Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 20
Filtrar
1.
PLoS One ; 12(9): e0185329, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28945824

RESUMO

The molecular receptive range (MRR) of a mammalian odorant receptor (OR) is the set of odorant structures that activate the OR, while the distribution of these odorant structures across odor space is the tuning breadth of the OR. Variation in tuning breadth is thought to be an important property of ORs, with the MRRs of these receptors varying from narrowly to broadly tuned. However, defining the tuning breadth of an OR is a technical challenge. For practical reasons, a screening panel that broadly covers odor space must be limited to sparse coverage of the many potential structures in that space. When screened with such a panel, ORs with different odorant specificities, but equal tuning breadths, might appear to have different tuning breadths due to chance. We hypothesized that ORs would maintain their tuning breadths across distinct odorant panels. We constructed a new screening panel that was broadly distributed across an estimated odor space and contained compounds distinct from previous panels. We used this new screening panel to test several murine ORs that were previously characterized as having different tuning breadths. ORs were expressed in Xenopus laevis oocytes and assayed by two-electrode voltage clamp electrophysiology. MOR256-17, an OR previously characterized as broadly tuned, responded to nine novel compounds from our new screening panel that were structurally diverse and broadly dispersed across an estimated odor space. MOR256-22, an OR previously characterized as narrowly tuned, responded to a single novel compound that was structurally similar to a previously known ligand for this receptor. MOR174-9, a well-characterized receptor with a narrowly tuned MRR, did not respond to any novel compounds in our new panel. These results support the idea that variation in tuning breadth among these three ORs is not an artifact of the screening protocol, but is an intrinsic property of the receptors.


Assuntos
Odorantes , Receptores Odorantes/fisiologia , Animais , Avaliação Pré-Clínica de Medicamentos , Fenômenos Eletrofisiológicos , Feminino , Humanos , Ligantes , Camundongos , Oócitos/metabolismo , Técnicas de Patch-Clamp , Receptores Odorantes/efeitos dos fármacos , Receptores Odorantes/genética , Proteínas Recombinantes/efeitos dos fármacos , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Olfato/efeitos dos fármacos , Olfato/genética , Olfato/fisiologia , Relação Estrutura-Atividade , Xenopus laevis
2.
Biosens Bioelectron ; 35(1): 335-341, 2012 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-22475887

RESUMO

We developed a nanovesicle-based bioelectronic nose (NBN) that could recognize a specific odorant and mimic the receptor-mediated signal transmission of human olfactory systems. To build an NBN, we combined a single-walled carbon nanotube-based field effect transistor with cell-derived nanovesicles containing human olfactory receptors and calcium ion signal pathways. Importantly, the NBN took advantages of cell signal pathways for sensing signal amplification, enabling ≈ 100 times better sensitivity than that of previous bioelectronic noses based on only olfactory receptor protein and carbon nanotube transistors. The NBN sensors exhibited a human-like selectivity with single-carbon-atomic resolution and a high sensitivity of 1 fM detection limit. Moreover, this sensor platform could mimic a receptor-meditated cellular signal transmission in live cells. This sensor platform can be utilized for the study of molecular recognition and biological processes occurring at cell membranes and also for various practical applications such as food screening and medical diagnostics.


Assuntos
Técnicas Biossensoriais/instrumentação , Odorantes/análise , Olfato/fisiologia , Sequência de Bases , Técnicas Biossensoriais/métodos , DNA Complementar/genética , Desenho de Equipamento , Células HEK293 , Humanos , Proteínas Imobilizadas , Nanotubos de Carbono , Receptores Odorantes/genética , Receptores Odorantes/fisiologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Transdução de Sinais
3.
PLoS One ; 4(10): e7647, 2009 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-19876403

