Detailed image of the human retina

What cell types are found in which human tissue, and where? Which genes are active in the individual cells, and which proteins are found there? Answers to these questions and more are to be provided by a specialised atlas -- in particular how the different tissues form during embryonic development and what causes diseases. In creating this atlas, researchers aim to map not only tissue directly isolated from humans, but also structures called organoids. These are three-dimensional clumps of tissue that are cultivated in the laboratory and develop in a way similar to human organs, but on a small scale.

"The advantage of organoids is that we can intervene in their development and test active substances on them, which allows us to learn more about healthy tissue as well as diseases," explains Barbara Treutlein, Professor of Quantitative Developmental Biology at the Department of Biosystems Science and Engineering at ETH Zurich in Basel.

To help produce such an atlas, Treutlein, together with researchers from the Universities of Zurich and Basel, has now developed an approach to gather and compile a great deal of information about organoids and their development. The research team applied this approach to the organoids of the human retina, which they derived from stem cells.

Many proteins visible simultaneously

At the heart of the methods the scientists used for their approach was the 4i technology: iterative indirect immunofluorescence imaging. This new imaging technique can visualise several dozen proteins in a thin tissue section at high resolution using fluorescence microscopy. The 4i technology was developed a few years ago by Lucas Pelkmans, a professor at the University of Zurich and coauthor of the study that has just been published in the scientific journal Nature Biotechnology. It is in this study that the researchers applied this method to organoids for the first time.

Typically, researchers use fluorescence microscopy to highlight three proteins in a tissue, each with a different fluorescent dye. For technical reasons, it is not possible to stain more than five proteins at a time. In 4i technology, three dyes are used, but these are washed from the tissue sample after measurements have been taken, and three new proteins are stained. This step was performed 18 times, by a robot, and the process took a total of 18 days. Lastly, a computer merges the individual images into a single microscopy image on which 53 different proteins are visible. They provide information on the function of the individual cell types that make up the retina; for example, rods, cones, and ganglion cells.

The researchers have supplemented this visual information of retinal proteins with information on which genes are read in the individual cells.

High spatial and temporal resolution

The scientists performed all these analyses on organoids that were of different ages and thus at different stages of development. In this way, they were able to create a time series of images and genetic information that describes the entire 39-week development of retinal organoids. "We can use this time series to show how the organoid tissue slowly builds up, where which cell types proliferate and when, and where the synapses are located. The processes are comparable to those of retinal formation during embryonic development," says Gray Camp, a professor at the University of Basel and a senior author of this study.

The researchers published their image information and more findings on retinal development on a publicly accessible website: EyeSee4is.

Further tissue types planned

So far, the scientists have been studying how a healthy retina develops, but in the future, they hope to deliberately disrupt development in retinal organoids with drugs or genetic modifications. "This will give us new insights into diseases such as retinitis pigmentosa, a hereditary condition that causes the retina's light-sensitive receptors to gradually degenerate and ultimately leads to blindness," Camp says. The researchers want to find out when this process begins and how it can be stopped.

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Eye color genes are critical for retinal health

Metabolic pathways consist of a series of biochemical reactions in cells that convert a starting component into other products. There is growing evidence that metabolic pathways coupled with external stress factors influence the health of cells and tissues. Many human diseases, including retinal or neurodegenerative diseases, are associated with imbalances in metabolic pathways.

Elisabeth Knust leads a team of researchers from the Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG) in Dresden, Germany, who describe an essential role for one such metabolic pathway in maintaining retinal health under conditions of stress. They studied the classic Drosophila genes cinnabar, cardinal, white, and scarlet, originally characterized decades ago and named due to their role in eye color pigmentation, in particular the formation of the brown pigment of the fly eye. These genes encode components of the kynurenine pathway, whose activity converts the amino acid tryptophan by various steps into other products. In this study, the authors have highlighted the function of this metabolic pathway in retinal health, independent of its role in pigment formation.

The Kynurenine pathway is an evolutionary conserved metabolic pathway that regulates a variety of biological processes. Its disruption can result in the buildup of either toxic or protective biomolecules or metabolites, which can worsen or improve, respectively, the health of the brain, including the retina. Knowledge on this important metabolic pathway was recently extended by the research team, led by Elisabeth Knust, Director Emerita at the MPI-CBG, in their publication in the journal Plos Genetics. Being aware of the remarkable conservation of this metabolic pathway and the genes that regulate it, they used flies as a model system to unravel the role of individual metabolites in retinal health. The researchers looked at four genes -- cinnabar, cardinal, white, and scarlet - named after abnormal eye colors following their loss in flies. "Since the Kynurenine pathway is conserved from flies to humans, we asked whether these genes regulate retinal health independent of their role in pigment formation," says Sarita Hebbar, one of the lead authors of the study.

