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The optical spike.

Among the signs of activity in excitable membranes, the action current (electrical spike) has been extensively studied. Recently, a new approach with optical methods has been rewarding. In nerves, a transient, rapid change of light scanning, birefringence and induced fluorescence can be observed during the passage of the action current. These optical effects are synchronous with the electrical spike and are therefore called the optical spikes. Birefringence decreases during excitation in the giant axon of the squid, the walking nerves of Maia, the vagus nerve of the rabbit, but it increases in the olfactory nerve of the pike, which contains 4 million nonmedullated nerve fibres. Light scattering increases or decreases depending on the angle of observation. Vitally stained nerves with fluorescent probes show an increase and a shift in the wavelength distribution of the fluorescent spike.

Action Potentials↗

Experimental studies on the olfactory marker protein. I. Presence of the olfactory marker protein in tufted and mitral cells.

Partial, unilateral olfactory nerve section was performed in mice, and the behavior of the olfactory marker protein (OMP) studied, after this experimental manipulation, with the peroxidase--antiperoxidase method. The protein, which in normal mice is present only in mature olfactory sensory neurons, after unilateral lesion of the fila olfactoria was observed in mitral and tufted cells of both olfactory bulbs. Positive elements, present at 5 days postoperative, increased in number up to 30 days and some could still be detected at 60 days. The functional implications of this finding are briefly discussed.

Animals↗

Suckling dysfunction caused by defects in the olfactory system in genetic arhinencephaly mice.

Mouse newborns find their mother's nipples and suckle milk by themselves. It has been argued which sense organ they use when locating their mother's nipples to suckle milk. Olfactory or tactile sensory systems are primary candidates. In the present study, we investigated the trigeminal-whisker sensory and olfactory systems in genetic arhinencephaly mouse embryos (Pdn/Pdn). Pdn/Pdn newborns do not suckle milk and die within 1 day after birth. Dysfunction of nipple-searching behavior was clear in Pdn/Pdn newborns. Pdn/Pdn newborns had a complete developmental failure in the olfactory nerve projection to the central nervous system and no olfactory bulb architecture. The trigeminal-whisker system was intact in this strain. From the results of these experiments, it was suggested that the olfactory system is essential for nipple-searching behavior and suckling milk and that the trigeminal-whisker system is not able to substitute for the lack of olfactory input in mouse newborns.

Animals↗

Optical mapping of spatiotemporal emergence of functional synaptic connections in the embryonic chick olfactory pathway.

In order to understand the functional maturation of the CNS, it is essential to first describe the functional maturation of sensory processing. We have approached this topic by following the ontogenetic patterning of neural circuit formation related to cranial and spinal sensory input using voltage-sensitive dye imaging. In previous studies, we have described the functional maturation of synapses in brainstem/midbrain neural circuits. Here, we elucidate the functional maturation of forebrain circuits by investigating neural networks related to the olfactory nerve (N. I) of chicken embryo. In the isolated N. I-olfactory bulb-forebrain preparation, application of electrical stimulation to N. I elicited excitatory postsynaptic potential (EPSP)-related slow optical signals in the olfactory bulb. The slow signal was mainly mediated by glutamate, and was easily fatigued with repetitive stimuli because of the immaturity of synapses in the embryonic CNS. Ontogenetically, the slow signal was detected from the 6-day embryonic stage, suggesting that functional synaptic connections between N. I and olfactory bulb emerge around this stage. In addition, from the 8-day embryonic stage, another response area was discriminated within the forebrain, which corresponded to the higher-ordered nucleus of the olfactory pathway. In comparison with our previous studies concerning the functional development of other cranial nerve-related sensory nuclei in the embryonic brainstem and midbrain, these results suggest that the olfactory pathway is functionally generated in the early stages of development when neural networks related to other visceral and somatic sensory inputs are also in the process of developing.

Aging↗

FMRFamide-like immunoreactivity in the brain and pituitary of the goldfish, Carassius auratus.

