(Image caption: On these images, the cerebral activation detected by ultrasound imaging is shown in red. During odor presentation, specific areas are activated in the olfactory bulb but not in the piriform cortex. Credit: © Mickael Tanter / Hirac Gurden)

Ultrasound tracks odor representation in the brain

A new ultrasound imaging technique has provided the first ever in vivo visualization of activity in the piriform cortex of rats during odor perception. This deep-seated brain structure plays an important role in olfaction, and was inaccessible to functional imaging until now. This work also sheds new light on the still poorly known functioning of the olfactory system, and notably how information is processed in the brain. This study is the result of a collaboration between the team led by Mickael Tanter at the Institut Langevin (CNRS/INSERM/ESPCI ParisTech/UPMC/Université Paris Diderot) and that led by Hirac Gurden in the Laboratoire Imagerie et Modélisation en Neurobiologie et Cancérologie (CNRS/Université Paris-Sud/Université Paris Diderot). Their findings are published in NeuroImage.

How can the perception of the senses help represent the external environment? How, for example, does the brain process food-or perfume-related olfactory data? Although the organization of the olfactory system is well known - it is similar in organisms ranging from insects to mammals - its functioning remains unclear. To answer these questions, the scientists focused on the two brain structures that act as major olfactory relays: the olfactory bulb and the piriform cortex. In the rat, the olfactory bulb is located between the eyes, just behind the nasal bone. The piriform cortex, meanwhile, is deep-seated in the brain of rodents, which made it impossible to obtain any functional images in a living animal until now.

Yet the neurofunctional ultrasound imaging technique developed by Mickael Tanter’s team, called fUS(functional Ultrasound), allows the monitoring of neuronal activity in the piriform cortex. It is based on the transmission of ultrasonic plane waves into the brain tissue. After data processing, the echoes returned by the structures crossed by these waves can provide images with unequalled spatial and temporal resolution: 80 micrometers and a few tens of milliseconds. The contrast on these images is due to variations in the brain’s blood flow. Indeed, the activity of nerve cells requires an input of energy: it is therefore coupled to an influx of blood into the zone concerned. By recording volume variations in the blood vessels irrigating the different brain structures, it is there fore possible to determine the location of activated neurons.

Several imaging techniques, such as MRI, are already based on the link between blood volume and neuronal activity. But fUS offers advantages in terms of cost, ease of use and resolution. Furthermore, it provides easier access to the deepest structures that are often located several centimeters beneath the cranium.

The recordings performed by Hirac Gurden’s team using this technique made it possible to observe the spatial distribution of activity within the olfactory bulb. When an odor was perceived, blood volume increased in clearly defined areas: each odor thus corresponded to a specific pattern of activated neurons. In addition to these findings, and for the first time, the images revealed an absence of spatial distribution in the piriform cortex. At this level, two different odors triggered the same activation throughout the region.

The cellular mechanisms responsible for the disappearance of a spatial signature are not yet clearly defined, but these findings lead to the formulation of several hypotheses. The piriform cortex could be a structure that serves not only to process olfactory stimuli but rather to integrate and memorize different types of data. By making abstraction of the strict odor-induced patterns, it would be possible to make associations and achieve a global concept. For example, based on the perception of the hundreds of odorant molecules found in coffee, the piriform cortex would be able to recognize a single odor, that of
coffee.

This work opens new perspectives for both imaging and neurobiology. The researchers will now be focusing on the effects of learning on cortical activity in order to elucidate its role and the specificities of the olfactory system.

How Mosquitoes Are Drawn to Human Skin and Breath

Female mosquitoes, which can transmit deadly diseases like malaria, dengue fever, West Nile virus and filariasis, are attracted to us by smelling the carbon dioxide we exhale, being capable of tracking us down even from a distance. But once they get close to us, they often steer away toward exposed areas such as ankles and feet, being drawn there by skin odors.

