秦岭蝮嗅觉系统和犁鼻系统的显微结构观察

用光镜观察了秦岭蝮Gloydius qinlingensis嗅觉系统和犁鼻系统的组织结构.结果显示秦岭蝮嗅觉系统主要包括嗅器和嗅球,犁鼻系统主要包括犁鼻器和副嗅球,并且嗅器和犁鼻器已经完全分离形成两个独立的囊,犁鼻器位于嗅器的内侧.嗅器粘膜上皮进一步分化为嗅上皮和呼吸上皮,背侧嗅上皮下的固有层内有丰富的Bowmans腺,腹侧呼吸上皮内有大量的杯状细胞,其固有层未见有Bowmans腺.鼻腔的中段出现了发达的犁鼻器,犁鼻上皮明显比嗅上皮厚,其固有层内未见有犁鼻腺,在犁鼻腔内还有蘑菇体.     

Ex Vivo Preparations of the Intact Vomeronasal Organ and Accessory Olfactory Bulb

The mouse accessory olfactory system (AOS) is a specialized sensory pathway for detecting nonvolatile social odors, pheromones, and kairomones. The first neural circuit in the AOS pathway, called the accessory olfactory bulb (AOB), plays an important role in establishing sex-typical behaviors such as territorial aggression and mating. This small (<1 mm³) circuit possesses the capacity to distinguish unique behavioral states, such as sex, strain, and stress from chemosensory cues in the secretions and excretions of conspecifics. While the compact organization of this system presents unique opportunities for recording from large portions of the circuit simultaneously, investigation of sensory processing in the AOB remains challenging, largely due to its experimentally disadvantageous location in the brain. Here, we demonstrate a multi-stage dissection that removes the intact AOB inside a single hemisphere of the anterior mouse skull, leaving connections to both the peripheral vomeronasal sensory neurons (VSNs) and local neuronal circuitry intact. The procedure exposes the AOB surface to direct visual inspection, facilitating electrophysiological and optical recordings from AOB circuit elements in the absence of anesthetics. Upon inserting a thin cannula into the vomeronasal organ (VNO), which houses the VSNs, one can directly expose the periphery to social odors and pheromones while recording downstream activity in the AOB. This procedure enables controlled inquiries into AOS information processing, which can shed light on mechanisms linking pheromone exposure to changes in behavior.     

4种两栖爬行动物嗅器和犁鼻器的显微结构比较

用光镜观察了4种两栖爬行动物嗅器和犁鼻器的组织结构.结果显示,北方山溪鲵(Batrachuperus tibetanus)鼻囊内开始分化出犁鼻器,犁鼻器位于嗅器的腹外侧,但犁鼻器还不发达;隆肛蛙(Feirana quadranus)犁鼻器与嗅器虽然共同位于鼻囊内,但犁鼻器较为发达且其周围有发达的犁鼻腺,犁鼻器通过一细小管道与嗅器相通;秦岭蝮(Gloydius qinlingensis)和菜花烙铁头(Trimeresurus jerdonii)犁鼻腔与鼻腔已经完全分离形成两个独立的囊,而且鼻腔又进一步分化为嗅部与呼吸部.说明犁鼻器从有尾两栖动物开始出现,至无尾两栖类开始分化,到蛇类高度发达且成为一个独立器官.犁鼻器的形成是脊椎动物适应陆地生活的直接结果,是四足动物的特征之一.     

黑斑蛙嗅器和犁鼻器的组化及超微结构特征

嗅感受器主要感知外界环境中化学信号分子.本文采用银染、NADPH-组化染色和电镜技术来观察黑斑侧褶蛙(Petophylax nigromaculatus)的嗅器和犁鼻器的功能差异及细胞组成.银染法可对嗅上皮和犁鼻上皮的细胞进行分类及区分.其中,支持细胞胞核深染成黑色,嗅细胞胞核银染为花斑状.细胞计数显示,犁鼻上皮的嗅神经细胞含量百分比显著高于嗅上皮.组化结果显示,黑斑侧褶蛙嗅上皮和犁鼻上皮对NADPH-d表达模式差异显著,前者表达明显高于后者.电镜结果显示,黑斑侧褶蛙嗅上皮和犁鼻上皮的支持细胞由两种类型的细胞组成,分别为纤毛型和颗粒型支持细胞.     

