Muscle paralysis eased by light-sensitive stem cells

A genetic tweak can make light work of some nervous disorders. Using flashes of light to stimulate modified neurons can restore movement to paralysed muscles. A study demonstrating this, carried out in mice, lays the path for using such “optogenetic” approaches to treat nerve disorders ranging from spinal cord injury to epilepsy and motor neuron disease.

Optogenetics has been hailed as one of the most significant recent developments in neuroscience. It involves genetically modifying neurons so they produce a light-sensitive protein, which makes them “fire”, sending an electrical signal, when exposed to light.

So far optogenetics has mainly been used to explore how the brain works, but some groups are exploring using it as therapy. One stumbling block has been fears about irreversibly genetically manipulating the brain.

In the latest study, a team led by Linda Greensmith of University College London altered mouse stem cells in the lab before transplanting them into nerves in the leg – this means they would be easier to remove if something went wrong.

"It’s a very exciting approach that has a lot of potential," says Ziv Williams of Harvard Medical School in Boston.

Muscles in action

Greensmith’s team inserted an algal gene that codes for a light-responsive protein into mouse embryonic stem cells. They then added signalling molecules to make the stem cells develop into motor neurons, the cells that carry signals to and from the spinal cord to the rest of the body. They implanted these into the sciatic nerve – which runs from the spinal cord to the lower limbs – of mice whose original nerves had been cut.

After waiting five weeks for the implanted neurons to integrate with the muscle, Greensmith’s team anaesthetised the mice, cut open their skin and shone pulses of blue light on the nerve. The leg muscles contracted in response. “We were surprised at how well this worked,” says Greensmith.

Most current approaches being investigated to help people who are paralysed involve electrically stimulating their nerves or muscles. But this can be painful because they may still have working pain neurons. Plus, the electricity makes the muscles contract too forcefully, making them tire quickly.

Using the optogenetic approach, however, allows the muscle fibres to be stimulated more gently, because the light level can be increased with each pulse. “It gives a very smooth contraction,” says Greensmith.

Breathing restoration

To make the technique practical for use in people, the researchers are developing a light-emitting diode in the form of a cuff that would go around the nerve, which could be connected to a miniature battery pack under the skin.

They are also trying to develop an alternative to using embryonic stem cells, as these would require the recipient to take drugs to stop their immune system attacking the transplanted neurons. Instead the team is working with induced pluripotent stem cells, cells that have been reprogrammed to behave like embryonic stem cells, but can be made from a small sample of the intended recipient’s own skin.

The team’s first goal is to help people with motor neuron disease who lose the ability to control their breathing muscles. “Walking involves contracting about 40 different muscles in complex sequences,” says Greensmith. “Breathing is very simple – one muscle contracts and relaxes.”

They plan to test the restoration of breathing ability in pigs, and are developing a pacemaker that could repeatedly illuminate the phrenic nerve in the chest, which controls the diaphragm.

Other groups are exploring different therapeutic applications of optogenetics, including treatments for epilepsy and Parkinson’s disease

Bioengineer Studying How the Brain Controls Movement

A University of California, San Diego research team led by bioengineer Gert Cauwenberghs is working to understand how the brain circuitry controls how we move. The goal is to develop new technologies to help patients with Parkinson’s disease and other debilitating medical conditions navigate the world on their own. Their research is funded by the National Science Foundation’s Emerging Frontiers of Research and Innovation program.

"Parkinson’s disease is not just about one location in the brain that’s impaired. It’s the whole body. We look at the problems in a very holistic way, combine science and clinical aspects with engineering approaches for technology," explains Cauwenberghs, a professor at the Jacobs School of Engineering and co-director of the Institute for Neural Computation at UC San Diego. "We’re using advanced technology, but in a means that is more proactive in helping the brain to get around some of its problems—in this case, Parkinson’s disease—by working with the brain’s natural plasticity, in wiring connections between neurons in different ways."

Outcomes of this research are contributing to the system-level understanding of human-machine interactions, and motor learning and control in real world environments for humans, and are leading to the development of a new generation of wireless brain and body activity sensors and adaptive prosthetics devices. Besides advancing our knowledge of human-machine interactions and stimulating the engineering of new brain/body sensors and actuators, the work is directly influencing diverse areas in which humans are coupled with machines. These include brain-machine interfaces and telemanipulation.

