Neuroscience

Researchers at Johns Hopkins have uncovered a protein switch that can either increase or decrease memory-building activity in brain cells, depending on the signals it detects. Its dual role means the protein is key to understanding the complex network of signals that shapes our brain’s circuitry, the researchers say. A description of their discovery appears in the July 31 issue of the Journal of Neuroscience.

“What’s interesting about this protein, AGAP3, is that it is effectively double-sided: One side beefs up synapses in response to brain activity, while the other side helps bring synapse-building back down to the brain’s resting state,” says Richard Huganir, Ph.D., a professor and director of the Solomon H. Snyder Department of Neuroscience at the Johns Hopkins University School of Medicine and co-director of the Brain Science Institute at Johns Hopkins. “The fact that it links these two opposing activities indicates AGAP3 may turn out to be central to controlling the strength of synapses.”

Huganir has long studied how connections between brain cells, known as synapses, are strengthened and weakened to form or erase memories. The new discovery came about when he and postdoctoral fellow Yuko Oku, Ph.D., investigated the chain reaction of signals involved in one type of synaptic strengthening.

In a study of the proteins that interact with one of the known proteins from that chain reaction, the previously unknown AGAP3 turned up. It contained not only a site designed to bind another protein involved in the chain reaction that leads from brain stimulation to learning, but also a second site involved in bringing synapse-building activity down to normal levels after a burst of activity.

Although it might seem the two different functions are behaving at cross-purposes, Oku says, it also could be that nature’s bundling of these functions together in a single protein is an elegant way of enabling learning and memory while preventing dangerous overstimulation. More research is needed, Oku says, to figure out whether AGAP3’s two sites coordinate by affecting each other’s activity, or are effectively free agents.

The steady accumulation of a protein in healthy, aging brains may explain seniors’ vulnerability to neurodegenerative disorders, a new study by researchers at the Stanford University School of Medicine reports.

The study’s unexpected findings could fundamentally change the way scientists think about neurodegenerative disease.

The pharmaceutical industry has spent billions of dollars on futile clinical trials directed at treating Alzheimer’s disease by ridding brains of a substance called amyloid plaque. But the new findings have identified another mechanism, involving an entirely different substance, that may lie at the root not only of Alzheimer’s but of many other neurodegenerative disorders — and, perhaps, even the more subtle decline that accompanies normal aging.

The study, published Aug. 14 in the Journal of Neuroscience, reveals that with advancing age, a protein called C1q, well-known as a key initiator of immune response, increasingly lodges at contact points connecting nerve cells in the brain to one another. Elevated C1q concentrations at these contact points, or synapses, may render them prone to catastrophic destruction by brain-dwelling immune cells, triggered when a catalytic event such as brain injury, systemic infection or a series of small strokes unleashes a second set of substances on the synapses.

“No other protein has ever been shown to increase nearly so profoundly with normal brain aging,” said Ben Barres, MD, PhD, professor and chair of neurobiology and senior author of the study. Examinations of mouse and human brain tissue showed as much as a 300-fold age-related buildup of C1q.

The finding was made possible by the diligence and ingenuity of the study’s lead author, Alexander Stephan, PhD, a postdoctoral scholar in Barres’ lab. Stephan screened about 1,000 antibodies before finding one that binds to C1q and nothing else. (Antibodies are proteins, generated by the immune system, that adhere to specific “biochemical shapes,” such as surface features of invading pathogens.)

Comparing brain tissue from mice of varying ages, as well as postmortem samples from a 2-month-old infant and an older person, the researchers showed that these C1q deposits weren’t randomly distributed along nerve cells but, rather, were heavily concentrated at synapses. Analyses of brain slices from mice across a range of ages showed that as the animals age, the deposits spread throughout the brain.

“The first regions of the brain to show a dramatic increase in C1q are places like the hippocampus and substantia nigra, the precise brain regions most vulnerable to neurodegenerative diseases like Alzheimer’s and Parkinson’s disease, respectively,” said Barres. Another region affected early on, the piriform cortex, is associated with the sense of smell, whose loss often heralds the onset of neurodegenerative disease.

Other scientists have observed moderate, age-associated increases (on the order of three- or four-fold) in brain levels of the messenger-RNA molecule responsible for transmitting the genetic instructions for manufacturing C1q to the protein-making machinery in cells. Testing for messenger-RNA levels — typically considered reasonable proxies for how much of a particular protein is being produced — is fast, easy and cheap compared with analyzing proteins.

But in this study, Barres and his colleagues used biochemical measures of the protein itself. “The 300-fold rise in C1q levels we saw in 2-year-old mice — equivalent to 70- or 80-year-old humans — knocked my socks off,” Barres said. “I was not expecting that at all.”

C1q is the first batter on a 20-member team of immune-response-triggering proteins, collectively called the complement system. C1q is capable of clinging to the surface of foreign bodies such as bacteria or to bits of our own dead or dying cells. This initiates a molecular chain reaction known as the complement cascade. One by one, the system’s other proteins glom on, coating the offending cell or piece of debris. This in turn draws the attention of omnivorous immune cells that gobble up the target.

The brain has its own set of immune cells, called microglia, which can secrete C1q. Still other brain cells, called astrocytes, secrete all of C1q’s complement-system “teammates.” The two cell types work analogously to the two tubes of an Epoxy kit, in which one tube contains the resin, the other a catalyst.

Previous work in Barres’ lab has shown that the complement cascade plays a critical role in the developing brain. A young brain generates an excess of synapses, creating a huge range of options for the potential formation of new neural circuits. These synapses strengthen or weaken over time, in response to their heavy use or neglect. The presence of feckless connections contributes noise to the system, so the efficiency of the maturing brain’s architecture is improved if these underused synapses are pruned away.