A Quantitative Study of Olfactory, Non-Olfactory, and Vomeronasal Epithelia in the Nasal Fossa of the Bat Megaderma lyra

In complexity, the mammalian nasal fossa is unparalleled among vertebrates. Although total nasal epithelial surface areas (SA) have been reported for numerous mammals, detailed quantitative reports on individual structures exist for few mammals. Here, we examine mucosal distribution in the nasal fossa of the greater false vampire bat, Megaderma lyra (Megadermatidae, Chiropera). The SA of the left nasal fossa of one adult Megaderma was measured in serial histological sections; the development of the nasal fossa was assessed using three fetal specimens. The nasal fossa of Megaderma has seven ethmoturbinals and multiple smaller interturbinals between them, all bearing olfactory mucosa. Nearly half of the total olfactory SA of the nasal fossa lines these turbinals; the remainder lines recesses and parts of the medial (septal) and lateral walls of the nasal fossa. The maxilloturbinal is diminutive, and the nasoturbinal is absent. Volumetric measurements of the fetal and adult vomeronasal organ suggest age-related reduction. Thirty-five percent of the nasal fossa is lined with olfactory mucosa, within the range reported previously for chiropterans. In Megaderma the frontal recess contributes little to total nasal SA (2% of all olfactory SA). This represents a significant departure in morphology compared to other mammals, including some bats, in which the frontal recess is much larger. The significance of the emphasis in olfactory SA distribution to central or more peripheral (paranasal) spaces could be investigated using a large sample of phylogenetically diverse mammals, such as bats. This study emphasizes the need for more histological detail to further such studies.     

GABAergic Neurons in the Embryonic Olfactory Pit/Vomeronasal Organ: Maintenance of Functional GABAergic Synapses in Olfactory Explants

In previous work, we showed a robust γ-aminobutyric acid (GABAergic) synaptic input onto embryonic luteinizing hormone-releasing hormone (LHRH) neurons maintained in olfactory explants. In this study, we identify GABAergic neurons in olfactory pit (OP) of embryonic micein vivoand study, using patch-pipet whole-cell current and voltage clamp techniques, synaptic interactions of these neurons in explant cultures.In vivo,glutamate decarboxylase (GAD, the enzyme which synthesizes GABA) mRNA was first detected in nasal regions on Embryonic Day (E) 11.5. From E12.5 to E13.5, robust GAD expression was localized to cells primarily in the ventral aspect of the OP. GAD mRNA was not detected over dorsally located cells in olfactory sensory or respiratory epithelium. In addition, GAD mRNA was not observed in cells along olfactory axons. GAD mRNA was dramatically reduced in the OP/vomeronasal organ by E16.5. Using antibodies against both GABA and GAD, immunopositive axonal-like tracts were detected in the nasal septum on E12.5. GABAergic staining decreased by E13.5. To examine synaptic interactions of these GABAergic cells, embryonic olfactory explants were generated and maintained in serum-free media. As explants spread, neuron-like cells migrated into the periphery, sometimes forming ganglion-like clusters. Cells were recorded, marked intracellularly with Lucifer Yellow and post-fixation, immunocytochemically examined. Forty-six cells, typically multipolar, were GABAergic, had resting potentials around −50 mV, and exhibited spontaneous action potentials which were generated by spontaneous depolarizing GABAergic (GABA_A) synaptic activity. OP neurons depolarized in response to GABA by increasing Cl⁻conductance. The biophysical properties of OP-derived GABAergic neurons were distinct from those reported for olfactory receptor neurons but similar to embryonic LHRH neurons. However, unlike LHRH neurons, GABAergic neurons did not migrate large distances in olfactory explants or appear to leave the olfactory pitin vivo.     

