Hologram-like 3-D brain helps researchers decode migraine pain

Wielding a joystick and wearing special glasses, pain researcher Alexandre DaSilva rotates and slices apart a large, colorful, 3-D brain floating in space before him.

Despite the white lab coat, it appears DaSilva’s playing the world’s most advanced virtual video game. The University of Michigan dentistry professor is actually hoping to better understand how our brains make their own pain-killing chemicals during a migraine attack.

The 3-D brain is a novel way to examine data from images taken during a patient’s actual migraine attack, says DaSilva, who heads the Headache and Orofacial Pain Effort at the U-M School of Dentistry and the Molecular and Behavioral Neuroscience Institute.

Different colors in the 3-D brain give clues about chemical processes happening during a patient’s migraine attack using a PET scan, or positron emission tomography, a type of medical imaging.

"This high level of immersion (in 3-D) effectively places our investigators inside the actual patient’s brain image," DaSilva said.

The 3-D research occurs in the U-M 3-D Lab, part of the U-M Library.

First steps of synapse building captured in live zebra fish embryos

Using spinning disk microscopy on barely day-old zebra fish embryos, University of Oregon scientists have gained a new window on how synapse-building components move to worksites in the central nervous system.

What researchers captured in these see-through embryos — in what may be one of the first views of early glutamate-driven synapse formation in a living vertebrate — were orderly movements of protein-carrying packets along axons to a specific site where a synapse would be formed.

Washbourne addresses:

► The basic importance of the findings

► The connection to diseases, including autism

The discovery, in research funded by the National Institutes of Health, is described in a paper placed online ahead of publication in the April 25 issue of the open-access journal Cell Reports. It is noteworthy because most synapses formed in vertebrates use glutamate as a neurotransmitter, and breakdowns in the process have been tied to conditions such as autism, schizophrenia and mental retardation.

The zebra fish has become one of the leading research models for studying early development, in general, and human-disease states.

In this case, researchers used immunofluorescence labeling to highlight the area they put under the microscopes. The embryos they studied were barely 24-hours old and a millimeter in length, but neurons in their spinal cord were already forming connections called synapses. Images were taken every 30 seconds over two hours.

"If we zoom out a bit and look at development in the human, the majority of synapse formation occurs in the cortex after birth and continues for the first two years in a baby’s life," said Philip Washbourne, a professor of biology and member of the UO’s Institute of Neuroscience.

Previous studies, done in vitro, contradicted each other, with one, in 2000, identifying a single packet of building blocks arriving at a pre-synaptic terminal. The other, in 2004, identified two protein packets. After watching the process unfold live, with imaging over long time spans, Washbourne said: “We now see at least three, and maybe more, such deliveries.”

"Axons are long processes — think of them as highways — of neurons. In humans, these can be a meter long, from spinal cord to your big toe," he said. It’s in the cell body where all the proteins are made, and they have to be transported out. Is it done by a single bus or by several cars? These results point to additional layers of complexity in the established mechanisms of synaptogenesis."

The new research also showed that sequence also is crucial. Two different pre-synaptic packages of molecules repeatedly arrived in the same order. A key building block — the protein synapsin — always arrived third. As these delivery vehicles traveled the axonal highway, another protein, a cyclin-dependent kinase known as Cdk5, acts as a stoplight at the synapse-construction site, where phosphorylation occurs. More research is needed on Cdk5, Washbourne said.

"Understanding how all this happens will inform us to what’s going wrong in neurodevelopment that leads to diseases," Washbourne said. "We have indications that the glue that gets all this going includes a gene that has been linked to autism, so knowing how these molecules start the process of synapse formation — and what goes wrong in people with mutations in these genes — might allow for a therapeutic targeting to correct the mutations and manipulate the stop signs."

Neural Activity in Bats Measured In Flight

Animals navigate and orient themselves to survive – to find food and shelter or avoid predators, for example. Research conducted by Dr. Nachum Ulanovsky and research student Michael Yartsev of the Weizmann Institute’s Neurobiology Department, published today in Science, reveals for the first time how three-dimensional, volumetric, space is perceived in mammalian brains. The research was conducted using a unique, miniaturized neural-telemetry system developed especially for this task, which enabled the measurement of single brain cells during flight.

The question of how animals orient themselves in space has been extensively studied, but until now experiments were only conducted in two-dimensional settings. These have found, for instance, that orientation relies on “place cells” – neurons located in the hippocampus, a part of the brain involved in memory, especially spatial memory. Each place cell is responsible for a spatial area, and it sends an electrical signal when the animal is located in that area. Together, the place cells produce full representations of whole spatial environments. Unlike the laboratory experiments, however, the navigation of many animals in the real world, including humans, is carried out in three dimensions. But attempts to expand the scope of experiments from two to three dimensions had encountered difficulties.