In a 2007 paper in Cell, Barres’ group reported that the complement system is essential to synaptic pruning in normal, developing brains. Then in 2012, in Neuron, in a collaboration with the lab of Harvard neuroscientist Beth Stevens, PhD, they showed that it is specifically microglia — the brain’s in-house immune cells — that attack and ingest complement-coated synapses.

Barres now believes something similar is happening in the normal, aging brain. C1q, but not the other protein components of the complement system, gradually becomes highly prevalent at synapses. By itself, this C1q buildup doesn’t trigger wholesale synapse loss, the researchers found — although it does seem to impair their performance. Old mice whose capacity to produce C1q had been eliminated performed subtly better on memory and learning tests than normal older mice did.

Still, this leaves the aging brain’s synapses precariously perched on the brink of catastrophe. A subsequent event such as brain trauma, a bad case of pneumonia or perhaps a series of tiny strokes that some older people experience could incite astrocytes — the second tube in the Epoxy kit — to start secreting the other complement-system proteins required for synapse destruction.

Most cells in the body have their own complement-inhibiting agents. This prevents the wholesale loss of healthy tissue during an immune attack on invading pathogens or debris from dead tissue during wound healing. But nerve cells lack their own supply of complement inhibitors. So, when astrocytes get activated, their ensuing release of C1q’s teammates may set off a synapse-destroying rampage that spreads “like a fire burning through the brain,” Barres said.

“Our findings may well explain the long-mysterious vulnerability specifically of the aging brain to neurodegenerative disease,” he said. “Kids don’t get Alzheimer’s or Parkinson’s. Profound activation of the complement cascade, associated with massive synapse loss, is the cardinal feature of Alzheimer’s disease and many other neurodegenerative disorders. People have thought this was because synapse loss triggers inflammation. But our findings here suggest that activation of the complement cascade is driving synapse loss, not the other way around.”

The synapses in the brain act as key communication points between approximately one hundred billion neurons. They form a complex network connecting various centres in the brain through electrical impulses.

New research from Lund University suggests that it is precisely here, in the synapses, that Huntington’s disease might begin.

The researchers looked into the brains of mice with real-time imaging methods, following some of the very first stages of the disease through advanced microscopes. What they discovered was an unprecedented degradation of synaptic activity. Long before the well documented nerve cell death, synapses that are important for communication between brain centres that control memory and learning begin to wither. This process has never been mapped before and could be an important step towards understanding the serious non-motor symptoms that affect Huntington patients long before the movement disorders start to show.
“With the naked eye, we have now been able to follow the step by step events when these synapses start to break down. If we are to halt or reverse this process in the future, it is necessary to understand exactly what happens in the initial phase of the disease. Now we know more”, says Professor Jia-Yi Li, the research group leader.

Huntington’s disease has long been characterized by the involuntary writhing movements faced by patients. But in fact, Huntington’s has a very broad and highly individual symptomatology. Depression, memory loss and sleep disorders are all common early on in the disease.
“Many patients testify that these symptoms affect quality of life significantly more than the involuntary jerky movements. Therefore, it is extremely important that we achieve progress in this field of research. Our goal now is to find new therapies that can increase the lifespan of these synapses and maintain their vital function”, explains postdoc Reena, who lead the imaging experiments.

Johns Hopkins researchers suggest neural stem cells may regenerate after anti-cancer treatment

Scientists have long believed that healthy brain cells, once damaged by radiation designed to kill brain tumors, cannot regenerate. But new Johns Hopkins research in mice suggests that neural stem cells, the body’s source of new brain cells, are resistant to radiation, and can be roused from a hibernation-like state to reproduce and generate new cells able to migrate, replace injured cells and potentially restore lost function.

“Despite being hit hard by radiation, it turns out that neural stem cells are like the special forces, on standby waiting to be activated,” says Alfredo Quiñones-Hinojosa, M.D., a professor of neurosurgery at the Johns Hopkins University School of Medicine and leader of a study described online today in the journal Stem Cells. “Now we might figure out how to unleash the potential of these stem cells to repair human brain damage.”

The findings, Quiñones-Hinojosa adds, may have implications not only for brain cancer patients, but also for people with progressive neurological diseases such as multiple sclerosis (MS) and Parkinson’s disease (PD), in which cognitive functions worsen as the brain suffers permanent damage over time.

In Quiñones-Hinojosa’s laboratory, the researchers examined the impact of radiation on mouse neural stem cells by testing the rodents’ responses to a subsequent brain injury. To do the experiment, the researchers used a device invented and used only at Johns Hopkins that accurately simulates localized radiation used in human cancer therapy. Other techniques, the researchers say, use too much radiation to precisely mimic the clinical experience of brain cancer patients.

In the weeks after radiation, the researchers injected the mice with lysolecithin, a substance that caused brain damage by inducing a demyelinating brain lesion, much like that present in MS. They found that neural stem cells within the irradiated subventricular zone of the brain generated new cells, which rushed to the damaged site to rescue newly injured cells. A month later, the new cells had incorporated into the demyelinated area where new myelin, the protein insulation that protects nerves, was being produced.

“These mice have brain damage, but that doesn’t mean it’s irreparable,” Quiñones-Hinojosa says. “This research is like detective work. We’re putting a lot of different clues together. This is another tiny piece of the puzzle. The brain has some innate capabilities to regenerate and we hope there is a way to take advantage of them. If we can let loose this potential in humans, we may be able to help them recover from radiation therapy, strokes, brain trauma, you name it.”