哺乳动物主要嗅觉系统和犁鼻系统信息识别的编码模式

哺乳动物具有两套嗅觉系统, 即主要嗅觉系统和犁鼻系统。前者对环境中的大多数挥发性化学物质进行识别, 后者对同种个体释放的信息素进行识别。本文从嗅觉感受器、嗅球、嗅球以上脑区三个水平综述了这两种嗅觉系统对化学信息识别的编码模式。犁鼻器用较窄的调谐识别信息素成分, 不同于嗅上皮用分类性合并受体的方式识别气味; 副嗅球以接受相同受体输入的肾丝球所在区域为单位整合信息, 而主嗅球通过对肾丝球模块的特异性合并编码信息; 在犁鼻系统, 信息素的信号更多地作用于下丘脑区域, 引起特定的行为和神经内分泌反应。而在主要嗅觉系统, 嗅皮层可能采用时间模式编码神经元群, 对气味的最终感受与脑的不同区域有关。犁鼻系统较主要嗅觉系统的编码简单, 可能与其执行的功能较少有关。     

Frequency-dependent Food Selection by Domestic Cats: A Comparative Study

Preferences for common food types (‘apostatic selection’) have been demonstrated in a wide variety of vertebrate predators, yet there are few examples of preferences for rare food types (‘anti-apostatic selection’). Anti-apostatic selection is predicted to occur when, among other things, there are nutritional benefits to be gained from the consumption of a mixed diet. We tested this hypothesis by examining the frequency-dependent food preferences of domestic cats (Felis silvestris catus) with different nutritional histories. Subjects were classified as being either nutritionally ‘experienced’ (farm and rescue shelter cats, with a history of scavenging for nutritionally variable foods) or nutritionally ‘inexperienced’ (cats reared indoors on high-quality, nutritionally complete diets). We tested for frequency dependence by allowing individuals and groups of cats from the two groups to select from high-density mixtures of two types of artificial food pellet. In experiments on individual cats, nutritionally experienced subjects showed significant anti-apostatic selection, whereas inexperienced cats produced only a weak anti-apostatic trend. In experiments on groups of cats, both inexperienced and experienced groups showed significant anti-apostatic selection. The apparent inconsistency between individual and group results could be explained in terms of the additional anti-apostatic effects that result from variation among individuals in group foraging situations (i.e. when the effects of individuals are pooled). Because other behavioural explanations, such as perceptual contrast and sampling effects, were unlikely to have influenced our results, we conclude that the differences in selection between experienced and inexperienced individuals were probably due to the differing extent to which the consumption of a mixed diet was beneficial. These experiments may offer some insight into the success of the domestic cat in urban areas: although obligate carnivores, they appear to possess flexible feeding strategies which will tend to allow them to select a reasonably balanced diet from nutritionally variable resources in, for example, refuse bins.     

Chemocommunication between bacteria and the higher vertebrate animals

Between bacteria and the higher vertebrate animals there are close chemocommunicational connections that are realized via signal molecules secreted by bacteria, on the hand, and vertebrate hormones and hormone-like substances, on the other hand. The review presents data on regulatory effects of biogenic amines (catecholamines and serotonin), peptide hormones, and immunoregulator of the higher vertebrates on the vitally important functions of bacterial cells, their virulence and survivability. It has been shown that some bacterial signal molecules, such as N-acylated derivatives of homoserine lactones, also are able to regulate fundamental cellular processes in the higher vertebrates. Deciphering of molecular mechanisms of information exchange between bacteria and the higher vertebrates is both of theoretical significance for studies on pathways of evolution of chemosignal systems in proand eukaryote organism and of practical significance for development of new approaches for treatment of bacterial infections.     

