Dodging dots helps explain brain circuitry

A neuroscience study provides new insight into the primal brain circuits involved in collision avoidance, and perhaps a more general model of how neurons can participate in networks to process information and act on it.

In the study, Brown University neuroscientists tracked the cell-by-cell progress of neural signals from the eyes through the brains of tadpoles as they saw and reacted to stimuli including an apparently approaching black circle. In so doing, the researchers were able to gain a novel understanding of how individual cells contribute in a broader network that distinguishes impending collisions.

The basic circuitry involved is present in a wide variety of animals, including people, which is no surprise given how fundamental collision avoidance is across animal behavior.

“Imagine yourself walking in a forest while keeping a conversation with your friend,” said Arseny Khakhalin, neuroscience postdoctoral scholar at Brown and lead author of the study in the European Journal of Neuroscience. “You can totally keep the conversation going, and at the same time avoid tree trunks and shrubs without even thinking about them consciously. That’s because you have a whole region in your brain that is dedicated, among other things, to this task.”

Turning tail

To learn how collision avoidance works, Khakhalin studied the task using tadpoles as a model organism, because as senior author and neuroscience professor Carlos Aizenman put it, they are “sufficiently complex to produce interesting behavior, but have nervous systems sufficiently simple to address in an integrated experimental approach.”

They started with the avoidance behavior. With tadpoles in a dish atop a screen, they projected digital black dots, representing virtual objects, of varying widths, at varying speeds and angles of approach. They also just flashed dots in place. The tadpoles would flee approaching dots as long as they reached a certain threshold angular size, but rarely reacted to the dots that merely blinked onto the scene but weren’t moving toward them. The response confirmed that tadpoles can distinguish approaching rather than merely proximate visual stimuli.

The researchers then sought to determine how the tadpoles process different stimuli. To do that they held the tadpoles in place while presenting a variety of simple animations via a fiber optic cable held next to an eye. The animations included a flashed circle, an apparently approaching circle (it became larger and larger), and a couple of “in between” animations, such as a circle that was faded in, rather than simply flashed into being.

While the tadpoles watched the animations, the researchers tracked their tail movements with a high-speed camera (to determine if the tadpoles were executing a fleeing maneuver) and recorded electrical signals along the visual processing circuitry: at the optic nerve leading from the retina to the brain’s optic tectum region, at “excitatory” and “inhibitory” synaptic inputs of neurons in the optic tectum, and at the outputs of the tectal neurons.

What the scientists found was that the tectum, rather than the retina, appears to be where the tadpoles determine that something is approaching rather than merely present. How did they know? The strongest difference between responses to the apparently approaching circle, versus responses to other stimuli, such as flashed or faded circles, was detected at the stage of output from tectal neurons.

Moreover, the difference in activity related to approaching vs. flashed circles increased as the signal propagated from the optic nerve, through tectum input, and to tectum output.

“The tectum is the first place that responded to approaching stimuli not just differently, but stronger,” Khakhalin said.

Inhibition moderates the conversation

An implication of the experiments was that when individual neurons in the tectum are uniquely activated by an apparently approaching stimulus, they collectively generate a signal to send to downstream parts of the brain that can get the tail moving to avoid the collision.

That’s indeed what excitatory neurons do, but the researchers wanted to know what role the inhibitory neurons were playing, especially because the balance of inhibitory and excitatory activity in the tectum varied with different stimuli.

To find out, they chemically blocked inhibitory neurons in the tectum in some tadpoles, chemically enhanced their activity in others and left still other tadpoles unaltered as controls. They found that when they altered the degree of inhibition in either direction, the output selectivity for an oncoming stimulus was lost. When inhibition was blocked, the individual excitatory cells lost their selectivity, too. When inhibition was enhanced, the individual excitatory cells retained their selectivity but could not project a signal collectively.

Khakhalin said the evidence seems to support the idea of inhibitory cells as facilitators of network function. They were not necessarily responsible for making the tectum selective. Instead, their ability to moderate excitation allowed the network of cells to function so that an organized signal from the individual excitatory neurons could emerge from the tectum.

The team was able to use these findings to create a conceptual model of the collision stimulus circuitry.

Khakhalin’s hypothesis of how it works is that inhibitory/excitatory balance allows the tectum to build up a necessary degree of excitement about the stimulus of interest (e.g. something has been getting bigger) while still allowing enough “calm” to consider the next moment wave of input (it just got bigger again).

