Intraspecies cell-cell communication in yeast.

Although yeasts are unicellular microorganisms that can live independently, they can also communicate with other cells, in order to adapt to the environment. Two yeast species, the budding yeast Saccharomyces cerevisiae and the fission yeast Schizosaccharomyces pombe, engage in various kinds of intraspecies cell-cell communication using peptides and chemical molecules that they produce, constituting a sort of 'language'. Cell-cell communication is a fundamental biological process, and its ultimate purpose is to promote survival by sexual reproduction and acquisition of nutrients from the environment. This review summarizes what is known about intraspecies cell-cell communication mediated by molecules including mating pheromones, volatile gases, aromatic alcohols and oxylipins in laboratory strains of S. cerevisiae and S. pombe.

Streptococcal peptides that signal Enterococcus faecalis cells carrying the pheromone-responsive conjugative plasmid pAM373.

Pheromone-mediated conjugative transfer of enterococcal plasmids can contribute to the dissemination of genes involved in antibiotic resistance, fitness, and virulence among co-residents of mixed microbial communities. We have previously shown that intergeneric signaling by the Streptococcus gordonii strain Challis heptapeptide s.g.cAM373 (SVFILAA) induces an aggregation substance-mediated mating response and facilitates plasmid transfer from Enterococcus faecalis cells carrying the pheromone-responsive plasmid pAM373 to both pheromone-producing and non-pheromone-producing oral streptococcal recipients. To further investigate the streptococcal pheromone-like peptides, s.g.cAM373-like sequences were identified in the signal sequences of streptococcal CamG lipoproteins and their abilities to induce a mating response in E. faecalis/pAM373 cells were examined. Synthetic heptamers with the consensus sequence (A/S)-(I/V)-F-I-L-(A/V/T)-(S/A) induced AS-mediated clumping. The conserved pheromone ABC transporter encoded by S. gordonii genome loci SGO_RS02660 and SGO_RS02665 was identified and confirmed to be required for s.g.cAM373 activity. Functional assays of culture supernatants from representative oral and blood isolates of S. gordonii showed that in addition to strains encoding s.g.cAM373, strain SK120, encoding the newly identified pheromone s.g.cAM373-V (SVFILVA), was able to induce enterococcal clumping, whereas strains SK6, SK8, SK9, and SK86 which encoded s.g.cAM373-T (SVFILTA) did not elicit a detectable mating response. Absence of pheromone activity in supernatants of heterologous hosts encoding its CamG precursor suggested that s.g.cAM373-T was not effectively processed and/or transported. Overall, these studies demonstrated the distribution of active pheromone peptides among strains of S. gordonii, and support a potential role for enterococcal-streptococcal communication in contributing to genetic plasticity in the oral metagenome.

Tissue-specific insulin signaling mediates female sexual attractiveness.

Individuals choose their mates so as to maximize reproductive success, and one important component of this choice is assessment of traits reflecting mate quality. Little is known about why specific traits are used for mate quality assessment nor about how they reflect it. We have previously shown that global manipulation of insulin signaling, a nutrient-sensing pathway governing investment in survival versus reproduction, affects female sexual attractiveness in the fruit fly, Drosophila melanogaster. Here we demonstrate that these effects on attractiveness derive from insulin signaling in the fat body and ovarian follicle cells, whose signals are integrated by pheromone-producing cells called oenocytes. Functional ovaries were required for global insulin signaling effects on attractiveness, and manipulations of insulin signaling specifically in late follicle cells recapitulated effects of global manipulations. Interestingly, modulation of insulin signaling in the fat body produced opposite effects on attractiveness, suggesting a competitive relationship with the ovary. Furthermore, all investigated tissue-specific insulin signaling manipulations that changed attractiveness also changed fecundity in the corresponding direction, pointing to insulin pathway activity as a reliable link between fecundity and attractiveness cues. The cues themselves, cuticular hydrocarbons, responded distinctly to fat body and follicle cell manipulations, indicating independent readouts of the pathway activity from these two tissues. Thus, here we describe a system in which female attractiveness results from an apparent connection between attractiveness cues and an organismal state of high fecundity, both of which are created by lowered insulin signaling in the fat body and increased insulin signaling in late follicle cells.