Uncovering the underlying causes of Parkinson’s disease

A breakthrough investigation by UTS researchers into the underlying causes of Parkinson’s disease has brought us a step closer to understanding how to manage the condition.

The team, led by UTS postdoctoral fellow Dr Dominic Hare and Professor Philip Doble, has produced the first empirical evidence that an imbalance of iron and dopamine in the substantia nigra pars compacta (SNc) region of the brain is the root cause of the neurodegenerative condition.

Caused by the slow loss of neurons in the SNc that control autonomous movement, Parkinson’s disease causes persistent shaking, gastrointestinal problems and a variety of other ailments.

More than 80,000 Australians suffer from the illness, most over the age of 60.

Hare’s findings, before only assumptions in the scientific community, finally validate the theory that iron and dopamine react to create free radicals in the brain that slowly destroy neuron pathways and bring about the onset of Parkinson’s.

"When these two chemicals react, it forms a toxic species of dopamine that essentially reacts like bleach in the brain," said Hare.

To conduct their research Hare and his team used a unique tagging technique using antibodies labelled with gold nanoparticles that acted as proxies for dopamine molecules. This enabled the team to monitor and “co-localise” metals with other molecules and proteins in the brain.

And the findings of this work, said Hare, were revelatory.

"What we found is those particular cells (in the SNc) have what you could call an ‘anti-Goldilocks effect’ – they have just the right amount of iron and just the right amount of dopamine to cause damage," said Dr Hare.

"When we give mice a toxin that mimics the effects of Parkinson’s disease, these cells degenerate."

Hare theorises that this effect is likely a natural result of aging, when the brain’s ability to securely store iron diminishes and allows iron molecules to “leak” into critical areas such as the SNc.

Finding ways to design drugs that can get into the brain and eliminate surplus iron – an initiative that is already well underway in the process of treating other illnesses like cancer and Alzheimer’s disease – is now the next step forward in research.

Preventative measures to halt the build-up of iron in the brain as humans undergo the aging process are also touted by Hare as an important next step, and is something he is now working on.

"I think the real hope is, while we might not necessarily find a cure, prevention is actually not that far away," said Hare.

"So it’s a case where you can wake up and say, ‘my Parkinson’s is flaring up again’, take a tablet and go about your business."

(Image caption: In this microscope photo of motor neurons created in the laboratory of Su-Chun Zhang, green marks the nucleus and red marks the nerve fibers. Zhang and co-workers at the Waisman Center have identified a misregulation of protein in the nucleus as the likely first step in the pathology of ALS. Credit: Hong Chen, Su-Chun Zhang/Waisman Center)

Study helps unravel the tangled origin of ALS

By studying nerve cells that originated in patients with a severe neurological disease, a University of Wisconsin-Madison researcher has pinpointed an error in protein formation that could be the root of amyotrophic lateral sclerosis.

Also called Lou Gehrig’s disease, ALS causes paralysis and death. According to the ALS Association, as many as 30,000 Americans are living with ALS.

After a genetic mutation was discovered in a small group of ALS patients, scientists transferred that gene to animals and began to search for drugs that might treat those animals. But that approach has yet to work, says Su-Chun Zhang, a neuroscientist at the Waisman Center at UW-Madison, who is senior author of the new report, published April 3 in the journal Cell Stem Cell.

Zhang has been using a different approach — studying diseased human cells in lab dishes. Those cells, called motor neurons, direct muscles to contract and are the site of failure in ALS.

About 10 years ago, Zhang was the first in the world to grow motor neurons from human embryonic stem cells. More recently, he updated that approach by transforming skin cells into iPS (induced pluripotent stem) cells that were transformed, in turn, into motor neurons.

IPS cells can be used as “disease models,” as they carry many of the same traits as their donor. Zhang says the iPS approach offers a key advantage over the genetic approach, which “can only study the results of a known disease-causing gene. With iPS, you can take a cell from any patient, and grow up motor neurons that have ALS. That offers a new way to look at the basic disease pathology.”

