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PubMed1997

Immunohistochemical demonstration of cytokeratin in human embryonic neurons arising from placodes.

Sensory neurons of the olfactory, trigeminal, facial, vestibulo-cochlear, glossopharyngeal and vagal nerves, and neurons migrating along the olfactory nerve to the brain have special anlagen, made up of placodes located in the epithelial layer. To investigate the characteristic phenotype of placode-derived neurons, immunohistochemical analysis of intermediate filaments was conducted on formalin-fixed human embryonic tissues. Neurons arising from placodes including luteinizing-hormone releasing hormone (LHRH) neurons migrating from the olfactory placode to the brain had immunoreactivity to antibodies specific to cytokeratin, AE1 and CAM5.2 during the embryonic stage. However, this immunoreactivity disappeared during the late embryonic to the post-embryonic stage and was not observed in the roots of these nerves in the post-natal stage. Immunoreactivity was detected in both the somata and processes, and the distribution differed from that described in rodent brain neurons. With this exception, no other human peripheral neurons, including spinal dorsal root ganglia, had immunoreactivity with anti-cytokeratin antibodies throughout the entire developmental stage. Although the cephalic neural crest also directly generates neurons to most of the cranial sensory ganglia, we could not find any evidence that it contributed to the genesis of cytokeratin-positive embryonic neurons. We concluded that cytokeratin is an intermediate filament common to human embryonic neurons of cephalic placodal origin and that this immunohistochemical marker may be useful in analyzing the developmental sequence of several congenital diseases involving the cranial nerves, such as Moebius syndrome and Goldenhar syndrome.

Adult↗
PubMedDate not supplied

Carnosine, nerve growth factor receptor and tyrosine hydroxylase expression during the ontogeny of the rat olfactory system.

The localizations of carnosine, nerve growth factor (NGF) receptor and tyrosine hydroxylase (TH) were studied in the embryonic and postnatal rat olfactory bulb and epithelium by means of single- and double-immunostaining methods. Tyrosine hydroxylase ontogeny was also evaluated at the mRNA level by in situ hybridization. All these molecules were expressed in the olfactory bulb but with different developmental patterns and cellular localization: carnosine immunoreactivity is seen from embryonic day 17 in primary olfactory neurons scattered in the nasal cavity and in fibres projecting from them to the olfactory bulb. Nerve growth factor-receptor immunoreactivity associated with small glial-like cells is visible in some glomeruli starting from the second day of postnatal life. At postnatal day 10 NGF-receptor immunoreactivity is extended to all glomeruli. Periglomerular neurons expressing TH mRNA and protein are present prenatally and their number sharply increases during the early postnatal development. Double-staining methods show that TH and NGF-receptor immunoreactivity do not overlap in cell bodies and processes. In addition, NGF-receptor immunoreactivity is not colocalized with carnosine. These findings definitely exclude NGF-receptor expression in periglomerular and primary olfactory neurons, suggesting that at least part of NGF-receptor expression in the olfactory bulb is associated with glial cells. In addition, they provide the first immunohistochemical data on carnosine ontogeny and confirm at the mRNA level previous studies on the ontogeny of TH protein.

Animals↗
PubMed2002

Atrial natriuretic peptide type C induces a cell-cycle switch from proliferation to differentiation in brain-derived neurotrophic factor- or nerve growth factor-primed olfactory receptor neurons.

With the discovery of postnatal stem cells within the brain, it has become important to understand how extracellular factors might affect the maturation of neuronal precursors in the postnatal brain. Neurotrophic factors are known to play a role in neuronal development but display pleiotrophic effects, in part because of their physiological interactions with other factors. One factor positioned to interact with neurotrophins in the brains of postnatal animals is atrial C-type natriuretic peptide (CNP). In this study, we used olfactory receptor neurons (ORNs) as a model, because their precursors demonstrate the most robust and functional postnatal neurogenesis of those systems thus far described. We examined the effects of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) and the interactions of these neurotrophins and CNP in postnatal olfactory neuronal precursors. Results obtained using mice with targeted deletion of the gene for BDNF indicated that BDNF is a neuroproliferation-inducing and survival factor for ORN precursors. These roles were confirmed in vitro using primary cultures of ORNs. NGF was found to be a proliferation-inducing factor but not a survival factor. The addition of CNP to either BDNF- or NGF-treated neuronal precursors resulted in an inhibition of proliferation and the promotion of maturation. These effects were accompanied by changes in cell-cycle proteins that suggest possible mechanisms for these effects. Thus, CNP may function in the postnatal brain to regulate the exit from the cell cycle in neuronal precursor cells.

Animals↗
PubMed1996

Luteinizing hormone-releasing hormone and innervation pathways in human prenatal nasal submucosa: factors of importance in evaluating Kallmann's syndrome.