His findings may not be all good news, however. Neural stem cells have been linked to brain tumor development, Quiñones-Hinojosa cautions. The radiation resistance his experiments uncovered, he says, could explain why glioblastoma, the deadliest and most aggressive form of brain cancer, is so hard to treat with radiation.

In a breakthrough that could have wide-ranging applications in molecular medicine, Stanford University researchers have created a bioengineered peptide that enables imaging of medulloblastomas, among the most devastating of malignant childhood brain tumors, in lab mice.

The researchers altered the amino acid sequence of a cystine knot peptide — or knottin — derived from the seeds of the squirting cucumber, a plant native to Europe, North Africa and parts of Asia. Peptides are short chains of amino acids that are integral to cellular processes; knottin peptides are notable for their stability and resistance to breakdown.

The team used their invention as a “molecular flashlight” to distinguish tumors from surrounding healthy tissue. After injecting their bioengineered knottin into the bloodstreams of mice with medulloblastomas, the researchers found that the peptide stuck tightly to the tumors and could be detected using a high-sensitivity digital camera.

The findings are described in a study published online Aug. 12 in the Proceedings of the National Academy of Sciences.

“Researchers have been interested in this class of peptides for some time,” said Jennifer Cochran, PhD, an associate professor of bioengineering and a senior author of the study. “They’re extremely stable. For example, you can boil some of these peptides or expose them to harsh chemicals, and they’ll remain intact.”

That makes them potentially valuable in molecular medicine. Knottins could be used to deliver drugs to specific sites in the body or, as Cochran and her colleagues have demonstrated, as a means of illuminating tumors.

For treatment purposes, it’s critical to obtain accurate images of medulloblastomas. In conjunction with chemotherapy and radiation therapy, the tumors are often treated by surgical resection, and it can be difficult to remove them while leaving healthy tissue intact because their margins are often indistinct.

“With brain tumors, you really need to get the entire tumor and leave as much unaffected tissue as possible,” Cochran said. “These tumors can come back very aggressively if not completely removed, and their location makes cognitive impairment a possibility if healthy tissue is taken.”

The researchers’ molecular flashlight works by recognizing a biomarker on human tumors. The bioengineered knottin is conjugated to a near-infrared imaging dye. When injected into the bloodstreams of a strain of mice that develop tumors similar to human medullublastomas, the peptide attaches to the brain tumors’ integrin receptors — sticky molecules that aid in adhesion to other cells.

But while the knottins stuck like glue to tumors, they were rapidly expelled from healthy tissue. “So the mouse brain tumors are readily apparent,” Cochran said. “They differentiate beautifully from the surrounding brain tissue.”

The new peptide represents a major advance in tumor-imaging technology, said Melanie Hayden Gephart, MD, neurosurgery chief resident at the Stanford Brain Tumor Center and a lead author of the paper.

"The most common technique to identify brain tumors relies on preoperative, intravenous injection of a contrast agent, enabling most tumors to be visualized on a magnetic resonance imaging scan," Gephart said. These MRI scans are used like in a computer program much like an intraoperative GPS system to locate and resect the tumors.

“But that has limitations,” she added. “When you’re using the contrast in an MRI scan to define the tumor margins, you’re basically working off a preoperative snapshot. The brain can sometimes shift during an operation, so there’s always the possibility you may not be as precise or accurate as you want to be. The great potential advantage of this new approach would be to illuminate the tumor in real time — you could see it directly under your microscope instead of relying on an image that was taken before surgery.”

Though the team’s research focused on medulloblastomas, Gephart said it’s likely the new knottins could prove useful in addressing other cancers.

“We know that integrins exist on many types of tumors,” she said. “The blood vessels that tumors develop to sustain themselves also contain integrins. So this has the potential for providing very detailed, real-time imaging for a wide variety of tumors.”

And imaging may not be the only application for the team’s engineered peptide.

“We’re very interested in related opportunities,” Cochran said. “We envision options we didn’t have before for getting molecules into the brain.” In other words, by substituting drugs for dye, the knottins might allow the delivery of therapeutic compounds directly to cranial tumors — something that has proved extremely difficult to date because of the blood/brain barrier, the mechanism that makes it difficult for pathogens, as well as medicines, to traverse from the bloodstream to the brain.

“We’re looking into it now,” Cochran said.

A little serendipity was involved in the peptide’s development, said Sarah Moore, a recently graduated bioengineering PhD student and another lead author of the study. Indeed, the propinquity of Cochran’s laboratory to co-author Matthew Scott’s lab at Stanford’s James H. Clark Center catalyzed the project. “Our labs are next to each other,” Moore said. “We had the peptide, and Matt had ideal models of pediatric brain tumors  —mice that develop tumors in a similar manner to human medulloblastomas. Our partnership grew out of that.”

Scott, PhD, professor of bioengineering and of developmental biology, credits the design of the Clark Center as a contributor to the project. The building is home to Stanford’s Bioengineering Department, a collaboration between the School of Engineering and the School of Medicine, and Stanford Bio-X, an initiative that encourages communication among researchers in diverse scientific disciplines.

“So in a very real sense, our project wasn’t an accident,” Scott said. “In fact, it’s exactly the kind of work the Clark Center was meant to foster. The lab spaces are wide and open, with very few walls and lots of glass. We have a restaurant that only has large tables — no tables for two, so people have to sit together. Everything is designed to increase the odds that people will meet and talk. It’s a form of social engineering that really works.”

