'Darting' mice may hold clues to ADHD, autism, bipolar disorder

Mice inserted with a rare human genetic variation in the dopamine transporter could lead to improvements in the diagnosis and treatment of brain disorders. Credit: Image courtesy of Vanderbilt University Medical Center

A darting mouse may hold an important clue in the development of Attention Deficit Hyperactivity Disorder (ADHD), autism and bipolar disorder, according to a study by a Vanderbilt University-led research team recently published in the Proceedings of the National Academy of Sciences.

The transgenic mouse, into which was inserted a rare human genetic variation in the dopamine transporter (DAT), could lead to improvements in the diagnosis and treatment of these all-too-common brain disorders, said Randy Blakely, Ph.D., the report's senior author.
The mutation, which has been found in people with ADHD, autism and bipolar disorder, affects the function of DAT, a protein that regulates the brain's supply of the neurotransmitter by removing excess dopamine from the synapse, or the space between nerve cells.

The DAT mutation causes the transporter to become "leaky" and spew out dopamine like "a vacuum cleaner in reverse," said Blakely, Allan D. Bass Professor of Pharmacology.

While mice with leaky DAT proteins have too much dopamine hanging around their synapses, surprisingly they aren't particularly hyperactive, possibly because DAT can still remove some of the dopamine.

But the mice exhibit an unusual "darting behavior." While their wild-type littermates are docile and quite unresponsive when researchers pick them up, those with the mutation "take off."

"Early on," Blakely said, "we could tell which ones carried the mutation by observing this response." Heightened anxiety does not appear to be the cause.

Blakely and his colleagues wonder whether this behavior is a form of "impulsivity." Rather than acting on their memories of being picked up a lot, the mice are opting for an inappropriate escape strategy.

Normal mice also stand up a lot to explore their cage. This "rearing" behavior is exacerbated by stimulant drugs. But not in these mice.

"We wonder whether this may be a sign that their behavior is driven less by searching for clues to appropriate behavior versus acting on innate impulses," Blakely said.

Other, better tests of impulsivity that evaluate premature decision-making can be applied in rodents and humans. "These tests are next on our docket," he said.

The actions of amphetamine and methylphenidate (Ritalin) are also affected by the mutation. In normal animals and people without ADHD, the stimulants flood the synapse with dopamine, eliciting hyperactivity.

But when given to the mutant animals, the drug demonstrates a "blunted" effect on both dopamine release and on locomotor activation compared to normal animals.

Blakely wonders whether stimulants like Adderall and Ritalin quell hyperactive and impulsive behaviors in some children with ADHD by reducing inappropriate dopamine leak. 
"These mice may give us much better clues as to how these drugs are acting," he said.
To that end, Blakely recently received a five-year, $2-million grant from the National Institutes of Health (NIH grant number MH109054) to pursue explorations of these mice.

"Dopamine has classically been implicated in reward and the ability to detect novelty and to respond to pleasure and to engage in effective social interactions," he continued. The darting mice thus might shed light on a much broader spectrum of behaviors.

"We've got a lot to do," he said, "a lot of needy people (to help)."

Source: Vanderbilt University Medical Center

Quest continues for peanut that won't cause allergic reaction

Peanuts (stock image). Scientists must eliminate peanut allergens below a certain threshold for patients to be safe, said Wade Yang, an assistant professor in food science and human nutrition and member of UF’s Institute of Food and Agricultural Sciences. Credit: © yurakp / Fotolia

University of Florida scientist has moved one step closer to his goal of eliminating 99.9 percent of peanut allergens by removing 80 percent of them in whole peanuts.

Scientists must eliminate peanut allergens below a certain threshold for patients to be safe, said Wade Yang, an assistant professor in food science and human nutrition and member of UF’s Institute of Food and Agricultural Sciences.

If Yang can cut the allergens from 150 milligrams of protein per peanut to below 1.5 milligrams, 95 percent of those with peanut allergies would be safe. It’s challenging to eliminate all peanut allergens, he said, because doing so may risk destroying peanuts’ texture, color, flavor and nutrition. But he said he’s using novel methods like pulsed light to reach an allergen level that will protect most people.

Yang, whose study is published online in this month’s issue of the journal Food and Bioprocess Technology, cautioned that he has done peanut allergen experiments only in a laboratory setting so far. He hopes to eventually conduct clinical trials on animals and humans.