RESUMO

Intranasal medications are used to treat various nasal disorders. However, their effects on olfaction remain unknown. Zicam (zinc gluconate; Matrixx Initiatives, Inc), a homeopathic substance marketed to alleviate cold symptoms, has been implicated in olfactory dysfunction. Here, we investigated Zicam and several common intranasal agents for their effects on olfactory function. Zicam was the only substance that showed significant cytotoxicity in both mouse and human nasal tissue. Specifically, Zicam-treated mice had disrupted sensitivity of olfactory sensory neurons to odorant stimulation and were unable to detect novel odorants in behavioral testing. These findings were long-term as no recovery of function was observed after two months. Finally, human nasal explants treated with Zicam displayed significantly elevated extracellular lactate dehydrogenase levels compared to saline-treated controls, suggesting severe necrosis that was confirmed on histology. Our results demonstrate that Zicam use could irreversibly damage mouse and human nasal tissue and may lead to significant smell dysfunction.


Assuntos
Gluconatos/farmacologia , Mucosa Nasal/efeitos dos fármacos , Neurônios Receptores Olfatórios/metabolismo , Receptores Odorantes/fisiologia , Administração Intranasal , Animais , Humanos , L-Lactato Desidrogenase/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Odorantes , Transtornos do Olfato/induzido quimicamente , Olfato , Tubulina (Proteína)/metabolismo
4.
J Neurosci ; 29(10): 3138-47, 2009 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-19279251

RESUMO

Leptin acts via its receptor (LepRb) on specific CNS neurons to signal the adequacy of long-term energy stores, thereby permitting the expenditure of resources on energy-intensive processes such as reproduction. The ventral premammillary nucleus of the hypothalamus (PMv), which has been implicated in the stimulation of gonadotropin release by olfactory cues, contains numerous LepRb neurons, suggesting a potential role for LepRb PMv neurons in transmitting both metabolic and odorant signals to the neuroendocrine reproductive system. Indeed, Fos immunoreactivity and electrophysiologic recordings revealed the direct activation of LepRb PMv neurons by leptin, and exposure to odors from mice of the opposite sex promoted Fos immunoreactivity (Fos-IR) in many LepRb PMv neurons. To determine the regions innervated by the LepRb PMv neurons, we used two novel cre-activated tract-tracing systems in Lepr(cre) animals; data from these systems and from standard tracing techniques revealed that LepRb PMv neurons project to a subset of the regions, including the preoptic area, that are innervated by the PMv as a whole. Furthermore, the retrograde accumulation in LepRb PMv neurons of a trans-synaptic tracer from GnRH neurons revealed the direct innervation of GnRH neurons by many LepRb PMv neurons. Thus, LepRb PMv neurons sense metabolic and sexual odorant cues and project to the rostral hypothalamus to directly innervate GnRH neurons. These results are consistent with a role for LepRb PMv neurons in regulating the reproductive axis in response to metabolic and odorant stimuli.


Assuntos
Hormônio Liberador de Gonadotropina/metabolismo , Hipotálamo/química , Neurônios/metabolismo , Receptores para Leptina/fisiologia , Receptores Odorantes/análise , Atrativos Sexuais/administração & dosagem , Animais , Feminino , Técnicas de Introdução de Genes , Hormônio Liberador de Gonadotropina/análise , Hipotálamo/efeitos dos fármacos , Hipotálamo/fisiologia , Técnicas In Vitro , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Rede Nervosa/química , Rede Nervosa/efeitos dos fármacos , Rede Nervosa/metabolismo , Neurônios/química , Neurônios/efeitos dos fármacos , Receptores para Leptina/genética , Receptores Odorantes/fisiologia , Atrativos Sexuais/fisiologia
5.
Chem Senses ; 34(2): 159-69, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19073950

RESUMO

Chemical signals are common in most crustacean social interactions and are often perceived via chemosensory (olfactory) organs on the first antenna. Intermolt courtship behaviors and mating were investigated in size-matched pairs of intermolt European lobsters (Homarus gammarus) where the olfactory receptors of either the male or the female were lesioned with distilled water (olfactory ablation) or seawater (control). Matings or advanced male courtship behaviors (mounting and turning) were common in seawater controls and olfactory-ablated females. In contrast, when male olfaction was ablated with distilled water, there was not a single mating, and the only male courtship behaviors seen were a few very brief and unsuccessful mounting attempts. Individual females mated up to 5 times with different males, showing that previously inseminated females were still attractive to males. Thus, male but not female olfaction is crucial for intermolt mating in H. gammarus, indicating the presence of a female sex pheromone during the entire female molt cycle, not only at the time of molting. Female sex discrimination may be based on other cues from the male in combination with typical male behaviors.


