Extra-bulbar primary olfactory projection in teleost fishes

Immunohistochemical investigations with anti-substance P antiserum demonstrate the existence of an extensive extrabulbar primary olfactory projection in several gymnotid teleost fish. This projection, never described before, originates in particular primary olfactory bundles which enter with the olfactory nerve into the olfactory bulb. While the bulk of the olfactory fibers end with glomeruli in the glomerular layer of the olfactory bulb, two particular bundles penetrate into the telencephalon and end, without forming glomeruli, in several telencephalic and diencephalic regions. A few fibers run as far as to the hypothalamus. In the light of these findings, the general notion that the primary olfactory projection is limited to the olfactory bulb and forms only glomeruli-like terminals, should be reconsidered.     

Immunocytochemistry of the olfactory marker protein.

The olfactory marker protein has been localized, by means of immunohistochemical techniques in the primary olfactory neurons of mice. The olfactory marker protein is not present in the staminal cells of the olfactory neuroepithelium, and the protein may be regarded as indicative of the functional stage of the neurons. Our data indicate that the olfactory marker protein is present in the synaptic terminals of the olfactory neurons at the level of the olfactory bulb glomeruli. The postsynaptic profiles of both mitral and periglomerular cells are negative.     

Blockade of neurotransmission in Drosophila mushroom bodies impairs odor attraction, but not repulsion

Olfaction can elicit a rich perceptual experience. It is not known, however, whether olfactory information is decomposed into various components and processed in distinct perceptual centers as in other sensory systems, such as vision, where neural representations of different visual sensations are segregated in different cortical regions, despite the fact that multiple structures of the primary olfactory cortex receive projections from the olfactory bulb. Here, we use Drosophila as a model to investigate whether different olfactory information may be processed in separate brain structures. Organizations of the peripheral olfactory system are remarkably similar from mammals to insects. As in vertebrates, the olfactory pathway in Drosophila follows similar convergence and divergence, and multiple high-order structures in the Drosophila brain, including the mushroom body (MB) and lateral horn (LH) of the protocerebrum, receive olfactory input. We specifically blocked neurotransmission in the MB while leaving the LH unaffected and examined its effect on olfactory avoidance and attraction behaviors. We show that blocking MB activity disrupted responses to attractive, but not repulsive, odors, and this finding suggests that attractive and repulsive olfactory information may be separately processed in higher olfactory centers of the Drosophila brain.     

Neurotrophins are not required for normal embryonic development of olfactory neurons

Neurons of the vertebrate olfactory epithelium (OE) regenerate continuously throughout life. The capacity of these neurons to regenerate and make new and precise synaptic connections in the olfactory bulb provides a useful model to study factors that may control or mediate neuronal regeneration. Expression and in vitro studies have suggested potential roles for the neurotrophins in the olfactory system. To directly examine whether neurotrophins are required for olfactory neuron development, we characterized in vivo the role of the neurotrophins in the primary olfactory system. For this, we generated mutant mice for TrkA, TrkB, TrkC, and also for BDNF and NT3 together with P2-IRES-tau-LacZ trangenic mice. Histochemical staining for beta-galactosidase at birth allowed in vivo analysis of the P2 subpopulation of olfactory neurons as well as their projections to the olfactory bulb. Our data indicate that Trk signaling is not required for normal embryonic development of the olfactory system.     

Ratings of overall olfactory function

The aim of this study was to investigate the accuracy of self-reported ratings of olfactory function in 83 healthy subjects. Such ratings were compared with quantitative measures of olfactory function, as well as with ratings of nasal patency. In experiment 1 subjects rated olfactory function and nasal patency before olfactory testing, whereas in experiment 2 the reverse was the case. No feedback regarding test results were provided until after completion of the testing. The principal findings were: (i) when ratings preceded measurements of olfactory function, there was no significant correlation between the two parameters. However, ratings of olfactory function correlated significantly with ratings of nasal airway patency. (ii) In contrast, when measurements of olfactory function preceded the ratings, this constellation switched. Now ratings of olfactory function correlated significantly with measured olfactory function, whereas there was no significant correlation between ratings of nasal airway patency and ratings of olfactory function. In conclusion, these data suggest that ratings of olfactory function are unreliable in healthy, untrained subjects. The ratings seem to reflect changes of nasal airway patency to a larger degree than measurable olfactory function. The results further indicate that this is mainly due to the limited attention the sense of smell receives in daily life.     