Dr. Shih-Wen Huang, professor emeritus in the Department of Pediatrics and Head of the Pediatric Allergy Clinic at UF Health, is familiar with the UF/IFAS research. Huang outlined more steps in the peanut allergen research.

The first is to see if the allergic antibody in the serum of peanut allergy patients will still bind with the residual allergy protein from the refined peanut products. The second is to see if the refined peanut extract would elicit skin-test reactions in peanut allergy patients.

The third step would be to conduct a double blind, placebo-controlled test to see if patients develop allergy symptoms after eating the refined products.

“I am pleased to see their work is progressing well,” Huang said. “However, more challenges are waiting until the final products are accepted from the public, especially the patients with peanut allergies.”

Two years ago, Yang was using his technique on peanut extract. He’s now testing it on the peanut itself. In his 2012 study, he removed up to 90 percent of the allergic potential from peanut protein extracts.

“This process proves that pulsed light can inactivate the peanut allergenic proteins and indicates that pulsed light has a great potential in peanut allergen mitigation,” Yang said.

About 1.9 million people, or 0.6 percent of U.S. residents, are allergic to peanuts, according to the National Institute of Allergy and Infectious Diseases, part of the National Institutes of Health.

Reactions can range from skin rashes to anaphylaxis, which can be fatal. Currently, the best way for those allergic to peanuts to stay safe is to avoid them, according to the NIH. Many people carry epinephrine injectors that help offset their allergy symptoms until they reach a hospital.

In the latest study, Yang and his colleagues applied the pulsed ultraviolet light technology to whole peanuts. That makes the findings more useful, because peanut processing usually starts from whole-peanut roasting, and roasted peanuts are then packaged to sell as whole peanuts or made into peanut butter, he said.

“The latest study moves one step closer to the actual production,” Yang said.

For the study, Yang used a pulsating light system – two lamps filled with xenon, two cooling blowers, one treatment chamber with a conveyor belt and a control module ─ to direct concentrated bursts of light to modify the peanut allergenic proteins. That way, human antibodies can’t recognize them as allergens and begin to release histamines.

Histamines create allergy symptoms such as itching, rashes and wheezing. The pulsing light reduced the allergenic potential of the major peanut proteins Ara h1-h3.

Source: University of Florida Institute of Food and Agricultural Sciences

Sharing that crowded holiday flight with countless hitchhiking dust mites

Predicted structure of the group 1 allergen protein from an American house dust mite. Arrow points to the location of a novel mutation discovered by the University of Michigan-led team.
Credit: Rubaba Hamid

As if holiday travel isn't stressful enough. Now University of Michigan researchers say we're likely sharing that already overcrowded airline cabin with countless tiny creatures including house dust mites.

"What people might not realize when they board a plane is that they can share the flight with a myriad of microscopic passengers-- including house dust mites--that take advantage of humanity's technological progress for their own benefit," said U-M biologist Pavel Klimov.

"House dust mites can easily travel on an airline passenger's clothes, skin, food and baggage," said Klimov, an assistant research scientist in the U-M Department of Ecology and Evolutionary Biology. "Like humans, they use air travel to visit new places, where they establish new populations, expand their ranges and interact with other organisms through various means."

Air travel likely explains some of the findings of a new genetic study conducted by Klimov and U-M visiting scholar Rubaba Hamid that looked at the connections between house dust mite populations in the United States and South Asia.

They found genetic mutations shared by mites in the U.S. and Pakistan that demonstrate the eight-legged creatures' propensity for intercontinental dispersal, according to a research paper scheduled for online publication Dec. 10 in the journal PLOS ONE.

"What we found suggests that mite populations are indeed linked through migration across continents, though geographic differences still can be detected," Hamid said. "Every time a mite successfully migrates to a new place, it brings its own genetic signature that can be detected in the resident population a long time after the migration event."

The study focused on two medically important mite species, the American and European house dust mite. Both species have global distributions, though the former is more abundant in the U.S.

Ancestors of the two species probably separated from each other nearly 81 million years ago--long before the origin of humans--when they inhabited bird nests. Today, house dust mites are blamed for causing allergic reactions in more than 65 million people worldwide and thrive in the mattresses, sofas and carpets of even the cleanest homes.

