Mice with ‘mohawks’ help scientists link autism to 2 biological pathways in brain

"Aha" moments are rare in medical research, scientists say. As rare, they add, as finding mice with Mohawk-like hairstyles.

But both events happened in a lab at NYU Langone Medical Center, months after an international team of neuroscientists bred hundreds of mice with a suspect genetic mutation tied to autism spectrum disorders.

Almost all the grown mice, the NYU Langone team observed, had sideways,”overgroomed” hair with a highly stylized center hairline between their ears and hardly a tuft elsewhere. Mice typically groom each other’s hair.

Researchers say they knew instantly they were on to something, as the telltale overgrooming — a repetitive motor behavior — had been linked in other experiments in mice to the brain condition that prevents children from developing normal social, behavioral, cognitive, and motor skills. People with autism, the researchers point out, exhibit noticeably dysfunctional behaviors, such as withdrawal, and stereotypical, repetitive movements, including constant hand-flapping, or rocking.

Now and for what NYU Langone researchers believe to be the first time, an autistic motor behavior has been traced to specific biological pathways that are genetically determined.

The findings, says senior study investigator Gordon Fishell, PhD, the Julius Raynes Professor of Neuroscience and Physiology at NYU Langone, could with additional testing in humans lead to new treatments for some autism, assuming the pathways’ effects as seen in mice are reversible.

In the study, to be published in the journal Nature online May 25, researchers knocked out production in mice of a protein called Cntnap4. This protein had been found in earlier studies in specialized brain cells, known as interneurons, in people with a history of autism.

Researchers found that knocking out Cntnap4 affected two highly specialized chemical messengers in the brain, GABA and dopamine. Both are so-called neurotransmitters, chemical signals released from one nerve cell to the next to stimulate similar sensations throughout the body. GABA, short for gamma-aminobutyric acid, is the main inhibitory neurotransmitter in the brain. It not only helps control brain impulses, but also helps regulate muscle tone. Dopamine is a well-known hormonal stimulant, highly touted for producing soothing, pleasing sensations.

Among the researchers’ key findings was that in Mohawk-coiffed mice, reduced Cntnap4 production led to depressed GABA signaling and overstimulation with dopamine. Researchers say the lost protein had opposite effects on the neurotransmitters because GABA is fast acting and quickly released, so interfering with its action decreases signaling, while dopamine’s signaling is longer-acting, so impairing its action increases its release.

"Our study tells us that to design better tools for treating a disease like autism, you have to get to the underlying genetic roots of its dysfunctional behaviors, whether it is overgrooming in mice or repetitive motor behaviors in humans," says Dr. Fishell. "There have been many candidate genes implicated in contributing to autism, but animal and human studies to identify their action have so far not led to any therapies. Our research suggests that reversing the disease’s effects in signaling pathways like GABA and dopamine are potential treatment options."

The U.S. Centers for Disease Control and Prevention estimate that one in 68 American children under age 8 has some form of autism, with five times as many boys as girls suffering from the spectrum of disorders.

As part of their study, researchers performed dozens of genetic, behavioral, and neural tests with growing mice to isolate and pinpoint where Cntnap4 acted in their brains, and how it affected chemical signaling among specific interneuron brain cells, which help relay and filter chemical signals between neurons in localized areas of the brain.

They found that Cntnap4 in mature interneurons strengthened GABA signaling, but did not do so in younger interneurons. When researchers traced where Cntnap4 acted in immature brain cells, Dr. Fishell says tests showed that it stimulated “a big bolus of dopamine.”

As part of testing to confirm the hereditary link among Cntnap4, the two pathways, and grooming behaviors, researchers exposed young mice with normal levels of Cntnap4, who did not groom each other, to mature mice with and without Cntnap4. Only mature mice deficient in Cntnap4 preened the hairstyle on other mice. Further tests in young mice without Cntnap4 showed that other, mature mice with normal amounts of Cntnap4 largely let them be, without any particular grooming or hairstyle.

Dr. Fishell and his team plan further analyses of how GABA and dopamine production changes as brain cells mature, and precisely what cellular mechanisms are involved in autism. Their goal is to control and rebalance any biological systems that go awry, as a possible future therapy for the disease.

Researchers examine how touch can trigger our emotions

While touch always involves awareness, it also sometimes involves emotion. For example, picking up a spoon triggers no real emotion, while feeling a gentle caress often does. Now, scientists in the Cell Press journal Neuron describe a system of slowly conducting nerves in the skin that respond to such gentle touch. Using a range of scientific techniques, investigators are beginning to characterize these nerves and to describe the fundamental role they play in our lives as a social species—from a nurturing touch to an infant to a reassuring pat on the back. Their work also suggests that this soft touch wiring may go awry in disorders such as autism.

The nerves that respond to gentle touch, called c-tactile afferents (CTs), are similar to those that detect pain, but they serve an opposite function: they relay events that are neither threatening nor tissue-damaging but are instead rewarding and pleasant.