Assuntos
Inseminação/fisiologia , Preferência de Acasalamento Animal/fisiologia , Nephropidae/fisiologia , Atrativos Sexuais/fisiologia , Olfato/fisiologia , Comunicação Animal , Animais , Feminino , Masculino , Receptores Odorantes/fisiologia , Atrativos Sexuais/análise , Fatores Sexuais
6.
Neuron ; 60(5): 767-74, 2008 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-19081373

RESUMO

The detection of diverse chemical structures by the vertebrate olfactory system is accomplished by the recognition of odorous ligands by their cognate receptors. In the present study, we used computational screening to discover novel high-affinity agonists of an olfactory G protein-coupled receptor that recognizes amino acid ligands. Functional testing of the top candidates validated several agonists with potencies higher than any of the receptor's known natural ligands. Computational modeling revealed molecular interactions involved in ligand binding and further highlighted interactions that have been conserved in evolutionarily divergent amino acid receptors. Significantly, the top compounds display robust activities as odorants in vivo and include a natural product that may be used to signal the presence of bacteria in the environment. Our virtual screening approach should be applicable to the identification of new bioactive molecules for probing the structure of chemosensory receptors and the function of chemosensory systems in vivo.


Assuntos
Avaliação Pré-Clínica de Medicamentos/métodos , Ligantes , Neurônios Receptores Olfatórios/fisiologia , Receptores Odorantes/antagonistas & inibidores , Receptores Odorantes/fisiologia , Olfato/fisiologia , Aminoácidos/química , Animais , Cálcio/metabolismo , Linhagem Celular Transformada , Desenho Assistido por Computador , Carpa Dourada , Humanos , Modelos Moleculares , Sondas Moleculares , Neurônios Receptores Olfatórios/efeitos dos fármacos , Ligação Proteica/efeitos dos fármacos , Ligação Proteica/fisiologia , Conformação Proteica , Curva ROC , Receptores Odorantes/química , Bibliotecas de Moléculas Pequenas , Relação Estrutura-Atividade
9.
Eur J Neurosci ; 22(12): 3147-60, 2005 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-16367781

RESUMO

A morphologically identifiable type of olfactory sensillum on the antenna of the American cockroach contains a pair of ON and OFF cells that responds oppositely to changes in the concentration of fruit odours. The odour of lemon oil was used to study the accuracy with which these cells can discriminate between rapid step-like, ramp-like and oscillating changes in odour concentration. The discharge rates of both cells are not only affected by the actual concentration at particular instants in time (instantaneous concentration) but also by the rate at which concentration changes. The impulse frequency of the fruit odour ON cell is high when odour concentration is high, but higher still when odour concentration is also rising. Conversely, the impulse frequency of the fruit odour OFF cell is high when odour concentration is low and higher still when odour concentration is also falling. Thus, the effect of odour concentration on the responses of both cells is reinforced by the rate of change. Sensitivity to the rate of concentration change becomes greater when the rate is low. Because of the high sensitivity to low rates of change, these cells are optimized to detect fluctuations in fruit odour concentration. Whereas the ON cell signals the arrival and presence of fruit odour, the OFF cell detects its termination and absence. These cells provide excitatory responses for both increase and decrease in fruit odour concentration and may therefore reinforce contrast information.