Why does the mosquito change its track and fly towards skin? How does it detect our skin? What are the odors from skin that it detects? And can we block the mosquito skin odor sensors and reduce attractiveness?

Recent research done by scientists at the University of California, Riverside can now help address these questions. They report on Dec. 5 in the journal Cell that the very receptors in the mosquito’s maxillary palp that detect carbon dioxide are ones that detect skin odors as well, thus explaining why mosquitoes are attracted to skin odor — smelly socks, worn clothes, bedding — even in the absence of CO₂.

“It was a real surprise when we found that the mosquito’s CO₂ receptor neuron, designated cpA, is an extremely sensitive detector of several skin odorants as well, and is, in fact, far more sensitive to some of these odor molecules as compared to CO₂,” said Anandasankar Ray, an associate professor in the Department of Entomology and the project’s principal investigator. “For many years we had primarily focused on the complex antennae of mosquitoes for our search for human-skin odor receptors, and ignored the simpler maxillary palp organs.”

Until now, which mosquito olfactory neurons were required for attraction to skin odor remained a mystery.  The new finding — that the CO₂-sensitive olfactory neuron is also a sensitive detector of human skin — is critical not only for understanding the basis of the mosquito’s host attraction and host preference, but also because it identifies this dual receptor of CO₂ and skin-odorants as a key target that could be useful to disrupt host-seeking behavior and thus aid in the control of disease transmission.

To test whether cpA activation by human odor is important for attraction, the researchers devised a novel chemical-based strategy to shut down the activity of cpA in Aedes aegypti, the dengue-spreading mosquito.  They then tested the mosquito’s behavior on human foot odor — specifically, on a dish of foot odor-laden beads placed in an experimental wind tunnel — and found the mosquito’s attraction to the odor was greatly reduced.

Next, using a chemical computational method they developed, the researchers screened nearly half a million compounds and identified thousands of predicted ligands. They then short-listed 138 compounds based on desirable characteristics such as smell, safety, cost and whether these occurred naturally. Several compounds either inhibited or activated cpA neurons of which nearly 85 percent were already approved for use as flavor, fragrance or cosmetic agents. Better still, several were pleasant-smelling, such as minty, raspberry, chocolate, etc., increasing their value for practical use in mosquito control.

Confident that they were on the right track, the researchers then zeroed in on two compounds: ethyl pyruvate, a fruity-scented cpA inhibitor approved as a flavor agent in food; and cyclopentanone, a minty-smelling cpA activator approved as a flavor and fragrance agent.  By inhibiting the cpA neuron, ethyl pyruvate was found in their experiments to substantially reduce the mosquito’s attraction towards a human arm. By activating the cpA neuron, cyclopentanone served as a powerful lure, like CO₂, attracting mosquitoes to a trap.

“Such compounds can play a significant role in the control of mosquito-borne diseases and open up very realistic possibilities of developing ways to use simple, natural, affordable and pleasant odors to prevent mosquitoes from finding humans,” Ray said.  “Odors that block this dual-receptor for CO₂ and skin odor can be used as a way to mask us from mosquitoes.  On the other hand, odors that can act as attractants can be used to lure mosquitoes away from us into traps.  These potentially affordable ‘mask’ and ‘pull’ strategies could be used in a complementary manner, offering an ideal solution and much needed relief to people in Africa, Asia and South America — indeed wherever mosquito-borne diseases are endemic.  Further, these compounds could be developed into products that protect not just one individual at a time but larger areas, and need not have to be directly applied on the skin.”

Currently, CO₂ is the primary lure in mosquito traps. Generating CO₂ requires burning fuel, evaporating dry ice, releasing compressed gas or fermentation of sugar — all of which is expensive, cumbersome, and impractical for use in developing countries.  Compounds identified in this study, like cyclopentanone, offer a safe, affordable and convenient alternative that can finally work with surveillance and control traps.

Sex, Smell And Science – The Genetics Of Olfaction

No two people smell exactly alike. That is, noses sense odors in individual ways. What one nose finds offensive, another may find pleasant, while another might not smell anything at all. Scientists have long known the way things smell to us is determined by our genes.