A previous study has demonstrated that luteinizing hormone-releasing hormone (LHRH) is localized in the human bilateral vomeronasal organs in the nasal septum during a 4-week period of intrauterine life (22). The purpose of the present study was to elucidate the location of LHRH-expressing cells outside the vomeronasal organs, with special emphasis on the submucosa of the medial wall and roof of the nasal cavity. An additional aim was to study the innervation pathways in the same regions. Both regions can be affected in Kallmann's syndrome, which is characterized by hypogonadotropic hypogonadism (lack of LHRH) and often associated with anosmia. Histological sections of craniofacial regions (49 normal human fetuses, 6-19 weeks) were examined by immunohistochemical techniques for LHRH and for neuronal tissue (protein gene product 9.5, PGP 9.5). LHRH reactions were only seen in the septal submucosa extending from the vomeronasal organs to the olfactory bulb. There was a close spatiotemporal association between the occurrence of LHRH and neuronal tissue. From the rhino-olfactory epithelium separate nerve tissue extended to the olfactory bulb. It is suggested that the medial region of the nasal placode giving rise to the septal wall is always affected in Kallmann's syndrome, and in cases in which the phenotypic features are associated with anosmia, also the more lateral part of the nasal placode, from which the rhino-olfactory region originates, is affected.

Cell Movement↗
PubMed1984

Vagus nerve stimulation modifies the electrical activity of the olfactory bulb.

Evoked potential and unit activity recording techniques were used to study the effects of the vagus nerve stimulation on the olfactory bulb. A biphasic potential was evoked in the olfactory bulb by a single pulse delivered to the vagus nerve. Half of the neurons studied decreased discharge frequency after single pulse or train stimulation. The interval during which neurons ceased activity corresponded to the duration of the negative wave of the evoked potential. Responsive neurons were marked with horseradish peroxidase applied iontophoretically. Responsive neurons were located in the periglomerular layer of the olfactory bulb. These results suggest the existence of a vagus nerve-olfactory bulb pathway. The functional significance of this pathway is discussed.

Afferent Pathways↗
PubMed1992

Neurofilaments are prominent in bullfrog olfactory axons but are rarely seen in those of the tiger salamander, Ambystoma tigrinum.

Bullfrog olfactory axons show variable numbers (0-29) of structurally typical neurofilaments (NFs) about 10 nm in diameter. In studies tracking these NFs through serial sections of axons in cross section, they were found to be discontinuous, with a calculated average length of about 118 microns. In contrast to olfactory axons in bullfrogs, those of the tiger salamander Ambystoma trigrinum rarely show NFs. To be certain that the absence of NFs is a specific characteristic of olfactory axons, pieces of salamander spinal cord, optic nerve, and sciatic nerve were examined and found to contain typical NFs. To minimize the possibility that NFs in salamander olfactory axons were degraded or poorly fixed during preparation for electron microscopy, samples were fixed by using a variety of fixative and buffer combinations. To exclude the possibility that proteases degraded NFs during processing, prior to fixation some pieces of olfactory nerve were incubated in physiological saline containing protease inhibitors. Regardless of the preparation method, NFs were generally not seen in salamander olfactory axons. Extracts of salamander olfactory nerve were subjected to SDS-polyacrylamide gel electrophoresis (PAGE) and immunoblotting studies with monoclonal antibodies to the three NF subunit proteins. The immunoblots showed negligible or trace amounts of NF-L (light) and NF-H (heavy), while an NF-M (medium) protein having a molecular mass (Mr) of 160 kD was present in abundance. Extracts of salamander spinal cord, on the other hand, showed all three subunit proteins (with Mrs of 230, 160, and 77 kD). If one assumes that cells assemble structural elements to provide for a given function, the findings suggest that NFs in olfactory neurons of bullfrogs provide a function that may be missing in olfactory neurons of the salamander; the evidence also suggests that the absence of NFs in the salamander may be due to a deficiency in two of the three NF subunit proteins.

Ambystoma↗
PubMed1991

Differential expression of mRNAs for the NGF family of neurotrophic factors in the adult rat central olfactory system.