Assuntos
Potenciais de Ação/fisiologia , Baratas/fisiologia , Odorantes , Neurônios Receptores Olfatórios/fisiologia , Receptores Odorantes/fisiologia , Órgãos dos Sentidos/citologia , Potenciais de Ação/efeitos dos fármacos , Animais , Relação Dose-Resposta a Droga , Modelos Lineares , Microscopia Eletrônica de Varredura/métodos , Neurônios Receptores Olfatórios/efeitos dos fármacos , Neurônios Receptores Olfatórios/ultraestrutura , Óleos de Plantas/farmacologia , Receptores Odorantes/efeitos dos fármacos , Fatores de Tempo
10.
Chem Senses ; 30(3): 241-51, 2005 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15741598

RESUMO

Recognition of the mother is of major importance for the survival of mammalian neonates. This recognition is based, immediately after birth, on the detection of odours that have been learned by the fetus in utero. If the ethological basis of a transnatal olfactory continuity is well established, little is known on the nature of its olfactory cues, and nothing about the presence of potential carrier proteins in the maternal fluids such as amniotic fluid, colostrum and milk. We have identified the components of the pig putative maternal pheromone in these fluids of the sow. We also used a ligand-oriented approach to functionally characterize carrier proteins for these compounds in the maternal fluids. Six proteins were identified, using binding assay, immunodetection and peptide mapping by mass spectrometry. These proteins are known to transport hydrophobic ligands in biological fluids. Among them, alpha-1 acid glycoprotein (AGP) and odorant-binding protein (OBP) have been described in the oral sphere of piglets as being involved in the detection of pig putative maternal pheromone components. These are the first chemical and biochemical data supporting a transnatal olfactory continuity between the fetal and the postnatal environments.


Assuntos
Animais Recém-Nascidos/fisiologia , Mucosa Olfatória/fisiologia , Feromônios/fisiologia , Receptores Odorantes/fisiologia , Líquido Amniótico , Animais , Sítios de Ligação , Colostro , Discriminação Psicológica , Feminino , Imunoquímica , Leite , Orosomucoide/fisiologia , Fotofluorografia , Órgãos dos Sentidos/fisiologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Suínos
11.
J Comp Physiol A ; 187(6): 477-87, 2001 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-11548994

RESUMO

Odorants evoke an outward current in cultured lobster olfactory receptor neurons voltage clamped at -60 mV. The reversal potential of the outward current is independent of the reversal potential of potassium, but shifts with imposed changes in the reversal potential of chloride. The slope of the current-voltage relationship is negative, suggesting that the current is mediated by the odorant suppressing a steady-state conductance. Anthracene-9-carboxylic acid, a specific chloride channel blocker, reversibly inhibits the steady-state conductance. Local application of odorants to the outer dendrites evokes a hyperpolarizing receptor potential in lobster olfactory receptor neurons current-clamped at -70 mV in situ. Consistent with the current characterized in the cultured cells, hyperpolarizing receptor potentials in some cells are voltage sensitive, blocked by anthracene-9-carboxylic acid and associated with a decrease in membrane conductance. These results support the hypothesis that odorants suppress a steady-state chloride conductance in lobster olfactory receptor neurons. Evidence that the chloride conductance can coexist with a 4-aminopyridine-blockable potassium conductance reported earlier in these cells suggests that two distinct mechanisms can mediate odorant-evoked inhibition in lobster olfactory receptor neurons.


Assuntos
Cloretos/química , Nephropidae/fisiologia , Receptores Odorantes/fisiologia , Olfato/fisiologia , Potenciais de Ação , Animais , Eletrofisiologia , Neurônios/fisiologia , Odorantes , Técnicas de Patch-Clamp
12.
Chem Senses ; 26(4): 385-98, 2001 May.
Artigo em Inglês | MEDLINE | ID: mdl-11369673

RESUMO

Many organisms use olfactory appendages bearing arrays of microscopic hairs to pick up chemical signals from the surrounding water or air. We report a morphometric and high speed kinematic analysis of the olfactory organs (lateral flagella of the antennules, which bear chemosensory aesthetasc hairs) of the spiny lobster, Panulirus argus. Panulirus argus sample specific locations by executing a rapid series of antennule flicks at one position, moving the antennule to a different spot and then performing another series of flicks. Odorant delivery to an aesthetasc depends on the water motion near it, which depends on its Reynolds number (Re, proportional to both the diameter and speed of the hair). High speed video enabled us to resolve that during a series of flicks, an antennule moves down rapidly (aesthetasc Re = 2) and up more slowly (Re = 0.5), pausing briefly ( approximately 0.54 s) before the next downstroke. The antennules of P. argus operate in a range of Re values and inter-aesthetasc spacings in which penetration of fluid between the hairs in an array is especially sensitive to changes in speed. Therefore, when antennules flick 'old' water is flushed out of the aesthetasc array during the leaky downstroke and is not picked up again during the less leaky upstroke, hence the antennules can take discrete samples. Thus, by operating in this critical Re range these antennules should be particularly effective at sniffing.