Aizenman said the paper illustrates broader approach that his lab is applying to fundamental neuroscience questions.

“It is part of a greater project to be able to take an entire behavior and break it down into all of its neuronal components, to build a model in which we can understand how activity in single neurons and in the connections between them can all synergize to produce a behavior,” he said.

Timing is everything: scientists control rapid re-wiring of brain circuits using patterned visual stimulation

In a new study, published in this week’s issue of the journal Science, researchers show for the first time how the brain re-wires and fine-tunes its connections differently depending on the relative timing of sensory stimuli. In most neuroscience textbooks today, there is a widely held model that explains how nerve circuits might refine their connectivity based on patterned firing of brain cells, but it has not previously been directly observed in real time. This “Hebbian Theory”, named after the McGill University psychologist Donald Olding Hebb who first proposed it in 1949 has been summarized as:

“Cells that fire together, wire together. Cells that fire out of sync, lose their link”

In other words, a nerve cell that fires at the same time as its nerve cell neighbors will cooperatively form strong, stable connections onto its partner cells. On the other hand, a nerve cell that fires out of synchrony with its neighbours, will end up destabilizing and withdrawing its connections. “For the first time, we have direct, real-time evidence from watching brain cells in an intact animal to support Hebb’s model, but, we also provide surprising, new details, fundamentally updating the model for the 21^st century,” says Dr. Edward Ruthazer, senior investigator on the study at the Montreal Neurological Institute and Hospital –The Neuro at McGill University and the McGill University Health Centre. 

The study, which used multiphoton laser-scanning microscopy to observe cells in the brains of intact animals, discovered that asynchronous firing, or “firing out of sync” not only caused brain cells to lose their ability to make other cells fire, but unexpectedly, also caused them to dramatically increase their elaboration of new branches in search of better matched partners. “The surprising and entirely unexpected finding is that even though nerve circuit remodeling from asynchronous stimulation actively weakens connections, there is a 60% increase in axon branches that are exploring the environment but these exploratory branches are not long-lived,” said Dr. Ruthazer.

IMAGES of nerves in action in transparent xenopus tadpoles: http://bit.ly/1lNuux0

Dr. Ruthazer’s lab charts the formation of brain circuitry during development in the hopes of better understanding the rules that control healthy brain wiring and of advancing treatments for injuries to the nervous system and therapies for neurodevelopmental disorders such as autism and schizophrenia. Astoundingly, nearly one out of every 100 Canadians suffers from one of these disorders, estimated to cost the Canadian economy over $10 billion annually in addition to inflicting a devastating impact on patients and their families.

In the developing brain, initially imprecise connections between nerve cells are gradually pruned away, leaving connections that are stronger and more specific. This refinement occurs in response to patterned stimulation from the environment. “The way we perceive the world as adults is directly impacted by what we saw when we were younger,” says Dr. Ruthazer.

Dr. Ruthazer’s team studies brain development in Xenopus tadpoles, which have the distinct advantage of being transparent, enabling the team to clearly see the nervous system inside. They have developed a model that allows them to watch nerve cell remodeling in vivo, in real time, and to measure the efficacy of connections between cells. Optic fibers were used to stimulate the eyes of the tadpoles with different light patterns, while imaging and recording nerve cell branch formation.  Asynchronous stimulation involved light flashes presented to each eye at different times, while synchronous stimulation involved simultaneous stimulation of both eyes.

Importantly, Dr. Ruthazer’s group also has begun to identify the molecular mechanisms underlying these changes in the nervous system. They show that the stabilization of the retinal nerve cell branches caused by synchronous firing involves signaling downstream of the synaptic activation of a neurotransmitter receptor called the N-methyl-D-aspartate receptor. In contrast, the enhanced exploratory growth that occurs with asynchronous activity does not appear to require the activation of this receptor.

Study reveals workings of working memory

Keep this in mind: Scientists say they’ve learned how your brain plucks information out of working memory when you decide to act.

Say you’re a busy mom trying to wrap up a work call now that you’ve arrived home. While you converse on your Bluetooth headset, one kid begs for an unspecified snack, another asks where his homework project has gone, and just then an urgent e-mail from your boss buzzes the phone in your purse. During the call’s last few minutes these urgent requests — snack, homework, boss — wait in your working memory. When you hang up, you’ll pick one and act.

