Why bright light keeps us awake

October 15, 2019

Science Daily/Salk Institute

Researchers are reporting a novel technique for tracing the activity of individual nerve fibers known as axons, and determining how neurons communicate. The team used this technique to uncover details about how the brain responds to light signals received by the retina in mice.

In recent decades, scientists have learned a great deal about how different neurons connect and send signals to each other. But it's been difficult to trace the activity of individual nerve fibers known as axons, some of which can extend from the tip of the toe to the head. Understanding these connections is important for figuring out how the brain receives and responds to signals from other parts of the body.

Researchers at the Salk Institute and UC San Diego are reporting a novel technique for tracing these connections and determining how neurons communicate. The team used this technique to uncover details about how the brain responds to light signals received by the retina in mice, published October 15, 2019, in Cell Reports.

"This study is a breakthrough because no one could figure out how to study these connections before," says Salk Professor Satchidananda Panda, co-corresponding author of the paper. "This new technique has enabled us to go well beyond the limitations of electron microscopy."

The new method makes use of several different laboratory techniques to understand a type of neuron called intrinsically photosensitive retinal ganglion cells (ipRGCs). These cells, which are found in the retina, in the back of the eye, express a protein called melanopsin that senses blue light.

The Salk and UCSD teams used a virus to deliver a protein called a mini-singlet oxygen-generating protein (mini-SOG) to the ipRGCs, so that the cells could be viewed in more detail under election microscopy. The system was designed to tether the mini-SOG to the membranes of the light-sensitive cells so that the entire neuron, including its long axons that reach out to different parts of the brain, can be easily tracked under both light and electron microscope.

"Thanks to development and application of new genetically introduced probes for correlated multiscale light and electron microscopic imaging, our Salk and UCSD-based research teams were able to follow the small processes emanating from nerve cells over centimeters, all the way from the retina to multiple places where they connect to brain regions critical to circadian rhythms, eye reflexes and vision," says Mark Ellisman, distinguished professor of neurosciences at UC San Diego and adjunct professor at Salk, who co-led the work. "We were able to obtain unprecedented three-dimensional information about the machinery required for these neuronal cells to signal the next neurons in the complex circuits."

Most of the previous work with mini-SOGs has been done in cell lines, and using them in mice, to map how neurons from the retina wire the brain, was a first, according the researchers. The method enabled them to glean new information about the connections between ipRGCs and different parts of the brain.

The ipRGCs are known to connect to many brain regions that regulate very different tasks. The cells tell one part of the brain how bright it is outside so that our pupil can rapidly close -- in less than a second. The same ipRGCs also connect to the master clock in the brain that regulates our sleep-wake cycle. "However, it takes several minutes of bright light to make us fully awake," Panda says. "How the same ipRGCs do these very different tasks with different time scales was not clear until now."

The investigators found that the difference has to do with the way that light detected by the retina reaches the brain. By delivering the mini-SOG to the eyes of the mice, they were able to trace the signal to the part of the brain that constricts the pupil in response to light.

"These connections were much stronger -- similar to water pouring out of a garden hose," Panda says. "Whereas the connection between the ipRGCs and the master clocks were weaker -- more like drip irrigation." Because the ipRGCs deliver the light signal to the circadian center through this slower drip system, it takes longer for any meaningful information to reach and reset the brain clock.

"This research helps explain why, when you get up in the night to get a drink of water and turn on the light for a few seconds, you're usually able to go right back to sleep," Panda says. "But if you hear a noise outside and end up walking around your house for half an hour with the lights on, it's much harder. There will be enough light signal reaching the master clock neurons in the brain that ultimately wakes up the rest of the brain."

Panda says that the new technique will be useful for studying other neural connections, as the researchers can essentially use the same viruses to express mini-SOGs in any neuron and ask how different neurons make connections to different appendages.

"These findings and methods open new opportunities for brain researchers studying the long-distance wiring of brains in normal and in animal models of human disease," adds Ellisman.

https://www.sciencedaily.com/releases/2019/10/191015131508.htm

Moving light-dark exposure could reduce disruption faced by night shift workers

March 27, 2018

Science Daily/The Physiological Society

New research shows that our brain clock can be shifted by light exposure, potentially to align it with night shift patterns. It highlights that a 'one size fits all' approach to managing sleep disruption in shift workers may not be appropriate. A personalized approach, with light-dark exposure scheduled and taking into account whether someone is a 'morning' or 'evening' person, could reduce shift workers' risk of health problems.

Our sleep-wake cycle, in part controlled by our brain clock, encompasses physical, mental and behavioural changes that follow a daily cycle. Light is the dominant environmental time cue which results in, for example, sleeping at night and being awake during the day.

Night time shift work disrupts the normal sleep-wake cycle and our internal circadian (24-hour) rhythms, and has been associated with significant health problems, such as a higher risk of heart disease and cancer. Alertness levels are often markedly impaired while working night shifts.

While it has been known that there are considerable differences in how the brain clock of different individuals responds to changing shift cycles, we have known very little about the mechanisms that underlie these differences between people. If someone was able to realign their brain clock to their shift pattern, then it would improve sleep and could lead to health benefits. While such realignment is rare, in some circumstances such as on offshore oil rig platforms, complete adaption has been observed.

This new research aims to understand the relationship between light exposure and how an individual's circadian rhythm is affected across a transition from day to night shift schedules. The researchers found that timing of light exposure is the primary factor in determining how the brain clock responds to night shift work, accounting for 71% of the variability in timing of the clock observed in the study. It also found that the extent to which an individual is a 'morning' or 'evening' type affects how the body responds, which shows that a personalised approach is important.

This study was led by the CRC for Alertness, Safety and Productivity and saw nursing and medical staff recruited from an Intensive Care Unit at a major hospital in Melbourne, Australia. Staff members were enrolled into the study when working a schedule of day or evening shifts, or days off, followed by at least 3 or 4 consecutive night shifts.

To examine how the sleep-wake cycle responds to the shift schedule, the timing of the brain clock was measured on the day schedule, and at the end of the night shifts. It was measured by monitoring urinary concentration of the major metabolite of melatonin, which is a hormone produced in the pineal gland known to be involved in the regulation of sleep cycles. Individual light exposure was measured using wrist actigraphs, worn for the duration of the study.

Prof Shantha Rajaratnam, from Monash University and the CRC for Alertness, Safety and Productivity, corresponding author for the study, said:

"We know that night time shift workers are more likely to suffer health problems due to disruption of their circadian clock, and the mismatch between the timing of the clock and their sleep-wake cycle. This research is important because if we can realign a person's clock to fit their shift pattern, then they will sleep better and this may result in improved health, safety and productivity.

"These results will drive development of personalised approaches to improve sleep-wake cycles of shift workers and other vulnerable people, and could potentially reduce the increased risk of disease due to circadian disruption."

https://www.sciencedaily.com/releases/2018/03/180327203014.htm