Hamid, Klimov and their colleagues examined genetic variation in the group 1 allergen gene from samples of the two mite species collected in the U.S. and Pakistan. The group 1 allergen gene encodes for the most important allergy-causing protein in house dust mites.

An inactive form of this protein is used in clinics worldwide as part of the standard skin-prick test for allergies. Though the test can be inaccurate if it does not include local genetic variants of the allergy-causing protein, geographical variation in group 1 allergen proteins has not been extensively studied in the U.S., Klimov said.

"We need to have a better idea about the diversity of allergenic proteins around the world, and particularly in the United States," he said.

In genetic sequences from American house dust mites (Dermatophagoides farinae), the 
researchers observed mutations at 14 positions along the length of the group 1 allergen gene.

All but one of the mutations are "silent," meaning they occur at the DNA level without changing the amino acid structure of the protein. Only mutations at the protein level have medical significance because they can change allergenic properties.

"The most unexpected result was the finding that a previously unknown mutation occurred at the active site of the protein at position 197," Klimov said. "This was a rare mutation, found in only a single population of house dust mite in South Asia.

"Our analysis indicates that this mutation might alter the enzyme activity of the protein. But allergenic properties, immune response and cross-reactivity of the protein are unknown at this time," he said. "Follow-up experiments to elucidate these issues are underway in our lab."

Source: University of Michigan

'Darting' mice may hold clues to ADHD, autism, bipolar disorder

Mice inserted with a rare human genetic variation in the dopamine transporter could lead to improvements in the diagnosis and treatment of brain disorders. Credit: Image courtesy of Vanderbilt University Medical Center

 A darting mouse may hold an important clue in the development of Attention Deficit Hyperactivity Disorder (ADHD), autism and bipolar disorder, according to a study by a Vanderbilt University-led research team recently published in the Proceedings of the National Academy of Sciences.

The transgenic mouse, into which was inserted a rare human genetic variation in the dopamine transporter (DAT), could lead to improvements in the diagnosis and treatment of these all-too-common brain disorders, said Randy Blakely, Ph.D., the report's senior author.

The mutation, which has been found in people with ADHD, autism and bipolar disorder, affects the function of DAT, a protein that regulates the brain's supply of the neurotransmitter by removing excess dopamine from the synapse, or the space between nerve cells.

The DAT mutation causes the transporter to become "leaky" and spew out dopamine like "a vacuum cleaner in reverse," said Blakely, Allan D. Bass Professor of Pharmacology.

While mice with leaky DAT proteins have too much dopamine hanging around their synapses, surprisingly they aren't particularly hyperactive, possibly because DAT can still remove some of the dopamine.

But the mice exhibit an unusual "darting behavior." While their wild-type littermates are docile and quite unresponsive when researchers pick them up, those with the mutation "take off."

"Early on," Blakely said, "we could tell which ones carried the mutation by observing this response." Heightened anxiety does not appear to be the cause.

Blakely and his colleagues wonder whether this behavior is a form of "impulsivity." Rather than acting on their memories of being picked up a lot, the mice are opting for an inappropriate escape strategy.

Normal mice also stand up a lot to explore their cage. This "rearing" behavior is exacerbated by stimulant drugs. But not in these mice.

"We wonder whether this may be a sign that their behavior is driven less by searching for clues to appropriate behavior versus acting on innate impulses," Blakely said.

Other, better tests of impulsivity that evaluate premature decision-making can be applied in rodents and humans. "These tests are next on our docket," he said.

The actions of amphetamine and methylphenidate (Ritalin) are also affected by the mutation. In normal animals and people without ADHD, the stimulants flood the synapse with dopamine, eliciting hyperactivity.

But when given to the mutant animals, the drug demonstrates a "blunted" effect on both dopamine release and on locomotor activation compared to normal animals.

Blakely wonders whether stimulants like Adderall and Ritalin quell hyperactive and impulsive behaviors in some children with ADHD by reducing inappropriate dopamine leak.

"These mice may give us much better clues as to how these drugs are acting," he said.

To that end, Blakely recently received a five-year, $2-million grant from the National Institutes of Health (NIH grant number MH109054) to pursue explorations of these mice.