"The evolutionary significance of such a system for a social species is yet to be fully determined," says first author Francis McGlone, PhD, of Liverpool John Moores University in England. "But recent research is finding that people on the autistic spectrum do not process emotional touch normally, leading us to hypothesize that a failure of the CT system during neurodevelopment may impact adversely on the functioning of the social brain and the sense of self."

For some individuals with autism, the light touch of certain fabrics in clothing can cause distress. Temple Grandin, an activist and assistant professor of animal sciences at Colorado State University who has written extensively on her experiences as an individual with autism, has remarked that her lack of empathy in social situations may be partially due to a lack of “comforting tactual input.” Professor McGlone also notes that deficits in nurturing touch during early life could have negative effects on a range of behaviors and psychological states later in life.

Further research on CTs may help investigators develop therapies for autistic patients and individuals who lacked adequate nurturing touch as children. Also, a better understanding of how nerves that relay rewarding sensations interact with those that signal pain could provide insights into new treatments for certain types of pain.

Professor McGlone believes that possessing an emotional touch system in the skin is as important to well-being and survival as having a system of nerves that protect us from harm. “In a world where human touch is becoming more and more of a rarity with the ubiquitous increase in social media leading to non-touch-based communication, and the decreasing opportunity for infants to experience enough nurturing touch from a carer or parent due to the economic pressures of modern living, it is becoming more important to recognize just how vital emotional touch is to all humankind.”

Can Chemicals Produced by Gut Microbiota Affect Children with Autism?

Children with autism spectrum disorders (ASD) have significantly different concentrations of certain bacterial-produced chemicals, called metabolites, in their feces compared to children without ASD. This research, presented at the annual meeting of the American Society for Microbiology, provides further evidence that bacteria in the gut may be linked to autism.

“Most gut bacteria are beneficial, aiding food digestion, producing vitamins, and protecting against harmful bacteria. If left unchecked, however, harmful bacteria can excrete dangerous metabolites or disturb a balance in metabolites that can affect the gut and the rest of the body, including the brain,” says Dae-Wook Kang of the Biodesign Institute of Arizona State University, an author on the study.

Increasing evidence suggests that children with ASD have altered gut bacteria. In order to identify possible microbial metabolites associated with ASD Kang and his colleagues looked for and compared the compounds in fecal samples from children with and without ASD. They found that children with ASD had significantly different concentrations of seven of the 50 compounds they identified.

“Most of the seven metabolites could play a role in the brain, working as neurotransmitters or controlling neurotransmitter biosynthesis,” says Kang. “We suspect that gut microbes may alter levels of neurotransmitter-related metabolites affecting gut-to-brain communication and/or altering brain function.”

Children with ASD had significantly lower levels of the metabolites homovanillate and N,N-dimethylglycine. Homovanillate is the breakdown product of dopamine (a major neurotransmitter), indicating an imbalance in dopamine catabolism (the breaking down in living organisms of more complex substances into simpler ones with the release of energy). N,N-dimethylglycine is a building block for proteins and neurotransmitters, and has been used to reduce symptoms of ASD and epileptic seizures.

The glutamine/glutamate ratio was significantly higher in children with ASD. Glutamine and glutamate are further metabolized to gamma-aminobutyric acid (GABA), an inhibitory neurotransmitter. An imbalance between glutamate and GABA transmission has been associated with ASD-like behaviors such as hyper-excitation.

Using next-generation sequencing technology, the researchers also were able to detect hundreds of unique bacterial species and confirmed that children with ASD harbored distinct and less diverse gut bacterial composition. 

“Correlations between gut bacteria and neurotransmitter-related metabolites are stepping stones for a better understanding of the crosstalk between gut bacteria and autism, which may provide potential targets for diagnosis or treatment of neurological symptoms in children with ASD,” says Kang.

(Image: Thinkstock)

Study confirms mitochondrial deficits in children with autism

Children with autism experience deficits in a type of immune cell that protects the body from infection. Called granulocytes, the cells exhibit one-third the capacity to fight infection and protect the body from invasion compared with the same cells in children who are developing normally.

The cells, which circulate in the bloodstream, are less able to deliver crucial infection-fighting oxidative responses to combat invading pathogens because of dysfunction in their tiny energy-generating organelles, the mitochondria.

The study is published online in the journal Pediatrics.

“Granulocytes fight cellular invaders like bacteria and viruses by producing highly reactive oxidants, toxic chemicals that kill microorganisms. Our findings show that in children with severe autism the level of that response was both lower and slower,”  said Eleonora Napoli, lead study author and project scientist in the Department of Molecular Biosciences in the UC Davis School of Veterinary Medicine. “The granulocytes generated less highly reactive oxidants and took longer to produce them.”

The researchers also found that the mitochondria in the granulocytes of children with autism consumed far less oxygen than those of the typically developing children — another sign of decreased mitochondrial function.