One of the more famous efforts in this area was conducted by the University of Arizona and NASA, in which they launched rats into space (aboard a space shuttle). However, although the rats moved around in zero gravity, they ran along a set of straight, one-dimensional lines. Other experiments with three-dimensional projections onto two-dimensional surfaces did not manage to produce volumetric data, either. The conclusion was that in order to understand movement in three-dimensional, volumetric space, it is necessary to allow animals to move through all three dimensions – that is, to research animals in flight.

Ulanovsky chose to study the Egyptian fruit bat, a very common bat species in Israel. Because these are relatively large, the researchers were able to attach the wireless measuring system in a manner that did not restrict the bats’ movements. Developing this sophisticated measuring system was a several-year effort. Ulanovsky, in cooperation with a US commercial company, created a wireless, lightweight (12 g, about 7% of the weight of the bat) device containing electrodes that measure the activity of individual neurons in the bat’s brain.

The next challenge the scientists faced was adapting the behavior of their bats to the needs of the experiment. Bats naturally fly toward their destination – for example, a fruit tree – in a straight line. In other words, their normal flight patterns are one-dimensional, while the experiment required their flights to fill a three-dimensional space.

The solution was to be found in a previous study in Ulanovsky’s group, which tracked wild fruit bats using miniature GPS devices. One of the discoveries was that when bats arrive at a fruit tree, they fly around it, utilizing the full volume of space surrounding the tree. To simulate this behavior in the laboratory – an artificial cave equipped with an array of bat-monitoring devices – the team installed an artificial “tree” made of metal bars and cups filled with fruit.

Measuring the activity of hippocampus neurons in the bats’ brains revealed that the representation of three-dimensional space is similar to that in two dimensions: Each place cell is responsible for identifying a particular spatial area in the “cave” and sends an electrical signal when the bat is located in that area. Together, the population of place cells provides full coverage of the cave – left and right, up and down.

A closer examination of the areas for which individual place cells are responsible provided an answer to a highly-debated question: Does the brain perceive the three dimensions of space as “equal,” that is, does it sense the height axis in the same way as that of length or width? The findings suggest that each place cell responds to a spherical volume of space, i.e., the perception of all three dimensions is uniform. The researchers note that for those non-flying animals that essentially move in flat space, the different axes might not be perceived at the same resolution. It may be that such animals are naturally more sensitive to changes along the length and width axes than that of height. This question is of particular interest when it comes to humans because on the one hand, humans evolved from apes that moved in three-dimensional space when swinging from branch to branch, but on the other hand, modern, ground-dwelling humans generally navigate in two-dimensional space.

The findings provide new insights into some basic functions of the brain: navigation, spatial memory and spatial perception. To a large extent, this is due to the development of innovative technology that allowed the first glimpse into the brain of a flying animal. Ulanovsky believes that this trend, in which research is becoming more “natural,” is the future wave of neuroscience.

Bat and Rat Brain Rhythms Differ When on the Move

To get a clear picture of how humans and other mammals form memories and find their way through their surroundings, neuroscientists must pay more attention to a broad range of animals rather than focus on a single model species, say two University of Maryland researchers, Katrina MacLeod and Cynthia Moss. Their new comparative study of bats and rats reports differences between the species that suggest the need to revise models of spatial navigation.

In a paper appearing in the April 19, 2013 issue of Science, the UMD researchers and two colleagues at Boston University reported significant differences between rats’ and bats’ brain rhythms when certain cells were active in a part of the brain used in memory and navigation.

These cells behaved as expected in rats, which mostly move along surfaces. But in bats, which fly, the continuous brain rhythm did not appear, said Moss, a professor in Psychology and Biology and the Institute for Systems Research.

The finding suggests that even though rats, bats, humans and other mammals share a common neural representation of space in a part of the brain that has been linked to spatial information and memory, they may have different cellular mechanisms to create or interpret those maps, said MacLeod, an assistant research scientist in Biology.

“To understand brains, including ours, we really must study neural activity in a variety of animals,” MacLeod said. “Common features across multiple species tell us ‘Aha, this is important,’ but differences can occur because of variances in the animals’ ecology, behavior, or evolutionary history.”

The research team focused on a brain region that contains specialized “grid cells,” so named because they form a hexagonal grid of activity related to the animal’s location as it navigates through space. This brain region, the medial entorhinal cortex, sits next to the hippocampus, the place that, in humans, forms memories of events such as where a car is parked. The medial entorhinal cortex acts as a hub of neural networks for memory and navigation.