Enhancement of the luteinising hormone surge by male olfactory signals is associated with anteroventral periventricular Kiss1 cell activation in female rats.

Olfactory stimuli play an important role in regulating reproductive functions in mammals. The present study investigated the effect of olfactory signals derived from male rats on kisspeptin neuronal activity and luteinising hormone (LH) secretion in female rats. Wistar-Imamichi strain female rats were ovariectomised (OVX) and implanted with preovulatory levels of 17ß-oestradiol (E2 ). OVX+E2 rats were killed 1 hour after exposure to either: clean bedding, female-soiled bedding or male-soiled bedding. Dual staining for Kiss1 mRNA in situ hybridisation and c-Fos immunohistochemistry revealed that the numbers of Kiss1-expressing cells and c-Fos-immunopositive Kiss1-expressing cells in the anteroventral periventricular nucleus (AVPV) were significantly higher in OVX+E2 rats exposed to male-soiled bedding than those of the other groups. No significant difference was found with respect to the number of c-Fos-immunopositive Kiss1-expressing cells in the arcuate nucleus and c-Fos-immunopositive Gnrh1-expressing cells between the groups. The number of c-Fos-immunopositive cells was also significantly higher in the limbic system consisting of several nuclei, such as the bed nucleus of the stria terminalis, the cortical amygdala and the medial amygdala, in OVX+E2 rats exposed to male-soiled bedding than the other groups. OVX+E2 rats exposed to male-soiled bedding showed apparent LH surges, and the peak of the LH surge and area under the curve of LH concentrations in the OVX+E2 group were significantly higher than those of the other two groups. These results suggest that olfactory signals derived from male rats activate AVPV kisspeptin neurones, likely via the limbic system, resulting in enhancement of the peak of the LH surge in female rats. Taken together, the results of the present study suggests that AVPV kisspeptin neurones are a target of olfactory signals to modulate LH release in female rats.

A comparison of the effects of male pheromone priming and optogenetic inhibition of accessory olfactory bulb forebrain inputs on the sexual behavior of estrous female mice.

Previous research has shown that repeated testing with a stimulus male is required for ovariectomized, hormone-primed female mice to become sexually receptive (show maximal lordosis quotients; LQs) and that drug-induced, epigenetic enhancement of estradiol receptor function accelerated the improvement in LQs otherwise shown by estrous females with repeated testing. We asked whether pre-exposure to male pheromones ('pheromone priming') would also accelerate the improvement in LQs with repeated tests and whether optogenetic inhibition of accessory olfactory bulb (AOB) projection neurons could inhibit lordosis in sexually experienced estrous female mice. In Experiment 1, repeated priming with soiled male bedding failed to accelerate the progressive improvement in LQs shown by estrous female mice across 5 tests, although the duration of each lordosis response and females' investigation of male body parts during the first test was augmented by such priming. In Experiment 2, acute optogenetic inhibition of AOB inputs to the forebrain during freely moving behavioral tests significantly reduced LQs, suggesting that continued AOB signaling to the forebrain during mating is required for maximal lordotic responsiveness even in sexually experienced females. Our results also suggest that pheromonal stimulation, by itself, cannot substitute for the full complement of sensory stimulation received by estrous females from mounting males that normally leads to the progressive improvement in their LQs with repeated testing.

No Evidence for Ionotropic Pheromone Transduction in the Hawkmoth Manduca sexta.