Now, two studies appearing in the journal Current Biology (1, 2) have identified “the genetic differences that underpin the differences in smell sensitivity and perception in different individuals.” And while some of these differences merely help determine our culinary preferences, others appear to play a subconscious role in how we choose our sexual partners.

For the first study, 200 people were tested to determine their sensitivity to 10 different chemical compounds commonly found in foods. The researchers found four of the ten odors had a genetic association. These were malt, apple, blue cheese, and a floral scent associated with violets.

The research team, led by Sara Jaeger, Jeremy McRae, and Richard Newcomb of Plant and Food Research in New Zealand, used a genome-wide association study. Their first task was to identify which test subjects could smell each chemical compound and which could not. They then searched the subjects’ genomes for areas of DNA that differed between these people.

“We were surprised how many odors had genes associated with them. If this extends to other odors, then we might expect everyone to have their own unique set of smells that they are sensitive to,” explained McRae

“These smells are found in foods and drinks that people encounter every day, such as tomatoes and apples. This might mean that when people sit down to eat a meal, they each experience it in their own personalized way.”

They further found there is no regional differentiation. A person in one part of the world is just as likely to be able to smell a particular compound as a person in another part of the world. In addition, sensitivity to one compound does not predict the ability to smell another compound.

The genes that determine our ability to perceive certain odors all lie in or near the genes that encode olfactory receptors. These receptors occur on the surface of sensory nerve cells in the upper part of the nose. A particular smell is perceived when these receptor molecules bind with a chemical compound wafting through the nose, causing nerve cells to send an impulse to the brain and producing our sensation of smell.

For the violet smell, caused by a naturally occurring chemical compound known as β-ionone, the researchers were able to pinpoint the exact mutation in gene OR5A1 that determines whether the smell is perceived as floral, sour or pungent, and whether it is found to be pleasant.

These findings might have future marketing value. According to Richard Newcomb, “Knowing the compounds that people can sense in foods, as well as other products, will have an influence on the development of future products. Companies may wish to design foods that better target people based on their sensitivity, essentially developing foods and other products personalized for their taste and smell.”

SEXY OR STINKY?

A separate study was conducted by Leslie Vosshall of the Rockefeller University Hospital. Humans have about 1,000 genes that influence smell, and around 400 of these are responsible for sensing a particular odor molecule.

Testing 391 human subjects, Vosshall studied olfactory responses to two closely related steroids, androstenone and androstadienone, which are found in male sweat. People generally have strong reactions to these steroids, finding them either sweet and florally or rank and noxious. The gene 0R7D4 determines the intensity of these odors as well as the perception of them being either pleasant or repulsive.

According to Vosshall’s report: “People who found the smell repulsive were more likely to have two functional copies of OR7D4; those who perceived it as a more mild smell tended to have one or two impaired copies of the gene.”

This study is part of the larger goal of understanding how genetic and neuronal factors influence behaviors.

A 2002 study published in Nature Genetics provided more insight into the effect of male pheromones on women. This study looked at the link between women’s preferences for the odors given off by men and a group of genes called the Major Histocompatibily Complex (MHC) which contribute to a persons’ immune response.

In this experiment, a group of 49 women were asked to smell 10 boxes. Some of the boxes held t-shirts worn by men with different MHC genes, and others contained familiar household odors such as bleach or cloves.

The t-shirts were worn by men who slept in them for two nights and avoided contact with other scents during that time, even to the point of avoiding other people. According to the report, “the women were then asked to rate each scent based on their familiarity, intensity, pleasantness and spiciness, as well as choose the one odor which they would choose if they had to smell it all the time.”

What the researchers found was the women did not choose the scents of men whose genes were similar to their own, nor did they choose those whose genes were too dissimilar. The women showed no preference for odors from men who had the same genes as their mothers, but did show a preference for odors from men who shared genes they inherited from their fathers.

