Olfactory nerve sparing technique for anterior skull base meningiomas: how I do it.

BACKGROUND: Olfactory function preservation is a desirable objective in anterior skull base (ASB) surgery. The "infracerebral-supraolfactory nerve" corridor is presented. METHOD: The technique for preserving the olfactory nerves (OlfNs) in anterior ASB meningioma removal involves the following points: deep knowledge of the ASB vascular and meningeal anatomy, precise preoperative planning, wide and sharp dissection of the OlfNs away from the frontal lobes, gravity-aided frontal lobe retraction, Gelfoam-assisted hemostasis on nervous structures, and access to the lesion through an infracerebral-supraolfactory nerve corridor. CONCLUSIONS: This technique may be a valid option for patients affected by anterior skull base meningiomas with intact preoperative olfactory function.

The Impact of Ovariectomy on Olfactory Neuron Regeneration in Mice.

Estrogen has been shown to affect differentiation and proliferation as a mitogen in various neural systems. Olfactory receptor cells are unique within the nervous system, and have the ability to regenerate even after an individual has reached maturity. Olfactory receptor cells also regenerate after experimentally induced degeneration. The purpose of this study is to observe the influence of estrogen depletion induced by ovariectomy on olfactory nerve regeneration. Female mice underwent bilateral ovariectomy at 8 weeks of age and received intraperitoneal administration of methimazole 1 week later. At 2, 4, and 6 weeks after methimazole administration, the olfactory mucosa was analyzed histochemically to determine olfactory epithelium (OE) thickness, olfactory marker protein distribution, and Ki-67 immunoreactivity. Furthermore, 2 weeks after ovariectomy, trkA protein distribution in the OE and nerve growth factor (NGF) levels in the olfactory bulb were determined by immunohistochemistry and enzyme-linked immunosorbent assay, respectively. Our results showed that in ovariectomized mice OMP, Ki-67, and trkA-immunopositive cells expression decreased at 2 weeks after methimazole injection, a time point at which regeneration is underway. At this same time point, although NGF production in the olfactory bulb had increased before methimazole administration, no differences were observed between the ovx and control groups. These results suggest that estrogen depletion induces a suppressive effect on regeneration of olfactory neurons, and that estrogen may have a potential use in the treatment of sensorineural olfactory dysfunction.

Insulin-like Signaling Promotes Glial Phagocytic Clearance of Degenerating Axons through Regulation of Draper.

Neuronal injury triggers robust responses from glial cells, including altered gene expression and enhanced phagocytic activity to ensure prompt removal of damaged neurons. The molecular underpinnings of glial responses to trauma remain unclear. Here, we find that the evolutionarily conserved insulin-like signaling (ILS) pathway promotes glial phagocytic clearance of degenerating axons in adult Drosophila. We find that the insulin-like receptor (InR) and downstream effector Akt1 are acutely activated in local ensheathing glia after axotomy and are required for proper clearance of axonal debris. InR/Akt1 activity, it is also essential for injury-induced activation of STAT92E and its transcriptional target draper, which encodes a conserved receptor essential for glial engulfment of degenerating axons. Increasing Draper levels in adult glia partially rescues delayed clearance of severed axons in glial InR-inhibited flies. We propose that ILS functions as a key post-injury communication relay to activate glial responses, including phagocytic activity.

First olfactory fiber as an anatomical landmark for frontal sinus surgery.