Mitochondria are the main intracellular source of oxygen free radicals, which are very reactive and can harm cellular structures and DNA. Cells can repair typical levels of oxidative damage. However, in the children with autism the cells produced more free radicals and were less able to repair the damage, and as a result experienced more oxidative stress. The free radical levels in the blood cells of children with autism were 1 ½ times greater than those without the disorder.

The study was conducted using blood samples of children enrolled in the Childhood Risk of Autism and the Environment (CHARGE) Study and included 10 children with severe autism age 2 to 5 and 10 age-, race- and sex-matched children who were developing typically.

In an earlier study the research team found decreased mitochondrial fortitude in another type of immune cell, the lymphocytes. Together, the findings suggest that deficiencies in the cells’ ability to fuel brain neurons might lead to some of the cognitive impairments associated with autism. Higher levels of free radicals also might contribute to autism severity.

“The response found among granulocytes mirrors earlier results obtained with lymphocytes from children with severe autism, underscoring the cross-talk between energy metabolism and response to oxidative damage,” said Cecilia Giulivi, professor in the Department of Molecular Biosciences in the UC Davis School of Veterinary Medicine and the study’s senior author.

“It also suggests that the immune response seems to be modulated by a nuclear factor named NRF2,” that controls antioxidant response to environmental factors and may hold clues to the gene-environment interaction in autism, Giulivi said.

(Image credit)

(Image caption: MRI images from a neurotypical control (left) and an adult with complete agenesis of the corpus callosum (right). The corpus callosum is indicated in red, fading as the fibers enter the hemispheres in order to suggest that they continue on. The anterior commissure is indicated by light aqua. The image illustrates the dramatic lack of inter hemispheric connections in callosal agenesis. Credit: Lynn Paul/Caltech)

Research Update: An Autism Connection

Building on their prior work, a team of neuroscientists at Caltech now report that rare patients who are missing connections between the left and right sides of their brain—a condition known as agenesis of the corpus callosum (AgCC)—show a strikingly high incidence of autism. The study is the first to show a link between the two disorders.

The findings are reported in a paper published April 22, 2014, in the journal Brain.

The corpus callosum is the largest connection in the human brain, connecting the left and right brain hemispheres via about 200 million fibers. In very rare cases it is surgically cut to treat epilepsy—causing the famous “split-brain” syndrome, for whose discovery the late Caltech professor Roger Sperry received the Nobel Prize. People with AgCC are like split-brain patients in that they are missing their corpus callosum—except they are born this way. In spite of this significant brain malformation, many of these individuals are relatively high-functioning individuals, with jobs and families, but they tend to have difficulty interacting with other people, among other symptoms such as memory deficits and developmental delays. These difficulties in social behavior bear a strong resemblance to those faced by high-functioning people with autism spectrum disorder.

"We and others had noted this resemblance between AgCC and autism before," explains Lynn Paul, lead author of the study and a lecturer in psychology at Caltech. But no one had directly compared the two groups of patients. This was a challenge that the Caltech team was uniquely positioned to do, she says, since it had studied patients from both groups over the years and had tested them on the same tasks.

"When we made detailed comparisons, we found that about a third of people with AgCC would meet diagnostic criteria for an autism spectrum disorder in terms of their current symptoms," says Paul, who was the founding president of the National Organization for Disorders of the Corpus Callosum.

The research was done in the laboratory of Ralph Adolphs, Bren Professor of Psychology and Neuroscience and professor of biology at Caltech and a coauthor of the study. The team looked at a range of different tasks performed by both sets of patients. Some of the exercises that involved certain social behaviors were videotaped and analyzed by the researchers to assess for autism. The team also gave the individuals questionnaires to fill out that measured factors like intelligence and social functioning.

"Comparing different clinical groups on exactly the same tasks within the same lab is very rare, and it took us about a decade to accrue all of the data," Adolphs notes.

One important difference between the two sets of patients did emerge in the comparison. People with autism spectrum disorder showed autism-like behaviors in infancy and early childhood, but the same type of behaviors did not seem to emerge in individuals with AgCC until later in childhood or the teen years.

"Around ages 9 through 12, a normally formed corpus callosum goes through a developmental ‘growth spurt’ which contributes to rapid advances in social skills and abstract thinking during those years," notes Paul. "Because they don’t have a corpus callosum, teens with AgCC become more socially awkward at the age when social skills are most important."

According to Adolphs, it is important to note that AgCC can now be diagnosed before a baby is born, using high-resolution ultrasound imaging during pregnancy. This latest development also opens the door for some exciting future directions in research.

"If we can identify people with AgCC already before birth, we should be in a much better position to provide interventions like social skills training before problems arise," Paul points out. "And of course from a research perspective it would be tremendously valuable to begin studying such individuals early in life, since we still know so little both about autism and about AgCC."

For example, the team would like to discern at what age subtle difficulties first appear in AgCC individuals, and at what point they start looking similar to autism, as well as what happens in the brain during these changes.

"If we could follow a baby with AgCC as it grows up, and visualize its brain with MRI each year, we would gain such a wealth of knowledge," Adolphs says.