Insect odorant receptors (ORs) are 7-transmembrane receptors with inverse membrane topology. They associate with the conserved ion channel Orco. As chaperon, Orco maintains ORs in cilia and, as pacemaker channel, Orco controls spontaneous activity in olfactory receptor neurons. Odorant binding to ORs opens OR-Orco receptor ion channel complexes in heterologous expression systems. It is unknown, whether this also occurs in vivo. As an alternative to this ionotropic transduction, experimental evidence is accumulating for metabotropic odor transduction, implicating that insect ORs couple to G-proteins. Resulting second messengers gate various ion channels. They generate the sensillum potential that elicits phasic-tonic action potentials (APs) followed by late, long-lasting pheromone responses. Because it is still unclear how and when Orco opens after odor-OR-binding, we used tip recordings to examine in vivo the effects of the Orco antagonist OLC15 and the amilorides MIA and HMA on bombykal transduction in the hawkmoth Manduca sexta. In contrast to OLC15 both amilorides decreased the pheromone-dependent sensillum potential amplitude and the frequency of the phasic AP response. Instead, OLC15 decreased spontaneous activity, increased latencies of phasic-, and decreased frequencies of late, long-lasting pheromone responses Zeitgebertime-dependently. Our results suggest no involvement for Orco in the primary transduction events, in contrast to amiloride-sensitive channels. Instead of an odor-gated ionotropic receptor, Orco rather acts as a voltage- and apparently second messenger-gated pacemaker channel controlling the membrane potential and hence threshold and kinetics of the pheromone response.

Revisiting fifty years of research on pheromone signaling in ciliates.

Among protists, pheromones have been identified in a great variety of algal species for their activity in driving gamete-gamete interactions for fertilization. Analogously in ciliates, pheromones have been identified for their activity in inducing the sexual phenomenon of conjugation. Although this identification was pioneered by Kimball more than fifty years ago, an effective isolation and chemical characterization of ciliate pheromones has remained confined to species of Blepharisma, Dileptus and Euplotes. In Euplotes species, in which the molecular structures have been determined, pheromones form species-specific families of structurally homologous helical, cysteine-rich, highly-stable proteins. Being structurally homologous, they can bind cells in competition with one another, raising interesting functional analogies with the families of growth factors and cytokines that regulate cell differentiation and development in higher organisms. In addition to inducing conjugation by binding cells in heterologous fashion, Euplotes pheromones act also as autocrine growth factors by binding to, and promoting the vegetative reproduction of the same cells from which they originate. This autocrine activity is most likely primary, providing a concrete example of how the original function of a molecule can be obscured during evolution by the acquisition of a new one.

Plant-mediated pheromone emission by a hemipteran seed feeder increases the apparency of an unreliable but rewarding host.

The defensive chemistry and persistence of plant tissues determine their suitability and apparency - the likelihood of being discovered - to insect herbivores. As consumers of plant tissues with transient apparency, florivores and seed-feeders must frequently migrate between host plants to synchronize colonization with plant phenology. Aggregation pheromones could provide information-based solutions to finding ephemeral hosts, but little is known about plant-influenced variation in this form of chemical communication. Combining analytical chemistry, de novo synthesis and field ecology, we investigated the change in colonization of two sympatric host plants, Nicotiana attenuata and Nicotiana obtusifolia, which differ in apparency-related life history traits, by a heteropteran seed-feeder, Corimelaena extensa. We identified a novel pheromone released by C. extensa males - (5Z,8Z)-tetradeca-5,8-dienal - and performed field assays with the synthetic pheromone, showing that it stimulates the formation of feeding aggregations on the post-fire annual N. attenuata. Corimelaena extensa pheromone emission was 40-fold higher when feeding on N. attenuata compared with the perennial N. obtusifolia, as were adult fecundity and seed capsule content of the putative biosynthetic precursor, linoleic acid. Higher pheromone emission increases the apparency and colonization of the ephemeral, high-quality host N. attenuata. This plant-specific variation in insect signaling could facilitate host-finding by seed-feeders migrating between plant patches.

Yeast CUP1 protects HeLa cells against copper-induced stress

As an essential trace element, copper can be toxic in mammalian cells when present in excess. Metallothioneins (MTs) are small, cysteine-rich proteins that avidly bind copper and thus play an important role in detoxification. Yeast CUP1 is a member of the MT gene family. The aim of this study was to determine whether yeast CUP1 could bind copper effectively and protect cells against copper stress. In this study, CUP1 expression was determined by quantitative real-time PCR, and copper content was detected by inductively coupled plasma mass spectrometry. Production of intracellular reactive oxygen species (ROS) was evaluated using the 2',7'-dichlorofluorescein-diacetate (DCFH-DA) assay. Cellular viability was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and the cell cycle distribution of CUP1 was analyzed by fluorescence-activated cell sorting. The data indicated that overexpression of yeast CUP1 in HeLa cells played a protective role against copper-induced stress, leading to increased cellular viability (P<0.05) and decreased ROS production (P<0.05). It was also observed that overexpression of yeast CUP1 reduced the percentage of G1 cells and increased the percentage of S cells, which suggested that it contributed to cell viability. We found that overexpression of yeast CUP1 protected HeLa cells against copper stress. These results offer useful data to elucidate the mechanism of the MT gene on copper metabolism in mammalian cells.