Grid cells were first noticed in rats navigating their environment, but recent work by Nachum Ulanovsky (Moss’s former postdoctoral researcher at UMD) and his research team at the Weizmann Institute in Rehovot, Israel, has shown these cells exist in bats as well.

In rats, grid cells fire in a pattern called a theta wave when the animals spatially navigate. Theta waves are fairly low-frequency electrical oscillations that also have been observed at the cellular level in the medial entorhinal cortex. The prominence of theta waves in rats suggested they were important. As a result, neuroscientists, trying to understand the relationship between theta waves and grid cells, have developed models of the brain based on the assumption that theta waves are key to spatial navigation in mammals.

However, Moss said, “recordings from the brains of bats navigating in space contain a surprise, because the expected theta rhythms aren’t continuously present as they are in the rodent.”

The new Science study doubles down on the lack of theta in bats by reporting that theta rhythms also are not present at the cellular level. “The bat neurons don’t ‘ring’ the way the rat neurons do,” says MacLeod. “This raises a lots of questions as to whether theta rhythms are actually doing what the spatial navigation theory proposes in rats or even humans.”

Scientists probe the source of a pulsing signal in the sleeping brain

New findings clarify where and how the brain’s “slow waves” originate. These rhythmic signal pulses, which sweep through the brain during deep sleep at the rate of about one cycle per second, are assumed to play a role in processes such as consolidation of memory. For the first time, researchers have shown conclusively that slow waves start in the cerebral cortex, the part of the brain responsible for cognitive functions. They also found that such a wave can be set in motion by a tiny cluster of neurons.

"The brain is a rhythm machine, producing all kinds of rhythms all the time," says Prof. Arthur Konnerth of the Technische Universitaet Muenchen (TUM). "These are clocks that help to keep many parts of the brain on the same page." One such timekeeper produces the so-called slow waves of deep sleep, which are thought to be involved in transmuting fragments of a day’s experience and learning into lasting memory. They can be observed in very early stages of development, and they may be disrupted in diseases such as Alzheimer’s.

Previous studies, relying mainly on electrical measurements, have lacked the spatial resolution to map the initiation and propagation of slow waves precisely. But using light, Konnerth’s Munich-based team – in collaboration with researchers at Stanford and the University of Mainz – could both stimulate slow waves and observe them in unprecedented detail. One key result confirmed that the slow waves originate only in the cortex, ruling out other long-standing hypotheses. “The second major finding,” Konnerth says, “was that out of the billions of cells in the brain, it takes not more than a local cluster of fifty to one hundred neurons in a deep layer of the cortex, called layer 5, to make a wave that extends over the entire brain.”

New light on a fundamental neural mechanism

Despite considerable investigation of the brain’s slow waves, definitive answers about the underlying circuit mechanism have remained elusive. Where is the pacemaker for this rhythm? Where do the waves start, and where do they stop? This study – based on optical probing of intact brains of live mice under anesthesia – now provides the basis for a detailed, comprehensive view.

"We implemented an optogenetic approach combined with optical detection of neuronal activity to explore causal features of these slow oscillations, or Up-Down state transitions, that represent the dominating network rhythm in sleep," explains Prof. Albrecht Stroh of the Johannes Gutenberg University Mainz. Optogenetics is a novel technique that enabled the researchers to insert light-sensitive channels into specific kinds of neurons, to make them responsive to light stimulation. This allowed for selective and spatially defined stimulation of small numbers of cortical and thalamic neurons.

Access to the brain via optical fibers allowed for both microscopic recording and direct stimulation of neurons. Flashes of light near the mouse’s eyes were also used to stimulate neurons in the visual cortex. By recording the flux of calcium ions, a chemical signal that can serve as a more spatially precise readout of the electric activity, the researchers made the slow waves visible. They also correlated optical recordings with more conventional electrical measurements. As a result, it was possible to watch individual wave fronts spread – like ripples from a rock thrown into a quiet lake – first through the cortex and then through other brain structures.

A new picture begins to emerge: Not only is it possible for a tiny local cluster of neurons to initiate a slow wave that will spread far and wide, recruiting multiple regions of the brain into a single event – this appears to be typical. “In spontaneous conditions,” Konnerth says, “as it happens with you and me and everyone else every night in deep sleep, every part of the cortex can be an initiation site.” Furthermore, a surprisingly simple communication protocol can be seen in the slow wave rhythm. During each one-second cycle a single neuron cluster sends its signal and all others are silenced, as if they are taking turns bathing the brain in fragments of experience or learning, building blocks of memory. The researchers view these findings as a step toward a better understanding of learning and memory formation, a topic Konnerth’s group is investigating with funding from the European Research Council. They also are testing how the slow waves behave during disease.