视觉和嗅觉信号对果蝇食物搜寻行为的协同作用

为了探索视觉和嗅觉信号在昆虫食物搜寻过程中的作用, 本研究利用杨梅和橘子为引诱物, 在实验室条件下测定了嗅觉和视觉信号诱集到的黑腹果蝇Drosophila melanogaster数量, 分析了嗅觉经历对果蝇嗅觉和视觉食物搜寻的影响。发现同源性嗅觉和视觉信号存在的杨梅诱集到的果蝇数量显著大于单一的视觉信号和嗅觉信号, 但异源性嗅觉和视觉信号组合诱集到的果蝇数量和单独的嗅觉信号相似。嗅觉信号预处理不仅能够显著增加嗅觉信号诱集到的果蝇数量, 其中杨梅嗅觉信号对杨梅预处理果蝇的吸引能力与视觉和嗅觉信号存在的杨梅相似, 而且异源性嗅觉和视觉信号组合诱集到的预处理果蝇数量也不低于视觉和嗅觉信号存在的杨梅。另外杨梅嗅觉信号预处理也能够显著增强杨梅视觉信号诱集到的果蝇数量。但嗅觉预处理并不会改变同源性视觉和嗅觉信号组合诱集到的果蝇数量。本研究表明, 果蝇同时利用视觉和嗅觉信号进行食物搜寻, 因此同源性视觉和嗅觉信号在果蝇诱集过程中具有协同作用。另外果蝇具有较强的记忆和学习能力, 能够将记忆中的嗅觉信号应用于食物搜寻。本研究结果不仅有利于我们了解果蝇在自然状态下的食物搜寻机制, 而且有利于开发更有效的果蝇新型诱捕器。     

The glucose transporter GLUT1 and the tight junction protein occludin in nasal olfactory mucosa.

The nervous cells in the brain and the peripheral nerves are isolated from the external environment by the blood-brain, blood-cerebrospinal fluid and blood-nerve barriers. The glucose transporter GLUT1 mediates the specific transfer of glucose across these barriers. The olfactory system is unique in that its sensory cells, olfactory receptor neurons, are embedded in the nasal olfactory epithelium and send their axons directly to the olfactory bulb of the brain. Only the apical parts of the olfactory receptor neurons are exposed to the lumen, and these serve as sensors for smell. Immunohistochemical examination showed that the tight junction protein occludin was present in the junctions of the olfactory epithelium. Endothelial cells in the blood vessels in the lamina propria of the olfactory mucosa were also positive for occludin. These observations suggest that the olfactory system is guarded from both the external environment and the blood. GLUT1 was abundant in these occludin-positive endothelial cells, suggesting that GLUT1 may serve in nourishing the cells of the olfactory system. Taken together, GLUT1 and occludin may serve as part of the machinery for the specific transfer of glucose in the olfactory system while preventing the non-specific entry of substances.     

Heterogeneity in olfactory neurons in mouse revealed by differential expression of glycoconjugates

Cell surface glycoconjugates have been implicated in the growth and guidance of subpopulations of primary olfactory axons. While subpopulations of primary olfactory neurons have been identified by differential expression of carbohydrates in the rat there are few reports of similar subpopulations in the mouse. We have examined the spatiotemporal expression pattern of glycoconjugates recognized by the lectin from Wisteria floribunda (WFA) in the mouse olfactory system. In the developing olfactory neuroepithelium lining the nasal cavity, WFA stained a subpopulation of primary olfactory neurons and the fascicles of axons projecting to the target tissue, the olfactory bulb. Within the developing olfactory bulb, WFA stained the synaptic neuropil of the glomerular and external plexiform layers. In adults, strong expression of WFA ligands was observed in second-order olfactory neurons as well as in neurons in several higher order olfactory processing centres in the brain. Similar, although distinct, staining of neurons in the olfactory pathway was detected with Dolichos biflorus agglutinin. These results demonstrate that unique subpopulations of olfactory neurons are chemically coded by the expression of glycoconjugates. The conserved expression of these carbohydrates across species suggests they play an important role in the functional organization of this region of the nervous system.     

