Study Suggests Disruptive Effects of Anesthesia on Brain Cell Connections Are Temporary

A study of juvenile rat brain cells suggests that the effects of a commonly used anesthetic drug on the connections between brain cells are temporary.

The study, published in this week’s issue of the journal PLOS ONE, was conducted by biologists at the University of California, San Diego and Weill Cornell Medical College in New York in response to concerns, arising from multiple studies on humans over the past decade, that exposing children to general anesthetics may increase their susceptibility to long-term cognitive and behavioral deficits, such as learning disabilities.

An estimated six million children, including 1.5 million infants, undergo surgery in the United States requiring general anesthesia each year and a least two large-scale clinical studies are now underway to determine the potential risks to children and adults.

“Since these procedures are unavoidable in most cases, it’s important to understand the mechanisms associated with the potentially toxic effects of anesthetics on the developing brain, and on the adult brain as well,” said Shelley Halpain, a professor of biology at UC San Diego and the Sanford Consortium for Regenerative Medicine, who co-headed the investigation. “Because the clinical studies haven’t been completed, preclinical studies, such as ours, are needed to define the effects of various anesthetics on brain structure and function.”

“There is concern now about cognitive dysfunction from surgery and anesthesia—how much these effects are either permanent or slowly reversible is very controversial,” said Hugh Hemmings, Jr., chair of anesthesiology at Weill Cornell and the study’s other senior author. “It has been suggested recently that some of the effects of anesthesia may be more lasting than previously thought. It is not clear whether the residual effects after an operation are due to the surgery itself, or the hospitalization and attendant trauma, medications and stress—or a combination of these issues.”

However, he added, “There is evidence that some of the delayed or persistent cognitive effects after surgery are not primarily due to anesthesia itself, but more importantly to brain inflammation resulting from the surgery. But this is not yet clear.”

The team of biologists examined one of the most commonly used general anesthetics, a derivative of ether called “isoflurane” used to maintain anesthesia during surgery.

“Previous studies in cultured neurons and in the intact brains of rodents provided evidence suggesting that exposure to anesthetics might render neurons more susceptible to cell death through a process called ‘apoptosis’,” said Halpain. “While overt cell death could certainly be one way to explain any long-lasting neurocognitive consequences of general anesthesia, we hypothesized that there could be other cellular mechanisms that disrupt neural circuits without inducing cell death per se.”

One such mechanism, she added, is known as “synaptotoxicity.” In this mechanism of neural-circuit disruption, the “synapses,” or junctions between neurons, become weakened or shrink away due to some factor that injures the neurons locally along their axons (the long processes of neurons that transmit signals) and dendrites (the threadlike extensions of neurons that receive nerve signals) without inducing the neurons themselves to die.

In the experiments at UC San Diego headed by Jimcy Platholi, a postdoctoral researcher in Halpain’s lab who is now at Weill Cornell, the scientists used neurons from embryonic rats taken from the hippocampus, a part of the mammalian forebrain essential for encoding newly acquired memories and ensuring that short-term memories are converted into long-term memories. The researchers cultured these brain cells in a laboratory dish for three weeks, allowing the neurons time to mature and to develop a dense network of synaptic connections and “dendritic spines”—specialized structures that protrude from the dendrites and are essential mediators of activity throughout neural networks.

“Evidence from animal studies indicates that new dendritic spines emerge and existing spines expand in size during learning and memory,” explained Halpain. “Therefore, the overall numbers and size of dendritic spines can profoundly impact the strength of neural networks. Since neural network activity underlies all brain function, changes in dendritic spine number and shape can influence cognition and behavior.”

Using neurons in culture, rather than intact animal brains, allowed the biologists to take images of the synapses at high spatial resolution using techniques called fluorescence light microscopy and confocal imaging. They also used time-lapse microscopy to observe structural changes in individual dendritic spines during exposure to isoflurane. Karl Herold, a research associate in the Hemmings laboratory and a co-author of the study, performed some of the image analysis.

“Imaging of human brain synapses at this level of detail is impossible with today’s technology and it remains very challenging even in laboratory rodents,” said Halpain. “It was important that we performed our study using rodent neurons in a culture dish, so that we could really drill down into the subcellular and molecular details of how anesthetics work.”

The researchers wondered whether brief exposure to isoflurane would alter the numbers and size of dendritic spines, so they applied the anesthetic to the cultured rat cells at concentrations and durations (up to 60 minutes) that are frequently used during surgery.

“We observed detectable decreases in dendritic spine numbers and shape within as little as 10 minutes,” said Halpain. “However this spine loss and shrinkage was reversible after the anesthetic was washed out of the culture.”

“Our study was reassuring in the sense that the effects are not irreversible and this fits in with known clinical effects,” said Hemmings. “For the most part, we find that the effects are reversible.”

“We clearly see an effect—a very marked effect on the dendritic spines—from use of this drug that was reversible, suggesting that it is not a toxic effect, but something more relevant to the pharmacological actions of the drug,” he added. “Connecting what we found to the cognitive effects of isoflurane will require much more detailed analysis.”

The team plans to follow up its study with future experiments to probe the molecular mechanisms and long-lasting consequences of isoflurane’s effects on neuron synapses and examine other commonly-used anesthetics for surgery.

A Trace of Memory: Researchers Watch Neurons in the Brain During Learning and Memory Recall

A team of neurobiologists led by Simon Rumpel at the Research Institute of Molecular Pathology (IMP) in Vienna succeeded in tracking single neurons in the brain of mice over extended periods of time. Advanced imaging techniques allowed them to establish the processes during memory formation and recall. The results of their observations are published this week in PNAS Early Edition.