Molecular coevolution of a sex pheromone and its receptor triggers reproductive isolation in Schizosaccharomyces pombe.

The diversification of sex pheromones is regarded as one of the causes of prezygotic isolation that results in speciation. In the fission yeast Schizosaccharomyces pombe, the molecular recognition of a peptide pheromone by its receptor plays an essential role in sexual reproduction. We considered that molecular coevolution of a peptide-mating pheromone, M factor, and its receptor, Map3, might be realized by experimentally diversifying these proteins. Here, we report the successful creation of novel mating-type pairs by searching for map3 suppressor mutations that rescued the sterility of M-factor mutants that were previously isolated. Several strong suppressors were found to also recognize WT M factor. The substituted residues of these Map3 suppressors were mapped to F204, F214, and E249, which are likely to be critical residues for M-factor recognition. These critical residues were systematically substituted with each of the other amino acids by in vitro mutagenesis. Ultimately, we successfully obtained three novel mating-type pairs constituting reproductive groups. These novel mating-type pairs could not conjugate with WT maters. Furthermore, no flow of chromosomally integrated drug-resistance genes occurred between the novel and the WT mating pairs, showing that each experimentally created reproductive group [e.g., M factor(V5H) and Map3(F214H)] was isolated from the WT group. In conclusion, we have succeeded in creating an artificial reproductive group that is isolated from the WT group. In keeping with the biological concept of species, the artificial reproductive group is a new species.

Expression analysis of two P450 monooxygenase genes of the tobacco cutworm moth (Spodoptera litura) at different developmental stages and in response to plant allelochemicals.

Cytochrome P450 monooxygenases (P450s) of insects are known to be involved in the metabolism or detoxification of plant allelochemicals and insecticides. Spodoptera litura (Lepidoptera, Noctuidae) is a polyphagous moth responsible for severe yield losses in many crops. In this study, two full-length P450 genes, CYP6B48 and CYP6B58, were cloned from S. litura. The cDNA sequences encode proteins with 503 and 504 amino acids, respectively. Phylogenetic analysis revealed that CYP6B48 and CYP6B58 belong to the CYP6B subfamily of P450s. Quantitative real-time PCR analyses showed that CYP6B48 and CYP6B58 were expressed only at larval stage, but not at pupal and adult stages. The highest levels of transcripts were found in the midguts and fat bodies of the larvae. No expression was detected in the ovary or hemolymph. Feeding with diets containing cinnamic acid, quercetin, or coumarin did not affect expression of CYP6B48. In contrast, diet supplemented with xanthotoxin dramatically increased the levels of CYP6B48 transcript in the midgut and fat bodies. Larvae fed with flavone had high levels of transcript of CYP6B48 in the midgut, whereas only slightly elevated levels were found in the fat bodies. Effects of the tested allelochemicals on CYP6B58 expression were minor. Hence, our findings show that S. litura responds to specific allelochemicals such as xanthotoxin with the accumulation of CYP6B48 transcripts, suggesting that specific signals in the food control the insect's ability to convert toxic allelochemicals to less harmful forms at the transcriptional level.

Plasma membrane aminoglycerolipid flippase function is required for signaling competence in the yeast mating pheromone response pathway.

