–506, July–August, 1990.     

V-ATPase expression in the mouse olfactory epithelium

The vacuolar proton-pumping ATPase (V-ATPase) is responsible for the acidification of intracellular organelles and for the pH regulation of extracellular compartments. Because of the potential role of the latter process in olfaction, we examined the expression of V-ATPase in mouse olfactory epithelial (OE) cells. We report that V-ATPase is present in this epithelium, where we detected subunits ATP6V1A (the 70-kDa "A" subunit) and ATP6V1E1 (the ubiquitous 31-kDa "E" subunit isoform) in epithelial cells, nerve fiber cells, and Bowman's glands by immunocytochemistry. We also located both isoforms of the 56-kDa B subunit, ATP6V1B1 ("B1," typically expressed in epithelia specialized in regulated transepithelial proton transport) and ATP6V1B2 ("B2") in the OE. B1 localizes to the microvilli of the apical plasma membrane of sustentacular cells and to the lateral membrane in a subset of olfactory sensory cells, which also express carbonic anhydrase type IV, whereas B2 expression is stronger in the subapical domain of sustentacular cells. V-ATPase expression in mouse OE was further confirmed by immunoblotting. These findings suggest that V-ATPase may be involved in proton secretion in the OE and, as such, may be important for the pH homeostasis of the neuroepithelial mucous layer and/or for signal transduction in CO₂ detection. proton secretion; vacuolar H⁺-ATPase; immunofluorescence; pH homeostasis; olfaction     

Glands in the olfactory epithelium of marine fish

Specific features of the morphohistochemical and ultrastructural organization of the secretory system of olfactory organs in marine fish from the Pacific region were studied. Multicellular alveolar and tubular glands were revealed in the olfactory epithelium of Pleurogrammus azonus, Myoxocephalus yaok, Enophrys diceraus, Alcichthys elongatus, Gymnocanthus pistilliger, Hemilepidotus gilberti, Hemitripterus villosus, Podothecus veternus, Liparis dubius, Clupea pallasi, as well as in salmonids (Oncorhynchus nerka, O. keta, O. gorbuscha, O. masou, O. kisutch, O. tschawytscha) and in tetraodontids (Takifugu rubripes). The morphofunctional characteristic of the described glands is close to the Bowman’s glands of land animals. The structure, localization, and density of location of olfactory glands are determined by specific features of the ecology of the species. These structures are most numerous in bottom and near-bottom representatives of marine ichthyofauna.     

Fine structural changes and localization of phosphatases in the epithelium of the duodenal crypt of X-irradiated mice

Summary The ultrastructural localization of acid phosphatase has been studied in the stem cells of duodenal crypt of X-irradiated mice. After a transitional increase of the cytochemically detectable activity of this enzyme, in the first hours after irradiation, the reaction appears less and less important in the cells. These observations are in agreement with the morphological alterations of the ultrastructures provoked by the irradiation. The meaning of these modifications is discussed.This work was done thanks to the contract C.E.N./A.I.E.A. No. 347/RB and thanks to grants from the Fonds de la Recherche Scientifique Fondamentale Collective.     

Fine structural demonstration of acid phosphatase in rabbit germinal epithelium prior to induced ovulation

The germinal or surface epithelium covering rabbit Graafian follicles contains occasional small, dark, lysosome-like bodies. After an ovulatory dose of human chorionic gonadotropin (HCG) such bodies gradually increase in size and number. At 8 hr after HCG there is a maximal accumulation in the apical follicle cells; then the dense bodies decrease and just prior to ovulation, 9.5 hr after HCG, only few of them remain in the attenuated surface epithelium. Most of the growing membrane-surrounded bodies probably represent lysosomes, since electron microscopy combined with cytochemistry revealed that many of them contain the lysosomal "marker" enzyme, acid phosphatase. The role of sex steroids and prostaglandins regarding lysosomal growth and LABILization is discussed. The close temporal relation between disappearance of the apical surface epithelial lysosomes and disintegration of the underlying tunica albuginea gives further support to our working hypothesis that at least part of the "ovulatory enzymes" emanate from the surface epithelium.     

Biopsies of human olfactory epithelium

It has been shown that olfactory epithelium can be safely biopsied from the living, intact human being. Observations of the ultrastructure of this epithelium shows changes that can then be correlated with the etiology and degree of olfactory loss, allowing a greater understanding of both normal transduction and of the pathology of dysfunction. Examples of the common forms of olfactory dysfunction are presented and discussed. Additionally, the technique will allow additional immuno-histochemical and molecular study of the tissue, will increase the understanding of both normal and pathological function and should translate to new therapeutic regimens.     