OBJECTIVES/HYPOTHESIS: Access to the frontal sinuses is technically challenging owing to their anterosuperior location, diverse anatomy, close proximity to critical structures and the need to work in a relatively narrow space with angled-lens endoscopes and instruments. This study attempts to study the relationship of the first olfactory fiber with the frontal sinus posterior wall, assessing its fidelity as a surgical landmark during frontal sinus surgery. STUDY DESIGN: Anatomic study. METHODS: Fifteen cadaveric specimens were studied. Measurements were obtained bilaterally using the data from individual CT scans. Median A-P was defined as the anteroposterior (A-P) diameter measured just lateral to the intersinus septum, paramedian A-P was measured 5 mm lateral to the septum, and maximum A-P was defined as the maximum anteroposterior diameter on axial images. A surgical navigation device was used to calculate the distance between the first olfactory fiber and the posterior table of the frontal sinus. RESULTS: The mean distance between the first olfactory fiber and the posterior wall of the frontal sinus was (4.03 ± 2.7) mm on the right side and (4.2 ± 2.9) mm on the left. This distance strongly correlated with the maximum A-P diameter of the sinus. CONCLUSIONS: In a cadaveric model, the first olfactory fiber was found to be an average of 4.0 mm posterior to the frontal sinus. The significant variability of this distance should be considered when using the first olfactory fiber to establish the posterior boundary of a frontal sinusotomy. Drilling no further posterior than 7 mm rostral to the first olfactory fiber would be safe in 91% of patients. LEVEL OF EVIDENCE: NA Laryngoscope, 126:1039-1045, 2016.

Unilateral Tailored Fronto-Orbital Approach for Giant Olfactory Groove Meningiomas: Technical Nuances.

OBJECTIVE: Giant olfactory groove meningiomas (maximum diameter ≥6 cm) remain a surgical challenge. Historically, extensive anterior and antero-lateral approaches have been the primary approaches for removal of such large tumors with limitations and morbidity pertaining to each approach. Herein, the authors describe a minimally invasive, unilateral, tailored fronto-orbital approach for resection of these complex lesions with an emphasis on preservation of the anterior cerebral arteries and olfactory nerves. METHODS: A 4-stage approach using neuronavigation is performed: 1) predefined corridor, 2) identification of the ipsilateral anterior cerebral artery, 3) postdefined corridor, and 4) tumor base. The details of this approach are described below in a stepwise fashion and supplemented by a sample of 3 cases utilizing this technique. RESULTS: In the 3 representative cases in which this technique was used, gross total resection was achieved without injury to any of the adjacent neurovascular structures. Significant sellar extension can be resected through a second stage endoscopic endonasal approach. CONCLUSION: Giant olfactory groove meningiomas (≥6 cm) can be safely and completely resected with this 4-stage, unilateral fronto-orbital technique. Furthermore, early identification and preservation of the adjacent critical neurovascular structures can be achieved. This technique avoids the inherent limitations and morbidity associated with the more classic pterional and bifrontal approaches respectively while minimizing normal tissue disruption.

Reduced nasal transport of insulin-like growth factor-1 to the mouse cerebrum with olfactory bulb resection.

Although the olfactory nerve is involved in nasal transport of insulin-like growth factor-1 (IGF-1) to the brain, to our knowledge there have been no direct assessments of the effects of olfactory nerve damage on this transport. To determine whether olfactory bulb resection resulted in reduced transport of nasally administered human recombinant IGF-1 (hIGF-1) to the cerebrum, we measured the uptake of nasally administered iodine-125 hIGF-1 ((125)I-hIGF-1) in the cerebrum as a percentage of that in the blood in male ICR mice subjected to left olfactory bulb resection (model mice) and in sham-operated male ICR mice (control mice). Phosphorylated extracellular signal-regulated kinase (ERK) 1/2 (Thr202/Tyr204)/(Thr185/Tyr187) as a percentage of total ERK 1/2 in the left cerebrum was also assessed by using enzyme-linked immunosorbent assay after nasal administration of hIGF-1. Uptake of nasally administered (125)I-hIGF-1 in the cerebrum as a percentage of that in the blood was significantly lower in the model group than in the control group 30min after nasal administration of hIGF-1. Unilateral olfactory bulb resection prevented nasally administered hIGF-1 from increasing the phosphorylation of ERK 1/2 in the mouse cerebrum in vivo. These findings suggest that olfactory bulb damage reduces nasal transport of hIGF-1 to the brain in vivo.

Preservation of olfaction in surgery of olfactory groove meningiomas.