When you do that, according to Brown University psychology researchers whose findings appear in the journal Neuron, you’ll employ brain circuitry that links a specific chunk of the striatum called the caudate and a chunk of the prefrontal cortex centered on the dorsal anterior premotor cortex. Selecting from working memory, it turns out, uses similar circuits to those involved in planning motion.

In lab experiments with 22 adult volunteers, the researchers used magnetic resonance imaging to track brain activity during a carefully designed working memory task. They also measured how quickly the subjects could choose from working memory — a phenomenon the scientists called “output gating.”

“In the immediacy of what we’re doing we have this small working memory capacity where we can hang on to a few things that are going to be useful in a few moments, and that’s where output gating is crucial,” said study senior author David Badre, professor of cognitive, linguistic, and psychological sciences at Brown.

From the perspective of cognition, said lead author and postdoctoral scholar Christopher Chatham, input gating — choosing what goes into working memory — and output gating allow people to maintain a course of action (e.g., finish that Bluetooth call) while being flexible enough to account for context in planning what’s next.

Of cognition and wingdings

In their experiments Badre, Chatham, and co-author Michael Frank, associate professor of cognitive, linguistic, and psychological sciences, provided their volunteers with four different versions of a similar working memory task. The versions distinguished output gating from input gating so that the anatomical action observed in the MRI could reliably associate with output gating behavior.

In each round, volunteers saw a sequence of characters — either letters of the alphabet or wingdings (typographical symbols like stars and snowflakes). Before or after the sequence, the volunteers were also given a context cue in the form of a numeral that told them which kind of character would be relevant at end of the task (e.g., “1” might mean a wingding while “2” might mean a letter). The last step for volunteers was to select between groups of characters on the screen that included whichever contextually relevant character they had seen in the sequence (e.g., if the subject had seen a “1” and later a snowflake during the sequence, they should select the group that included a snowflake).

When the context numeral came first, say a “2,” volunteers would “input gate” only letters into their working memory. When it came time to make a selection, they’d simply “output gate” the correct letter from the letters in working memory. If the context came last, people would have to input gate everything they saw into working memory, making all the real thinking a matter of output gating. If the context cue came last, they would carry a higher load of characters in working memory. To address this disparity, the experimenters created two more conditions in which a global context indicator, “3,” required people to keep everything they saw in working memory whether it came before the sequence or after.

With this experimental design the researchers could measure performance and monitor brain activity with subjects who had distinct moments of input and output gating, regardless of the character load in working memory.

People accomplished the tasks with a range of speeds, which the researchers regarded as a proxy for the amount of cognitive work volunteers had to do. People were slowest in making a selection when they got the context cue last and then had to gate just one specific symbol out of memory (e.g., they saw the sequence, then saw a 1, and then had to choose the option with a wingding they had seen). People were fastest at making a selection when they were given the context first and then had to pick the one character of that kind that they saw (e.g., they saw a “2,” then the sequence in which only letters mattered, and then had to choose the option with a letter they had seen).

In analyzing the results, Chatham and his co-authors found that the caudate and the dorsal anterior premotor cortex, contributed distinctly to the reaction times they saw. These separate roles in the partnership agree with computational models of how the brain works.

“The division of labor that’s specifically posited by these computational models is one in which there is a basically a context being represented in the prefrontal cortex that determines the overall efficiency of going from stimulus to response – like a route,” Chatham said. “The striatum is involved in the actual gating of that flow of information,” he said, “like traffic lights along the route.”

So the cortex interprets the context, while the striatum implements the gating. When the context is unhelpfully general and the gating is very specific, for example, the task takes a lot of time.

The findings help advance studies of how cognition works in the brain and could help psychiatrists analyze behavior in people where those areas of the brain have been injured, the researchers said. It also highlights how similar brain circuits can execute different functions – motion and working memory gating.

Addiction: Can You Ever Really Completely Leave It Behind?

A new study in Biological Psychiatry suggests the answer is no

It is often said that once people develop an addiction, they can never completely eliminate their attraction to the abused substance. New findings provide further support for this notion by suggesting that even long-term abstinence from cocaine does not result in a complete normalization of brain circuitry.

Scientists are currently trying to answer some of the ‘chicken and egg’ questions surrounding the abuse of drugs. In particular, one of those questions is whether individuals who abuse psychostimulants like cocaine are more impulsive and show alterations in brain reward circuits as a consequence of using the drug, or whether such abnormalities existed prior to their drug use. In the former case, one might expect brain alterations to normalize following prolonged drug abstinence.