Distribution of Kroeyerina elongata (Kroyeriidae: Siphonostomatoida, Copepoda) in the olfactory sacs of the blue shark, Prionace glauca

The infection pattern of Kroeyerina elongata (Kroyeriidae, Copepoda) in the olfactory sacs of the blue shark, Prionace glauca, was investigated using 4,722 copepods from 54 olfactory sacs. Copepod prevalence and mean intensity of infection per olfactory sac were 94.0 and 91.1%, respectively, and the most intensely infected olfactory sac and shark hosted 218 and 409 copepods, respectively. There were significant linear relationships between the number of female and total copepods per left olfactory sac and shark fork length as well as between the numbers of female, male, and total copepods per shark and mean olfactory sac width and cumulative olfactory sac width. Female copepods typically outnumbered males within olfactory sacs (mean intensity = 65.7 and 26.3, respectively), and no statistical differences were detected between the numbers of copepods inhabiting the left and right olfactory sacs. Copepods were not evenly distributed within olfactory sacs. Typically, female copepods occupied olfactory chambers located centrally along the length of the olfactory sac, while males infected lateral olfactory chambers nearest the naris. The orientation of most copepods (84.6%) suggested positive rheotaxis relative to the path of water through the olfactory sac. Within olfactory chambers, most mature females (68.2%) infected the first third of the peripheral excurrent channel and the adjacent fringe of olfactory lamellae, while most males (91.7%) infected the olfactory lamellae, and the 4 larval females collected were attached within the lamellar field and grasped by males. Based on the observed infection patterns and the pattern of water flow throughout the olfactory sac, a hypothesis regarding the life cycle of K. elongata is advanced wherein infective copepodids are swept into the olfactory sac from the surrounding sea and initially colonize the olfactory lamellae. Copepodids feed and mature among the olfactory lamellae, and adult males search for mates and copulate with young females among the olfactory lamellae. Inseminated females move to the peripheral excurrent channels to mature and produce ovisacs. Hatching ovisacs release free-swimming nauplii into the excurrent water flow to be swept into the milieu, where they can molt into infective copepodids that may infect new hosts.     

Receptor cells of different types and quantitative relations between them in the organ of smell of larval and adult specimens of acipenserid fishes]

Three types of olfactory cells: rod-like and cone-like (flagellar olfactory cells) and filamentous (microvillar olfactory cells), which have been described previously in adult Acipenseridae were found in the olfactory organ of the ten-days larval sturgeons (Acipenser güldenst?dti), sevrugas (Acipenser stellatus) and sterlets (Acipenser ruthenus). The flagellar olfactory receptors appeared to predominate in both ten-days larvae and adults of the anadromous sturgeons and sevrugas, while the microvillar olfactory receptors predominate in freshwater sterlets in ten-days larvae as well in adults. The facts obtained confirm the idea that the rod-like, cone-like and filamentous olfactory cells are independent types of olfactory receptors. The different ratios of these cells in the olfactory organs of anadromous and fresh-water Acipenseridae may be a result of their ecological adaptations.     

Olfactory ability in the healthy population: reassessing presbyosmia

Age-associated loss of olfactory function, or presbyosmia, has been described in many studies of olfactory ability. Presbyosmia has been ascribed to idiopathic causes despite recognition that many neurodegenerative diseases also induce loss of olfactory function and increase in incidence in the aged population. Often this olfactory loss is unnoticed or unreported by affected individuals. More effective olfactory function in women compared with men is another common feature of many studies of olfactory function. Here we report on normative data from an Australian population study (n = 942) that has been divided into 2 subpopulations and reassessed as (included) a population of healthy, nonmedicated, nonsmokers with no history of nasal problems (n = 485) and (excluded) a population of participants who were either medicated, smokers or had a history of nasal problems (n = 457). The "included" data set shows a strong relationship between self-reporting of olfactory sensitivity and olfactory function score. The included data set shows a small but significant decline in olfactory ability after 65 years of age and better olfactory function in females compared with males. Data from the excluded population show a marked decline in olfactory ability after 65 years of age, no difference between males and females, and a weak relationship between self-reporting of olfactory function and actual olfactory function. The power of this approach is that it provides a normative data set against which many factors such as medication schedules and pathological conditions can be compared.     

