Endonasal anatomy of the olfactory neural network: Surgical implications.

OBJECTIVES/HYPOTHESIS: Define the anatomic distribution of the olfactory filaments within specific mucosal regions of the nasal cavity. STUDY DESIGN: Cadaveric study. METHODS: Seventeen cadaveric specimens (34 sides) were dissected to study the anatomical distribution and density of olfactory fila within different regions of the nasal cavity. Olfactory fila were dissected retrogradely to their point of entry into the anterior cranial fossa through the cribriform plate. Anatomic relationships among various components of the olfactory system and their corresponding arterial supply were determined subjectively. RESULTS: The highest density of olfactory fila was found at the mucosa of the ethmoid roof and superior turbinates. Olfactory fila were found at regions not previously considered to be part of the olfactory system: lateral wall of the nose, ethmoidal bullae, and between the os sphenoidale and arc of the posterior choana. Furthermore, at the septum, 20% of the olfactory fila crossed contralaterally before exiting the nose. The anterior ethmoidal arteries were the primary blood supply to the olfactory epithelium. CONCLUSIONS: This study suggests that olfactory filaments extend beyond previously established boundaries. These findings may have clinical implications regarding oncologic resections and could serve as the foundation for the development of techniques that better preserve olfactory function. LEVEL OF EVIDENCE: NA Laryngoscope, 2473-2477, 2018.

Neurophysiological Identification of Cranial Nerves During Endoscopic Endonasal Surgery of Skull Base Tumors: Pilot Study Technical Report.

INTRODUCTION: Intraoperative identification of cranial nerves is crucial for safe surgery of skull base tumors. Currently, only a small number of published papers describe the technique of trigger electromyography (t-EMG) in endoscopic endonasal removal of such tumors. OBJECTIVE: To assess the effectiveness of t-EMG in preventing intraoperative cranial nerve damage in endoscopic endonasal surgery of skull base tumors. MATERIALS AND METHODS: Nine patients were operated on using the endoscopic endonasal approach within a 1-year period. The tumors included large skull base chordomas and trigeminal neurinomas localized in the cavernous sinus. During the surgical process, cranial nerve identification was carried out using monopolar and bipolar t-EMG methods. Assessment of cranial nerve functional activity was conducted both before and after tumor removal. RESULTS: We mapped 17 nerves in 9 patients. Third, fifth, and sixth cranial nerves were identified intraoperatively. There were no cases of postoperative functional impairment of the mapped cranial nerves. In one case we were unable to get an intraoperative response from the fourth cranial nerve and observed its postoperative transient plegia (the function was normal before surgery). CONCLUSION: t-EMG allows surgeons to control the safety of cranial nerves both during and after skull base tumor removal.

[АNTINOCICEPTIVE PROTECTION ON A STAGE OF CONCLUDING OF SURGICAL INTERVENTION AND EARLY POSTOPERATIVE PERIOD].

Efficacy of antinociceptive defense at the terminal period of operation and in early (6 h) postoperative period, using additional injection of phentanil, paracetamol and nalbufin in anesthesiological support, and applying sevofluran in 107 patients, оperated on facial skull, in 2 stage of operative risk in accordance to ASA, was a nalyzed. Insufficient antinociceptive protection at the end of operation and in early postoperative period while using phentanil and nonsteroidal antiinflammatory medicines only for anesthesia, was established, basing on analysis of hemodynamic indices, pain syndrome severity and indices of metabolic stress. Application of paracetamol have promoted raising of the antinociceptive protection efficacy during short period (up to 2 h) only. Prescription of nalbufin have had guaranteed enhanced efficacy and duration of antinociceptive protection in early postoperative period, that's why its wide application is recommended.

A new method of identifying the posterior inferior nasal nerve: implications for posterior nasal neurectomy.