To address these questions, Krishna Patel at Institute of Living/Hartford Hospital and colleagues compared neural responses between three groups of people who were asked to complete a task that resembles bidding on eBay items. The 3 groups consisted of 47 healthy controls, 42 currently drug-abusing cocaine users, and 35 former cocaine users who had been abstinent an average of 4 years. They also compared all three groups on their levels of impulsivity and reward responding.

They found that active users showed abnormal activation in multiple brain regions involved with reward processing, and that the abstinent individuals who were previously cocaine dependent manifested differences in a subset of those regions. Both current and former cocaine users displayed similarly elevated impulsivity measures compared to healthy controls, which may indicate that these individuals had a pre-existing risk for addiction. Indeed, the degree of impulsivity correlated with several of the brain activation abnormalities.

These findings suggest that prolonged abstinence from cocaine may normalize only a subset of the brain abnormalities associated with active drug use.

"The knowledge that some neural changes associated with addiction persist despite long periods of abstinence is important because it supports clinical wisdom that recovery from addiction is a lifelong process," says Dr. John Krystal, Editor of Biological Psychiatry. "Further, it is the start of a deeper question: How do these persisting changes develop and how can they be reversed?"

The authors agree that further studies will be needed to investigate such questions, including the continued attempt to determine the extent to which differences in former cocaine users reflect aspects of pre-existing features, exposure to cocaine, or recovery.

(Image: Shutterstock)

Brain Wiring Quiets the Voice Inside Your Head

Researchers find nerve circuits connecting motion and hearing

During a normal conversation, your brain is constantly adjusting the volume to soften the sound of your own voice and boost the voices of others in the room. This ability to distinguish between the sounds generated from your own movements and those coming from the outside world is important not only for catching up on water cooler gossip, but also for learning how to speak or play a musical instrument.

Now, researchers have developed the first diagram of the brain circuitry that enables this complex interplay between the motor system and the auditory system to occur.

The research, which appears Sept. 4 in The Journal of Neuroscience, could lend insight into schizophrenia and mood disorders that arise when this circuitry goes awry and individuals hear voices other people do not hear.

"Our finding is important because it provides the blueprint for understanding how the brain communicates with itself, and how that communication can break down to cause disease," said Richard Mooney, Ph.D., senior author of the study and professor of neurobiology at Duke University School of Medicine. "Normally, motor regions would warn auditory regions that they are making a command to speak, so be prepared for a sound. But in psychosis, you can no longer distinguish between the activity in your motor system and somebody else’s, and you think the sounds coming from within your own brain are external."

Researchers have long surmised that the neuronal circuitry conveying movement — to voice an opinion or hit a piano key — also feeds into the wiring that senses sound. But the nature of the nerve cells that provided that input, and how they functionally interacted to help the brain anticipate the impending sound, was not known.

In this study, Mooney used a technology created by Fan Wang, Ph.D., associate professor of cell biology at Duke, to trace all of the inputs into the auditory cortex — the sound-interpreting region of the brain. Though the researchers found that a number of different areas of the brain fed into the auditory cortex, they were most interested in one region called the secondary motor cortex, or M2, because it is responsible for sending motor signals directly into the brain stem and the spinal cord.

"That suggests these neurons are providing a copy of the motor command directly to the auditory system," said David M. Schneider, Ph.D., co-lead author of the study and a postdoctoral fellow in Mooney’s lab. "In other words,they send a signal that says ‘move,’ but they also send a signal to the auditory system saying ‘I am going to move.’"

Having discovered this connection, the researchers then explored what type of influence this interaction was having on auditory processing or hearing. They took slices of brain tissue from mice and specifically manipulated the neurons that led from the M2 region to the auditory cortex. The researchers found that stimulating those neurons actually dampened the activity of the auditory cortex.

"It jibed nicely with our expectations," said Anders Nelson, co-lead author of the study and a graduate student in Mooney’s lab. "It is the brain’s way of muting or suppressing the sounds that come from our own actions."

Finally, the researchers tested this circuitry in live animals, artificially turning on the motor neurons in anesthetized mice and then looking to see how the auditory cortex responded. Mice usually sing to each other through a kind of song called ultrasonic vocalizations, which are too high-pitched for a human to hear. The researchers played back these ultrasonic vocalizations to the mice after they had activated the motor cortex and found that the neurons became much less responsive to the sounds.

"It appears that the functional role that these neurons play on hearing is they make sounds we generate seem quieter," said Mooney. "The question we now want to know is if this is the mechanism that is being used when an animal is actually moving. That is the missing link, and the subject of our ongoing experiments."