The Legal Status of Cats in New Zealand: A Perspective on the Welfare of Companion,Stray, and Feral Domestic Cats (Felis catus)

Pinpointing and safeguarding the welfare status of domestic cats is problematic, especially in New Zealand where cats are introduced predators with significant impact on indigenous fauna. Usually the identification of welfare status depends on conservational, legal, and public attitudes that are often contrasting. Cats may rapidly transgress definitions placed on them, confounding attempts to categorize them. In 1 generation, cats can move from a human-dependent state (“stray” or “companion”) to wild (“feral”). Often this categorization uses arbitrary behavioral and or situational parameters; consequent treatment and welfare protection for these cats are similarly affected. Terminology used to describe cats is not equitable across research. However, the New Zealand Animal Welfare (Companion Cats) Code of Welfare 2007 seeks to create a new definition of the terms companion, stray, and feral. It distinguishes between cats who live within and without human social constructs. This legislation mandates that cats in human environments or indirectly dependent on humans cannot be classified as feral. Such definitions may prove vital when safeguarding the welfare of free-living domestic cats and cat colonies.     

猫嗅球外丛层和僧帽细胞层年龄相关形态学变化

分别用Nissl法及免疫组织化学ABC法标记青、老年猫嗅球中嗅觉二级神经元和外丛层胶质细胞,显微镜下观察其分布并计数,对嗅觉二级神经元胞体直径和外丛层厚度进行测量,比较其年龄相关性变化,研究神经元与胶质细胞之间的关系,探讨老年性嗅觉功能衰退的相关神经机理。结果显示,老年猫嗅觉二级神经元胞体直径和分布密度均有不同程度的显著性下降(P<0.05);外丛层厚度变化不明显(P>0.05);外丛层胶质细胞特别是星形胶质细胞显著性增生(P<0.05)。表明在衰老过程中嗅觉二级神经元有丢失,并呈现功能下降,可能是老年性嗅觉功能衰退的原因之一。同时外丛层胶质细胞增生以进一步保护神经元,延缓其衰老。     

Photoperiod Mediated Changes in Olfactory Bulb Neurogenesis and Olfactory Behavior in Male White-Footed Mice (Peromyscus leucopus)

Brain plasticity, in relation to new adult mammalian neurons generated in the subgranular zone of the hippocampus, has been well described. However, the functional outcome of new adult olfactory neurons born in the subventricular zone of the lateral ventricles is not clearly defined, as manipulating neurogenesis through various methods has given inconsistent and conflicting results in lab mice. Several small rodent species, including Peromyscus leucopus, display seasonal (photoperiodic) brain plasticity in brain volume, hippocampal function, and hippocampus-dependent behaviors; plasticity in the olfactory system of photoperiodic rodents remains largely uninvestigated. We exposed adult male P. leucopus to long day lengths (LD) and short day lengths (SD) for 10 to 15 weeks and then examined olfactory bulb cell proliferation and survival using the thymidine analog BrdU, olfactory bulb granule cell morphology using Golgi-Cox staining, and behavioral investigation of same-sex conspecific urine. SD mice did not differ from LD counterparts in granular cell morphology of the dendrites or in dendritic spine density. Although there were no differences due to photoperiod in habituation to water odor, SD mice rapidly habituated to male urine, whereas LD mice did not. In addition, short day induced changes in olfactory behavior were associated with increased neurogenesis in the caudal plexiform and granule cell layers of the olfactory bulb, an area known to preferentially respond to water-soluble odorants. Taken together, these data demonstrate that photoperiod, without altering olfactory bulb neuronal morphology, alters olfactory bulb neurogenesis and olfactory behavior in Peromyscus leucopus.     

Quantifying Neurite Growth Mediated by Interactions among Secretory Vesicles, Microtubules, and Actin Networks