Scientists believe there are two reasons for preferring a mate whose MHC genes are different than one’s own. One is that it would tend to create offspring with more genetic diversity and thus more robust immune systems. The other is it helps to avoid inbreeding.

Of course, when people choose their mates, there are a number of social factors that come into play as well. However, studies have shown married people tend to have different types of genes than their spouses.

So, the next time you like the way a person smells, keep in mind it may mean you have complementary genes.

It Just Smells

If you play sounds of many different frequencies at the same time, they combine to produce neutral “white noise.” Neuroscientists say they have created an analogous generic scent by blending odors. Such “olfactory white” might rarely, if ever, be found in nature, but it could prove useful in research, other scientists say.

Using just a few hundred types of biochemical receptors, each of which respond to just a few odorants, the human nose can distinguish thousands of different odors. Yet humans can’t easily identify the individual components of a mixture, even when they can identify the odors alone, says Noam Sobel, a neuroscientist at the Weizmann Institute of Science in Rehovot, Israel. Now, he and his colleagues suggest, various blends made up of a large number of odors all begin to smell the same—even when the blends share no common components.

…

Although many scents—such as coffee, wine, roses, and dirty socks—are complex blends containing hundreds of components, they are very distinctive. At least two factors are responsible, Sobel says: The individual odorants are often chemically related, and often one or more of them is vastly more intense than the rest.

The team’s findings are “a clever piece of work that shows the olfactory system works exactly as we would predict from our current understanding of it,” says Tim Jacob, a neuroscientist at Cardiff University in the United Kingdom. “That is, if you stimulate every olfactory ‘channel’ to the same extent, the brain cannot characterize or identify a particular smell,” he notes.

“Olfactory white is a neat idea, and it draws interesting parallels to white light and white noise,” says Jay Gottfried, an olfactory neuroscientist at Northwestern University’s Feinberg School of Medicine in Chicago, Illinois. The new study “definitely adds new information about how the brain interprets odors,” he notes.

Even though olfactory white is not likely to be encountered in nature, the concept could be useful, Gottfried says. “Researchers have found that white noise is a useful stimulus in experiments to probe auditory responses,” he notes, and scientists probing the human sense of smell might find similar uses for olfactory white.

How Cells in the Nose Detect Odors

Now a team of scientists, led by neurobiologists at the University of California, Riverside, has an explanation. Focusing on the olfactory receptor for detecting carbon dioxide in Drosophila (fruit fly), the researchers identified a large multi-protein complex in olfactory neurons, called MMB/dREAM, that plays a major role in selecting the carbon dioxide receptors to be expressed in appropriate neurons.

Study results appear in the Nov. 15 issue of Genes & Development.  The research is featured on the cover of the issue.

According to the researchers, a molecular mechanism first blocks the expression of most olfactory receptor genes (~60) in the fly’s antennae. This mechanism, which acts like a brake, relies on repressive histones —proteins that tightly wrap DNA around them. All insects and mammals are equipped with this mechanism, which keeps the large families of olfactory receptor genes repressed.

“How, then, do you release this brake so that only the carbon dioxide receptor is expressed in the carbon dioxide neuron while the remaining receptors are repressed?” said Anandasankar Ray, an assistant professor of entomology, whose lab conducted the research. “Our lab, in collaboration with a lab at Stanford University, has found that the MMB/dREAM multi-protein complex can act on the genes of the carbon dioxide receptors and de-repress the braking mechanism — akin to taking the foot off the brake pedal. This allows these neurons to express the receptors and respond to carbon dioxide.”

Ray explained that one way to understand the mechanism in operation is to consider a typewriter. When none of the keys are pressed, a spring mechanism or “brake” can be imagined to hold the type bars away from the paper. When a key is pressed, however, the brake on that key is overcome and the appropriate letter is typed onto the paper. And just as typing only one letter in one spot is important for each letter to be recognized, expressing one receptor in one neuron lets different sensor types to be generated in the nose.