INTRODUCTION: Olfaction is commonly considered as secondary among the sensory functions, perhaps reflecting a lack of interest in sparing olfaction after surgery for the olfactory groove meningiomas (OGM). However, considering the repercussions of olfaction for the quality of life, the assessment of post-operative olfaction should be necessary. We retrospectively reviewed the olfactory outcome in patients with OGM and investigated the factors associated with sparing the post-operative olfaction. METHODS: Between 1993 and 2012, 40 patients with OGM underwent surgical resection and estimated the olfactory function using the Korean version of "Sniffin'Sticks" test (KVSS). Variable factors, such as tumor size, degree of preoperative edema, tumor consistency, preoperative olfactory function, surgical approaches, patient's age, and gender were analyzed with attention to the post-operative olfactory function. RESULTS: Anatomical and functional preservation of olfactory structures were achieved in 26 patients (65%) and 22 patients (55%), respectively. Among the variable factors, size of tumor was significant related to the preservation of post-operative olfaction. (78.6% in size<4 cm and 42.3% in size>4 cm, p=0.035). Sparing the olfaction was significantly better in patients without preoperative olfactory dysfunction (84.6%) compared with ones with preoperative olfactory dysfunction (40.7%, p=0.016). The frontolateral approach achieved much more excellent post-operative olfactory function (71.4%) than the bifrontal approach (36.8%, p=0.032). CONCLUSIONS: If the tumor was smaller than 4 cm and the patients did not present olfactory dysfunction preoperatively, the possibility of sparing the post-operative olfaction was high. Among the variable surgical approaches, frontolateral route may be preferable sparing the post-operative olfaction.

Sensory experience selectively regulates transmitter synthesis enzymes in interglomerular circuits.

Sensory experience influences brain organization and function. A particularly striking example is in the olfactory bulb where reduction of odorant sensory signals profoundly down-regulates dopamine in glomerular neurons. There are two large populations of glomerular inhibitory interneurons: (1) GABAergic periglomerular (PG) cells, whose processes are limited to a single glomerulus, regulate intraglomerular processing and (2) DAergic-GABAergic short axon (SA) cells, whose processes contact multiple glomeruli, regulate interglomerular processing. The inhibitory neurotransmitter GABA is synthesized from L-glutamic acid by the enzyme glutamic acid decarboxylase (GAD) of which there are two major isoforms: GAD65 and GAD67. GAD65 is expressed in uniglomerular PG cells. GAD67 is expressed by SA cells, which also co-express the rate-limiting enzyme for dopamine synthesis, tyrosine hydroxylase (TH). Deafferentation or sensory deprivation decreases TH expression but it is not known if sensory input alters GAD isoforms. Here we report that either deafferentation or reduction of sensory input by nares occlusion significantly reduced GAD67 protein and the number of SA cells expressing GAD67. However, neither manipulation altered GAD65 protein or the number of GAD65 PG cells. These findings show that sensory experience strongly impacts transmitter regulation in the circuit that controls neural processing across glomeruli but not in the circuit that regulates intraglomerular processing.

[Expression of odorant receptor genes on the olfactory epithelium following olfactory nerve disconnection].

OBJECTIVE: To constitute the animal model of unilateral olfactory nerve transection and observe the expression level and distribution of odorant receptors. METHODS: Thirty-two rats were divided into two groups: the olfactory nerve transection group (20) and the control group (12). The former group received the operation to transect the left olfactory nerve following the left olfactory bulb was exposed under microscope and the latter group did not give any disposal. At every stage of five days, two weeks, four weeks and six weeks after the operation, five rats from the nerve transection group and three from the control group were anaesthetized simultaneously, and olfactory epithelium were taken out after transcardial perfusion, then paraffin imbedding. Coronal sections were sliced for HE staining to observe the thickness changes of the olfactory epithelium, and for in situ hybridization (ISHs) to investigate the expression of olfactory receptor genes (Olr287, Olr226, Olr1493 and Olr1654) in the epithelium, also to evaluate the changes of the expression level and location of the selected receptors during the regeneration of olfactory epithelium. RESULTS: HE staining showed that 5 days after the operation cell quantity and thickness of the olfactory epithelium decreased obviously, which increased gradually 2 or 4 weeks after operation. After 6 weeks' recovery, the thickness of the epithelium could reach the control level. The pattern of cell staining by ISH showed a specific spatial distribution along the anteroposterior (AP) and dorsoventral (DV) axis. Evidence suggested that odorant receptors were distributed in continuous and multiple overlapping bands in the normal or nerve transected-recovered epithelium rather than in the conventionally accepted three or four zones. The data also demonstrated that the distribution of sensory neuron types, as identified and defined by odorant receptor expression, was restored to normal or nearly so by 6 weeks after operation. Likewise, the numbers of probe-labeled neurons in the nerve transected-recovered had an obvious decrease 5 days after olfactory nerve transection. Reactive cells (x(-) +/- s) of Olr1493 in the operated side was (53.9 +/- 19.9), compared with (419.0 +/- 21.2) in the unoperated side, there was statistic significance between them (t = 63.960, P < 0.01). Reactive cells increased gradually according to the regeneration of the epithelium, and were nearly equivalent to the normal side 6 weeks later without significant differentiation (t = 2.600, P > 0.05), according to the absolute positive cells in the operated and unoperated side of (417.8 +/- 32.4) and (445.3 +/- 10.0) respectively. CONCLUSION: The regeneration of the sensory neurons and receptors, both the number and the distribution, can recover to normal after olfactory nerve transection.