The class 4 P-type ATPases ("flippases") maintain membrane asymmetry by translocating phosphatidylethanolamine and phosphatidylserine from the outer leaflet to the cytosolic leaflet of the plasma membrane. In Saccharomyces cerevisiae, five related gene products (Dnf1, Dnf2, Dnf3, Drs2, and Neo1) are implicated in flipping of phosphatidylethanolamine, phosphatidylserine, and phosphatidylcholine. In MAT A: cells responding to α-factor, we found that Dnf1, Dnf2, and Dnf3, as well as the flippase-activating protein kinase Fpk1, localize at the projection ("shmoo") tip where polarized growth is occurring and where Ste5 (the central scaffold protein of the pheromone-initiated MAPK cascade) is recruited. Although viable, a MAT A: dnf1∆ dnf2∆ dnf3∆ triple mutant exhibited a marked decrease in its ability to respond to α-factor, which we could attribute to pronounced reduction in Ste5 stability resulting from an elevated rate of its Cln2⋅Cdc28-initiated degradation. Similarly, a MAT A: dnf1∆ dnf3∆ drs2∆ triple mutant also displayed marked reduction in its ability to respond to α-factor, which we could attribute to inefficient recruitment of Ste5 to the plasma membrane due to severe mislocalization of the cellular phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate pools. Thus proper remodeling of plasma membrane aminoglycerolipids and phosphoinositides is necessary for efficient recruitment, stability, and function of the pheromone signaling apparatus.

When the nose must remain responsive: glutathione conjugation of the mammary pheromone in the newborn rabbit.

In insects, xenobiotic-metabolizing enzymes were demonstrated to regulate pheromones inactivation, clearing them from the olfactory periphery and keeping receptors ready for stimulation renewal. Here, we investigate whether similar processes could occur in mammals, focusing on the pheromonal communication between female rabbits and their newborns. Lactating rabbits emit in their milk a volatile aldehyde, 2-methylbut-2-enal, that elicits searching-grasping in neonates; called the mammary pheromone (MP), it is critical for pups which are constrained to find nipples within the 5 min of daily nursing. For newborns, it is thus essential to remain sensitive to this odorant during the whole nursing period to display several actions of sucking. Here, we show that the MP is enzymatically conjugated to glutathione in newborn olfactory epithelium (OE), in accordance with the high mRNA expression of glutathione transferases evidenced by quantitative reverse transcription-PCR. This activity in the nose is higher than in the liver and in OE of newborns compared with weanlings (no more responsive to the pheromone). Therefore, the results pinpoint the existence of a high level of MP-glutathione conjugation activity in the OE of young rabbits, especially in the developmental window where the perceptual sensitivity toward the MP is crucial for survival.

Órgano vomeronasal: estudio anatómico de prevalencia y su función / The vomeronasal organ: anatomic study of prevalence and its role

Introducción: El órgano vomeronasal (OVN) descrito por Jacobson en mamíferos distintos al ser humano, es una incógnita tanto en lo que se refiere a su localización así como a su función en la raza humana. Se considera como un vestigio del olfato, que en los animales mamíferos parece influir en los hábitos sexuales (feromonas) y sociales. Hasta la fecha han sido escasos los estudios concluyentes al respecto en humanos. Objetivo: Conocer la prevalencia del órgano vomeronasal en nuestras consultas. Material y método: Presentamos un estudio prospectivo de prevalencia de la frecuencia de aparición de dicho órgano en 150 sujetos distribuidos por edad y sexo, explorados por endoscopia nasosinusal rígida. Por otro lado, analizamos la influencia sobre la libido (normal-disminuida-aumentada) en el posoperatorio de 35 septoplastías, a los 15 días tras retirada de taponamiento nasal y a los 30 días y lo comparamos con un grupo de 40 pacientes intervenidos timpanoplastías. Resultados: Estudiados 150 sujetos, encontramos la presencia del órgano vomeronasal en el 39,33% (59), de los cuales el 72,88% (43) fue unilateral (23 derecha y 20 izquierda) y el 27,12% (16) bilateral. En 91 (60,67%) no hallamos dicha estructura. La libido de los 35 pacientes intervenidos de septoplastía estaba disminuida, a los 15 días, en el 77,14% (27) frente al 40% (16) de las timpanoplastías, normal en el 17,14% (6) frente al 50% (20) de las cirugías otológicas, y en 2 (5,7%) poseptoplastía había aumentado, frente al 10% (4) del otro grupo. A los 30 días, en el 77,14% (27) de las septoplastías se había normalizado frente al 90% (36) del grupo otológico, en 2 (5,71%) de la cirugía nasal continuaba disminuida frente al 10% (4) del grupo de las timpanoplastías y en 6 (17,14%) tras septoplastía había aumentado. A todos los pacientes se les aplicó el mismo test no normalizado. Conclusión: El órgano vomeronasal de Jacobson continúa siendo un gran desconocido. Es una estructura que, al parecer, no es constante, al menos a la exploración endoscópica nasosinusal. Es difícil valorar si la cirugía en sí misma o el trauma psicológico posquirúrgico son los que afectan la libido de los pacientes tras la cirugía.