The distribution of FMRFamide-like immunoreactivity was determined in the brain and the pituitary of the goldfish, Carassius auratus. Immunoreactive perikarya were observed in the olfactory nerve, nucleus entopeduncularis, nucleus anterioris periventricularis, nucleus posterioris periventricularis, lateral part of the nucleus lateralis tuberis pars posterioris, midbrain tegmentum, and medulla oblongata. Immunoreactive fibers were widely distributed in the brain, in particular in the ventral telencephalon and the hypothalamus. A few immunoreactive nerve fibers were observed in the pituitary. The findings are discussed in relation to male sexual behavior and the involvement of FMRFamide-like peptide in pituitary functions in the goldfish.

Animals↗

Olfactory responses of chum salmon to amino acids are independent of large differences in salt concentrations between fresh and sea water.

In chum salmon captured at the coastal sea and the natal river, the magnitudes of the olfactory nerve responses to the amino acids after perfusion of the olfactory epithelium with artificial pond water (APW) were similar to those after perfusion with artificial sea water (ASW), although the concentrations of Na+, Cl- and Ca2+ in ASW were 986, 430 and 27 times higher than those in APW, respectively. The findings suggest that the permeability of these ions across the apical membranes of olfactory cells do not essentially contribute to the transduction mechanism in the salmon.

Amino Acids↗

Astroglial ciliary neurotrophic factor mRNA expression is increased in fields of axonal sprouting in deafferented hippocampus.

Evidence that ciliary neurotrophic factor promotes axonal sprouting and regeneration in the periphery raises the possibility that this factor is involved in reactive axonal growth in the brain. In situ hybridization was used in the present study to determine whether ciliary neurotrophic factor mRNA expression is increased in association with axonal sprouting in deafferented adult rat hippocampus. In untreated rats, ciliary neurotrophic factor cRNA labeling density was high in the olfactory nerve, pia mater, and aspects of the ventricular ependyma and was relatively low within areas of white matter (fimbria, internal capsule) and select neuronal fields (hippocampal cell layers, habenula). After an entorhinal cortex lesion, hybridization was markedly increased in fields of anterograde degeneration, including most prominently the ipsilateral dentate gyrus outer molecular layer and hippocampal stratum lacunosum moleculare. Labeling in these fields was increased by 3 days postlesion, was maximal at 5 days, and returned to normal levels by 14 days. Double labeling demonstrated that, in both control and experimental tissue, ciliary neurotrophic factor mRNA was colocalized with glial fibrillary acidic protein immunoreactivity in astroglia, but it was not colocalized with markers for oligodendrocytes or microglia. These results demonstrate that astroglial ciliary neurotrophic factor expression is increased in fields of axonal and terminal degeneration and that increased expression is coincident with 1) increased insulin-like growth factor-1 and basic fibroblast growth factor expression and 2) the onset of reactive axonal growth. The synchronous expression of these glial factors in fields of deafferentation suggests the possibility of additive or synergistic interactions in the coordination of central axonal growth.

Afferent Pathways↗

Raman spectroscopy of nerve fibers. A study of membrane lipids under steady state conditions.

The molecular structures of different nerve fibers kept in good physiological conditions were studied by laser Raman spectroscopy. For myelinated nerves like the rat sciatic nerve, the Raman spectrum is dominated by bands due to the lipid component of the myelin sheath. The temperature dependence of these bands does not reveal any thermotropic phase transition between 0 and 40 degrees C. There is, however, with temperature, a linear increase in the intermolecular disorder that is accompanied by an increase in the number of gauche bonds of the phospholipid acyl chains. For unmyelinated nerves such as the lobster leg nerve, the C-H stretching region of the Raman spectrum is covered by bands arising from the protein component of the axoplasm. However, for the garfish olfactory nerve that has a high density of excitable membranes, phospholipid bands are observed and can be used as intrinsic structural probes of the excitable membranes. The relative intensity of these bands is also temperature dependent.

Animals↗

Axonal transport in the garfish optic nerve: comparison with the olfactory system.