Use of thallium transport to visualize functional olfactory nerve regeneration in vivo.

OBJECTIVE: To image olfactory nerve regeneration in vivo using a high-resolution gamma cam- era and radiography after nasal administration of thallium-201 (olfacto-scintigraphy). METHODS: Six Wistar rats were trained to avoid the smell of cycloheximide as a test of olfactory function. The olfactory nerve fibers of 3 rats were then carefully transected bilaterally with a Teflon knife, avoiding damage to the olfactory bulbs. The remaining 3 rats underwent sham operations and were used as controls. Steel wires were implanted in the left olfactory bulb of each rat for locating the bulbs with plain X-rays. The rats were assessed 2, 14, 28, and 42 d after the olfactory nerve transection or sham operation for their ability to detect odours and for transport of 201Tl to the olfactory bulb area 8 h after nasal administration of 201Tl. RESULTS: Both transport of 201Tl to the olfactory bulb area (p < 0.04) and ability to detect odours (p < 0.04) significantly increased with a time course after olfactory nerve transection. CONCLUSION: 201Tl transport to the olfactory bulb may be useful to visually assess olfactory ability in vivo. We plan to test olfacto-scintigraphy clinically by nasal administration of 201Tl in patients with posttraumatic olfactory loss.

Effects of systemic transplantation of adipose tissue-derived stem cells on olfactory epithelium regeneration.

OBJECTIVE/HYPOTHESIS: The purpose of the study was to investigate the effect of intravenous adipose tissue-derived stem cell (ADSC) transplantation on olfactory epithelium regeneration following transection of the olfactory nerve in rats. STUDY DESIGN: This was a experimental study using primary cultures of mesenchymal stem cells derived from animal adipose tissue with histological analysis of animal olfactory tissue. METHODS: All rats underwent unilateral transection of the olfactory nerve to induce degeneration of olfactory epithelium, and then were observed for regeneration according to time sequences. ADSCs were cultivated from neck adipose tissue of rats, and systemically injected into the experimental group. The control group was injected with phosphate buffered solution, instead of ADSCs. After 30 days, regeneration of olfactory epithelium was observed with olfactory marker protein (OMP) and proliferating cell nuclear antigen. To observe the characteristics of the transplanted ADSCs, olfactory epithelium was stained with von Willebrant factor and OMP. RESULTS: After olfactory nerve transection, mature olfactory cells disappeared in 5 days, but gradually regained their thickness with increased cell numbers at approximately 10 to 15 days. By 30 days post-transection, the thickness and cellular composition of epithelium was almost restored to baseline levels pretransection. However, OMP expressions remained decreased compared with day 0 or 3. Systemically injected ADSCs were transplanted into the olfactory epithelium and survived beyond 4 weeks. The ADSCs promoted regeneration of olfactory epithelium in the animal model and differentiated into olfactory receptor neurons and endothelial cells. CONCLUSIONS: Our findings suggest the feasibility of ADSC transplantation as a treatment for head trauma-related olfactory dysfunction.