Once the researchers have pinned down the basics of the circuitry, they could begin to investigate whether altering this circuitry could induce auditory hallucinations or perhaps even take them away in models of schizophrenia.

Re-learning how to see: researchers find crucial on-off switch in visual development

A new discovery by a University of Maryland-led research team offers hope for treating “lazy eye” and other serious visual problems that are usually permanent unless they are corrected in early childhood.

Amblyopia afflicts about three percent of the population, and is a widespread cause of vision loss in children. It occurs when both eyes are structurally normal, but mismatched – either misaligned, or differently focused, or unequally receptive to visual stimuli because of an obstruction such as a cataract in one eye.

During the so-called “critical period” when a young child’s brain is adapting very quickly to new experiences, the brain builds a powerful neural network connecting the stronger eye to the visual cortex. But the weaker eye gets less stimulation and develops fewer synapses, or points of connection between neurons. Over time the brain learns to ignore the weaker eye. Mild forms of amblyopia such as “lazy eye” result in problems with depth perception. In the most severe form, deprivation amblyopia, a cataract blocks light and starves the eye of visual experiences, significantly altering synaptic development and seriously impairing vision.

Because brain plasticity declines rapidly with age, early diagnosis and treatment of amblyopia is vital, said neuroscientist Elizabeth M. Quinlan, an associate professor of biology at UMD. If the underlying cause of amblyopia is resolved early enough, the child’s vision can recover to normal levels. But if the treatment comes after the end of the critical period and the loss of synaptic plasticity, the brain cannot relearn to see with the weaker eye.

“If a child is born with a cataract and it is not removed very early in life, very little can be done to improve vision,” Quinlan said. “The severe amblyopia that results is the most difficult to treat. For that reason, science has the most to gain by a better understanding of the underlying mechanisms.”

Quinlan, who specializes in studying how communication through the brain’s circuits changes over the course of a lifetime, wanted to find out what process controls the timing of the critical period of synaptic plasticity. If researchers could find the neurological on-off switch for the critical period, she reasoned, clinicians could use the information to successfully treat older children and adults.

Researchers in Quinlan’s University of Maryland lab teamed up with the laboratory of Alfredo Kirkwood at Johns Hopkins University to address two questions: What are the age boundaries of the critical period for synaptic plasticity, when it comes to determining eye dominance? And what developmental processes are involved?

Experiments in rodents suggested the timing of the critical period is controlled by a specific class of inhibitory neurons, which come into play after a visual stimulus activates excitatory neurons that link the eye to the visual cortex. The inhibitory neurons act as signal controllers, affecting the interactions between excitatory neurons and synapses.

“The generally accepted view has been that as the inhibitory neurons develop, synaptic plasticity declines, which was thought to occur at about five weeks of age in rodents,” roughly equivalent to five years of age in humans, Quinlan said. But in earlier experiments, Quinlan and Kirkwood found no correlation between the development of these inhibitory neurons and the loss of plasticity. In fact, they found the visual circuitry in rodents was highly adaptable at ages beyond five weeks.

In their latest research the UMD-led team looked “one synapse upstream from these inhibitory neurons,” Quinlan said, studying the control of that synapse by a protein called NARP (Neuronal Activity-Regulated Pentraxin). Working with two sets of mice – one group genetically similar to wild mice and another that lacked the NARP gene - the researchers covered one eye in each animal to simulate conditions that produce amblyopia.

The mice that were genetically similar to wild mice developed amblyopia, with characteristic dominance of the normal eye over the deprived eye. But the mice that lacked NARP did not develop amblyopia, regardless of age or the length of time one eye was deprived of stimulation.

The study, published in the current issue of the peer-reviewed journal Neuron, demonstrated that only one specific class of synapses was affected by the absence of NARP. Without NARP, the mice simply had no critical period in which the brain circuitry was weakened in response to the impaired blocking vision in one eye, Quinlan said. Except for the lack of this plasticity, their vision was normal.

“It’s remarkable how specific the deficit is,” Quinlan said. Without the NARP protein, “these animals develop normal vision. Their brain circuitry just isn’t plastic. We can completely turn off the critical period for plasticity by knocking out this protein.”

Since there are indications that NARP levels vary with age, the discovery raises hope that a treatment targeting NARP levels in humans could allow correction of amblyopia late in life, without affecting other aspects of vision.