Introduction: The vomeronasal organ (OVN) described by Jacobson in mammals other than humans is unknown both in terms of its location and its role in the human race. It is viewed as a vestige of smell, that mammals in the animal seems to influence the sexual habits (pheromone) and social. To date, few studies have been inconclusive on this in humans. Aim: To determine the prevalence of vomeronasal organ in our medical consultations. Materials and methods: We report a prospective prevalence study of the occurrence of such a body in 150 subjects distributed by age and sex explored by endoscopic sinus rigid. On the other hand, we analyze the effect on the libido (normally less-plus) in the postoperative 35 septoplasty, 15 days after the withdrawal of nasal pack and 30 days and compared with a group of 40 tympanoplasty surgery. Results: Studied 150 subjects, we found the presence of the vomeronasal organ in 39.33% (59), of which 72.88% (43) had unilateral (23 right and 20 left) and 27.12% (16) bilaterally. In 91 (60.67%) did not find such a structure. The libido of the 35 patients who underwent septoplasty was decreased at 15 days, at 77.14% (27) versus 40% (16) of tympanoplasty, normal in 17.14% (6) compared to 50% (20) of otologic surgery, and in 2 (5.7%) postseptoplasty had increased, compared to 10% (4) the other group. At 30 days, in 77.14% (27) of the septoplasty group the libido was normalized against 90% (36) in the otologic group. In 2 cases (5.71%) of nasal surgery group was still decreased versus 10% (4) of cases of the tympanoplasty group, and in 6 (17.14%) postseptoplasty was increased. All patients were administered the same test is not standardized. Conclusion: The vomeronasal organ of Jacobson remains the great unknown. It is a structure that apparently is not constant, at least in the endoscopic sinus exploration. With regard to their role, it is difficult to assess whether the psychological trauma after surgery or the surgery by itself is responsible of the libido changes.

Fatal attraction: sexually cannibalistic invaders attract naive native mantids.

Overlap in the form of sexual signals such as pheromones raises the possibility of reproductive interference by invasive species on similar, yet naive native species. Here, we test the potential for reproductive interference through heterospecific mate attraction and subsequent predation of males by females of a sexually cannibalistic invasive praying mantis. Miomantis caffra is invasive in New Zealand, where it is widely considered to be displacing the only native mantis species, Orthodera novaezealandiae, and yet mechanisms behind this displacement are unknown. We demonstrate that native males are more attracted to the chemical cues of introduced females than those of conspecific females. Heterospecific pairings also resulted in a high degree of mortality for native males. This provides evidence for a mechanism behind displacement that has until now been undetected and highlights the potential for reproductive interference to greatly influence the impact of an invasive species.

In situ tip-recordings found no evidence for an Orco-based ionotropic mechanism of pheromone-transduction in Manduca sexta.