Effects of transplanting olfactory ensheathing cells on recovery of olfactory epithelium after olfactory nerve transection in rats.

BACKGROUND: Olfactory ensheathing cells (OECs) can remyelinate injured spinal cord and the peripheral nerve system, but little is known about its effect on the transected olfactory nerve. We investigated recovery of olfactory epithelium after transplanting allogeneic OECs in transected rat olfactory nerves. MATERIAL/METHODS: Olfactory ensheathing cells from the olfactory bulb were cultured in DMEM/F-12 medium and purified with cytosine arabinoside (Ara-c). Forty Sprague-Dawley rats were divided into 2 groups; the left olfactory nerve was transected in all animals. In the transplant group, DiI-labeled OECs were injected into the gap between the dura and the cribriform plate (n=24); DMEM/F-12 medium was injected in control animals (n=16). Rats were subsequently killed for histologic examination. Olfactory evoked potentials (OEPs) were used to evaluate nerve conduction. RESULTS: After transecting the olfactory nerve, there was no horseradish peroxidase staining in the olfactory bulb; some OEPs disappeared. Five days after surgery, there was no horseradish peroxidase staining in the olfactory bulb of any animal. Apoptotic cells appeared in the epithelium; the thickness and cell number of the olfactory epithelium were decreased. Two weeks later, the thickness and cell number of the olfactory epithelium increased gradually. Some horseradish peroxidase staining in the olfactory bulb and OECs was detected; more growth associated protein-43 marked olfactory receptor neurons were visible. Six weeks after surgery, the cell number was greater in the transplant group (P<0.05); there was no statistically significant between-group difference regarding olfactory epithelium thickness. CONCLUSIONS: Transplanted OECs may be used to treat transected olfactory nerves.

Odor detection ability and thallium-201 transport in the olfactory nerve of traumatic olfactory-impaired mice.

Although olfactory nerve damage is a contributing factor in the diagnosis of posttraumatic olfactory loss, at present, there are no methods to directly assess injury to these nerves. We have shown that following olfactory nerve injury in mice, thallium-201 (201 Tl) transport from the nasal cavity to the olfactory bulb decreases. To determine if olfactory function after nerve injury could be assessed with nasal administration of 201 Tl, we measured the correlation between odor detection ability (ODA) and the rate of transport of 201 Tl in olfactory nerves. Both ODA and 201 Tl transport were measured after bilateral olfactory nerve transection for a 4-week period. Cycloheximide solution was used for ODA against tap water. 201 Tl transport was measured as the ratio of radioactivity in the nasal cavity and olfactory bulb with gamma spectrometry. There was a significant correlation between ODA and the rate of 201 Tl transport in the olfactory nerve. These findings suggest that olfactory function after nerve injury can be objectively evaluated with the nasal administration of 201 Tl.

Navigational abilities of homing pigeons deprived of olfactory or trigeminally mediated magnetic information when young.

Anatomical evidence and conditioning experiments have recently suggested that magnetoreceptors are located in the upper beak of homing pigeons, where they are innervated by the ophthalmic branch of the trigeminal nerve. These findings have raised the issue of whether the trigeminally mediated magnetoreception is involved in the navigational mechanisms of homing pigeons. Recent data have shown that, in inexperienced pigeons, section of the ophthalmic branch of the trigeminal nerve does not impair navigational abilities, whereas the navigational performance of inexperienced pigeons is disrupted after section of the olfactory nerve. Nevertheless, the issue of whether the stimuli available during development of the navigational mechanism can influence the types of cues used in determining the direction of displacement remains unresolved. To address this issue, we surgically deprived young pigeons of either olfactory or trigeminally mediated magnetic information, and then later tested their navigational abilities subsequent to an intensive training flight program of up to 10 km in different directions. The birds deprived of trigeminally mediated magnetic information when young developed navigational abilities at the same level as intact control pigeons, whereas the olfactory deprived pigeons displayed randomly scattered initial orientation and poor homing performance. Our data show that olfactory cues are needed for the development of navigational abilities from unfamiliar locations and that the lack of magnetic information does not affect the development of homing abilities.