Fast and slow axonal transports were studied in the optic nerve of the garfish and compared with previous studies on the olfactory nerve. The composition of fast-transport proteins was very similar in the two nerves. Although the velocity of fast transport was slightly lower in the optic nerve, there was a linear increase in velocity with temperature in both nerves. As in the olfactory nerve, only a single wave of slow-transport protein radioactivity moves along the nerve. The velocity of slow transport also increased linearly with temperature, but the coefficient was less than in the olfactory system. The composition of slow transport in the optic nerve was significantly different from that in the olfactory nerve, a finding reflecting the different cytoskeletal constituents of the two types of axons. The slow wave could be differentiated into several subcomponents, with the order of velocities being a 105-kilodalton protein and actin greater than tubulins and clathrin greater than fodrin much greater than neurofilaments. It can be concluded that the temperature dependence of fast and slow axonal transport in different nerves reflects the influence of temperature on the individual polypeptides constituting the various transport phases. The garfish optic nerve preparation may be advantageous for studies of axonal transport in retinal ganglion cell axons, because its great length avoids the complications of having to study transport in the optic tract or in material accumulating at the tectum.

Actins↗

Rapid axoplasmic transport of free leucine.

Axoplasmic transport of free 3H-leucine has been studied in vivo in the pike olfactory nerve following application of labeled leucine to the olfactory mucosa. A considerable amount of free 3H-leucine is transported at constant velocity along the axon in the form of a distinct peak. The maximum transport velocity for free 3H-leucine is the same as for rapidly transported 3H-protein (130 and 135 mm/day, respectively, at 19 degrees C). Microtubule inhibitors block or significantly reduce the amount of free 3H-leucine transported, but do not influence the transport velocity. Disruption of the oxygen supply abolishes free 3H-leucine transport, so that this phenomenon cannot be explained by diffusion. The amount of free leucine in the rapidly moving peak decreases with time and distance along the axon and is not detectable after 5 h or more. The transported 3H-leucine is not derived from the circulation or from proteolysis of rapidly transported proteins. This study may help to resolve the controversy over the axoplasmic transport of free amino acids since it shows that free 3H-leucine is transported rapidly but does not travel by rapid axoplasmic transport to the end of axons longer than about 30 mm.

Animals↗

Developmental expression of reactivity to monoclonal antibodies generated against olfactory epithelia.

The developmental expression of immunocytochemical reactivity to 3 monoclonal antibodies (Mabs Neu 4, Neu 5, and Neu 9) that were generated against adult rat olfactory epithelium was examined in olfactory tissues of embryonic rats. Tissues examined included the nasal olfactory epithelium, nerve, and olfactory bulb, as well as vomeronasal epithelium and nerve. Reactivity patterns of these Mabs in adult rats have been described previously (Hempstead and Morgan, 1985a). All 3 Mabs show reactivity on the cell surfaces of neurons, axons, and dendrites of the olfactory epithelium proper. Neu 5 alone shows reactivity on the dendritic knobs, site of transduction of the olfactory stimuli. These reactivities appear early, suggesting developmentally significant roles for the antigens to these Mabs. For Neu 5 and Neu 9 initial reactivity occurs on outgrowing olfactory axons at E13. Dendritic and perikaryal reactivities begin appearing at E14. For Neu 4 initial reactivity occurs simultaneously on olfactory neuronal perikarya, axons, and dendrites at E14. Reactivity also occurs on cells that migrate from the olfactory epithelium and are associated with the olfactory nerves. Within the developing olfactory bulb, Neu 5 behaves as a general cell-surface marker. Neu 4 and Neu 9, however, show enhanced reactivity in the glomerular layer after the onset of synaptogenesis. Reactivity is also seen in the nasal respiratory epithelium and in the vomeronasal epithelia and nerve. Neu 5 and several antibodies to rat neural cell adhesion molecules (N-CAMs) show similar, although not identical, immunohistochemical staining patterns. They also react with the same bands in Western blots of brain membrane preparations. Western blots of Neu 5-reactive material also show developmental and spatial correlations of apparent molecular-weight distributions expected of N-CAM-like components as well.

Animals↗

Errors in lamina growth of primary olfactory axons in the rat and mouse olfactory bulb.