Scott said he is gratified by the collaboration that led to the team’s breakthrough, and observed that the peptide has proved a direct boon to his own work. About 15 percent of Scott’s mice develop the tumors requisite for medulloblastoma research. The problem, he said, is that the cancers are cryptic in their early stages.

“By the time you know the mice have them, many of the things you want to study — the genesis and development of the tumors — are past,” Scott said. “We needed ways to detect these tumors early, and we needed methods for following the steps of tumor genesis.”

Ultimately, Scott concluded, the development of the new peptide can be attributed to Stanford’s long-established traditions of openness and relentless inquiry.

“You find not just a willingness, but an eagerness to exchange ideas and information here,” Scott said. “It transcends any competitive instinct, any impulse toward proprietary thinking. It is what makes Stanford — well, Stanford.”

A team of neuroscientists has identified a modification to a protein in laboratory mice linked to conditions associated with Alzheimer’s Disease. Their findings, which appear in the journal Nature Neuroscience, also point to a potential therapeutic intervention for alleviating memory-related disorders.

The research centered on eukaryotic initiation factor 2 alpha (eIF2alpha) and two enzymes that modify it with a phosphate group; this type of modification is termed phosphorylation. The phosphorylation of eIF2alpha, which decreases protein synthesis, was previously found at elevated levels in both humans diagnosed with Alzheimer’s and in Alzheimer’s Disease (AD) model mice.

"These results implicate the improper regulation of this protein in Alzheimer’s-like afflictions and offer new guidance in developing remedies to address the disease," said Eric Klann, a professor in New York University’s Center for Neural Science and the study’s senior author.

The study’s co-authors also included: Douglas Cavener, a professor of biology at Pennsylvania State University; Clarisse Bourbon, Evelina Gatti, and Philippe Pierre of Université de la Méditerranée in Marseille, France; and NYU researchers Tao Ma, Mimi A. Trinh, and Alyse J. Wexler.

It has been known for decades that triggering new protein synthesis is vital to the formation of long-term memories as well as for long-lasting synaptic plasticity — the ability of the neurons to change the collective strength of their connections with other neurons. Learning and memory are widely believed to result from changes in synaptic strength.

In recent years, researchers have found that both humans with Alzheimer’s Disease and AD model mice have relatively high levels of eIF2alpha phosphorylation. But the relationship between this characteristic and AD-related afflictions was unknown.

Klann and his colleagues hypothesized that abnormally high levels of eIF2alpha phosphorylation could become detrimental because, ultimately, protein synthesis would diminish, thereby undermining the ability to form long-term memories.

To explore this question, the researchers examined the neurological impact of two enzymes that phosphorylate eIF2alpha, kinases termed PERK and GCN2, in different populations of AD model mice — all of which expressed genetic mutations akin to those carried by humans with AD. These were: AD model mice; AD model mice that lacked PERK; and AD model mice that lacked GCN2.

Specifically, they looked at eIF2alpha phosphorylation and the regulation of protein synthesis in the mice’s hippocampus region — the part of the brain responsible for the retrieval of old memories and the encoding of new ones. They then compared these levels with those of postmortem human AD patients.

Here, they found both increased levels of phosphorylated eIF2alpha in the hippocampus of both AD patients and the AD model mice. Moreover, in conjunction with these results, they found decreased protein synthesis, known to be required for long-term potentiation — a form of long-lasting synaptic plasticity—and for long-term memory.

To test potential remedies, the researchers examined phosphorylation of eIF2alpha in mice lacking PERK, hypothesizing that removal of this kinase would return protein synthesis to normal levels. As predicted, mice lacking PERK had levels of phosphorylated eIF2alpha and protein synthesis similar to those of normal mice.

They then conducted spatial memory tests in which the mice needed to navigate a series of mazes. Here, the AD model mice lacking PERK were able to successfully maneuver through the mazes at rates achieved by normal mice. By contrast, the other AD model mice lagged significantly in performing these tasks.

The researchers replicated these procedures on AD model mice lacking GCN2. The results here were consistent with those of the AD model mice lacking PERK, demonstrating that removal of both kinases diminished memory deficits associated with Alzheimer’s Disease.

Bad language could be good for you, a new study shows. For the first time, psychologists have found that swearing may serve an important function in relieving pain.

The study, published in the journal NeuroReport, measured how long college students could keep their hands immersed in cold water. During the chilly exercise, they could repeat an expletive of their choice or chant a neutral word. When swearing, the 67 student volunteers reported less pain and on average endured about 40 seconds longer.

Although cursing is notoriously decried in the public debate, researchers are now beginning to question the idea that the phenomenon is all bad. “Swearing is such a common response to pain that there has to be an underlying reason why we do it,” says psychologist Richard Stephens of Keele University in England, who led the study. And indeed, the findings point to one possible benefit: “I would advise people, if they hurt themselves, to swear,” he adds.

How swearing achieves its physical effects is unclear, but the researchers speculate that brain circuitry linked to emotion is involved. Earlier studies have shown that unlike normal language, which relies on the outer few millimeters in the left hemisphere of the brain, expletives hinge on evolutionarily ancient structures buried deep inside the right half.

One such structure is the amygdala, an almond-shaped group of neurons that can trigger a fight-or-flight response in which our heart rate climbs and we become less sensitive to pain. Indeed, the students’ heart rates rose when they swore, a fact the researchers say suggests that the amygdala was activated.

That explanation is backed by other experts in the field. Psychologist Steven Pinker of Harvard University, whose book The Stuff of Thought (Viking Adult, 2007) includes a detailed analysis of swearing, compared the situation with what happens in the brain of a cat that somebody accidentally sits on. “I suspect that swearing taps into a defensive reflex in which an animal that is suddenly injured or confined erupts in a furious struggle, accompanied by an angry vocalization, to startle and intimidate an attacker,” he says.