Effect of Ninjin-yoei-to (Rensheng-Yangrong-Tang) on olfactory behavior after olfactory nerve transection.

Ninjin-yoei-to (NYT), a Japanese traditional medicine, is used to treat athrepsia due to surgery, anorexia, cold constitution, and anemia. There are reports of the effects of NYT on the nervous system; however, there have been no behavioral studies of the effect of NYT on olfactory function. The olfactory system undergoes continuous replacement of sensory neurons. Morphologic and behavioral studies have shown that the olfactory system recovers after bilateral olfactory nerve transection (BNX). However, in the humans, olfactory function does not always recover. In this study, we examined the effect of oral NYT on behavioral recovery after BNX. Fourteen mice were subjected to BNX. The regular diet was mixed with 2% NYT (NYT diet). Mice were separated into two groups; seven mice were fed the regular diet (control group), and seven mice were fed the NYT diet (NYT group). NYT was administered beginning 7 days prior to BNX and continuing for 35 days after BNX. Mice in both groups had free access to food and water. Olfactory function was evaluated by testing each mouse's ability to avoid cotton balls treated with acetic acid. After BNX, mice lost their ability to avoid cotton balls treated with acetic acid. In the control group, the time for behavioral recovery after BNX was 28 days. In the NYT group, the time for behavioral recovery after BNX was 21 days. NYT hastened behavioral recovery after BNX. NYT may have therapeutic benefits for patients with olfactory disorders.

Microanatomy and surgical relevance of the olfactory cistern.

All surgical approaches to the anterior skull base involve the olfactory cistern and have the risk of damaging the olfactory nerve. The purpose of this study was to describe the microanatomical features of the olfactory cistern and discuss its surgical relevance. In this study, the olfactory cisterns of 15 formalin-fixed adult cadaveric heads were dissected using a surgical microscope. The results showed that the olfactory cistern was situated in the superficial part of the olfactory sulcus, which separated the gyrus retus from the orbital gyrus. In coronal section, the cistern was triangular in shape; its anterior part enveloped the olfactory bulbs and was high and broad; its posterior part was medial-superior to internal carotid artery and was also much broader. There were one or several openings in the inferior wall of the posterior part in 53.4% of the cisterns. The olfactory cistern communicated with the surrounding subarachnoind cisterns through these openings. The middle part of the olfactory cistern gradually narrowed down posteriorly. Most cisterns were spacious with a few fibrous trabeculas and bands between the olfactory nerves and cistern walls. However 23% of the cisterns were narrow with the cistern walls tightly encasing the olfactory nerve. There were two or three of arterial loops in each olfactory sulcus, from which long, fine olfactory arteries originated. The olfactory arteries coursed along the olfactory nerve and gave off many terminal branches to provide the main blood supply to the olfactory nerve in most cisterns, but the blood supply was in segmental style in a few cisterns. Moreover, the veins of the cistern appeared to be more segmental than the olfactory arteries in most cisterns. These results suggested that most olfactory cisterns are spacious with relatively independent blood supply, and it is reasonable to separate the olfactory tract with its independent blood supply from the frontal lobe by 1-2 cm in the subfrontal approach, the pterional approach, or anterior interhemispheric approach. However, in the minority of cases, separation of the olfactory tract is not safe because of the anterior origin of the olfactory arteries or segmental blood supply. It is difficult to separate the olfactory nerve without any damage to the olfactory nerve, even with very skilled hands.

Thallium transport and the evaluation of olfactory nerve connectivity between the nasal cavity and olfactory bulb.