脊椎动物中的一种解剖学新模型——黄鳝外周嗅觉系统

除单鼻型的圆口类外, 脊椎动物的左、右两侧嗅觉器官和嗅神经皆互为独立地分布于头前端, 而且它们的前鼻孔(外鼻孔)、嗅腔、嗅觉副囊腔(部分鱼具嗅觉副囊)与后鼻孔(或内鼻孔)也都互为相通, 且多呈开放状态。它们还通常具有一个体积相对较大且较稳定的嗅腔, 而嗅上皮则多位于嗅腔的一侧。此外, 鱼类的嗅囊与鼻窝之间通常也无明显间隙。然而, 运用常规的解剖学方法发现, 黄鳝(Monopterus albus)外周嗅觉系统(嗅觉器官和嗅神经)在解剖结构上已发生如下重大变化: (1)虽然具有前、后鼻孔, 但两者互不相通, 而嗅腔仅靠前鼻孔通至外界; (2)两侧嗅囊的末端及两侧嗅神经的前段均分别发生了合并。此外, 在该鱼上还发现:(1)嗅囊为一柔软而扁塌的长管囊结构, 其唯一的开口(即位于前鼻孔球上的前鼻孔)却常呈关闭状, 故此时该嗅腔实际上是一个体积被压扁到最小且暂时被封闭的空间; (2)嗅囊纵向地贴附于长鼻窝的内侧壁上, 它仅占鼻窝的一小部分空间, 故鼻窝显得相对很宽敞; (3)嗅觉副囊不与嗅腔相通, 而与鼻窝共同经后鼻孔通至外界; (4)两侧嗅囊的末端相向地穿越鼻窝内侧壁, 进入筛骨与额骨之间的“筛-额横管”, 在那里发生嗅囊合并;(5)嗅囊壁周缘几乎都内衬着嗅上皮, 且具数个褶窝(说明该嗅囊有扩张的可能)。因此, 黄鳝的这套解剖学特征不同于包括鱼类在内的所有脊椎动物的外周嗅觉系统。研究所发现的黄鳝这套形态学特征不仅为脊椎动物外周嗅觉系统的研究提供了一个独特的解剖学新模型, 同时也为动物进化研究提供了一个有关前、后鼻孔互不相通的进化特例。此外, 研究还依据上述发现提出嗅囊扩张-压缩假说以解释气味媒质进出于黄鳝这种特殊嗅腔的动力学机制。       

Electrophoretic changes in olfactory system proteins in masu salmon during parr-smolt transformation

Changes in cytosolic proteins of the olfactory system (olfactory epithelium, olfactory nerve and olfactory bulb) and the telencephalon of masu salmon Oncorhynchus masou were analysed by two-dimensional polyacrylamide gel electrophoresis during parr-smolt transformation; parr, pre-smolt and full-smolt stages. In the olfactory system, several protein spots appeared and disappeared in the course of smolting. One protein spot in particular with an estimated molecular weight of 27 kDa and isoelectric point of 5.6 (M27) disappeared in common with the olfactory system during smolting. The disappearance of M27 was also observed in the telencephalon. These proteins, which appeared and disappeared, may reflect the changes in olfactory function during smolting. Simultaneously, the present study confirmed a salmonid olfactory specific protein of 24 kDa (N24), which existed in the olfactory system but not in the telencephalon, as a single neutral protein spot in masu salmon.     

Nasal Administration of Cholera Toxin as a Mucosal Adjuvant Damages the Olfactory System in Mice

Cholera toxin (CT) induces severe diarrhea in humans but acts as an adjuvant to enhance immune responses to vaccines when administered orally. Nasally administered CT also acts as an adjuvant, but CT and CT derivatives, including the B subunit of CT (CTB), are taken up from the olfactory epithelium and transported to the olfactory bulbs and therefore may be toxic to the central nervous system. To assess the toxicity, we investigated whether nasally administered CT or CT derivatives impair the olfactory system. In mice, nasal administration of CT, but not CTB or a non-toxic CT derivative, reduced the expression of olfactory marker protein (OMP) in the olfactory epithelium and olfactory bulbs and impaired odor responses, as determined with behavioral tests and optical imaging. Thus, nasally administered CT, like orally administered CT, is toxic and damages the olfactory system in mice. However, CTB and a non-toxic CT derivative, do not damage the olfactory system. The optical imaging we used here will be useful for assessing the safety of nasal vaccines and adjuvants during their development for human use and CT can be used as a positive control in this test.     

Transregulation of erbB expression in the mouse olfactory bulb.

Previously, we have shown that erbB-3 expression is restricted to the ensheathing cells of the olfactory nerve layer, while erbB-4 is found in the periglomerular and mitral/tufted cells of the olfactory bulb and in cells coming out from the rostral migratory stream of the subependymal layer. In the present work, we have treated adult mice with zinc sulfate intranasal irrigation and analyzed erbB-3 and erbB-4 expression in the deafferented olfactory bulb. Following treatment, olfactory axons undergo degeneration, as indicated by the loss of OMP expression in the deafferented olfactory bulb. The thickness of the olfactory nerve layer is reduced, but the specific intensity of erbB-3 labeling in the remaining olfactory nerve layer is increased with respect to control. Interestingly, following deafferentation, erbB-4 immunoreactivity decreases specifically in cell types that normally make synaptic contacts with primary olfactory neurons in the glomeruli, i.e. periglomerular and mitral/tufted cells. Partial lesion of the olfactory epithelium allows regenerative axon growth of olfactory neurons to the olfactory bulb. Following olfactory axon regeneration, erbB-3 and erbB-4 immunoreactivity in the olfactory bulb is similar to control. Thus, like tyrosine hydroxylase, the down regulation of erbB-4 expression in the periglomerular cells is reversible.     