Neurite growth is a fundamental process of neuronal development, which requires both membrane expansions by exocytosis and cytoskeletal dynamics. However, the specific contribution of these processes has not been yet assessed quantitatively. To study and quantify the growth process, we construct a biophysical model in which we relate the overall neurite outgrowth rate to the vesicle dynamics. By considering the complex motion of vesicles in the cell soma, we demonstrate from biophysical consideration that the main step of finding the neurite initiation site relies mainly on a two-dimensional diffusion/sequestration/fusion at the cell surface and we obtain a novel formula for the flux of vesicles at the neurite base. In the absence of microtubules, we show that a nascent neurite initiated by vesicular delivery can only reach a small length. By adding the microtubule dynamics to the secretory pathway and using stochastic analysis and simulations, we study the complex dynamics of neurite growth. Within this model, depending on the coupling parameter between the microtubules and the neurite, we find different regimes of growth, which describe dendritic and axonal growth. To validate one aspect of our model, we demonstrate that the experimental flux of TI-VAMP but not Synaptobrevin 2 vesicles contributes to the neurite growth. We conclude that although vesicles can be generated randomly in the cell body, the search for the neurite position using the microtubule network and diffusion is quite fast. Furthermore, when the TI-VAMP vesicular flow is large enough, the interactions between the microtubule bundle and the neurite control the growth process. In addition, all of these processes intimately cooperate to mediate the various modes of neurite growth: the model predicts three different growing modes including, in addition to the stable axonal growth and the stochastic dendritic growth, a fast oscillatory regime. Finally our study demonstrates that cytoskeletal dynamics is necessary to generate long protrusion, while vesicular delivery alone can only generate small neurite.     

Experimental Verification of the Effects on Normal Domestic Cats by Feeding Prescription Diet for Decreasing Stress

The objective of this study was to evaluate the effects of diet on the feline stress response by measuring plasma and urinary cortisol. A study diet was developed with a unique combination of nutrients that supports the management of stressful situations. The specific formulation of the diet included alpha-casozepine, which is believed to have an anxiolytic effect, and tryptophan supplementation. Tryptophan is the precursor for the synthesis of the neurotransmitter serotonin. Twenty-one indoor cats were fed with the study diet (n = 10) or a control diet (n = 11) for 8 weeks, after which physiological responses were evaluated. The study diet significantly increased the ratio of plasma tryptophan to large neutral amino acids and decreased urinary cortisol concentrations after being consumed daily for 8 weeks, but there was no effect on plasma cortisol levels following a stressful event (veterinary examination and blood draw). Further studies, such as behavioral analyses, are needed to clarify the effects of the study diet.     

Mate Attraction in a Burrowing Wolf-Spider (Araneae, Lycosidae) is not Olfactory Mediated

Spiders are mostly solitary living animals, and males actively search for females by using chemical cues. Compared with the overwhelming evidence demonstrating the role of spider contact sex-pheromones, support for the role of olfactory sex-pheromones in spider communication is scarce. In Lycosa tarantula (L., 1758), a Mediterranean burrowing wolf spider inhabiting dry and open habitats and dependent on chemical cues for mate searching, we investigated the role of olfactory sex-pheromones in mate attraction. We conducted both laboratory and field experiments, using multiple experimental approaches (two-choice olfactometer in the laboratory and pit-fall traps and two-choice open arenas in the field) and controlling for male sexual arousal. Our results support the hypothesis that mate attraction in this species is not olfactory-mediated. In the two-choice olfactometer, males did not orient preferentially to the side where the stimulus was concealed. Stimuli did not influence response latency or the relative time spent in the experimental side of the olfactometer. In the field, the effectiveness of experimental pit-fall traps baited with potential olfactory sex-pheromones did not differ from that of control traps. In the two-choice open arenas, males did not show any clear preference for the experimental half containing the potential olfactory sex-pheromone, nor did they show any sexual response. These results demonstrate that olfactory-mediated cues are not important during mate attraction in this species, and enforce the idea that habitat preferences and life style might explain the relative role of contact and olfactory-mediated chemical cues during spider mate searching.     