The mechanisms of insect odor transduction are still controversial. Insect odorant receptors (ORs) are 7TM receptors with inverted membrane topology. They colocalize with a conserved coreceptor (Orco) with chaperone and ion channel function. Some studies suggest that insects employ exclusively ionotropic odor transduction via OR-Orco heteromers. Other studies provide evidence for different metabotropic odor transduction cascades, which employ second messenger-gated ion channel families for odor transduction. The hawkmoth Manduca sexta is an established model organism for studies of insect olfaction, also due to the availability of the hawkmoth-specific pheromone blend with its main component bombykal. Previous patch-clamp studies on primary cell cultures of M. sexta olfactory receptor neurons provided evidence for a pheromone-dependent activation of a phospholipase Cß. Pheromone application elicited a sequence of one rapid, apparently IP3-dependent, transient and two slower Ca(2+)-dependent inward currents. It remains unknown whether additionally an ionotropic pheromone-transduction mechanism is employed. If indeed an OR-Orco ion channel complex underlies an ionotropic mechanism, then Orco agonist-dependent opening of the OR-Orco channel pore should add up to pheromone-dependent opening of the pore. Here, in tip-recordings from intact pheromone-sensitive sensilla, perfusion with the Orco agonist VUAA1 did not increase pheromone-responses within the first 1000 ms. However, VUAA1 increased spontaneous activity of olfactory receptor neurons Zeitgebertime- and dose-dependently. We conclude that we find no evidence for an Orco-dependent ionotropic pheromone transduction cascade in M. sexta. Instead, in M. sexta Orco appears to be a slower, second messenger-dependent pacemaker channel which affects kinetics and threshold of pheromone-detection via changes of intracellular Ca(2+) baseline concentrations.

Behavioral transition from attack to parenting in male mice: a crucial role of the vomeronasal system.

Sexually naive male mice show robust aggressive behavior toward pups. However, the proportion of male mice exhibiting pup-directed aggression declines after cohabitation with a pregnant female for 2 weeks after mating. Subsequently, on becoming fathers, they show parental behavior toward pups, similar to maternal behavior by mothers. To elucidate the neural mechanisms underlying this behavioral transition, we examined brain regions differentially activated in sexually naive males and fathers after exposure to pups, using c-Fos expression as a neuronal activation marker. We found that, after pup exposure, subsets of neurons along the vomeronasal neural pathway-including the vomeronasal sensory neurons, the accessory olfactory bulb, the posterior medial amygdala, the medioposterior division of the bed nucleus of stria terminalis, and the anterior hypothalamic area-were more strongly activated in sexually naive males than in fathers. Notably, c-Fos induction was not observed in the vomeronasal sensory neurons of fathers after pup exposure. Surgical ablation of the vomeronasal organ in sexually naive males resulted in the abrogation of pup-directed aggression and simultaneous induction of parental behavior. These results suggest that chemical cues evoking pup-directed aggression are received by the vomeronasal sensory neurons and activate the vomeronasal neural pathway in sexually naive male mice but not in fathers. Thus, the downregulation of pup pheromone-induced activation of the vomeronasal system might be important for the behavioral transition from attack to parenting in male mice.

Olfactory signals mediate social buffering of conditioned fear responses in male rats.

In social animals, the presence of an affiliative conspecific alleviates acute stress responses, and this is called social buffering. We previously reported that social buffering mitigates the fear responses of male rats to auditory conditioned stimuli that had been paired with foot shocks. Subsequent studies revealed that signals that are perceived by the main olfactory epithelium are important for social buffering. Because olfactory signals are the signal perceived by the main olfactory epithelium, we hypothesized that we could induce the social buffering of conditioned fear responses by presenting olfactory signals that were derived from a conspecific. In order to test this hypothesis, we exposed fear-conditioned subjects to a conditioned stimulus either in a clean test box or in a test box that was odorized by keeping a conspecific in it as an odor donor beforehand. When the subjects were tested in the clean test box, they showed behavioral fear responses and enhanced Fos expression in the paraventricular nucleus. In contrast, the presence of conspecific olfactory signals blocked these fear responses and Fos expression. These results suggested that olfactory signals suppress conditioned fear responses. Fos expression in the posteromedial region of the olfactory peduncle and amygdala suggested that this suppression involves the same neural mechanisms as those of social buffering. Taken together, we concluded that olfactory signals mediate the social buffering of conditioned fear responses.