In the adult olfactory nerve pathway of rodents, each primary olfactory axon forms a terminal arbor in a single glomerulus in the olfactory bulb. During development, axons are believed to project directly to and terminate precisely within a glomerulus without any exuberant growth or mistargeting. To gain insight into mechanisms underlying this process, the trajectories of primary olfactory axons during glomerular formation were studied in the neonatal period. Histochemical staining of mouse olfactory bulb sections with the lectin Dolichos biflorus-agglutinin revealed that many olfactory axons overshoot the glomerular layer and course into the deeper laminae of the bulb in the early postnatal period. Single primary olfactory axons were anterogradely labelled either with the lipophilic carbocyanine dye, 1,1'-dioctodecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI), or with horse-radish peroxidase (HRP) by localized microinjections into the nerve fiber layer of the rat olfactory bulb. Five distinct trajectories of primary olfactory axons were observed in DiI-labelled preparations at postnatal day 1.5 (P1.5). Axons either coursed directly to and terminated specifically within a glomerulus, branched before terminating in a glomerulus, bypassed glomeruli and entered the underlying external plexiform layer, passed through the glomerular layer with side branches into glomeruli, or branched into more than one glomerulus. HRP-labelled axon arbors from eight postnatal ages were reconstructed by camera lucida and were used to determine arbor length, arbor area, and arbor branch number. Whereas primary olfactory axons display errors in laminar targeting in the mammalian olfactory bulb, axon arbors typically achieve their adult morphology without exuberant growth. Many olfactory axons appear not to recognize appropriate cues to terminate within the glomerular layer during the early postnatal period. However, primary olfactory axons exhibit precise targeting in the glomerular layer after P5.5, indicating temporal differences in either the presence of guidance cues or the ability of axons to respond to these cues.

Aging↗

Expression of a member of tumor necrosis factor receptor superfamily, TROY, in the developing olfactory system.

TROY is a recently identified member of the tumor necrosis factor (TNF) receptor superfamily. We have previously reported that TROY induces the activation of nuclear factor kappaB via TNF receptor-associated factor 2, 5, and 6, and is strongly expressed in the developing central nervous system, including the olfactory bulb. In this study, we investigated the detailed cellular characterization of TROY-expressing cells in the developing olfactory system of mice using in situ hybridization and immunohistochemistry. Both mRNA and protein of TROY were first detected in the olfactory nerve layer (ONL) of the olfactory bulb at embryonic day 13.5. During late embryogenesis, TROY expression was most intense in the inner ONL (ONL-i). In the postnatal olfactory bulb, TROY-expressing cells were also detected in the glomerular layer (GL), in addition to the ONL-i. The double-immunofluorescence method demonstrated that TROY was expressed in olfactory ensheathing cells (OECs) of the ONL-i, which were positive for neuropeptide Y (NPY), but neither S-100 nor p75 low-affinity nerve growth factor receptor. Some TROY-expressing cells in the ONL-i were observed with the astrocyte-like phenotype (GFAP+/NPY-). In addition, TROY was also detected in GFAP+ glial cells of the GL. Thus, TROY was expressed in some specific subsets of glial cells in the olfactory bulb, including OECs, suggesting that TROY may play some roles in the developing and adult olfactory system.

Animals↗

Carnosine-like immunoreactivity in the olfactory bulb of the rat: an electron microscopic study.

Carnosine-immunoreactive primary olfactory nerve terminals are demonstrated in the glomerular layer of the rat olfactory bulb by immunoelectron microscopy. Asymmetrical synapses between dendrites of mitral/tufted cells and that of periglomerular cells could be observed. In the accessory olfactory system, carnosine-like immunoreactivity is also detected in the vomeronasal neurons.

Animals↗

Interaction of olfactory ensheathing cells with astrocytes may be the key to repair of tract injuries in the spinal cord: the 'pathway hypothesis'.

Transplantation of cultured adult olfactory ensheathing cells has been shown to induce anatomical and functional repair of lesions of the adult rat spinal cord and spinal roots. Histological analysis of olfactory ensheathing cells, both in their normal location in the olfactory nerves and also after transplantation into spinal cord lesions, shows that they provide channels for the growth of regenerating nerve fibres. These channels have an outer, basal lamina-lined surface apposed by fibroblasts, and an inner, naked surface in contact with the nerve fibres. A crucial property of olfactory ensheathing cells, in which they differ from Schwann cells, is their superior ability to interact with astrocytes. When confronted with olfactory ensheathing cells the superficial astrocytic processes, which form the glial scar after lesions, change their configuration so that their outer pial surfaces are reflected in continuity with the outer surfaces of the olfactory ensheathing cells. The effect is to open a door into the central nervous system. We propose that this formation of a bridging pathway may be the crucial event by which transplanted olfactory ensheathing cells allow the innate growth capacity of severed adult axons to be translated into regeneration across a lesion so that functionally valuable connections can be established.