In the new report, Zhang, Waisman scientist Hong Chen, and colleagues have pointed a finger at proteins that build a transport structure inside the motor neurons. Called neurofilament, this structure moves chemicals and cellular subunits to the far reaches of the nerve cell. The cargo needing movement includes neurotransmitters, which signal the muscles, and mitochondria, which process energy.

Motor neurons that control foot muscles are about three feet long, so neurotransmitters must be moved a yard from their origin in the cell body to the location where they can signal the muscles, Zhang says. A patient lacking this connection becomes paralyzed; tellingly, the first sign of ALS is often paralysis in the feet and legs.

Scientists have known for some time that in ALS, “tangles” along the nerve’s projections, formed of misshapen protein, block the passage along the nerve fibers, eventually causing the nerve fiber to malfunction and die. The core of the new discovery is the source of these tangles: a shortage of one of the three proteins in the neurofilament.

The neurofilament combines structural and functional roles, Zhang says. “Like the studs, joists and rafters of a house, the neurofilament is the backbone of the cell, but it’s constantly changing. These proteins need to be shipped from the cell body, where they are produced, to the most distant part, and then be shipped back for recycling. If the proteins cannot form correctly and be transported easily, they form tangles that cause a cascade of problems.”

Finding neurofilament tangles in an autopsy of an ALS patient “will not tell you how they happen, when or why they happen,” Zhang says. But with millions of cells — all carrying the human disease — to work with, Zhang’s research group discovered the source of the tangles in the protein subunits that compose the neurofilaments. “Our discovery here is that the disease ALS is caused by misregulation of one step in the production of the neurofilament,” he says.

Beyond ALS, Zhang says “very similar tangles” appear in Alzheimer’s and Parkinson’s diseases. “We got really excited at the idea that when you study ALS, you may be looking at the root of many neurodegenerative disorders.”

While working with motor neurons sourced in stem cells from patients, Zhang says he and his colleagues saw “quite an amazing thing. The motor neurons we reprogrammed from patient skin cells were relatively young, and we found that the misregulation happens very early, which means it is the most likely cause of this disease. Nobody knew this before, but we think if you can target this early step in pathology, you can potentially rescue the nerve cell.”

In the experiment just reported, Zhang found a way to rescue the neural cells living in his lab dishes. When his group “edited” the gene that directs formation of the deficient protein, “suddenly the cells looked normal,” Zhang says.

Already, he reports, scientists at the Small Molecule Screening and Synthesis Facility at UW-Madison are looking for a way to rescue diseased motor neurons. These neurons are made by the millions from stem cells using techniques that Zhang has perfected over the years.

Zhang says “libraries” of candidate drugs, each containing a thousand or more compounds, are being tested. “This is exciting. We can put this into action right away. The basic research is now starting to pay off. With a disease like this, there is no time to waste.”

New hope for treating ALS

Patient stem cells help identify common problem, leading to clinical trials

Harvard stem cell scientists have discovered that a recently approved medication for epilepsy might be a meaningful treatment for amyotrophic lateral sclerosis (ALS), also known as Lou Gehrig’s disease, a uniformly fatal neurodegenerative disorder. The researchers are now collaborating with Massachusetts General Hospital (MGH) to design an initial clinical trial testing the safety of the treatment in ALS patients.

The investigators all caution that a great deal of work needs to be done to assure the safety and efficacy of the treatment in ALS patients before physicians should start offering it.

The work, laid out in two related advance online publications in April by Cell Stem Cell and Cell Reports, is the long-term fruit of studies by Harvard Stem Cell Institute (HSCI) principal faculty member Kevin Eggan, who in a 2008 Science paper first raised the possibility of using ALS patient-derived stem cells to better understand the disease and identify therapeutic targets for new drugs.

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Meditation as object of medical research

Mindfulness meditation produces personal experiences that are not readily interpretable by scientists who want to study its psychiatric benefits in the brain. At a conference near Boston April 5, 2014, Brown University researchers will describe how they’ve been able to integrate mindfulness experience with hard neuroscience data to advance more rigorous study.

Mindfulness is always personal and often spiritual, but the meditation experience does not have to be subjective. Advances in methodology are allowing researchers to integrate mindfulness experiences with brain imaging and neural signal data to form testable hypotheses about the science — and the reported mental health benefits — of the practice.