Assuntos
Flagelos/fisiologia , Nephropidae/anatomia & histologia , Nephropidae/fisiologia , Receptores Odorantes/fisiologia , Órgãos dos Sentidos/anatomia & histologia , Olfato/fisiologia , Animais , Fenômenos Biomecânicos , Flagelos/química , Receptores Odorantes/química , Órgãos dos Sentidos/fisiologia , Gravação em Vídeo/instrumentação , Movimentos da Água
13.
Brain Res ; 879(1-2): 73-87, 2000 Oct 06.
Artigo em Inglês | MEDLINE | ID: mdl-11011008

RESUMO

In 1999 we reported that odorants evoke in the Helix pedal ganglion (PG) activity patterns which are largely odorant-specific and related to the nature of odor and its behavioral output. Notably, some activities (for example, approximately 1.5 and approximately 3 Hz), nonspecific to odorants, were consistently evoked in PG. The present contribution goes farther in a deeper survey of the intrinsic and odorant-evoked activities of PG with special weight on the nonspecific fluctuations. We address the following questions. (i) What are the features of the activities? (ii) Are they comparable to the activities found in the motor systems of the other invertebrates? (iii) To what functions can they be related? Three main frequency components represented by power peaks at <1 Hz, 1-2 Hz and 2-8 Hz seem to feature the response activities of PG. (a) The aversive odorants induce odorant-specific patterns represented by peak power frequencies at <1 Hz. (b) The oscillation at approximately 1 Hz, which exists intrinsically in the Helix PG, can be specifically enhanced by appetitive odors. Activities induced in the procerebrum (PC), the visceral ganglion (VG) and PG by appetitive odorants, such as ethanol and apple, peak at 1.3-2 Hz, whereas those induced by aversive ones, such as formic acid and onion at <1 Hz. (c) The 2-8 Hz components always accompany the odorant-evoked activities of the PG either as the second or third strongest component, or in the form of conspicuous, long-lasting approximately 3 Hz oscillations. (d) The nonspecific odor-evoked 1-2 Hz and approximately 3 Hz activities, and the intrinsic approximately 1 Hz activity of the PG seem to be interrelated by a degree of mutual exclusion. We may therefore consider these activities as elementary, slow components that are involved in the processing of signals in this ganglion. It can be inferred from the findings in other invertebrates that the 1-3 Hz spontaneous discharge is strongly connected with motor activity that involves the feedback mechanism of the procerebro-cerebro-buccal or -procerebro-cerebro-pedal circuit. Our approach differs from most others reported so far in the following aspects: (i) use of gross steel electrodes for recording population activities; (ii) lengthy stimulation (10 min); (iii) long observation during and after stimulation; (iv) power spectral presentation of temporal evolution of activity patterns; (v) estimation of peak power frequency by Frequency-Amplitude Plot (FAP) (obtained from signals averaged in the frequency domain; a method based on systems theory).