But cursing is more than just aggression, explains Timothy Jay, a psychologist at the Massachusetts College of Liberal Arts who has studied our use of profanities for the past 35 years. “It allows us to vent or express anger, joy, surprise, happiness,” he remarks. “It’s like the horn on your car, you can do a lot of things with that, it’s built into you.”

In extreme cases, the hotline to the brain’s emotional system can make swearing harmful, as when road rage escalates into physical violence. But when the hammer slips, some well-chosen swearwords might help dull the pain.

There is a catch, though: The more we swear, the less emotionally potent the words become, Stephens cautions. And without emotion, all that is left of a swearword is the word itself, unlikely to soothe anyone’s pain.

Self-perceived social status predicts hippocampal function and stress hormones

A mother’s perceived social status predicts her child’s brain development and stress indicators, finds a study at Boston Children’s Hospital. While previous studies going back to the 1950s have linked objective socioeconomic factors — such as parental income or education — to child health, achievement and brain function, the new study is the first to link brain function to maternal self-perception.

In the study, children whose mothers saw themselves as having a low social status were more likely to have increased cortisol levels, an indicator of stress, and less activation of their hippocampus, a structure in the brain responsible for long-term memory formation (required for learning) and reducing stress responses.

Findings were published online August 6th by the journal Developmental Science, and will be part of a special issue devoted to the effects of socioeconomic status on brain development.

"We know that there are big disparities among people in income and education," says Margaret Sheridan, PhD, of the Labs of Cognitive Neuroscience at Boston Children’s Hospital, the study’s first author. "Our results indicate that a mother’s perception of her social status ‘lives’ biologically in her children."

Sheridan, senior investigator Charles Nelson, PhD, of Boston Children’s Hospital and colleagues studied 38 children aged 8.3 to 11.8 years. The children gave saliva samples to measure levels of cortisol, and 19 also underwent functional MRI of the brain, focusing on the hippocampus.

Mothers, meanwhile, rated their social standing on a ladder on a scale of 1 to 10, comparing themselves with others in the United States. Findings were as follows:

• After controlling for gender and age, the mother’s self-perceived social status was a significant predictor of cortisol levels in the child. This finding is consistent with studies in animals. “In animal research, your stress response is related to your relative standing in the hierarchy,” Sheridan says.
• Similarly, the mother’s perceived social status significantly predicted the degree of hippocampal activation in their children during a learning task.
• In contrast, actual maternal education or income-to-needs ratio (income relative to family size) did not significantly predict cortisol levels or hippocampal activation.

The findings suggest that while actual socioeconomic status varies, how people perceive and adapt to their situation is an important factor in child development. Some of this may be culturally determined, Sheridan notes. She is currently participating in a much larger international study of childhood poverty, the Young Lives Project, that is looking at objective and subjective measures of social status along with health measures and cognitive function. The study will capture much wider extremes of socioeconomic status than would a U.S.-based study.

What the current study didn’t find was evidence that stress itself alters hippocampal function; no relationship was found between cortisol and hippocampal function, as has been seen in animals, perhaps because of the small number children having brain fMRIs. “This needs further exploration,” says Sheridan. “There may be more than one pathway leading to differences in long-term memory, or there may be an effect of stress on the hippocampus that comes out only in adulthood.”

NIH-funded scientists show new genetically engineered proteins may be important tool for the President’s BRAIN Initiative

Scientists used fruit flies to show for the first time that a new class of genetically engineered proteins can be used to watch electrical activity in individual brain cells in live brains. The results, published in Cell, suggest these proteins may be a promising new tool for mapping brain cell activity in multiple animals and for studying how neurological disorders disrupt normal nerve cell signaling. Understanding brain cell activity is a high priority of the President’s Brain Research through Advancing Innovative Neurotechnologies (BRAIN) Initiative.

Brain cells use electricity to control thoughts, movements and senses.  Ever since the late nineteenth century, when Dr. Luigi Galvani induced frog legs to move with electric shocks, scientists have been trying to watch nerve cell electricity to understand how it is involved in these actions. Usually they directly monitor electricity with cumbersome electrodes or toxic voltage-sensitive dyes, or indirectly with calcium detectors. This study, led by Michael Nitabach, Ph.D., J.D., and Vincent Pieribone, Ph.D., at the Yale School of Medicine, New Haven, CT, shows that a class of proteins, called genetically encoded fluorescent voltage indicators (GEVIs), may allow researchers to watch nerve cell electricity in a live animal.

Dr. Pieribone and his colleagues helped develop ArcLight, the protein used in this study. ArcLight fluoresces, or glows, as a nerve cell’s voltage changes and enables researchers to watch, in real time, the cell’s electrical activity. In this study, Dr. Nitabach and his colleagues engineered fruit flies to express ArcLight in brain cells that control the fly’s sleeping cycle or sense of smell. Initial experiments in which the researchers simultaneously watched brain cell electricity with a microscope and recorded voltage with electrodes showed that ArcLight can accurately monitor electricity in a living brain. Further experiments showed that ArcLight illuminated electricity in parts of the brain that were previously inaccessible using other techniques. Finally, ArcLight allowed the researchers to watch brain cells spark and fire while the flies were awakening and smelling. These results suggest that in the future neuroscientists may be able to use ArcLight and similar GEVIs in a variety of ways to map brain cell circuit activity during normal and disease states.

An innovative series of experiments could help to unlock the mysteries of how the brain makes sense of the hustle and bustle of human activity we see around us every day.