INTRODUCTION: Posterior nasal neurectomy is an effective way of treating recalcitrant rhinitis. The aim of this study is to describe the anatomic relationship between the posterior inferior nasal nerve (PINN) and the structures that might be important for posterior nasal neurectomy. MATERIALS AND METHODS: An anatomic study was conducted in a university hospital dissection laboratory with 15 formalin-fixed, sagittally cut adult cadaver heads. The distance between PINN and (1) nasal sill, (2) maxillary sinus ostium, (3) posterior fontanel, (4) torus tubarius, and (5) crista ethmoidalis was measured and the location of PINN with respect to the sphenopalatine artery was assessed to define the exact location of PINN. RESULTS: The mean distance between PINN and nasal sill (56.4 mm), maxillary sinus ostium (27 mm), posterior fontanel (12.5 mm), torus tubarius (13 mm), and crista ethmoidalis (8 mm) was determined. PINN was found consistently posterior to the sphenopalatine artery where the inferior turbinate attaches to the lateral nasal wall. CONCLUSION: Instead of finding PINN around the sphenopalatine foramen, PINN can be located more easily posterior to the sphenopalatine artery where the inferior turbinate attaches to the lateral nasal wall without cauterizing the sphenopalatine artery.

The membrane proteome of sensory cilia to the depth of olfactory receptors.

In the nasal cavity, the nonmotile cilium of olfactory sensory neurons (OSNs) constitutes the chemosensory interface between the ambient environment and the brain. The unique sensory organelle facilitates odor detection for which it includes all necessary components of initial and downstream olfactory signal transduction. In addition to its function in olfaction, a more universal role in modulating different signaling pathways is implicated, for example, in neurogenesis, apoptosis, and neural regeneration. To further extend our knowledge about this multifunctional signaling organelle, it is of high importance to establish a most detailed proteome map of the ciliary membrane compartment down to the level of transmembrane receptors. We detached cilia from mouse olfactory epithelia via Ca(2+)/K(+) shock followed by the enrichment of ciliary membrane proteins at alkaline pH, and we identified a total of 4,403 proteins by gel-based and gel-free methods in conjunction with high resolution LC/MS. This study is the first to report the detection of 62 native olfactory receptor proteins and to provide evidence for their heterogeneous expression at the protein level. Quantitative data evaluation revealed four ciliary membrane-associated candidate proteins (the annexins ANXA1, ANXA2, ANXA5, and S100A5) with a suggested function in the regulation of olfactory signal transduction, and their presence in ciliary structures was confirmed by immunohistochemistry. Moreover, we corroborated the ciliary localization of the potassium-dependent Na(+)/Ca(2+) exchanger (NCKX) 4 and the plasma membrane Ca(2+)-ATPase 1 (PMCA1) involved in olfactory signal termination, and we detected for the first time NCKX2 in olfactory cilia. Through comparison with transcriptome data specific for mature, ciliated OSNs, we finally delineated the membrane ciliome of OSNs. The membrane proteome of olfactory cilia established here is the most complete today, thus allowing us to pave new avenues for the study of diverse molecular functions and signaling pathways in and out of olfactory cilia and thus to advance our understanding of the biology of sensory organelles in general.

Therapeutic effectiveness of endoscopic vidian neurectomy for the treatment of vasomotor rhinitis.

CONCLUSION: Our results indicate that vidian neurectomy may be recommended as an effective method for the treatment of vasomotor rhinitis (VMR). OBJECTIVE: The aim of this work was to study the feasibility and effectiveness of vidian neurectomy treatment under the nasal endoscope for VMR. METHODS: The study included 45 patients with VMR. They were all assigned to functional endoscopic surgery with vidian neurectomy. RESULTS: The obtained data showed that, using the rhinoconjunctivitis quality of life questionnaire, vidian neurectomy treatment relieved the symptoms of VMR in 82.2% of the patients. Vidian neurectomy also led to the reduction of expression of several cytokines, including vasoactive intestinal polypeptide, calcitonin gene-related peptide, substance P, interleukin (IL)-4, and IL-5.

Endoscopic approach to the trigeminal nerve: an anatomic study.