Assuntos
Gânglios dos Invertebrados/fisiologia , Caracois Helix/fisiologia , Odorantes , Receptores Odorantes/fisiologia , Álcoois , Amônia , Animais , Potenciais Evocados , Formiatos , Frutas , Técnicas In Vitro , Cebolas
14.
J Neurosci ; 19(17): 7426-33, 1999 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-10460249

RESUMO

Here, we provide the first evidence for functional expression of a human olfactory receptor protein (OR17-40) and show that recombinant olfactory receptors can be functionally expressed in heterologous systems. A mixture of 100 different odorants (Henkel 100) elicited a transient increase in intracellular [Ca(2+)] in human embryonic kidney 293 (HEK293) cells stably or transiently transfected with the plasmid pOR17-40. By subdividing the odorant mixture into progressively smaller groups, we identified a single component that represented the only effective substance: helional. Only the structurally closely related molecule heliotroplyacetone also activated the receptor. Other compounds, including piperonal, safrole, and vanillin, were completely ineffective. Mock-transfected cells and cells transfected with other receptors showed no change in intracellular [Ca(2+)] in response to odor stimulation. We were also able to functionally express OR17-40 in Xenopus laevis oocytes. Coexpression of a "reporter" channel allowed measurement of the response of oocytes injected with the cRNA of the human receptor to the odor mixture Henkel 100. The effective substances were the same (helional, heliotropylacetone) as those identified by functionally expressing the receptor in HEK293 cells and were active at the same, lower micromolar concentration. These findings open the possibility of now characterizing the sensitivity and specificity of many, if not all, of the hundreds of different human olfactory receptors.


Assuntos
Receptores Odorantes/fisiologia , Trifosfato de Adenosina/farmacologia , Sequência de Aminoácidos , Animais , Cálcio/metabolismo , Linhagem Celular , Membrana Celular/fisiologia , DNA Complementar , Feminino , Humanos , Rim , Potenciais da Membrana/efeitos dos fármacos , Modelos Neurológicos , Dados de Sequência Molecular , Odorantes , Oócitos/fisiologia , Estrutura Secundária de Proteína , RNA Complementar/metabolismo , Receptores Odorantes/química , Receptores Odorantes/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Mapeamento por Restrição , Transfecção , Xenopus laevis
15.
Chem Senses ; 22(4): 467-76, 1997 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-9279469

RESUMO

Odorant receptors of zebrafish and C elegans were functionally expressed in vertebrate kidney cells (HEK293) using the eucaryotic expression vector pSMyc. Receptor-encoding cDNA cloned into this vector was expressed as a fusion protein with the N-terminal membrane import sequence of the guinea-pig serotonin receptor followed by a myc tag. Immunocytochemical evidence indicates that this strategy directs a protein with the predicted immunoreactivity and approximate molecular weight to the plasma membrane. Fish food extract (TetraMin) evoked a transient increase in intracellular [Ca2+] in HEK293 cells transiently transfected with plasmids containing cDNA for three fish odorant receptors and converted to stable cell lines. The effect of the extract was concentration dependent and limited to the fraction of the extract < 5 kDa. Pretreating the transfected cells with the PLC inhibitor U73122 reduced the odor-evoked signal. Fish food extract also evoked a transient increase in intracellular [Ca2+] in HEK293 cells transiently transfected with plasmids containing cDNA for single fish odorant receptors. Diacetyl evoked a transient increase in intracellular [Ca2+] in HEK293 cells transiently transfected with plasmids encoding the cDNA of ODR10, an odorant receptor of C. elegans suggested in other work to be specific for diacetyl. These results strongly imply that odorant receptors can be functionally expressed in HEK293 cells using this novel expression protocol.


Assuntos
Caenorhabditis elegans/genética , Receptores Odorantes/genética , Receptores Odorantes/fisiologia , Peixe-Zebra/genética , Animais , Sequência de Bases , Western Blotting , Cálcio/metabolismo , Linhagem Celular , Clonagem Molecular , DNA Complementar/genética , DNA Complementar/fisiologia , Expressão Gênica/genética , Expressão Gênica/fisiologia , Vetores Genéticos , Humanos , Imuno-Histoquímica , Ligantes , Dados de Sequência Molecular , Receptores Odorantes/imunologia , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/fisiologia , Transdução de Sinais/fisiologia , Transfecção/genética , Transfecção/fisiologia
16.
J Exp Biol ; 200(Pt 15): 2073-81, 1997 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-9255949