Very little is known about the psychological processes which enable us to pick out a potential mugger from a busy street or to spot an old friend approaching us across a crowded room. Such judgements of social intention, which we make countless times each day, enable us to respond in appropriate ways to the dynamic and complex world around us.

George Mather, Professor of Vision Science at the University of Lincoln, UK, and one of the world’s foremost experts on human visual perception, will lead a new research project investigating the mechanisms behind this crucial ability to perceive and interpret the intentions of other people from the way they move.

Numerous experiments have explored the way we use visual signals to extract meaning from our environment, but most have been based on static images, such as photos of different facial expressions.

Other studies into the perception of moving images have relied on very simple animated scenes, like moving patterns of regularly-spaced lines or random dots, devoid of the richness and nuances of scenes from the ‘real world’.

There remains limited scientific understanding of how the human visual system makes sense of the flurry of movement we see around us in modern societies: for example, whether a person approaching us is sprinting or strolling, whether that means they are angry or calm, and how we should react in response.

Professor Mather aims to bridge this gap in the academic literature through a series of world-first experiments. He has been awarded a grant of £287,000 by the UK’s Economic & Social Research Council (ESRC) for a three-year study. The aim is to shed new light on the process by which the human visual system identifies and decodes ‘dynamic cues of social intention’.

Professor Mather said: “It’s true that actions speak louder than words. Perception of movement is fundamental to many of our everyday social interactions. But simply judging speed is in itself a very complex task. When you see somebody walking across your field of view, how do you know how fast they are going? That information can be very useful because it might tell you something about their intentions but it’s surprisingly difficult to make an accurate judgement. A basic problem is that the further away a moving object is, the slower it moves in the image received by the eye. We don’t really understand at the moment how the human visual system is able to compensate for different viewing conditions.”

Motion perception has been a consistent theme of Professor Mather’s research career. In previous studies he has shown that the brain can deduce socially meaningful information from very simple depictions of human movement, such as collections of dots denoting the major joints of the body.

The research in this latest project will answer fundamental questions about how the brain combines ‘low-level’ information about image motion with ‘high level’ knowledge of the social world to make meaningful assessments of the speed and nature of human movements.

If you forget where you put your car keys and you can’t seem to remember things as well as you used to, the problem may well be with the GluN2B subunits in your NMDA receptors.

And don’t be surprised if by tomorrow you can’t remember the name of those darned subunits.

They help you remember things, but you’ve been losing them almost since the day you were born, and it’s only going to get worse. An old adult may have only half as many of them as a younger person.

Research on these biochemical processes in the Linus Pauling Institute at Oregon State University is making it clear that cognitive decline with age is a natural part of life, and scientists are tracking the problem down to highly specific components of the brain. Separate from some more serious problems like dementia and Alzheimer’s disease, virtually everyone loses memory-making and cognitive abilities as they age. The process is well under way by the age of 40 and picks up speed after that.

But of considerable interest: It may not have to be that way.

“These are biological processes, and once we fully understand what is going on, we may be able to slow or prevent it,” said Kathy Magnusson, a neuroscientist in the OSU Department of Biomedical Sciences, College of Veterinary Medicine, and professor in the Linus Pauling Institute. “There may be ways to influence it with diet, health habits, continued mental activity or even drugs.”

The processes are complex. In a study just published in the Journal of Neuroscience, researchers found that one protein that stabilizes receptors in a young animal – a good thing conducive to learning and memory – can have just the opposite effect if there’s too much of it in an older animal.

But complexity aside, progress is being made. In recent research, supported by the National Institutes of Health, OSU scientists used a genetic therapy in laboratory mice, in which a virus helped carry complementary DNA into appropriate cells and restored some GluN2B subunits. Tests showed that it helped mice improve their memory and cognitive ability.

The NMDA receptor has been known of for decades, Magnusson said. It plays a role in memory and learning but isn’t active all the time – it takes a fairly strong stimulus of some type to turn it on and allow you to remember something. The routine of getting dressed in the morning is ignored and quickly lost to the fog of time, but the day you had an auto accident earns a permanent etching in your memory.

Within the NMDA receptor are various subunits, and Magnusson said that research keeps pointing back to the GluN2B subunit as one of the most important. Infants and children have lots of them, and as a result are like a sponge in soaking up memories and learning new things. But they gradually dwindle in number with age, and it also appears the ones that are left work less efficiently.

“You can still learn new things and make new memories when you are older, but it’s not as easy,” Magnusson said. “Fewer messages get through, fewer connections get made, and your brain has to work harder.”

Until more specific help is available, she said, some of the best advice for maintaining cognitive function is to keep using your brain. Break old habits, do things different ways. Get physical exercise, maintain a good diet and ensure social interaction. Such activities help keep these “subunits” active and functioning.

Gene therapy such as that already used in mice would probably be a last choice for humans, rather than a first option, Magnusson said. Dietary or drug options would be explored first.

“The one thing that does seem fairly clear is that cognitive decline is not inevitable,” she said. “It’s biological, we’re finding out why it happens, and it appears there are ways we might be able to slow or stop it, perhaps repair the NMDA receptors. If we can determine how to do that without harm, we will.”

Study could help yield new drugs for brain disorders

Johns Hopkins biophysicists have discovered that full activation of a protein ensemble essential for communication between nerve cells in the brain and spinal cord requires a lot of organized back-and-forth motion of some of the ensemble’s segments. Their research, they say, may reveal multiple sites within the protein ensemble that could be used as drug targets to normalize its activity in such neurological disorders as epilepsy, schizophrenia, Parkinson’s and Alzheimer’s disease.