"Dopamine has classically been implicated in reward and the ability to detect novelty and to respond to pleasure and to engage in effective social interactions," he continued. The darting mice thus might shed light on a much broader spectrum of behaviors.

"We've got a lot to do," he said, "a lot of needy people (to help)."

Source: Vanderbilt University Medical Center

Genetic factors behind surviving or dying from Ebola shown in mouse study

In an emerging disease research lab at the University of Washington, Chris Williams, a research scientist who specializes in microbiology laboratory robotics, programs a piece of equipment that can be programmed to performs many lab tasks. Credit: Brian Donohue

A newly developed mouse model suggests that genetic factors are behind the mild-to-deadly range of reactions to the Ebola virus.

People exposed to Ebola vary in how the virus affects them. Some completely resist the disease, others suffer moderate to severe illness and recover, while those who are most susceptible succumb to bleeding, organ failure and shock.

In earlier studies of populations of people who have contracted Ebola, these differences are not related to any specific changes in the Ebola virus itself that made it more or less dangerous; instead, the body's attempts to fight infection seems to determine disease severity.

In the Oct. 30 edition of Science, scientists describe strains of laboratory mice bred to test the role of an individual's genetic makeup in the course of Ebola disease. Systems biologists and virologists Angela Rasmussen and Michael Katze from the Katze Laboratory at the University of Washington Department of Microbiology led the study in collaboration with the National Institutes of Health's Rocky Mountain Laboratories in Montana and University of North Carolina at Chapel Hill.

Research on Ebola prevention and treatment has been hindered by the lack of a mouse model that replicates the main characteristics of human Ebola hemorrhagic fever. The researchers had originally obtained this genetically diverse group of inbred laboratory mice to study locations on mouse genomes associated with influenza severity.

The research was conducted in a highly secure, state-of-the-art biocontainment safety level 4 laboratory in Hamilton, Mont. The scientists examined mice that they infected with a mouse form of the same species of Ebola virus causing the 2014 West Africa outbreak. The study was done in full compliance with federal, state, and local safety and biosecurity regulations. This type of virus has been used several times before in research studies. Nothing was done to change the virus.

Interestingly, conventional laboratory mice previously infected with this virus died, but did not develop symptoms of Ebola hemorrhagic fever.

In the present study, all the mice lost weight in the first few days after infection. Nineteen percent of the mice were unfazed. They not only survived, but also fully regained their lost weight within two weeks. They had no gross pathological evidence of disease. Their livers looked normal.

Eleven percent were partially resistant and less than half of these died. Seventy percent of the mice had a greater than 50 percent mortality. Nineteen percent of this last group had liver inflammation without classic symptoms of Ebola, and thirty-four percent had blood that took too long to clot, a hallmark of fatal Ebola hemorrhagic fever in humans. Those mice also had internal bleeding, swollen spleens and changes in liver color and texture.

The scientists correlated disease outcomes and variations in mortality rates to specific genetic lines of mice.

"The frequency of different manifestations of the disease across the lines of these mice screened so far are similar in variety and proportion to the spectrum of clinical disease observed in the 2014 West African outbreak," Rasmussen said.

While acknowledging that recent Ebola survivors may have had immunity to this or a related virus that saved them during this epidemic, Katze said, "Our data suggest that genetic factors play a significant role in disease outcome."

In general, when virus infection frenzied the genes involved in promoting blood vessel inflammation and cell death, serious or fatal disease followed. On the other hand, survivors experienced more activity in genes that order blood vessel repair and the production of infection-fighting white blood cells.

The scientists note that certain specialized types of cells in the liver could also have limited virus reproduction and put a damper on systemic inflammation and blood clotting problems in resistant mice. Susceptible mice had widespread liver infection, which may explain why they had more virus in their bodies and poorly regulated blood coagulation. The researchers also noticed that spleens in the resistant and susceptible mice took alternate routes to try to ward off infection.

"We hope that medical researchers will be able to rapidly apply these findings to candidate therapeutics and vaccines," Katze said. They believe this mouse model can be promptly implemented to find genetic markers, conduct meticulous studies on how symptoms originate and take hold, and evaluate drugs and that have broad spectrum anti-viral activities against all Zaire ebolaviruses, including the one responsible for the current West African epidemic.