The cellular localization of mRNAs for nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 (NT3), in the rat central olfactory system was evaluated with in situ hybridization of 35S-labeled cRNA probes. In the main olfactory bulb, low levels of NGF and BDNF mRNA expression were detected. NGF mRNA was restricted to the glomerular region while BDNF mRNA was predominantly localized to the granule cell layer. No cellular hybridization to NT3 cRNA was seen. The accessory olfactory bulb did not express detectable levels of mRNA for any of the three related neurotrophic factors. Areas which receive olfactory bulb afferents expressed comparatively high levels of both NGF and BDNF mRNA. Cell labeling with cRNAs for NGF and BDNF occurred throughout the cellular layers of the anterior olfactory nucleus and in layers 2 and 3 of rostral piriform cortex. BDNF mRNA expression in these areas appeared more robust than that of NGF mRNA, while NT3 mRNA was not detectable. In contrast, tenia tecta exhibited dense labeling with the cRNAs for all three neurotrophic factors. The localization of NGF mRNA to primary target neurons of the olfactory nerve in the periglomerular region of the main olfactory bulb suggests that bulb cells may influence the ingrowth and continual turnover of olfactory sensory afferents. However, as there is a strong correlation between the distribution of neurotrophic factor mRNAs within rostral olfactory structures and the distribution of centrifugal cholinergic afferents, it is more likely that bulb-derived NGF, and possibly BDNF, act on the cholinergic neurons of the basal forebrain.

Animals↗
PubMed1992

Regulation of the low affinity receptor for nerve growth factor, p75NGFR, in the olfactory system of neonatal and adult rat.

Using MAb192, a monoclonal antibody to the rat low affinity receptor for nerve growth factor (p75NGFR), we determined the expression of p75NGFR in rat neonatal and adult olfactory system. In neonates and adults, we observed discrete p75NGFR-immunoreactivity (p75NGFR-ir) in the glomerular layer of the main olfactory bulb. The intensity and organization of glomerular p75NGFR-ir increased with age. This was in keeping with the general ontogeny of the main olfactory bulb. Generally, granule cells, mitral cells and periglomerular cells of the main olfactory bulb were not specifically stained. However, in early neonates, granule cells close to the lateral olfactory tract exhibited p75NGFR-ir. Additional specific staining was found in the olfactory receptor neurons of neonatal and adult olfactory neuroepithelium, the olfactory fascicles and in the glomeruli of the accessory olfactory bulb. The intensity, but not the organization, of specific staining in the accessory olfactory bulb increased as the animal matured. We believe that p75NGFR-ir in the olfactory system is associated with its unique capacity to regenerate its peripheral input to the main olfactory bulb. The presence of p75NGFR-ir in the accessory olfactory bulb would suggest a broader role for this protein. Here we discuss the implications of these findings with regards to nerve growth factor, other trophic molecules, and their receptors. The data presented provide a foundation for studies involving manipulation of regenerative phenomena while monitoring the expression of neurotrophic factors and their receptors.

Aging↗
PubMed2003

[Olfactory ensheathing cells enhance functional recovery of injured sciatic nerve].

OBJECTIVE: To investigate the effect of olfactory ensheathing cells (OECs) on functional recovery after sciatic nerve injury. METHODS: Upon silicone-tubulization of transected sciatic nerve in 30 adult rats. Thirty rats were divided into two groups(SAL group and OECs group); saline and OECs were injected into the silicone chamber in SAL group and in OECs group respectively. The status of functional recovery of injured sciatic nerve was observed by electrophysiological analysis, axon morphometry analysis. RESULTS: In OECs group on the 30th and the 90th days after sciatic nerve transection: 1. The latent period of CMAP shortened by 0.60 ms and 0.56 ms; the nerve conduction velocity promoted by 6.42 m/s and 5.36 m/s; the amplitude enhanced by 3.92 mv and 5.84 mv, respectively; 2. The HRP positive cells in lateral nucleus of spinal anterior horn increased by 11.63% and 25.01%; 3. The number of nerve fibers increased by 1,047/mm2 and 1,422/mm2 and the thickness of myelim sheath increased by 0.43 micron and 0.63 micron, respectively. CONCLUSION: The olfactory ensheathing cells are capable of promoting the functional recovery after peripheral nerve injury.

Action Potentials↗
PubMed2003

Presynaptic cyclic nucleotide-gated ion channels modulate neurotransmission in the mammalian olfactory bulb.

Cyclic nucleotide-gated channels (CNGCs) on the dendritic cilia of olfactory receptor neurons (ORNs) are critical for sensory transduction in the olfactory system. Do CNGCs also play a role in the axons and/or nerve terminals of ORNs? We find that the cyclic nucleotides cAMP and cGMP can both facilitate and depress synaptic transmission between olfactory nerve fibers and their targets in olfactory bulb glomeruli. Cyclic nucleotides increase intracellular Ca(2+) in ORN terminals and enhance spontaneous transmitter release; at higher concentrations, cyclic nucleotides depress evoked transmission by altering olfactory nerve excitability. Cyclic nucleotides have no effect on transmission or nerve excitability, however, in mice lacking olfactory CNGCs. Taken together, our results identify a novel role for presynaptic CNGCs in modulating neurotransmission.

Animals↗