A team of Brown University researchers, led by junior Juan Santoyo, will present their research approach at 2:45 p.m on Saturday, April 5, 2014, at the 12th Annual International Scientific Conference of the Center for Mindfulness at the University of Massachusetts Medical School. Their methodology employs a structured coding of the reports meditators provide about their mental experiences. That can be rigorously correlated with quantitative neurophysiological measurements.

“In the neuroscience of mindfulness and meditation, one of the problems that we’ve had is not understanding the practices from the inside out,” said co-presenter Catherine Kerr, assistant professor (research) of family medicine and director of translational neuroscience in Brown’s Contemplative Studies Initiative. “What we’ve really needed are better mechanisms for generating testable hypotheses – clinically relevant and experience-relevant hypotheses.”

Now researchers are gaining the tools to trace experiences described by meditators to specific activity in the brain.

“We’re going to [discuss] how this is applicable as a general tool for the development of targeted mental health treatments,” Santoyo said. “We can explore how certain experiences line up with certain patterns of brain activity. We know certain patterns of brain activity are associated with certain psychiatric disorders.”

Structuring the spiritual

At the conference, the team will frame these broad implications with what might seem like a small distinction: whether meditators focus on their sensations of breathing in their nose or in their belly. The two meditation techniques hail from different East Asian traditions. Carefully coded experience data gathered by Santoyo, Kerr, and Harold Roth, professor of religious studies at Brown, show that the two techniques produced significantly different mental states in student meditators.

“We found that when students focused on the breath in the belly their descriptions of experience focused on attention to specific somatic areas and body sensations,” the researchers wrote in their conference abstract. “When students described practice experiences related to a focus on the nose during meditation, they tended to describe a quality of mind, specifically how their attention ‘felt’ when they sensed it.”

The ability to distill a rigorous distinction between the experiences came not only from randomly assigning meditating students to two groups – one focused on the nose and one focused on the belly – but also by employing two independent coders to perform standardized analyses of the journal entries the students made immediately after meditating.

This kind of structured coding of self-reported personal experience is called “grounded theory methodology.” Santoyo’s application of it to meditation allows for the formation of hypotheses.

For example, Kerr said, “Based on the predominantly somatic descriptions of mindfulness experience offered by the belly-focused group, we would expect there to be more ongoing, resting-state functional connectivity in this group across different parts of a large brain region called the insula that encodes visceral, somatic sensations and also provides a readout of the emotional aspects of so-called ‘gut feelings’.”

Unifying experience and the brain

The next step is to correlate the coded experiences data with data from the brain itself. A team of researchers led by Kathleen Garrison at Yale University, including Santoyo and Kerr, did just that in a paper in Frontiers in Human Neuroscience in August 2013. The team worked with deeply experienced meditators to correlate the mental states they described during mindfulness with simultaneous activity in the posterior cingulate cortex (PCC). They measured that with real-time functional magnetic resonance imaging.

They found that when meditators of several different traditions reported feelings of “effortless doing” and “undistracted awareness” during their meditation, their PCC showed little activity, but when they reported that they felt distracted and had to work at mindfulness, their PCC was significantly more active. Given the chance to observe real-time feedback on their PCC activity, some meditators were even able to control the levels of activity there.

“You can observe both of these phenomena together and discover how they are co-determining one another,” Santoyo said. “Within 10 one-minute sessions they were able to develop certain strategies to evoke a certain experience and use it to drive the signal.”

Toward therapies

A theme of the conference, and a key motivator in Santoyo and Kerr’s research, is connecting such research to tangible medical benefits. Meditators have long espoused such benefits, but support from neuroscience and psychiatry has been considerably more recent.

In a February 2013 paper in Frontiers in Human Neuroscience, Kerr and colleagues proposed that much like the meditators could control activity in the PCC, mindfulness practitioners may gain enhanced control over sensory cortical alpha rhythms. Those brain waves help regulate how the brain processes and filters sensations, including pain, and memories such as depressive cognitions.

Santoyo, whose family emigrated from Colombia when he was a child, became inspired to investigate the potential of mindfulness to aid mental health beginning in high school. Growing up in Cambridge and Somerville, Mass., he observed the psychiatric difficulties of the area’s homeless population. He also encountered them while working in food service at Cambridge hospital.