Fine structural aspects on the fate of rat black thyroids induced by minocycline

The fate of the black thyroid induced by minocycline treatment (100 mg/kg daily for 21 days) in the rat was studied by light and electron microscopy after 6 months. The black discoloration of the thyroid gland remained and numerous dense bodies containing highly electron-dense deposits were seen in most of the follicular epithelial cells. It appears that the turnover rate of follicular epithelial cells is very low and the electron-dense deposits are largely retained, though debris derived from degenerate follicular epithelial cells containing the dense deposits may be phagocytosed by macrophage-like cells in the interfollicular connective tissue. Brown-black granules are also found in the extremely attenuated follicular epithelial cells of cold follicles.     

Fine structural changes in the epididymal epithelium of moles (Talpa europaea) throughout the year.

The epididymis of the European mole (Talpa europaea) was studied by light and electron microscopy. In the sexually active animal, spermatozoa mature during their passage through the epididymis and the structure of the cells lining the duct suggests a clear regional division into initial, middle and terminal segments. Numerous intra-epithelial vesicles were present in the distal part of the middle segment of sexually active moles and the lining epithelium in the terminal segment appeared to be secretory. Variation in the sensitivity of different regions of the epididymis to androgens was apparent: the principal cells of the initial segment were morphologically active only during the peak of the breeding season in spring, while the cells of the terminal segment became active earlier and remained so for longer. During sexual regression, many autophagic vacuoles were found in the principal cells, and these became transformed into lipofuscin pigment granules. Cells heavily laden with these granules appeared concurrently in the lining epithelium. It is suggested that such cells may be involved in the regression of principal cells by means of heterophagic activity. A similar situation was also observed, but to a lesser extent, at the beginning of the breeding season. Outside the breeding season, the organelles of the principal cells were poorly developed throughout the epididymis, and lipofuscin pigment granules remained in the principal and basal cells of adults. Such granules were seldom seen in immature animals.     

Response patterns of single neurons in the tortoise olfactory epithelium and olfactory bulb

The responses to odor stimulation of 40 single units in the olfactory mucosa and of 18 units in the olfactory bulb of the tortoise (Gopherus polyphemus) were recorded with indium-filled, Pt-black-tipped microelectrodes. The test battery consisted of 27 odorants which were proved effective by recording from small bundles of olfactory nerve. Two concentrations of each odorant were employed. These values were adjusted for response magnitudes equal to those for amyl acetate at –2.5 and –3.5 log concentration in olfactory twig recording. Varying concentrations were generated by an injection-type olfactometer. The mucosal responses were exclusively facilitory with a peak frequency of 16 impulses/sec. 19 mucosal units responded to at least one odorant and each unit was sensitive to a limited number of odorants (1–15). The sensitivity pattern of each unit was highly individual, with no clear-cut types, either chemical or qualitative, emerging. Of the 18 olfactory bulb units sampled, all responded to at least one odorant. The maximum frequency observed during a response was 39 impulses/sec. The bulbar neurons can be classified into two types. There are neurons that respond exclusively with facilitation and others that respond with facilitation to some odorants and with inhibition to others. Qualitatively or chemically similar odorants did not generate similar patterns across bulbar units.     

Coupling between sensory neurons in the olfactory epithelium

Coupling of olfactory sensory neurons (OSNs) in the olfactory epithelium of Necturus maculosus was demonstrated by dye-transfer with Lucifer yellow CH; however, the incidence of dye-transfer was low. Immunocytochemistry and Western blot analysis indicated that connexin 43, a gap junction channel subunit, was widely expressed by cells in the olfactory epithelium. Electrical coupling by presumptive gap junctions was assessed using electrophysiological recordings, heptanol block, tracer-uptake through hemi-junctions, and tracer-injection into tissue whole-mounts. Coupling, which involved pairs of OSNs only, was detected in approximately 3-10% of the OSN population; there was no evidence that OSNs were coupled into extended neural syncitia. These results suggest that coupling of OSNs by gap junctions is unlikely to have a general role in olfactory responses by mature (odor responsive) OSNs. Instead, the incidence of inter-neuronal coupling was small, similar to the fraction of immature OSNs, suggesting a possible role of gap junctions in the continual turnover and development of OSNs or possibly their senescence.