Source: University of Washington Health Sciences/UW Medicine

Teeth, sex and testosterone reveal secrets of aging in wild mouse lemurs

A brown mouse lemur in the wild. Mouse lemurs, weighing a mere 30 to 80 grams, are the world's smallest primates. Credit: Jukka Jernvall

Mouse lemurs can live at least eight years in the wild -- twice as long as some previous estimates, a long-term longitudinal study finds.

PLOS ONE published the research on brown mouse lemurs (Microcebus rufus) led in Madagascar by biologist Sarah Zohdy, a post-doctoral fellow in Emory's Department of Environmental Sciences and the Rollins School of Public Health. Zohdy conducted the research while she was a doctoral student at the University of Helsinki.

"It's surprising that these tiny, mouse-sized primates, living in a jungle full of predators that probably consider them a bite-sized snack, can live so long," Zohdy says. "And we found individuals up to eight years of age in the wild with no physical symptoms of senescence like some captive mouse lemurs start getting by the age of four."

It is likely that starvation, predation, disease and other environmental stressors reduce the observed rate of senescence in the wild, Zohdy notes, but a growing body of evidence also suggests that captive conditions may affect mental and physical function.

"We focused on wild mouse lemurs because we want to know what happens naturally when a primitive primate is exposed to all of the extrinsic and intrinsic mortality factors that shaped them as a species," Zohdy says. "Comparing longevity data of captive and wild mouse lemurs may help us understand how the physiological and behavioral demands of different environments affect the aging process in other primates, including humans."

The study determined ages of wild mouse lemurs in Madagascar's Ranomafana National Park through a dental mold method that had not previously been used with small mammals. In addition to the high-resolution tooth-wear analysis for aging, fecal samples underwent hormone analysis.

The researchers found no difference between the longevity of male and female mouse lemurs, unlike most vertebrates where males tend to die first.

"And even more interestingly, we found no difference in testosterone levels between males and females," Zohdy says. Mouse lemurs are female dominant, which may explain why their testosterone levels are on a par with males.

"While elevated male testosterone levels have been implicated in shorter lifespans in several species, this is one of the first studies to show equivalent testosterone levels accompanying equivalent lifespans," Zohdy says.

A co-author of the study is primatologist Patricia Wright of the Centre ValBio Research Station in Madagascar and Stony Brook University. Other institutions involved in the study include Colorado State University, Duke University and the University of Arizona, Tucson.
Mouse lemurs, found only on the island of Madagascar, are the world's smallest primates. They are among nearly 100 species of lemurs that arrived in Madagascar some 65 million years ago, perhaps floating over from mainland Africa on mats of vegetation.

Mouse lemurs weigh a mere 30 to 80 grams but in captivity they live six times longer than mammals of similar body size, such as mice or shrews. Captive gray mouse lemurs (Microcebus murinus) can live beyond age 12. By age four, however, they can start exhibiting behavioral and neurologic degeneration. In addition to slowing of motor skills and activity levels, reduced memory capacity and sense of smell, the captive four-year-olds can start developing gray hair and cataracts, Zohdy says.

The wild brown mouse lemurs in the study were trapped, marked and released during the years 2003 to 2010. A total of 420 dental impressions were taken from the lower-right mandibular tooth rows of 189 unique individuals. Over the course of seven years, 270 age estimates were calculated. For 23 individuals captured three or more times during the duration of the study, the regression slopes of wear rates were calculated and the mean slope was used to calculate ages for all individuals.

"We found that wild brown mouse lemurs can live at least eight years," Zohdy says. "In the population that we studied, 16 percent lived beyond four years of age. And we found no physical signs of senescence, such as graying hair or cataracts, in any wild individual."

Limitations of the study include the inability to document gradual physiological symptoms of senescence in the wild. "Our results do not provide information about wild brown mouse lemurs that can be directly compared to senescence in captive gray mouse lemurs," Zohdy says. "Further research, using identical measures of senescence, will help to reveal whether patterns of physiological senescence occur consistently across the genus and in both captive and wild conditions."

Another confounding factor Zohdy cites is "the Sleeping Beauty effect," the fact that wild mouse lemurs hibernate for half the year, possibly boosting their life span.
"We now know that mouse lemurs can live a relatively long time in the wild," she says, "but we don't know the exact mechanisms behind why they live so long."

Source: Emory Health Sciences