Animals↗

ELECTRICAL ACTIVITY AND HISTOLOGICAL CHANGE IN THE DEGENERATING OLFACTORY EPITHELIUM.

Electrical activity and histological changes were studied in the degenerating olfactory epithelium of the bullfrog after the olfactory nerve had been sectioned. After nerve section, the electrical responses to odors disappeared in the olfactory epithelium in 8 days in the summer, in 11 days in the early autumn, and in 16 days in the early winter. In the degenerating olfactory epithelium a striking decrease in the number of olfactory cells was found, but not of supporting cells. The ratio of the number of olfactory cells to that of supporting cells was found to decrease from 5 or 6 to below 2 after the nerve section. At a ratio below 2, the electrical responses to odor disappeared. The histological changes in the bullfrog are compared with those in the mouse and rabbit. The localization of the olfactory pigment and the electrical activity of the supporting cell are discussed. It was concluded that all three types of responses to odors originate from the activity of the olfactory cell.

Animals↗

Male hamster copulatory responses to a high molecular weight fraction of vaginal discharge: effects of vomeronasal organ removal.

The importance of the vomeronasal (accessory olfactory) system for the copulatory responses of male hamsters to a high molecular weight fraction (HMF) of vaginal discharge was assessed in animals that had their vomeronasal organs (VNO) removed. These organs were extirpated bilaterally using an oral approach through the palate so as to eliminate the peripheral afferents to the accessory olfactory bulb (AOB) with minimal or no damage to the main olfactory system. The selective peripheral deafferentation procedure was verified by applying horseradish peroxidase intranasally following intraperitoneal injections of epinephrine to facilitate the vomeronasal pumping mechanism that draws fluids into the VNO. Heavy, bilateral anterograde labeling was evident in the olfactory nerve afferents within the main olfactory bulb of males that had their VNO removed and of animals that received sham surgery. Sham-operated males also had heavy, bilateral labeling in the vomeronasal nerve afferents within the AOB, whereas no such labeling occurred among animals with bilateral removal of the VNO. In sham-operated animals, both the HMF and the unfractionated discharge significantly increased the incidence of intromission attempts toward anesthetized males (surrogate females) whose hindquarters were scented with these stimuli. The unfractionated discharge also produced a significant elevation of overt copulatory behavior in males with selective peripheral deafferentation of the vomeronasal system, whereas the HMF did not facilitate copulatory behavior in these animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Reproduction phase-related expression of GnRH-like immunoreactivity in the olfactory receptor neurons, their projections to the olfactory bulb and in the nervus terminalis in the female Indian major carp Cirrhinus mrigala (Ham.).

The reproductive biology of the Indian major carp Cirrhinus mrigala is tightly synchronized with the seasonal changes in the environment. While the ovaries show growth from February through June, the fish spawn in July-August to coincide with the monsoon; thereafter the fish pass into the postspawning and resting phases. We investigated the pattern of GnRH immunoreactivity in the olfactory system at regular intervals extending over a period of 35 months. Although no signal was detected in the olfactory organ of fish collected from April through February following year, distinct GnRH-like immunoreactivity appeared in the fish collected in March. Intense immunoreactivity was noticed in several olfactory receptor neurons (ORNs) and their axonal fibers as they extend over the olfactory nerve, spread in the periphery of the olfactory bulb (OB), and terminate in the glomerular layer. Strong immunoreactivity was seen in some fascicles of the medial olfactory tracts extending from the OB to the telencephalon. Some neurons of the ganglion cells of nervus terminalis showed GnRH immunostaining during March; no immunoreactivity was detected at other times of the year. Plexus of GnRH immunoreactive fibers extending throughout the bulb represented a different component of the olfactory system; the fiber density showed a seasonal pattern that could be related to the status of gonadal maturity. While it was highest in the prespawning phase, significant reduction in the fiber density was noticed in the fish of spawning and the following regressive phases. Taken together the data suggest that the GnRH in the olfactory system of C. mrigala may play a major role in translation of the environmental cues and influence the downstream signals leading to the stimulation of the brain-pituitary-ovary axis.

Animals↗