To find this out, the scientists used a combination of genetics, dietary changes, and biochemical analysis of metabolites to study different mutations of the fruit fly, Drosophila melanogaster. Sofia Traikov, a co-author, developed a method for the biochemical analysis of the metabolites of the Kynurenine pathway. This allowed the researchers to link different metabolite levels to the health state of the retina. They found that one metabolite, 3-hydroxykynurenine (3OH-K), is damaging to the retina. More importantly, they could show that the degree of degeneration is influenced by the balance between toxic 3OH-K and protective metabolites, such as Kynurenic Acid (KYNA), and not just by their absolute amounts. Sarita continues: "We also fed two of these metabolites to normal (non-mutant) flies and found that 3OH-K enhanced stress-induced retinal damage, whereas KYNA protected the retina from stress-related damage." This means that retinal health in certain conditions can be improved by altering the ratio of metabolites of the Kynurenine pathway.

Furthermore, by targeting these four genes and therefore four distinct steps within the pathway, the researchers were able to demonstrate that not only the accumulation of 3OH-K as such, but also its location in the cell and hence its availability in further reactions, is important for retinal health.

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Eye-opening discovery about adult brain's ability to recover vision

A discovery about how some visually impaired adults could start to see offers a new vision of the brain's possibilities. The finding that the adult brain has the potential to partially recover from inherited blindness comes from a collaboration between researchers in the University of California, Irvine School of Biological Sciences and the School of Medicine. Their paper appears in Current Biology.

The team was examining treatment for Leber congenital amaurosis, known as LCA. The term refers to a group of inherited retinal diseases distinguished by severe visual impairment at birth. The condition, which stems from mutations in any of over two dozen genes, causes degeneration or dysfunction in the retina's photoreceptors.

Administering chemical compounds that target the retina, called synthetic retinoids, can restore a notable amount of vision in children with LCA. The UCI team wanted to find out if the treatment could make a difference for adults who have the condition.

"Frankly, we were blown away by how much the treatment rescued brain circuits involved in vision," said Sunil Gandhi, professor of neurobiology and behavior and the corresponding author. Gandhi is a fellow of UCI's Center for the Neurobiology of Learning and Memory and a member of the Center for Translational Vision Research. "Seeing involves more than intact and functioning retinae. It starts in the eye, which sends signals throughout the brain. It's in the central circuits of the brain where visual perception actually arises." Until now, scientists believed that the brain must receive those signals in childhood so that central circuits could wire themselves correctly.

Working with rodent models of LCA, the collaborators were surprised by what they found. "The central visual pathway signaling was significantly restored in adults, especially the circuits that deal with information coming from both eyes," Gandhi said. "Immediately after the treatment, the signals coming from the opposite-side eye, which is the dominant pathway in the mouse, activated two times more neurons in the brain. What was even more mind-blowing was that the signals coming from the same-side eye pathway activated five-fold more neurons in the brain after the treatment and this impressive effect was long-lasting. The restoration of visual function at the level of the brain was much greater than expected from the improvements we saw at the level of the retinae. The fact that this treatment works so well in the central visual pathway in adulthood supports a new concept, which is that there is latent potential for vision that is just waiting to be triggered."

The finding opens exciting research possibilities. "Whenever you have a discovery that breaks with your expectations about the possibility for the brain to adapt and rewire, it teaches you a broader concept," Gandhi said. "This new paradigm could aid in the development of retinoid therapies to more completely rescue the central visual pathway of adults with this condition."

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Cell fusion ‘awakens’ regenerative potential of human retina

Fusing human retinal cells with adult stem cells could be a potential therapeutic strategy to treat retinal damage and visual impairment, according to the findings of a new study published in the journal eBioMedicine. The hybrid cells act by awakening the regenerative potential of human retinal tissue, previously only thought to be the preserve of cold-blood vertebrates.

Cell fusion events -- the combination of two different cells into one single entity -- are known to be a possible mechanism contributing to tissue regeneration. Though rare in humans, the phenomenon has been consistently detected in the liver, brain, and gastrointestinal tract.

A team led by ICREA Research Professor Pia Cosma at the Centre for Genomic Regulation (CRG) in Barcelona and funded by Fundació "la Caixa" has now found that cell fusion events also take place in the human retina.

The researchers tested whether cell fusion events could differentiate into cells that turn into neurons, which would show potential for tissue regeneration. The team fused Müller glia, cells that play a secondary but important role in maintaining the structure and function of the retina, with adult stem cells derived from human adipose tissue or bone marrow.

"We were able to carry out cell fusion in vitro,creating hybrid cells. Importantly, the process was more efficient in the presence of a chemical signal transmitted from the retina in response to damage, resulting in rates of hybridisation increasing twofold. This gave us an important clue for the role of cell fusion in the retina," says Sergi Bonilla, postdoctoral researcher at the CRG at the time of publication and first author of the study.