Most of our behavior – and thus our personality – is shaped by previous experience. To store the memory of these experiences and to be able to retrieve the information at will is therefore considered one of the most basic and important functions of the brain. The current model in neuroscience poses that memory is stored as long-lasting anatomical changes in synapses, the specialized structures by which nerve cells connect and signal to each other.

At the Research Institute of Molecular Pathology (IMP) in Vienna, Simon Rumpel and Kaja Moczulska used mice to study the effects of learning and memorizing on the architecture of synapses. They employed an advanced microscopic technique called in vivo two-photon imaging that allows the analysis of structures as small as a thousandth of a millimetre in the living brain.

Using this technology, the neurobiologists tracked individual neurons over the course of several weeks and analysed them repeatedly. They focussed their attention on dendritic spines that decorate the neuronal processes and correspond to excitatory synapses. The analyses were combined with behavioral experiments in which the animals underwent classic auditory conditioning. The results showed that the learning experience triggered the formation of new synaptic connections in the auditory cortex. Several of these new structures persisted over time, suggesting a long-lasting trace of memory and confirming an important prediction of the current model.

Apart from the changes during memory formation, the IMP-scientists were interested in the act of remembering. Earlier studies had shown that memory recall is associated with molecular processes similar to the initial formation of memory. These similarities have been suggested to reflect remodelling of memory traces during recall.

To test this hypothesis, previously trained mice were exposed to the auditory cue a week after conditioning while tracking dendritic spines in the auditory cortex. The results showed that although some molecular processes indeed resembled those during memory formation, the anatomical structure of the synapses did not change. These findings suggest that memory retrieval does not lead to a modification of the memory trace per se. Instead, the molecular processes triggered by memory formation and recall could reflect the stabilization of previously altered or recently retrieved synaptic connections.

The primary goal of elucidating the processes during memory formation and recall is to increase our basic knowledge. Insights gained from these studies might however help us to understand diseases of the nervous system that affect memory. They may also, in the future, provide the basis for treatments that offer relief to traumatized patients.

What Color is Your Night Light? It May Affect Your Mood

Study Finds Red Light Least Harmful, While Blue Light is Worst

When it comes to some of the health hazards of light at night, a new study suggests that the color of the light can make a big difference.

In a study involving hamsters, researchers found that blue light had the worst effects on mood-related measures, followed closely by white light.

But hamsters exposed to red light at night had significantly less evidence of depressive-like symptoms and changes in the brain linked to depression, compared to those that experienced blue or white light.

The only hamsters that fared better than those exposed to red light were those that had total darkness at night.

The findings may have important implications for humans, particularly those whose work on night shifts makes them susceptible to mood disorders, said Randy Nelson, co-author of the study and professor of neuroscience and psychology at The Ohio State University.

“Our findings suggest that if we could use red light when appropriate for night-shift workers, it may not have some of the negative effects on their health that white light does,” Nelson said.

The study appears in the Aug. 7, 2013, issue of The Journal of Neuroscience.

The research examined the role of specialized photosensitive cells in the retina — called ipRGCs — that don’t have a major role in vision, but detect light and send messages to a part of the brain that helps regulate the body’s circadian clock. This is the body’s master clock that helps determine when people feel sleepy and awake.

Other research suggests these light-sensitive cells also send messages to parts of the brain that play a role in mood and emotion.

“Light at night may result in parts of the brain regulating mood receiving signals during times of the day when they shouldn’t,” said co-author Tracy Bedrosian, a former graduate student at Ohio State who is now a postdoctoral researcher at the Salk Institute. “This may be why light at night seems to be linked to depression in some people.”

What people experience as different colors of light are actually lights of different wavelengths. The ipRGCs don’t appear to react to light of different wavelengths in the same way.

“These cells are most sensitive to blue wavelengths and least sensitive to red wavelengths,” Nelson said. “We wanted to see how exposure to these different color wavelengths affected the hamsters.”

In one experiment, the researchers exposed adult female Siberian hamsters to four weeks each of nighttime conditions with no light, dim red light, dim white light (similar to that found in normal light bulbs) or dim blue light.

They then did several tests with the hamsters that are used to check for depressive-like symptoms. For example, if the hamsters drink less-than-normal amounts of sugar water — a treat they normally enjoy — that is seen as evidence of a mood problem.

Results showed that hamsters that were kept in the dark at night drank the most sugar water, followed closely by those exposed to red light. Those that lived with dim white or blue light at night drank significantly less of the sugar water than the others.

After the testing, the researchers then examined the hippocampus regions of the brains of the hamsters.

Hamsters that spent the night in dim blue or white light had a significantly reduced density of dendritic spines compared to those that lived in total darkness or that were exposed to only red light. Dendritic spines are hairlike growths on brain cells that are used to send chemical messages from one cell to another. 

A lowered density of these dendritic spines has been linked to depression, Nelson said.

“The behavior tests and changes in brain structure in hamsters both suggest that the color of lights may play a key role in mood,” he said.

“In nearly every measure we had, hamsters exposed to blue light were the worst off, followed by those exposed to white light,” he said. “While total darkness was best, red light was not nearly as bad as the other wavelengths we studied.”

Nelson and Bedrosian said they believe these results may be applicable to humans.

In addition to shift workers, others may benefit from limiting their light at night from computers, televisions and other electronic devices, they said. And, if light is needed, the color may matter.

“If you need a night light in the bathroom or bedroom, it may be better to have one that gives off red light rather than white light,” Bedrosian said.