棕色田鼠雄性幼体不同发育期犁鼻器和副嗅球的组织结构

通过对出生后不同发育时期雄性棕色田鼠犁鼻器和副嗅球进行组织学观察, 探讨棕色田鼠出生后犁鼻器和副嗅球的发育规律。实验以出生后当天(0 日龄) , 5 日龄, 15 日龄, 25 日龄以及成年棕色田鼠为研究对象,副嗅球采用Pischinger 氏染色法染色, 犁鼻器用H. E. 染色法染色后进行组织学观察。结果显示, 棕色田鼠出生时, 犁鼻器和副嗅球就已具有成体的基本结构, 随着动物个体的发育, 犁鼻上皮逐渐增厚, 犁鼻管变长, 犁鼻上皮中神经元密度增加; 腺体逐渐增大, 犁鼻管腔填充物增多, 犁鼻管背外侧的静脉血管逐日增大, 管腔周围出现越来越多的血管; 副嗅球长宽都增加, 僧帽细胞层和颗粒细胞层逐渐增长, 各层细胞密度变化稍有不同;出生后15 日内, 僧帽细胞层细胞密度增加, 15 日龄以后又开始降低, 25 日龄及成体的僧帽细胞层细胞密度与5日龄的相似; 颗粒细胞层细胞密度持续增高。实验结果提示, 棕色田鼠5 日龄时, 犁鼻器和副嗅球已具有了完整的结构, 到25 日龄时可能达到了功能上的成熟。     

Bartonella Infection among Cats Adopted from a San Francisco Shelter,Revisited

Bartonella infection among cats from shelters can pose a health risk to adopters. Bartonella henselae is the most common species, with B. clarridgeiae and B. koehlerae being less common. The lower rates of infection by the latter species may reflect their rarity or an inefficiency of culture techniques. To assess the incidence of infection, blood cultures, serology, and PCR testing were performed on 193 kittens (6 to 17 weeks old) and 158 young adult cats (5 to 12 months old) from a modern regional shelter. Classical B. henselae culture medium was compared to a medium supplemented with insect cell growth factors. Bartonella colonies were isolated from 115 (32.8%) animals, including 50 (25.9%) kittens and 65 (41.1%) young adults. Therefore, young adults were twice as likely to be culture positive as kittens. Enhanced culture methods did not improve either the isolation rate or species profile. B. henselae was isolated from 40 kittens and 55 young adults, while B. clarridgeiae was cultured from 10 animals in each group. B. koehlerae was detected in one young adult by PCR only. B. henselae genotype II was more commonly isolated from young adults, and genotype I was more frequently isolated from kittens. Kittens were 4.7 times more likely to have a very high bacterial load than young adults. A significantly higher incidence of bacteremia in the fall and winter than in the spring and summer was observed. Bartonella antibodies were detected in 10% (19/193) of kittens and 46.2% (73/158) of young adults, with culture-positive kittens being 9.4 times more likely to be seronegative than young adults.     

Human Classification of Context-Related Vocalizations Emitted by Familiar And Unfamiliar Domestic Cats: An Exploratory Study

ABSTRACT

Previous research has shown that human classification of contextspecific domestic cat “meow” vocalizations is relatively poor, although improves with experience and/or general affinity to cats. To investigate whether such classification further improves when recipients (humans) of the vocalizations reside with the vocalizing animal (cat), cat owners (n=10) were asked to listen to eight audio recordings of a single meow (4 from their own cat and 4 from an unfamiliar cat) produced during one of four possible contexts, and identify the context in which each meow was emitted. Contexts comprised food preparation, food-withholding, negotiating a barrier, and attention solicitation. In addition, participants were asked to rate 20 meow vocalizations (produced by unfamiliar cats in the four contexts) on scales measuring pleasantness and urgency, in order to investigate whether participants reached consensus on the emotional content of the vocalizations. Successful identification of both the context and the hypothesized emotional content of the vocalization would comprise the first steps in our understanding of whether human-directed cat “meow” vocalizations are fully advantageous. Forty percent of the participants identified the correct contexts at a level greater than chance when the vocalizations belonged to their own cat. However, no participants performed above chance when vocalizations belonged to an unfamiliar cat. Participants’ urgency ratings were not significantly influenced by the context in which the vocalization was produced. Pleasantness ratings, however, were significantly higher for the context of negotiating a barrier in comparison with attention solicitation. These results suggest that the domestic cat, as a species, does not have a context-specific repertoire of human-directed vocalizations. Successful context classification of cat meows however, was possible for some owners. Whether such success was due to individual learning ability or recognition of hypothesized emotional content of the call requires further investigation.