OBJECTIVE: To describe an endoscopic perspective of the surgical anatomy of the trigeminal nerve. METHODS: Nine adult cadaveric heads were dissected endoscopically. RESULTS: Opening the pterygopalatine fossa is important because many key anatomical structures (V2, pterygopalatine ganglion, vidian nerve) can be identified and traced to other areas of the trigeminal nerve. From the pterygopalatine ganglion, the maxillary nerve and vidian nerve can be identified, and they can be traced to the gasserian ganglion and internal carotid artery. An anteromedial maxillectomy increases the angle of approach from the contralateral nares due to an increase in diameter of the piriform aperture, and provides excellent access to the mandibular nerve, the petrous carotid, and the cochlea. CONCLUSIONS: Identification of key anatomical structures in the pterygopalatine fossa can be used to identify other areas of the trigeminal nerve, and an anteromedial maxillectomy is necessary to expose the ipsilateral mandibular nerve and contralateral cranial level of the trigeminal nerve.

Unmyelinated fibers of the anterior ethmoidal nerve in the rat co-localize with neurons in the medullary dorsal horn and ventrolateral medulla activated by nasal stimulation.

The anterior ethmoidal nerve (AEN) innervates the nasal passages and external nares, and serves as the afferent limb of the nasopharyngeal and diving responses. However, although 65% of the AEN is composed of unmyelinated fibers, it has not been determined whether this afferent signal is carried by unmyelinated or myelinated fibers. We used the transganglionic tracers WGA-HRP, IB4-HRP, and CTB-HRP to trace the central projections of the AEN of the rat. Interpretation of the labeling patterns suggests that AEN unmyelinated fibers project primarily to the ventral tip of the ipsilateral medullary dorsal horn (MDH) at the level of the area postrema. Other unmyelinated projections were to the ventral paratrigeminal nucleus and ventrolateral medulla, specifically the Bötzinger and RVLM/C1 regions. Myelinated AEN fibers projected to the ventral paratrigeminal and mesencephalic trigeminal nuclei. Stimulating the nasal passages of urethane-anesthetized rats with ammonia vapors produced the nasopharyngeal response that included apnea, bradycardia and an increase in arterial blood pressure. Central projections of the AEN co-localized with neurons within both MDH and RVLM/C1 that were activated by nasal stimulation. Within the ventral MDH the density of AEN terminal projections positively correlated with the rostral-caudal location of activated neurons, especially at and just caudal to the obex. We conclude that unmyelinated AEN terminal projections are involved in the activation of neurons in the MDH and ventrolateral medulla that participate in the nasopharyngeal response in the rat. We also found that IB4-HRP was a much less robust tracer than WGA-HRP.

Direct transport of VEGF from the nasal cavity to brain.

The aim of the present study was to assess the potential of delivering VEGF directly into the central nervous system (CNS) following intranasal administration. Adult Sprague-Dawley rats were randomized into two groups, given [(125)I]-VEGF intranasally or intravenously. VEGF was intranasally administered in both nares alternately, the single dose is 10 microl with time interval of 2 min for about 18.5 min. The intravenous (IV) group was treated with 100 microl [(125)I]-VEGF intravenously. Thirty minutes after administration, rats were killed following blood sample collections, then the brains were removed, and olfactory bulb, striatum corpora, cortex, thalamus, pons, cerebella, medulla, hippocampus, cervical cord and other tissues were collected, weighted, under auto gamma counting and autoradiography analysis. Cisternal sampling of cerebrospinal fluid (CSF) was performed in an additional group of animals. Both gamma counting and high resolution phosphor imaging of tissue sections showed that intranasal administration of [(125)I]-VEGF resulted in substantial delivery throughout the CNS. The highest CNS tissue concentration following IN delivery was found in the trigeminal nerve, followed by the optic nerve, olfactory bulbs, olfactory tubercle, striatum, medulla, frontal cortex, midbrain, pons, appendix cerebri, thalamus, hippocampus, cerebellum. Intranasal administration of [(125)I]-VEGF also targeted the deep cervical lymph nodes. CSF did not contain [(125)I]-VEGF following intranasal administration. Intravenous [(125)I]-VEGF resulted in blood and peripheral tissue exposure higher concentrations than that intranasal administration, but CNS concentrations were significantly lower. The results suggest intranasally delivered VEGF can bypass the blood-brain barrier via olfactory- and trigeminal-associated extracellular pathways to directly entry into the CNS. Intranasal administration of VEGF may provide an effective way for the treatments of CNS diseases.