The hybrid cells were injected into a growing retinal organoid, a model that closely resembles the function of the human retina. The researchers found that the hybrid cells successfully engrafted into the tissue and differentiated into cells that closely resemble ganglion cells, a type of neuron essential for vision.

"Our findings are important because they show that the Müller Glia in the human retina have the potential to regenerate neurons," says Pia Cosma. "Salamanders and fish can repair damage caused to the retina thanks to their Müller glia, which differentiate into neurons that rescue or replace damaged neurons. Mammalian Müller glia have lost this regenerative capacity, which means retinal damage or degradation can lead to visual impairment for life. Our findings bring us one step closer to recovering this ability."

The authors caution that much work remains to be done before the development of any potential treatments. One of the next steps is understanding why hybrid cells -- with four complete sets of chromosomes -- don't result in chromosomal instability and cancer development. The authors of the study believe the retina may have a mechanism regulating chromosome segregation similar to the liver, which contains tetraploid cells that act as a genetic reservoir, undergoing mitosis in response to stress and injury.

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In visual memory, size matters

Every day we encounter images on the wall, in newspapers, books, and electronic devices. Some become etched in our memory and some don't. The elements influencing whether we remember one image and not the other aren't yet known, but researchers have assumed that image size and memory aren't connected to one another, since we usually understand what appears in an image, whether it is large or small.

A new study led by Dr. Sharon Gilaie-Dotan, of Bar-Ilan University's School of Optometry and Vision Science and Gonda (Goldschmied) Multidisciplinary Brain Research Center, sought to determine whether large images are better remembered than small ones during natural daily behavior. Her assumption was based on the fact that large images require the visual system to utilize greater resources for processing them.

The results of the study, just published in the journal Proceedings of the National Academy of Sciences, show for the first time that in natural vision, visual memory of images is affected by the size of the image on the retina. These findings can have many implications, including on the use of different types of electronic screens and the quality of information processing when we rely on large vs. small screens.

Shaimaa Masarwa and Olga Kreichman, PhD students in Dr. Gilaie-Dotan's lab, examined what happens to visual memory when participants were asked to look at pictures without knowing anything about a memory task to come. Each participant was shown different pictures in different sizes, each presented to them just once.

One hundred eighty-two subjects participated in seven different experiments. Time and time again the researchers found that the large images were better remembered (1.5 times more) than the small images. This phenomenon was not dependent on specific stimuli, the order in which the images appeared, their resolution, or the amount of information they contained.

To understand whether this result was determined by size rather than amount of detail, the researchers also examined whether large, blurred images are better etched in memory than clear, small images, where the large images contained the same details as the small images. To their surprise, they found that even in this case, the participants remembered the large, blurry images better than the small, clear images.

They also found that most images were better remembered when they were presented as bigger relative to when they were presented as smaller.

"In areas of the brain that represent the retinal image, more resources will be directed to processing large images than to processing small images because the processing is determined by the area of the retina that the image stimulates," says Dr. Gilaie-Dotan. She points out that additional factors may contribute to remembering large images, such as different eye movements, and more attention and interest that large images elicit.

The study was conducted on young adults aged 18-40, ages in which vision is completely developed but has not yet begun to age. Different ages may be affected by the size of the stimuli in a different way, since both age and experience with screens are quite different between young and old.

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New type of nerve cell discovered in the retina

Scientists at the John A. Moran Eye Center at the University of Utah have discovered a new type of nerve cell, or neuron, in the retina.

In the central nervous system a complex circuitry of neurons communicate with each other to relay sensory and motor information; so-called interneurons serve as intermediaries in the chain of communication. Publishing in the Proceedings of the National Academy of Sciences of the United States of America, a research team led by Ning Tian, PhD, identifies a previously unknown type of interneuron in the mammalian retina.

The discovery marks a notable development for the field as scientists work toward a better understanding of the central nervous system by identifying all classes of neurons and their connections.

"Based on its morphology, physiology, and genetic properties, this cell doesn't fit into the five classes of retinal neurons first identified more than 100 years ago," said Tian. "We propose they might belong to a new retinal neuron class by themselves."

The research team named their discovery the Campana cell after its shape, which resembles a hand bell. Campana cells relay visual signals from both types of light-sensing rod and cone photoreceptors in the retina, but their precise purpose is the subject of ongoing research. Experiments showed Campana cells remain activated for an unusually long time -- as long as 30 seconds -- in response to a 10 millisecond light flash stimulation.

"In the brain, persistent firing cells are believed to be involved in memory and learning," said Tian. "Since Campana cells have a similar behavior, we theorize they could play a role in prompting a temporal 'memory' of a recent stimulation."

The published research study is: "An uncommon neuronal class conveys visual signals from rods and cones to retinal ganglion cells." Authors are: Brent K. Young, Charu Ramakrishnan, Tushar Ganjawala, Ping Wang, Karl Deisseroth, and Ning Tian.

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