The glutamate-binding segments (blue, yellow) of ionotropic glutamate receptors undergo a “rocking” motion during activation by glutamate (red). (The dotted line provides a point of reference.)

A summary of the results, published online in the journal Neuron on Aug. 7, shows that full activation of so-called ionotropic glutamate receptors is more complex than previously envisioned. In addition to the expected shape changes that occur when the receptor “receives” and clamps down on glutamate messenger molecules, the four segments of the protein ensemble also rock back and forth in relation to each other when fewer than four glutamates are bound.

“We believe that our study is the first to show the molecular architecture and behavior of a prominent neural receptor protein ensemble in a state of partial activation,” says Albert Lau, Ph.D., assistant professor of biophysics and biophysical chemistry at the Johns Hopkins University School of Medicine.

Glutamate receptors reside in the outer envelope of every nerve cell in the brain and spinal cord, Lau notes, and are responsible for changing chemical information — the release of glutamate molecules from a neighboring nerve cell — into electrical information, the flow of charged particles into the receiving nerve cell. There would be sharply reduced communication between nerve cells in our brains if these receptors were disabled, he added, and thought and normal brain function in general would be severely compromised. Malfunctioning receptors, says Lau, have been linked with numerous neurological disorders and are therefore potential targets for drug therapies.

Lau explained that each glutamate receptor is a united group of four protein segments that has a pocket for clamping down on glutamate like a Venus fly trap snaring a bug. Below the glutamate-binding segments are four other segments embedded in the cell’s outer envelope to form a channel for charged particles to flow through. When no glutamates are bound to the receptor, the channel is closed; full activation of the receptor and full opening of the channel occur when four glutamates are bound, each to a difference pocket.

Previously, Lau says, investigators thought that the level of receptor activation simply corresponded to the degree to which each glutamate-binding segment changed shape during the glutamate-binding process. Using a combination of computer modeling, biophysical “imaging” of molecular structure, biochemical analysis and electrical monitoring of individual cells, the researchers teased apart some of the steps in between zero activation and full activation. They were able to show that the four glutamate-binding segments, in addition to clamping down on glutamate, also rock back and forth in pairs when fewer than four glutamates are bound.

“It isn’t clear yet how this rocking motion affects receptor function, but we now know that activation depends on more than how much each glutamate-binding segment clamps down,” says Lau. Previous development of drugs targeting the receptor focused on the four glutamate-binding pockets. “Our discovery of this molecular motion could aid the development of drugs by revealing additional drug-binding sites on the receptor,” he adds.

Adults could be at greater risk of becoming anxious and vulnerable to poor mental health if they were deprived of certain hormones while developing in the womb according to new research by scientists at Cardiff and Cambridge universities.

New research in mice has revealed the role of the placenta in long-term programming of emotional behaviour and the first time scientists have linked changes in adult behaviour to alterations in placental function.

Insulin-like growth factor-2 has been shown to play a major role in foetal and placental development in mammals, and changes in expression of this hormone in the placenta and foetus are implicated in growth restriction in the womb.

"The growth of a baby is a very complex process and there are lots of control mechanisms which make sure that the nutrients required by the baby to grow can be supplied by the mother," according to Professor Lawrence Wilkinson, a behavioural neuroscientist from Cardiff University’s School of Psychology who led the research.

"We were interested in how disrupting this balance could influence emotional behaviours a long time after being born, as an adult," he added.

In order to explore how a mismatch between supply and demand of certain nutrients might affect humans, Professor Wilkinson and his colleagues Dr Trevor Humby, Mikael Mikaelsson - both also from Cardiff University – and Dr Miguel Constancia of Cambridge University, examined the behaviour of adult mice with a malfunctioned supply of a vital hormone.

Dr Humby added: “We achieved this by damaging a hormone called Insulin-like growth factor-2, important for controlling growth in the womb. What we found when we did this was an imbalance in the supply of nutrients controlled by the placenta, and that this imbalance had major effects on how subjects were during adulthood – namely, that subject became more anxious later in life.

"These symptoms were accompanied by specific changes in brain gene expression related to this type of behaviour. This is the first example of what we have termed ‘placental-programming’ of adult behaviour. We do not know exactly how these very early life events can cause long-range effects on our emotional predispositions, but we suspect that our research findings may indicate that the seeds of our behaviour, and possibly vulnerability to brain and mental health disorders, are sown much earlier than previously thought."

Although these studies were carried out in mice, the findings may have wider implications for human development. Further studies are planned to investigate the brain mechanisms linking early life events, placental dysfunction and the emotional state of adults.

"Pressing the button of the lift at your work place, or apartment building is an automatic action – a habit. You don’t even really look at the different buttons; your hand is almost reaching out and pressing on its own. But what happens when you use the lift in a new place? In this case, your hand doesn’t know the way, you have to locate the buttons, find the right one, and only then your hand can press a button. Here, pushing the button is a goal-directed action." It is with this example that Rui Costa, principal investigator at the Champalimaud Neuroscience Programme (CNP), explains how critical it is to be able to shift between habits and goal-direct actions, in a fast and accurate way, in everyday life.

To unravel the circuit that underlies this capacity, the capacity to “break habits”, was the goal of this study, carried out by Christina Gremel and Rui Costa, at NIAAA, National Institutes of Health, USA and the Champalimaud Foundation, in Portugal, that is published today (Date) in Nature Communications.