RESUMO

The responsiveness of chemoreceptor neurons depends on a combination of perireceptor and receptor events. Olfactory neurons of crustaceans are packaged into distinctive cuticular sensilla called aesthetascs. The cuticle of aesthetascs is thin and permeable, even though it does not contain any obvious surface pores or pore tubules. This suggests that this 'spongy' aesthetasc cuticle may act as a molecular sieve that restricts large odorant molecules from entering the sensilla and binding to the olfactory neurons. We examined whether this is so for the aesthetasc cuticle of the Caribbean spiny lobster Panulirus argus. We used a chromatographic column packed with aesthetasc cuticle and connected to a flow-through ultraviolet spectrophotometer to measure the elution times of ultraviolet-absorbent molecular mass markers between 165 and 2 x 10(6) Da. Molecules larger than approximately 8.5 kDa had similar elution times, indicating that they did not penetrate the cuticle. Molecules smaller than 8.5 kDa had longer elution times that were directly and inversely proportional to their molecular mass. These results suggest that aesthetasc cuticle excludes molecules larger than 8.5 kDa from having access to the olfactory receptor neurons. We conclude that the molecular sieving capacity of the aesthetasc cuticle of P. argus is a perireceptor mechanism that is a critical determinant of the types of molecules capable of stimulating its olfactory receptors.


Assuntos
Cromatografia/métodos , Nephropidae/anatomia & histologia , Nephropidae/fisiologia , Odorantes , Receptores Odorantes/fisiologia , Animais , Microscopia Eletrônica de Varredura , Peso Molecular , Tamanho da Partícula , Espectrofotometria Ultravioleta
17.
Nat Struct Biol ; 3(11): 934-9, 1996 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-8901871

RESUMO

Odorant binding protein (OBP) is the major odorant binding component of mammalian nasal mucosa. The two structures of bovine OBP reported in this paper (one crystallized as purified and one soaked in the presence of a selenium-containing odorant) show that: (i) the OBP dimer is composed of two compact domains related by an approximate two-fold axis of symmetry; (ii) between residues 122 and 123 the polypeptide chains cross from one domain to the other such that each domain is formed by residues from both monomers; (iii) purified OBP already contains two bound odorant molecules (one per monomer)-odorant binding occurs by replacement of these molecules with the added odorant; and (iv) the structure of the odorant binding site can explain OBP's extraordinarily broad odorant specificity.


Assuntos
Receptores Odorantes/química , Olfato/fisiologia , alfa-Globulinas/química , Animais , Sítios de Ligação , Bovinos , Cristalografia por Raios X , Dimerização , Odorantes , Estrutura Secundária de Proteína , Estrutura Terciária de Proteína , Receptores Odorantes/fisiologia , Selênio
18.
J Comp Physiol A ; 176(4): 527-40, 1995 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-7722958

RESUMO

1. The effects of the odorant compounds adenosine-5'-monophosphate (5'AMP), ammonium, betaine, L-cysteine, L-glutamate, DL-succinate, and taurine and of mixtures of these compounds on binding of taurine and 5'AMP to dendritic membrane from the olfactory organ of spiny lobsters (Panulirus argus) were quantified to evaluate the contribution of inhibition of odorant-receptor binding to the generation of physiological responses to mixtures. 2. Taurine binding sites belong to two affinity classes, while 5'AMP binding sites belong to a single affinity class. Binding of either taurine or 5'AMP was partially inhibited in an apparently noncompetitive, concentration dependent fashion by most odorant compounds, with 25-40% inhibition by 1 mM of odorant. Mixtures of two or more odorant compounds also inhibited binding of taurine or 5'AMP to its sites. However, the inhibition by mixtures was often significantly less than expected from the inhibition produced by a mixture's components assuming either a noncompetitive or competitive mechanism. 3. By including this binding inhibition between compounds into models for predicting physiological responses to mixtures from the responses to the components, the predictive power of the models is significantly improved. This result strongly suggests that binding inhibition can influence the physiological responsiveness of chemoreceptor cells to mixtures.