Recordings of the optical intrinsic signal from the middle turbinate in response to olfactory and trigeminal stimulation: a pilot study.

Responses from the middle turbinate elicited by olfactory and trigeminal stimuli were studied using the intrinsic optical signal (IOS) recording technique. Nasal cavity was illuminated by 617 nm light. Olfactory (H2S) or trigeminal (CO2) stimuli of 5-s duration were presented using a computer-controlled olfactometer; IOS responses were captured by a special camera. Averages across five individual IOS recordings were analyzed. When the nasal cavity was exposed to H2S, a significant change of the IOS was found; responses to CO2 were even more pronounced. The present results argue for the idea that the IOS is an indicator of intranasal chemosensory activation.

Fos expression in the brainstem nuclei evoked by nasal air-jet stimulation in rats.

BACKGROUND: Noxious stimulation of the nasal mucosa may induce protective reflexes in the upper airway in rats. Previously, we have reported that nasal air-jet stimulation increases the activities of the laryngeal muscles in decerebrate cats; however, the neuronal mechanism of this phenomenon still is not clarified. METHODS: After the application of nasal air-jet stimulation for 2 hours, we investigated the distribution of Fos-positive cells (FPCs) throughout the medulla compared with sham-operated rats using Fos immunoreactivity. RESULTS: FPCs in the spinal trigeminal nucleus, the parvocellular reticular nucleus, and the nucleus of the solitary tract were more frequent than the sham-operated rats. CONCLUSION: These results suggest that the afferents induced by air-jet stimulation were conveyed to these FPCs and that some of these cells might participate in the augmentation of laryngeal muscle activities during nasal air-jet stimulation.

The olfactory organ modulates gonadotropin-releasing hormone types and nest-building behavior in the tilapia Oreochromis niloticus.

Direct olfactory inputs to any of the known gonadotropin-releasing hormone (GnRH) containing neurons have not been demonstrated. Therefore, the rationale of this study was to examine whether olfactory inputs might in some way interact with the GnRH system(s) to synchronize reproductive behaviors. In order to establish this, we used anosmic mature male tilapia to investigate changes in reproductive behaviors, gonadal morphology, and GnRH1, GnRH2, and GnRH3 cellular morphology and change in GnRH mRNA levels by real-time polymerase chain reaction. Bilateral removal of the olfactory rosettes followed by occlusion of the nasal cavity (ORX) inhibited nest-building behavior, but had no effect on aggressive and sexual behaviors or gonadal morphology. ORX failed to alter the morphological features of GnRH1, GnRH2, and GnRH3 (cell number, size, GnRH optical density), but significantly decreased copies of GnRH1 and GnRH2 mRNAs. GnRH immunoreactive fibers were not evident in the olfactory nerve and rosettes. DiI application to the olfactory nerve labeled inputs primarily to the glomerular layer of the olfactory bulbs and extrabulbar inputs to the forebrain but not to GnRH neurons. These results provide evidence that the olfactory rosette is crucial for modulating nest-building behavior through second-order olfactory pathways interacting with GnRH1 and GnRH2 neuronal systems.

Evidence for the disproportionate mapping of olfactory airspace onto the main olfactory bulb of the hamster.

Olfactory receptor neurons (ORNs) project to the rodent main olfactory bulb (MOB) from spatially distinct air channels in the olfactory recesses of the nose. The relatively smooth central channels of the dorsal meatus map onto the dorsal MOB, whereas the highly convoluted peripheral channels of the ethmoid turbinates project to the ventral MOB. Medial and lateral components of each projection stream innervate the medial and lateral MOB, respectively. To ascertain whether such topography entails the disproportionate representation seen in other sensory maps, we used disector-based stereological techniques in hamsters to estimate the number of ORNs associated with each channel in the nose and the number of their targets (glomeruli and mitral and tufted cells) in corresponding divisions of the MOB. Each circumferential half of the MOB (dorsal/ventral, medial/lateral) contained about 50% of the 3,100 glomeruli and about 50% of the 160,000 mitral and tufted cells per bulb. We found equivalent numbers of ORNs with dendritic knobs in the medial and lateral channels (4.5 million each). However, the central channels had only 2 million knobbed ORNs, whereas the peripheral channels had 7 million. Thus, there is a disproportionate mapping of the central-peripheral axis of olfactory airspace onto the dorsal-ventral axis of the MOB, encompassing a greater than threefold variation in the average convergence of ORNs onto MOB secondary neurons. We hypothesize that the disproportionate projections help to optimize chemospecific processing by compensating, with differing sensitivity, for significant variation in the distribution and concentration of odorant molecules along the olfactory air channels during sniffing.

Solitary chemoreceptor cells in the nasal cavity serve as sentinels of respiration.

Inhalation of irritating substances leads to activation of the trigeminal nerve, triggering protective reflexes that include apnea or sneezing. Receptors for trigeminal irritants are generally assumed to be located exclusively on free nerve endings within the nasal epithelium, requiring that trigeminal irritants diffuse through the junctional barrier at the epithelial surface to activate receptors. We find, in both rats and mice, an extensive population of chemosensory cells that reach the surface of the nasal epithelium and form synaptic contacts with trigeminal afferent nerve fibers. These chemosensory cells express T2R "bitter-taste" receptors and alpha-gustducin, a G protein involved in chemosensory transduction. Functional studies indicate that bitter substances applied to the nasal epithelium activate the trigeminal nerve and evoke changes in respiratory rate. By extending to the surface of the nasal epithelium, these chemosensory cells serve to expand the repertoire of compounds that can activate trigeminal protective reflexes. The trigeminal chemoreceptor cells are likely to be remnants of the phylogenetically ancient population of solitary chemoreceptor cells found in the epithelium of all anamniote aquatic vertebrates.

A subpopulation of nervus terminalis neurons projects to the olfactory mucosa in Xenopus laevis.

Biocytin application to the normal or zinc sulfate-treated nasal cavity of Xenopus laevis was used to trace retrogradely neurons associated with the terminal nerve (TN). Immunocytochemistry was conducted to identify the relationship of gonadotropin-releasing hormone-immunoreactive (GnRH-ir) TN neurons with biocytin-labeled neurons. Neurons that accumulated biocytin were located in olfactory nerve fascicles close to the olfactory mucosa lining the caudal, medial, and rostral walls of the principal cavity. GnRH-ir fibers were observed only in the olfactory nerve fascicle projecting to the rostral edge of the principal cavity. In addition, GnRH-ir fibers did not contact biocytin-labeled TN neurons. We hypothesize that these two classes of neurons represent separate components of the TN.

Function of gonadotropin-releasing hormone in olfaction.

Gonadotropin-releasing hormone (GnRH) is present within neurons of the nervus terminalis, the zeroeth cranial nerve. In all vertebrate species, except in sharks where it is a separate nerve, the nervus terminalis consists of a chain of neurons embedded within olfactory or vomeronasal nerves in the nasal cavity. The function of the GnRH component of the nervus terminalis is thought to be neuromodulatory. Our research on GnRH effects on olfaction confirms this hypothesis. The processes of GnRH neural cell bodies located within chemosensory nerves project centrally into the ventral forebrain and peripherally into the lamina propria of the nasal chemosensory mucosa. GnRH receptors are expressed by chemosensory neurons as shown by RT-PCR/Southern blotting and GnRH agonist binding studies. Patch-clamp studies have shown that GnRH alters the responses of isolated chemosensory neurons to natural or electrophysiological stimulation through the modulation of voltage-gated and receptor-gated channels. Behavioral experiments demonstrate that interfering with the nasal GnRH system leads to deficits in mating behavior. These studies suggest that the function of the intranasal GnRH system is to modify olfactory information, perhaps at reproductively auspicious times. We speculate that the purpose of this altered olfactory sense is to make pheromones more detectable and salient.

Midline nasal tissue influences nestin expression in nasal-placode-derived luteinizing hormone-releasing hormone neurons during development.

Neurons differentiating into the luteinizing hormone-releasing hormone (LHRH) neuroendocrine phenotype are derived from the nasal placode. Cells within the vomeronasal organ anlage that turn on LHRH gene and peptide expression subsequently migrate into the forebrain where they influence reproductive function. The molecular and cellular cues regulating differentiation and migration of these cells are unknown. Discovery of developmental markers can indicate proteins directing or associated with differentiation. Analysis of such markers after manipulation of external cues can elucidate important extracellular differentiation signals. Embryonic LHRH neurons were examined in vivo for Mash-1 and nestin, two factors that delineate precursor populations in PNS and forebrain CNS cells. Nestin, but not Mash-1, was detected in early expressing LHRH cells in the vomeronasal organ anlage. These results were duplicated in LHRH neurons maintained in vitro in nasal explants. Such LHRH cells expressed nestin mRNA but not Mash-1 mRNA and were also negative for three other olfactory epithelial developmental transcription factors, Math4A, Math4C/neurogenin1, and NeuroD mRNA. Experimental manipulation of nasal explants revealed dual expression of nestin protein and LHRH in cells proximal to the vomeronasal organ anlage that was dependent upon midline cartilaginous/mesenchymal tissues. Prolonged nestin expression in LHRH cells after midline removal is consistent with nasal midline tissues modulating differentiation of LHRH neurons from the nasal placode.

NMDA receptor subunit NR1-immunoreactivity in the rat pons and brainstem and colocalization with Fos induced by nasal stimulation.

In the present study, we examined the distribution of neurons in the parabrachial nucleus (PB), the Kölliker-Fuse nucleus (KF), the spinal trigeminal nucleus caudalis (Sp5C), the nucleus of the solitary tract (NTS) and the ventrolateral medulla (VLM), which are activated by evoking the nasotrigeminal reflex and which exhibit immunoreactivity for the N-methyl-D-aspartate (NMDA) receptor subunit NR1. By stimulating the nasal mucosa with saline, we induced the expression of the immediate early gene c-fos and combined the immunocytochemical detection of the Fos protein with the detection of the NR1 subunit. Cell counts revealed that nasal stimulation, compared to anesthesia controls, resulted in highly significant increases (p < or = 0.001) of Fos-immunoreactive (-ir) neurons in the midlevel KF, the external lateral PB, and the Sp5C. In the central lateral PB, the rostral ventrolateral medulla including the Bötzinger/pre-Bötzinger complex, and in the ventrolateral and commissural NTS the increases were only moderately significant (p < or = 0.05). With respect to the numbers of NR1-/Fos-ir double-labeled neurons, significant increases were only observed in a subset of these pontomedullary nuclei. Increases were highly significant in the Sp5C (p < or = 0.001) and the midlevel KF (p < or = 0.01) and moderately significant (p < or = 0.05) in the external lateral PB, Bötzinger/pre-Bötzinger complex, and ventrolateral NTS. The present study revealed that nasotrigeminally activated neurons in mandatory and potential relay sites of the nasotrigeminal reflex circuit express the NR1 subunit. This finding strongly suggests that NMDA-type glutamate receptors are involved in the mediation of the nasotrigeminally evoked cardiovascular and respiratory responses.

Pseudo-cerebrospinal fluid rhinorrhea after skull base surgery.

A 41-year-old female underwent complete resection of a left petroclival meningioma via an anterior transpetrosal approach, during which the greater superficial petrosal nerve was divided. On the 14th day after the operation, she first noticed leakage of clear fluid from her right nostril whenever the ambient room temperature rose. This pseudo-cerebrospinal fluid rhinorrhea may have developed because of parasympathetic hypersensitivity due to division of the greater superficial petrosal nerve.