Little is known regarding how alkali metal ions are transported in the olfactory nerve following their intranasal administration. In this study, we show that an alkali metal ion, thallium is transported in the olfactory nerve fibers to the olfactory bulb in mice. The olfactory nerve fibers of mice were transected on both sides of the body under anesthesia. A double tracer solution (thallium-201, (201)Tl; manganese-54, (54)Mn) was administered into the nasal cavity the following day. Radioactivity in the olfactory bulb and nasal turbinate was analyzed with gamma spectrometry. Auto radiographic images were obtained from coronal slices of frozen heads of mice administered with (201)Tl or (54)Mn. The transection of the olfactory nerve fibers was confirmed with a neuronal tracer. The transport of intranasal administered (201)Tl/(54)Mn to the olfactory bulb was significantly reduced by the transection of olfactory nerve fibers. The olfactory nerve transection also significantly inhibited the accumulation of fluoro-ruby in the olfactory bulb. Findings indicate that thallium is transported by the olfactory nerve fibers to the olfactory bulb in mice. The assessment of thallium transport following head injury may provide a new diagnostic method for the evaluation of olfactory nerve injury.

Olfactory schwannoma.

We report a case of olfactory schwannoma with calcification. Intraoperative findings indicated that the tumour originated from the olfactory groove. Intraoperative findings of previous studies have not indicated a clear relationship between subfrontal schwannoma and the olfactory nerve, which seems strange, given the association between tumours and cranial nerves at other sites. We suggest this observation has not been reported because the growing olfactory schwannoma changes the local morphology, affecting the appearance of the olfactory nerve.

Afferent input modulates the chronic hypercapnia-induced increase in respiratory-related central pH/CO2 chemosensitivity in the cane toad (Bufo marinus).

The goal of this study was to examine the role of respiratory-related afferent input on the chronic hypercapnia (CHC)-induced increase in central respiratory-related pH/CO2 chemosensitivity in cane toads (Bufo marinus). Toads were exposed to CHC (3.5% CO2) for 10 days, following which in vitro brainstem-spinal cord preparations were used to assess central respiratory-related pH/CO2 chemosensitivity. Motor output from the vagus nerve root was used as an index of breathing (fictive breathing). Olfactory denervation (OD), prior to exposure to CHC, was used to remove the influence of CO2-sensitive olfactory chemoreceptors, which inhibit breathing. Exposure to chronic hyperoxic hypercapnia (CHH) was used to reduce the level of arterial chemoreceptor input compared with CHC alone. In vivo experiments examined the effects of CHC, CHH and OD on the acute hypercapnic ventilatory response of intact animals. In vitro, a reduction in artifical cerebral spinal fluid (aCSF) pH increased fictive breathing in preparations taken from control and CHC animals. CHC caused an increase in fictive breathing compared with controls. OD and CHH abolished the CHC-induced augmentation of fictive breathing. In vivo, CHC did not cause an augmentation of the acute hypercapnic ventilatory response. CHH reduced the in vivo acute hypercapnic ventilatory response compared with animals exposed to CHC. In vivo, OD reduced breathing frequency and increased breath amplitude in both control and CHC animals. The results suggest that afferent input from olfactory and arterial chemoreceptors, during CHC, is involved in triggering the CHC-induced increase in central respiratory-related pH/CO2 chemosensitivity.

[Impact of olfactory nerve transection on the apoptosis of mice olfactory receptor neurons].

OBJECTIVE: To analyze the impact of olfactory nerve transection on the apoptosis of mice olfactory receptor neurons (ORN), and discuss the reliability of this experimental model. METHODS: After olfactory nerve transection of mice, anterograde horseradish peroxidase (HRP) tracing was performed to confirm the completion of nerve transection. On 8 h, 2 d, 3 d and 5 d after surgery, TdT mediated deoxyuridine triphosphate-biotin nick end labelling (TUNEL) was used to observe the apoptosis of ORN, while relative semi-quantitative RT-PCR and immunohistochemistry were used to reflect the expression of olfactory marker protein (OMP, special marker of mature ORN) in olfactory epithelium. RESULTS: No HRP label was observed in olfactory bulb after olfactory nerve transaction. Both TUNEL-positive and OMP-positive cells were ORN. After the surgery, TUNEL-positive cells increased remarkably and peaked on 2 d after the surgery. Meanwhile OMP mRNA in olfactory epithelium began to decrease markedly till 5 d after the surgery, and the olfactory epithelium got thinner accordingly. CONCLUSIONS: This experimental model can be used reliably to sever mice olfactory nerve and manipulate simultaneous apoptosis of mice ORN.