Assuntos
Monofosfato de Adenosina/farmacologia , Bulbo Olfatório/fisiologia , Receptores Odorantes/fisiologia , Taurina/farmacologia , Animais , Sítios de Ligação , Cisteína/farmacologia , Relação Dose-Resposta a Droga , Nephropidae , Compostos de Amônio Quaternário/farmacologia
19.
Proc Natl Acad Sci U S A ; 92(1): 62-6, 1995 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-7816848

RESUMO

Chemical signals connect most of life's processes, including interorganismal relationships. Detection of chemical signals involves not only recognition of a spectrum of unique compounds or mixtures of compounds but also their spatial and temporal distribution. Both spectral and temporal signal processing determine what is a signal and what is background noise. Each animal extracts its unique information from the chemical world and uniquely contributes to it. Lobsters have provided important information on temporal signal processing. Marine chemical signals can be measured with high spatio-temporal resolution giving us a novel view of the lobster's environment. Lobster chemoreceptor cells have flicker fusion frequencies of 4 Hz and can integrate stimuli over 200 ms, closely corresponding to odor sampling behavior with 4-Hz "sniffs." Using this information, spatial odor gradients can be determined from temporal analysis of odor patches typical of turbulent dispersal. Lobsters appear to use this information to locate odor sources. Lobster social behavior depends greatly on chemical signals. Urine carries important information for courtship, dominance, and individual recognition. A novel gland in the nephropore is strategically located to release its products into the urine. Urine, in turn, is injected into the gill current, which jets water 1-2 m ahead of the animal. Lobsters control three different currents that carry chemical signals to and from them. The study of odor dynamics has only just begun. It will be exciting to see how signal dispersal, receptor temporal tuning, neural processing, and animal behavior interact to enhance signals for communication and detection and to reduce signals for chemical camouflage.


Assuntos
Células Quimiorreceptoras/fisiologia , Nephropidae/fisiologia , Receptores Odorantes/fisiologia , Comportamento Sexual Animal/fisiologia , Transdução de Sinais , Predomínio Social , Agressão/fisiologia , Animais , Meio Ambiente , Feminino , Masculino , Odorantes , Água do Mar , Fatores de Tempo
20.
Chem Senses ; 19(1): 71-86, 1994 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-8055260

RESUMO

Natural odors occur as turbulent plumes resulting in spatially and temporally variable odor signals at the chemoreceptor cells. Concentrations can fluctuate widely within discrete packets of odor and individual packets are very intermittent and unpredictable. Chemoreceptor cells display the temporally dynamic properties of adaptation and disadaptation, which serve to alter their responses to these fluctuating odor patterns. A computational model, modified from one previously published, was used to investigate the effect of adaptation and recovery of adaptation (disadaptation) on the spike output of model olfactory receptor cells under natural stimulus conditions. The response characteristics of model cells were based upon empirically determined dose-response, adaptation, disadaptation and flicker fusion properties of peripheral olfactory cells. The physiological properties of the model cell (adaptation and disadaptation rate and the dose-response relationship) could be modified independently, allowing assessment of the role of each in shaping the responses of the model cell. Complete adaptation and disadaptation time courses ranged from 500 ms (rapid cells) to 10 s (slow cells). The stimuli for the model cells were quantified odor plume recordings obtained under a variety of biologically relevant flow conditions. As expected, the rapidly adapting model cells displayed different response characteristics than the slowly adapting model cells to identical temporal odor profiles. Responses of the model cells depended upon their adaptation and disadaptation rates, and the frequency characteristics of the odor presentation. These results indicate that adaptation and disadaptation determine the range of concentration fluctuations over which a particular cell will respond. Thus, these properties function as an olfactory equivalent of a band-pass filter in electronics. This type of filtering has implications for the extraction of information from odor signals, such as the coding of temporal and intensity features.


Assuntos
Adaptação Fisiológica/fisiologia , Neurônios Aferentes/fisiologia , Receptores Odorantes/fisiologia , Animais , Simulação por Computador , Eletrodos , Análise de Fourier , Modelos Neurológicos , Nephropidae , Orientação/fisiologia , Percepção Espacial/fisiologia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA