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Spatio-temporal patterns of ensheathing cell differentiation in the rat olfactory system during development.

An immunocytochemical approach with specific glial markers was used to investigate the temporal and spatial patterns of differentiation of ensheathing glia wrapping axon fascicles along the primary olfactory pathway of the rat during development. The two glial markers tested, the proteins S-100 and glial fibrillary acidic protein, are known to be expressed at different stages of maturation in glial cells. The S-100 protein was first weakly expressed in cells accompanying the olfactory axons at embryonic day 14 (E14), while a first faint glial fibrillary acidic protein staining was detected along the olfactory axons at E15 and along the vomeronasal nerves at E16. A strong S-100 immunoreactivity was already present from E16 onwards along the axon fascicles through their course in both the nasal mesenchyme and the subarachnoid space before entering the olfactory nerve layer of the olfactory bulb. A gradual increase in glial fibrillary acidic protein expression was observed along this part of the developing olfactory pathway from E16 up to E20, when an adult-like pattern of staining intensity was seen. By contrast, most of the ensheathing cells residing in the olfactory nerve layer exhibited some delay in their differentiation timing and also a noticeable delayed maturation. It was only from E20 onwards that a weak to moderate S-100 expression was detected in an increasing number of cells throughout this layer, and only few of them appeared weakly glial fibrillary acidic protein positive at postnatal days 1 and 5. The immunocytochemical data indicate that there is a proximodistal gradient of differentiation of ensheathing cells along the developing olfactory pathway. The prolonged immaturity of ensheathing cells in the olfactory nerve layer, which coincides with the formation of the first glomeruli, might facilitate the sorting out of olfactory axons leading to a radical reorganization of afferents before they end in specific glomeruli.

Aging↗

[Evaluation of turnover of olfactory epithelium in mice by using anti-BrdU monoclonal antibody].

Among nerve cells of vertebrates, the olfactory elements are uncommon in their capacity to turnover and to be replaced after injury. An autoradiographical and morphological observation has shown that degenerated olfactory nerve cells are reconstituted by a new population of neurons which originate from basal cells. However, an autoradiographic method requires a special isotope institute and it takes a long time for the final specimen to observe. Recently, a rapid technique without the radioisotope has been alternatively developed in which a thymidine analogue, 5-bromo-2'-deoxyuridine (BrdU), is incorporated into replicating DNA and subsequently localized using a specific monoclonal antibody. In the present study, cell dynamics of olfactory mucosa in mice were investigated by means of immunohistological technique. The results were as follows. 1. The labelled elements were concentrated at the basal layer of the epithelium, which were observed 5 hrs after the first injection of BrdU. 2. At 15 days after administration of BrdU, the labelled elements were located in the mid-layer of the epithelium, where can be recognized as the compartment of nerve cells. 3. After 30 days, the labelled cells disappeared from the epithelium. It indicates that the period of turnover in the olfactory epithelium of mice is within 30 days. 4. Fifteen days after axotomy of the olfactory nerves, two stained patterns which were numerously or sparsely labelled regions were observed. The former is considered that immature neurons predominantly exist, and the latter is the area which mature neurons abundantly locate. It is considered that this immunohistological approach is useful for the observation of the turnover of the olfactory epithelium.

Animals↗

Post-traumatic olfactory dysfunction.

OBJECTIVES: This study demonstrates histopathologic and immunocytochemical changes in the olfactory bulb of a patient with post-traumatic olfactory dysfunction. These results are analyzed in light of current understanding of the pathophysiology of anosmia and dysosmia following head trauma. Emphasis is placed on potential mechanisms of human regeneration and recovery. STUDY DESIGN: The current study documents the history of a patient with the initial complaint of complete anosmia following minor head trauma. Two months after the injury the patient developed persistent, severe dysosmia with debilitating weight loss. Neurosurgical treatment, including removal of the olfactory bulbs and tracts, resulted in permanent resolution of dysosmia. METHODS: Histopathologic and immunocytochemical analysis of the olfactory bulbs was undertaken and compared with age-matched control tissue. RESULTS: Pathological analysis of the olfactory bulb revealed a marked reduction in the number of nerve processes with few intact olfactory glomeruli compared with an age-matched control. Specific immunohistochemical staining for the olfactory neuron-specific protein OMP, however, demonstrated the presence of intact axonal projections between the olfactory mucosa and the bulb. CONCLUSIONS: These results support the hypothesis that post-traumatic anosmia involves, at least in part, damage to peripheral olfactory nerve fibers with histological changes in the olfactory bulb. Potential mechanisms for the development of post-traumatic dysosmia are also discussed.

Adult↗

GABA(B) receptor-mediated stimulation of adenylyl cyclase activity in membranes of rat olfactory bulb.

Previous studies have shown that GABA(B) receptors facilitate cyclic AMP formation in brain slices likely through an indirect mechanism involving intracellular second messengers. In the present study, we have investigated whether a positive coupling of GABA(B) receptors to adenylyl cyclase could be detected in a cell-free preparation of rat olfactory bulb, a brain region where other Gi/Go-coupled neurotransmitter receptors have been found to stimulate the cyclase activity. The GABA(B) receptor agonist (-)-baclofen significantly increased basal adenylyl cyclase activity in membranes of the granule cell and external plexiform layers, but not in the olfactory nerve-glomerular layer. The adenylyl cyclase stimulation was therefore examined in granule cell layer membranes. The (-)-baclofen stimulation (pD2=4.53) was mimicked by 3-aminopropylphosphinic acid (pD2=4.60) and GABA (pD2=3.56), but not by (+)-baclofen, 3-aminopropylphosphonic acid, muscimol and isoguvacine. The stimulatory effect was counteracted by the GABA(B) receptor antagonists CGP 35348 (pA2=4.31), CGP 55845 A (pA2=7.0) and 2-hydroxysaclofen (pKi=4.22). Phaclofen (1 mM) was inactive. The (-)-baclofen stimulation was not affected by quinacrine, indomethacin, nordihydroguaiaretic acid and staurosporine, but was completely prevented by pertussis toxin and significantly reduced by the alpha subunit of transducin, a betagamma scavenger. The betagamma subunits of transducin stimulated the cyclase activity and this effect was not additive with that produced by (-)-baclofen. In the external plexiform and granule cell layers, but not in the olfactory nerve-glomerular layer, (-)-baclofen enhanced the adenylyl cyclase stimulation elicited by the neuropeptide pituitary adenylate cyclase activating polypeptide (PACAP) 38. Conversely, the adenylyl cyclase activity stimulated by either forskolin or Ca2+/calmodulin-(Ca2+/CaM) was inhibited by (-)-baclofen in all the olfactory bulb layers examined. These data demonstrate that in specific layers of rat olfactory bulb activation of GABA(B) receptors enhances basal and neurotransmitter-stimulated adenylyl cyclase activities by a mechanism involving betagamma subunits of Gi/Go. This positive coupling is associated with a widespread inhibitory effect on forskolin- and Ca2+/CaM-stimulated cyclic AMP formation.

Adenylate Cyclase Toxin↗

Neuromodulation by 5-hydroxytryptamine in the antennal lobe of the sphinx moth Manduca sexta.

Using intracellular recording techniques, we have begun to examine the effects of 5-hydroxytryptamine (5-HT) on antennal-lobe (AL) neurones in the brain of the adult moth Manduca sexta. 5-HT modulated the responses of local interneurones and projection neurones, which were recognized on the basis of well-established electrophysiological criteria, to primary synaptic input elicited by electrical stimulation of the ipsilateral antennal nerve. 5-HT applied at low concentration (10(-8) mol l-1) reduced the excitatory responses evoked by electrical stimulation of the antennal nerve, whereas at high concentration (10(-4) mol l-1), 5-HT enhanced the responses. At 10(-4) mol l1, 5-HT increased cell input resistance, led to broadening of action potentials and caused increased cell excitability in many AL neurones.

Action Potentials↗

FMRFamide-like immunoreactivity in the olfactory system responds to morphine treatment in the teleost Clarias batrachus: involvement of opiate receptors.

In view of the close relationship between the FMRF-related peptides and the central opiate-sensitive system, we investigated the effects of morphine, alone and in combination with naloxone, on the FMRFamide-like immunoreactivity in the olfactory system of the teleost, Clarias batrachus. In the olfactory system of normal and untreated fish, FMRFamide-like immunoreactivity was confined to the ganglion cells and fibers of the terminal nerve; the cells in the olfactory epithelium per se or the olfactory nerve were not immunoreactive. Intensely immunoreactive cells appeared in the olfactory epithelium following 2 h of intracranial morphine administration. FMRFamide-like immunoreactivity also appeared in the olfactory nerve fibers as they ran caudally and arborized in the glomerular layer of the bulb. However, immunoreactivity in the ganglion cells of the terminal nerve and the ensuing fibers was abolished, suggesting the transport/release of the immunoreactive material. Pretreatment with naloxone, a potent opiate receptor antagonist, reversed the effects of morphine, suggesting the involvement of opiate receptors in the regulation of the ganglion cells of the terminal nerve. The results provide initial immunocytochemical evidence in favor of a relationship between the opiates and FMRFamide-containing systems within the framework of the olfactory system.

Amino Acid Sequence↗

The role of nasal chemical senses in garter snake response to airborne odor cues from prey.

Garter snakes increase tongue-flick rates in the presence of airborne odorants (amyl acetate, limonene, earthworm wash vapor, fish water vapor, earthworms, goldfish) but not water vapor. Main olfactory (MO) nerve lesions result in loss of this tongue-flick rate elevation in the presence of airborne odorants. Vomeronasal (VN) nerve lesions result in a diminution of tongue-flick rate elevation. Garter snakes discriminate between the 2 arms of a maze containing airborne earthworm odor as compared with a blank control. Following MO nerve lesions, snakes fail to make this discrimination. Following VN nerve lesions or VN duct suture, snakes are initially able to make the discrimination but may eventually cease to prefer the prey-odor side of the maze and may stop eating prey rewards. These results support the idea that airborne odorants are detected by the main MO system and that such detection does not require a functional VN system.

Animals↗

Physical variables in the olfactory stimulation process.

Electrical recording from small twigs of nerve in a tortoise showed that olfactory, vomeronasal, and trigeminal receptors in the nose are responsive to various odorants. No one kind of receptor was most sensitive to all odorants. For controlled stimulation, odorant was caused to appear in a stream of gas already flowing through the nose. Of the parameters definable at the naris, temperature, relative humidity, and nature of inert gas had little effect on olfactory responses to amyl acetate, whereas odorant species, odorant concentration, and volume flow rate effectively determined the responses of all nasal chemoreceptors. An intrinsic variable of accessibility to the receptors, particularly olfactory, was demonstrated. Flow dependence of chemoreceptor responses is thought to reflect the necessity for delivery of odorant molecules to receptor sites. Since the olfactory receptors are relatively exposed, plateauing of the response with flow rate for slightly soluble odorants suggests an approach to concentration equilibrium in the overlying mucus with that in the air entering the naris. Accordingly, data for responses to amyl acetate were fitted with Beidler's (1954) taste equation for two kinds of sites being active. The requirement for finite aqueous solubility, if true, suggests substitution of aqueous solutions for gaseous solutions. A suitable medium was found and results conformed to expectations. Olfactory receptors were insensitive to variation of ionic strength, pH, and osmotic pressure.

Chemoreceptor Cells↗

Changes in glycosaminoglycans during regeneration of post-crush sciatic nerves of adult guinea pigs.

The glycosaminoglycans of sciatic nerves recovering from crush-injury were studied in adult guinea pigs and compared with those of non-injured mature neural tissues. The glycosaminoglycans were recovered from the 1,900 g supernatant and pellet of the tissue homogenates and assayed for hexuronate contents and susceptibilities to hyaluronidase, chondroitinase ABC, and nitrous acid. In the normal brain and central nerve tracts, the glycosaminoglycans were distributed both in the supernatant and pellet fractions; the brain showed a predominance of chondroitin sulphates but the tracts showed a predominance of heparan sulphates. Twice as much glycosaminoglycans were found in normal sciatic nerves, only in the pellet fraction and with heparan sulphate predominant. In the 2 weeks post-crush, progressive increase in hexuronate was observed, due mainly to additional chondroitin sulphate forms in the supernatant; the pellet fraction in the same period was however similar to the untreated controls in relative abundance of glycosaminoglycan classes and hexuronate content. At 4 weeks post-crush, although the total hexuronate returned to the control level, a significant proportion of glycosaminoglycans remained in the supernatant fraction. Evidence is thus provided for the need to modulate the glycosaminoglycan expression pattern in adult neural tissue to allow post-traumatic tissue remodelling and axonal regrowth.

Animals↗

Ultrastructural localization and identification of adrenergic and cholinergic nerve terminals in the olfactory mucosa.

Pharmacological and ultrastructural methods were used to demonstrate alpha-adrenergic regulation of secretory granule content of acinar cells of Bowman's glands and to localize and identify adrenergic and cholinergic axonal varicosities and terminals in the olfactory mucosa of the tiger salamander. The alpha-adrenergic agonist phenylephrine caused secretory granule depletion from Bowman's glands; the alpha-adrenergic antagonist phentolamine partially blocked this effect. These observations were quantified using light microscopic computer-assisted morphometric techniques. Both drugs caused morphological signs of electrolye/water transport. Adrenergic axonal varicosities were identified by the presence of small granular vesicles (SGVs, 45-60 nm in diameter) containing electron-dense material that was enhanced by 5-hydroxydopamine loading and chromaffin reaction fixation techniques. Throughout the lamina propria, small fascicles with axons containing SGVs as well as varicosities and terminals with SGVs were located adjacent to blood vessels, Bowman's gland acini, and melanocytes. Mean vesicle diameters at these sites were 54 +/- 7 nm, 50 +/- 9 nm, and 56 +/- 8 nm, respectively; varicosities were located approximately 0.1-1.0 microns from their presumed cellular targets. Axonal varicosities containing small agranular vesicles (AGVs, 65 +/- 8 nm in diameter), identified as cholinergic by their size and by the absence of electron-dense material after 5-hydroxydopamine loading and chromaffin reaction fixation, were located between adjacent acinar cells. In addition, adrenergic varicosities containing SGVs (56 +/- 6 nm in diameter) were found within 1 micron of blood vessels associated with Bowman's gland ducts and sustentacular cells near the base of the olfactory epithelium. These results characterize the ultrastructural basis for adrenergic and cholinergic regulation of vasomotor tone and secretion within the olfactory mucosa.

Adrenergic Fibers↗