Chemosensory age discrimination in the snake Boa constrictor (Serpentes: Boidae)

Many snakes are able to use their chemosensory system to detect scent of conspecifics, which is important in many social contexts. Age discrimination based on chemical cues may be especially important to ensure access to sexually mature potential partners. In this study, we used 24 individual Boa constrictor snakes (12 adults mature and 12 non-mature individuals) that had been captured in different areas of Ecuador, and were maintained in captivity at the Vivarium of Quito. We used tongue-flick experiments to examine whether these snakes were able to discriminate between scents from mature and non-mature individuals. Results showed that B. constrictor snakes used chemical cues to recognize conspecifics and that the scent of individuals of different ages elicited chemosensory responses of different magnitudes. The scents from adult conspecifics elicited the quickest and highest chemosensory responses (i.e., short latency times and high tongue-flick rates), although we did not find differential responses to scent of males and females. The magnitude of the responses was lower to scent of sub adult individuals, and then even lower to scent of juvenile snakes, but in all cases the scent of snakes was discriminated from a blank control. We discuss the potential chemical mechanisms that may allow age recognition and its implications for social and sexual behavior of this snake species.

Muchas serpientes son capaces de usar su sistema quimiosensorial para detectar el olor de individuos coespecíficos, lo que es importante en muchos contextos sociales. La discriminación de la edad basada en señales químicas puede ser especialmente importante para asegurar el acceso a parejas potenciales que sean sexualmente maduras. En este estudio, usamos 24 individuos de una especie de boa (Boa constrictor) (12 individuos adultos y 12 inmaduros) que habían sido capturados en diferentes partes de Ecuador y eran mantenidos en cautividad el Vivarium de Quito. Usamos experimentos de protusiones linguales para examinar si esta serpiente es capaz de discriminar entre el olor de individuos maduros y no maduros. Los resultados mostraron que B. constrictor usa señales químicas para reconocer co-específicos y que el olor de individuos de distinta edad provoca respuestas quimiosensoriales de diferente magnitud. El olor de individuos adultos provocó las respuestas más rápidas y elevadas (esto es, tiempos de latencia más cortos y tasas más altas de protusiones linguales), aunque no encontramos diferencias en las respuestas a olores de machos y hembras. La magnitud de las respuestas fue más baja a olores de sub adultos, e incluso más baja a olor de juveniles, pero en todos los casos el olor de una serpiente era discriminado de un control no oloroso. Discutimos los posibles mecanismos químicos que pueden permitir esta discriminación de la edad y sus implicaciones para el comportamiento social y sexual de esta serpiente.

Time of day changes in cyclic nucleotides are modified via octopamine and pheromone in antennae of the Madeira cockroach.

The cockroach Rhyparobia (Leucophaea) maderae expresses a circadian rhythm in pheromone-dependent mating activity that peaks at the late day/early night. In contrast, the circadian rhythm in olfactory sensitivity of the Madeira cockroach is at its minimum during this time. Until now, the reasons for this obvious discrepancy in phase were not understood. Previously, it was shown that cyclic nucleotides modulate olfactory sensitivity in a zeitgeber time (ZT)-dependent manner. In moths' olfactory receptor neurons, adapting pheromone concentrations elevate cGMP levels, which decrease pheromone sensitivity. In contrast, cAMP elevations sensitized pheromone responses. Thus, with immunoassay kits, it was determined whether cAMP and cGMP baseline levels vary in a ZT-dependent manner in antennal lysates of female R. maderae, revealing underlying circadian rhythms in olfactory sensitivity. Furthermore, it was examined whether adapting pheromone exposure elevates cGMP levels in cockroach antennae, possibly overshadowing underlying circadian rhythms in sensitivity via sensory adaptation. It was shown for the first time that cAMP and cGMP baseline levels oscillate in antiphase in a ZT-dependent manner in an insect's antenna, with the maximum in cAMP concentrations coinciding with maximal mating activity during the late day. Moreover, the cAMP baseline level oscillation expressed a circadian rhythm since it persisted under constant darkness in contrast to cGMP baseline levels. Furthermore, while excess exposure to male pheromones increased cGMP and decreased cAMP baseline levels, the stress hormone octopamine increased adenylyl cyclase activity at all ZTs tested. Therefore, it is suggested that cyclic nucleotide-dependent modulation of olfactory sensitivity due to olfactory overstimulation and stress could be responsible for previously measured phase discrepancies between rhythms in mating behavior and pheromone sensitivity.