“In low-income communities you always see a lot of untreated mental health disorders,” said Santoyo, who meditates regularly and helps to lead a mindfulness group at Brown. He is pursuing a degree in neuroscience and contemplative science. “The perspective of contemplative theory is that we learn about the mind by observing experience, not just to tickle our fancy but to learn how to heal the mind.”

It’s a long path, perhaps, but Santoyo and his collaborators are walking it with progress.

Positive, negative thinkers’ brains revealed

The ability to stay positive when times get tough – and, conversely, of being negative – may be hardwired in the brain, finds new research led by a Michigan State University psychologist.

The study, which appears in the Journal of Abnormal Psychology, is the first to provide biological evidence validating the idea that there are, in fact, positive and negative people in the world.

“It’s the first time we’ve been able to find a brain marker that really distinguishes negative thinkers from positive thinkers,” said Jason Moser, lead investigator and assistant professor of psychology.

For the study, 71 female participants were shown graphic images and asked to put a positive spin on them while their brain activity was recorded. Participants were shown a masked man holding a knife to a woman’s throat, for example, and told one potential outcome was the woman breaking free and escaping.

The participants were surveyed beforehand to establish who tended to think positively and who thought negatively or worried. Sure enough, the brain reading of the positive thinkers was much less active than that of the worriers during the experiment.

“The worriers actually showed a paradoxical backfiring effect in their brains when asked to decrease their negative emotions,” Moser said. “This suggests they have a really hard time putting a positive spin on difficult situations and actually make their negative emotions worse even when they are asked to think positively.”

The study focused on women because they are twice as likely as men to suffer from anxiety related problems and previously reported sex differences in brain structure and function could have obscured the results.

Moser said the findings have implications in the way negative thinkers approach difficult situations.

“You can’t just tell your friend to think positively or to not worry – that’s probably not going to help them,” he said. “So you need to take another tack and perhaps ask them to think about the problem in a different way, to use different strategies.”

Negative thinkers could also practice thinking positively, although Moser suspects it would take a lot of time and effort to even start to make a difference.

A Brain Region for Resisting Alcohol’s Allure

As recovering spring breakers are regretting binge drinking escapades, it may be hard for them to appreciate that there is a positive side to the nausea, sleepiness, and stumbling. University of Utah neuroscientists report that when a region of the brain called the lateral habenula is chronically inactivated in rats, they repeatedly drink to excess and are less able to learn from the experience. The study, published online in PLOS ONE on April 2, has implications for understanding behaviors that drive alcohol addiction.

While complex societal pressures contribute to alcoholism, physiological factors are also to blame. Alcohol is a drug of abuse, earning its status because it tickles the reward system in the brain, triggering the release of feel-good neurotransmitters. The dreaded outcomes of overindulging serve the beneficial purpose of countering the pull of temptation, but little is understood about how those mechanisms are controlled.

U of U professor of neurobiology and anatomy Sharif Taha, Ph.D., and colleagues, tipped the balance that reigns in addictive behaviors by inactivating in rats a brain region called the lateral habenula. When the rats were given intermittent access to a solution of 20% alcohol over several weeks, they escalated their alcohol drinking more rapidly, and drank more heavily than control rats.

“In people, escalation of intake is what eventually separates a social drinker from someone who becomes an alcoholic,” said Taha. “These rats drink amounts that are quite substantial. Legally they would be drunk if they were driving.”

The lateral habenula is activated by bad experiences, suggesting that without this region the rats may drink more because they fail to learn from the negative outcomes of overindulging. The investigators tested the idea by giving the rats a desirable, sweet juice then injecting them with a dose of alcohol large enough to cause negative effects.

“It’s the same kind of learning that mediates your response in food poisoning. You taste something and then you get sick, and then of course you avoid that food in future meals,” explained Taha.

Yet rats with an inactivated lateral habenula sought out the juice more than control animals, even though it meant a repeat of the bad experience.

“The way I look at it is the rewarding effects of drinking alcohol compete with the aversive effects,” explained Andrew Haack, who is co-first author on the study with Chandni Sheth, both neuroscience graduate students. “When you take the aversive effects away, which is what we did when we inactivated the lateral habenula, the rewarding effects gain more purchase, and so it drives up drinking behavior.”