"We developed a task where mice would shift between making the same action in a goal-directed or habitual manner. We could then, for the first time, directly examine brain areas controlling the capacity to break habits," explains the study’s lead author Christina Gremel from NIAAA. Evidence from previous studies has shown that two neighbouring regions of the brain are necessary for these different functions – the dorsal medial striatum is necessary for goal-directed actions and the dorsal lateral striatum is necessary for habitual actions. What was not known, and this new study reveals, is that a third region, the orbital frontal cortex (OFC), is critical for shifting between these two types of actions. As explained by Rui Costa, "when neurons in the OFC were inhibited, the generation of goal-directed actions was disrupted, while activation of these neurons, by means of a technique called optogenetics, selectively increased goal-directed actions."

For Costa, the results of this study suggest “something quite extraordinary – the same neural circuits function in a dynamic way, enabling the learning of automatic and goal-directed actions in parallel.”

These results have important implications for understanding neuropsychiatric disorders where the balance between habits and goal-directed actions is disrupted, such as obsessive-compulsive disorder.

The neural bases of behaviour, and their connection to neuropsychiatric disorders, are at the core of ongoing work by neuroscientists and clinicians at the Champalimaud Foundation.

Not only does practice make perfect, it also makes for more efficient generation of neuronal activity in the primary motor cortex, the area of the brain that plans and executes movement, according to researchers from the University of Pittsburgh School of Medicine. Their findings, published online today in Nature Neuroscience, showed that practice leads to decreased metabolic activity for internally generated movements, but not for visually guided motor tasks, and suggest the motor cortex is “plastic” and a potential site for the storage of motor skills.

The hand area of the primary motor cortex is known to be larger among professional pianists than in amateur ones. This observation has suggested that extensive practice and the development of expert performance induces changes in the primary motor cortex, said senior investigator Peter L. Strick, Ph.D., Distinguished Professor and chair, Department of Neurobiology, Pitt School of Medicine.

Prior imaging studies have shown that markers of synaptic activity, meaning the input signals to neurons, decrease in the primary motor cortex as repeated actions become routine and an individual develops expertise at a motor skill. The researchers found that markers of synaptic activity also display a marked decrease in monkeys trained to perform sequences of movements that are guided from memory — an internally generated task — rather than from vision. They wondered whether the change in synaptic activity indicated that neuron firing also declined. To examine this issue they recorded neuron activity and sampled metabolic activity, a measure of synaptic activity in the same animals.

All the monkeys were trained on two tasks and were rewarded when they reached out to touch an object in front of them. In the visually guided task, a visual target showed the monkeys where to reach and the end point was randomly switched from trial to trial. In the internally generated task the monkeys were trained to perform short sequences of movements without visual cues. They practiced the sequences until they achieved a level of skill comparable to an expert typist.

The researchers found neuron activity was comparable between monkeys that performed visually guided and internally generated tasks. However, metabolic activity was high for the visually guided task, but only modest during the internally generated task.

“This tells us that practicing a skilled movement and the development of expertise leads to more efficient generation of neuron activity in the primary motor cortex to produce the movement. The increase in efficiency could be created by a number of factors such as more effective synapses, greater synchrony in inputs and more finely tuned inputs,” Dr. Strick noted. “What is really important is that our results indicate that practice changes the primary motor cortex so that it can become an important substrate for the storage of motor skills. Thus, the motor cortex is adaptable, or plastic.

Neuropathic pain — pain that results from a malfunction in the nervous system — is a daily reality for millions of Americans. Unlike normal pain, it doesn’t go away after the stimulus that provoked it ends, and it also behaves in a variety of other unusual and disturbing ways. Someone suffering from neuropathic pain might experience intense discomfort from a light touch, for example, or feel as though he or she were freezing in response to a slight change in temperature.

A major part of the answer to the problem of neuropathic pain, scientists believe, is found in spinal nerve cells that release a signaling chemical known as GABA. These GABA neurons act as a sort of brake on pain impulses; it’s thought that when they die or are disabled the pain system goes out of control. If GABA neurons could be kept alive and healthy after peripheral nerve or tissue injury, it’s possible that neuropathic pain could be averted.

Now, University of Texas Medical Branch at Galveston researchers have found a way to, at least partially, accomplish this objective. The key, they determined, is stemming the biochemical assault by reactive oxygen species that are generated in the wake of nerve injury.

"GABA neurons are particularly susceptible to oxidative stress, and we hypothesized that reactive oxygen species contribute to neuropathic sensitization by promoting the loss of GABA neurons as well as hindering GABA functions," said UTMB professor Jin Mo Chung, senior author of a paper on the research now online in the journal Pain.

To test this hypothesis — and determine whether GABA neurons could be saved — the researchers conducted a series of experiments in mice that had been surgically altered to simulate the conditions of neuropathic pain. In one key experiment, mice treated with an antioxidant compound for a week after surgery were compared with untreated mice. The antioxidant mice showed less pain-associated behavior and were found to have far more GABA neurons than the untreated mice.

"So by giving the antioxidant we lowered the pain behavior, and when we look at the spinal cords we see the GABA neuron population is almost the same as normal," Chung said. "That suggested we prevented those neurons from dying, which is a big thing."

One complication, Chung noted, is a “moderate quantitative mismatch” between the behavioral data and the GABA-neuron counts. While the anti-oxidant mice displayed less pain behavior, their behavioral improvement wasn’t as substantial as their high number of GABA neurons would suggest. One possibility is that the surviving neurons were somehow impaired — a hypothesis supported by electrophysiological data.

Although no clinical trials are planned in the immediate future, Chung believes anti-oxidants have great potential as a treatment for neuropathic pain. “If this is true and it works in humans — well, any time you can salvage neurons, it’s a good thing,” he said. “Neuropathic pain is very difficult to treat, and I think this is a possibility, a good possibility.”