Investigation of breathing parameters during odor perception and olfactory imagery

Compared with visual and auditory imagery, little is known aboutolfactory imagery. There is evidence that respiration may bealtered by both olfactory perception and olfactory imagery.In order to investigate this relationship, breathing parameters(respiratory minute volume, respiratory amplitude, and breathingrate) in human subjects during olfactory perception and olfactoryimagery were investigated. Fifty-six subjects having normalolfactory function were tested. Nasal respiration was measuredusing a respiratory pressure sensor. Using an experimental blockdesign, we alternately presented odors or asked the subjectsto imagine a given smell. Four different pleasant odors wereused: banana, rose, coffee, and lemon odor. We detected a significantincrease in respiratory minute volume between olfactory perceptionand the baseline condition as well as between olfactory imageryand baseline condition. Additionally we found significant differencesin the respiratory amplitude between imagery and baseline conditionand between odor and imagery condition. Differences in the breathingrate between olfactory perception, olfactory imagery, and baselinewere not statistically significant. We conclude from our resultsthat olfactory perception and olfactory imagery both have effectson the human respiratory profile and that these effects arebased on a common underlying mechanism.     

Disruption of the olfactory placode and brain conditioned medium increase the number of LHRH immunostained neurons in explants

The olfactory placode gives rise to both olfactory receptor neurons, which remain as a component of the peripheral nervous system, and to luteinizing hormone-releasing hormone (LHRH) neurons, which migrate to the central nervous system. In this study, we used chick olfactory placode explants to ask several questions regarding LHRH neuronal differentiation. We found that explants of ectoderm from the fronto-nasal region of embryos as early as Hamilton & Hamburger (HH) stage 12 gave rise to LHRH neurons, that explants from all regions of the olfactory placode were able to generate LHRH neurons, that both brain conditioned medium and disruption of the olfactory placode increase the number of LHRH neurons observed in explants, and that the combination of these two manipulations results in the production of more LHRH neurons than either treatment alone. We conclude that LHRH neurons originate in the olfactory epithelium and that some of the same factors which influence olfactory receptor neuron development also affect LHRH neuronal development.     

Morphology of the olfactory system in the predatory mite Phytoseiulus Persimilis

The predatory mite Phytoseiulus persimilis locates its prey, the two-spotted spider mite, by means of herbivore-induced plant volatiles. The olfactory response to this quantitatively and qualitatively variable source of information is particularly well documented. The mites perform this task with a peripheral olfactory system that consists of just five putative olfactory sensilla that reside in a dorsal field at the tip of their first pair of legs. The receptor cells innervate a glomerular olfactory lobe just ventral of the first pedal ganglion. We have made a 3D reconstruction of the caudal half of the olfactory lobe in adult females. The glomerular organization as well as the glomerular innervation appears conserved across different individuals. The adult females have, by approximation, a 1:1 ratio of olfactory receptor cells to olfactory glomeruli.     

An olfactory bulb slice-based biosensor for multi-site extracellular recording of neural networks

Multi-site recording is the important component for studies of the neural networks. In order to investigate the electrophysiological properties of the olfactory bulb neural networks, we developed a novel slice-based biosensor for synchronous measurement with multi-sites. In the present study, the horizontal olfactory bulb slices with legible layered structures were prepared as the sensing element to construct a tissue-based biosensor with the microelectrode array. This olfactory bulb slice-based biosensor was used to simultaneously record the extracellular potentials from multi-positions. Spike detection and cross-correlation analysis were applied to evaluate the electrophysiological activities. The spontaneous potentials as well as the induced responses by glutamic acid took on different electrophysiological characteristics and firing patterns at the different sites of the olfactory bulb slice. This slice-based biosensor can realize multi-site synchronous monitoring and is advantageous for searching after the firing patterns and synaptic connections in the olfactory bulb neural networks. It is also helpful for further probing into olfactory information encoding of the olfactory neural networks.