The group’s findings may help explain results from previous clinical investigations demonstrating that men who were less sensitive to the negative effects of alcohol drank more heavily, and were more likely to become problem drinkers later in life.

The researches think the lateral habenula likely works in one of two ways. The region may regulate how badly an individual feels after over-drinking. Alternatively, it may control how well an individual learns from their bad experience. Future work will resolve between the two.

“If we can understand the brain circuits that control sensitivity to alcohol’s aversive effects, then we can start to get a handle on who may become a problem drinker,” said Taha.

Noisy brain signals: How the schizophrenic brain misinterprets the world

People with schizophrenia often misinterpret what they see and experience in the world. New research provides insight into the brain mechanisms that might be responsible for this misinterpretation. The study from the Montreal Neurological Institute and Hospital – The Neuro - at McGill University and McGill University Health Centre, reveals that certain errors in visual perception in people with schizophrenia are consistent with interference or ‘noise’ in a brain signal known as a corollary discharge. Corollary discharges are found throughout the animal kingdom, from bugs to fish to humans, and they are thought to be crucial for monitoring one’s own actions. The study, published in the April 2 issue of the Journal of Neuroscience, identifies a corollary discharge dysfunction in schizophrenia, which could aid with diagnosis and treatment of this difficult disorder. It was carried out in collaboration with researchers Veronica Whitford, Gillian O’Driscoll, and Debra Titone in the Department of Psychology, McGill University.

“A corollary discharge is a copy of a nervous system message that is sent to other parts of the brain, in order to make us aware that we are doing something,” said Dr. Christopher Pack, neuroscientist at The Neuro and lead investigator on the study. “For example, if we want to move our arm, the motor area of the brain sends a signal to the muscles to produce a movement. A copy of this command, which is the corollary discharge, is sent to other regions of the brain, to inform them of the impending movement. If you were moving your arm, and you didn’t have the corollary discharge signal, you might assume that someone else was moving your arm. Similarly, if you generated a thought, and you had an impaired corollary discharge, then you might assume that someone else placed the thought in your mind. Corollary discharges ensure that different areas of the brain are communicating with each other, so that we are aware that we are moving our own arm, talking, or thinking our own thoughts.”

Schizophrenia is a disorder that interferes with the ability to think clearly and to manage emotions. People with schizophrenia often attribute their own thoughts and actions to external sources, as in the case of auditory hallucinations. Other common symptoms include delusions and disorganized thinking and speech. 

Recent research has suggested that an impaired corollary discharge can account for some of these symptoms. However, the nature of the impairment was unknown. In their study, Dr. Pack and his colleagues (including Dr. Alby Richard, neurology resident at The Neuro) used a test called a perisaccadic localization task, to investigate corollary discharge activity. In this test, subjects are asked to make quick eye movements to follow a dot on a computer screen. At the same time they are also asked to localize visual stimuli that appear briefly on the screen from time to time. In order to perform this task accurately, subjects need to know where on the screen they are looking – in other words they use corollary discharges signals that arise from the brain structures that control the eye muscles.

The results showed that people with schizophrenia were less accurate in figuring out where they were looking. Consequently they made more mistakes in estimating the position of the stimuli that were flashed on the screen. “What is interesting and potentially clinically important is that the pattern of mistakes made by the patients correlated with the extent of their symptoms,” said Dr. Pack. “This is particularly interesting because the circuits that control eye movements include the best-understood structures in the brain. So we are optimistic that we can work backward from the behavioral data to the biological basis of the corollary discharge effects. We have already started to do this with computational modeling. Mathematically we can convert the corollary discharge of a healthy control into the corollary discharge of a patient with schizophrenia by adding noise and randomness. It is not that people with schizophrenia have no corollary discharge, or a corollary discharge with delayed or weaker amplitude. Rather the patients appear primarily to have a noisy corollary discharge signal. This visual test is very easy thing to do and quite sensitive to individual differences.“

The study shows that patients with schizophrenia make larger errors in localizing visual stimuli compared to controls. These results could be explained by a corollary discharge signal, which also predicts patient symptom severity, suggesting a possible basis for some of the most common symptoms of schizophrenia. This work was supported by The Natural Sciences and Engineering Research Council of Canada, The Brain & Behavior Research Foundation (NARSAD) and the EJLB Foundation.