{"id":3128,"date":"2015-01-26T09:00:48","date_gmt":"2015-01-26T08:00:48","guid":{"rendered":"http:\/\/news.embl.de\/?p=3128"},"modified":"2024-11-29T16:55:24","modified_gmt":"2024-11-29T15:55:24","slug":"1501-cell-control","status":"publish","type":"post","link":"https:\/\/www.embl.org\/news\/science\/1501-cell-control\/","title":{"rendered":"Cell control in a flash"},"content":{"rendered":"\n

By Adam Rutherford<\/span><\/h3>\n\n\n\n

Emerging in 2003, optogenetics was developed by several scientists across the globe, including Ernst Bamberg, Ed Boyden, Karl Deisseroth, Peter Hegemann, Gero Miesenb\u00f6ck, and Georg Nagel. The aim was to elicit control over ever-smaller neural pathways, trying to unpick the circuits of thought. But the simplicity of the idea betrays an impressive and imaginative technical feat. The principle is straightforward: a light-sensitive protein is incorporated into a specific population of cells, and a laser tuned to that protein\u2019s specific firing frequency is used to activate the cells with millisecond precision. The timing is crucial, as it mirrors the physiological response time, and the light triggers the neuron\u2019s action potential.<\/p>\n\n\n\n

Standard techniques of genetic modification, such as the cre\/lox system and lentiviral vectors, are used to target the gene encoding a phototransduction protein \u2013 often a microbial opsin \u2013 to specific neuronal cells. But the more fiddly aspect of this technique is that this has to be done in such a way that normal function is not upset. The use of optogenetics in the past decade has blossomed, and uncovered all sorts of revelations about brain function \u2013 for example, the circuits that modulate fear and the conditioning of cells involved in cocaine dependency. Speculation has been rife for some time that sooner rather than later, this is a tool that will bag a few Nobel Prizes.<\/p>\n\n\n\n

we are very excited about adding this new technique to our research arsenal<\/p><\/blockquote>\n\n\n\n

In touch<\/h3>\n\n\n\n

Paul Heppenstall<\/a>, a group leader at EMBL Monterotondo, has now <\/span>adapted the system for use outside the darkness of the brain. \u201cIt was the ability to look through the microscope and image neurons under the skin that got me excited,\u201d he explains. Heppenstall\u2019s team work on our perception of touch and pain, which means their targets are the cells at the periphery of the nervous system \u2013 neurons that react to mechanical or chemical stimuli, touch, heat and pressure. The specific populations for each of these are poorly understood, and not easy to disentangle.<\/p>\n\n\n\n

It was the ability to image neurons under the skin that got me excited<\/p><\/blockquote>\n\n\n\n

\u201cTraditionally we\u2019ve used electrophysiology of single cells, which is tedious and slow,\u201d says Heppenstall. \u201cIt takes several months to put the data together, and at the end, you might see something. Or you might not.\u201d<\/p>\n\n\n\n

Their new technique<\/a> involves identifying a cell-specific marker, a gene that is active in a discreet population of skin neurons. They do this by scanning the literature, making educated guesses and also through a bit of luck. Once they find such a gene, they attach an optogenetic control switch onto it using the cre\/lox system. Then they can activate the cells by shining blue light on a patch of skin and monitor the behaviour this evokes in mice, such as withdrawal for pain-sensing neurons and scratching for itch receptors. As a complementary experiment, they also use the same system to ablate specific populations of cells through a photosensitive dye. Once they\u2019ve ablated the cell, a stimulus can be applied and changes in behaviour studied.<\/p>\n\n\n\n

Shaping up<\/h3>\n\n\n\n

Optogenetics offers cell-specific and microsecond precision. What\u2019s odd is that this powerful technique has always been about brain cells. Even in the 1970s, Francis Crick pondered light-controlled cells, but only for neurons.<\/p>\n\n\n\n

I knew it would be great if it worked, but to tell you the truth, I was quite sceptical<\/p><\/blockquote>\n\n\n\n

At EMBL Heidelberg, group leader Stefano De Renzis<\/a> is set to break that stranglehold. \u201cThe biggest surprise for us was that it\u2019s a technique that works so well for studying Drosophila<\/i> morphogenesis,\u201d he explains. It was De Renzis\u2019 graduate student Giorgia Guglielmi who first suggested co-opting optogenetics from neuroscience to scrutinise basic fruit fly development. \u201cI knew it would be great if it worked, but to tell you the truth, I was quite sceptical. Giorgia who had just started her PhD in my lab was immediately enthusiastic to try something new.\u201d<\/p>\n\n\n\n

\"Surface
Optogenetically activated cells in a fruit fly embryo lose the regular hexagonal shape displayed here, and the embryo will fail to develop properly. IMAGE: EMBL\/STEFANO DE RENZIS<\/figcaption><\/figure><\/div>\n\n\n\n

Their work is centred on how proteins and other elements of the cell membrane regulate the shape of cells during Drosophila<\/i> embryogenesis. This is crucial to overall tissue patterning as cells mature and take on specific shapes and sizes. Differentiation of individual cells affects their neighbours and ultimately the whole embryo. But these changes are highly temporal and spatial, and addressing the underlying mechanisms is a question that has been hampered by an inability to command the whens and wheres of gene expression. And there\u2019s no action potential to trigger.<\/p>\n\n\n\n

Knocking down<\/h3>\n\n\n\n

\u201cWe were always looking for ways of controlling specific cells during development \u2013 to knock-down protein activity, but in a very temporal and spatial way,\u201d De Renzis says. \u201cIn the past we\u2019ve used knock-outs, and temperature-sensitive mutants, but it takes time for the clones to grow, which complicates the interpretation of results. With optogenetics we can do it in a fraction of the time, enabling us to look at immediate cell and tissue responses.\u201d<\/p>\n\n\n\n

In some ways, this seems a simpler proposition than eliciting complex behavioural changes in mammals, as has been the mainstay of optogenetics so far. The De Renzis team is looking at actin and membrane trafficking, and how they affect patterning and cell shape. They are quizzing the way the embryo invaginates \u2013 a process of folding in on itself to internalise mesodermal cells that run the length of the fruit fly embryo. This is regulated by molecules on the plasma membrane, and by simply turning these off using light in only a small population of cells, the effect is global<\/a>. The targeted cells fail to contract, and the knock-on effect is the arrest of invagination across the whole embryo.<\/p>\n\n\n\n

This first foray into using optogenetics in developmental biology is now submitted for publication and no doubt the technique is set to become a versatile tool in the future. \u201cTo control cellular activity experimentally has not been possible with conventional genetics so far,\u201d De Renzis says, \u201cat least not at very precise resolution. Now, we are very excited about adding this new technique to our research arsenal and we look forward to sharing it with other labs interested in shedding light on the cellular and molecular mechanisms driving the development of organisms.\u201d<\/p>\n","protected":false},"excerpt":{"rendered":"

From using light to control brain activity to illuminating fruit fly development and mice\u2019s sense of touch<\/p>\n","protected":false},"author":16,"featured_media":3269,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[2,17591],"tags":[362,55,43,69,79,74,67,363,514],"embl_taxonomy":[],"class_list":["post-3128","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-science","category-science-technology","tag-de-renzis","tag-development","tag-heidelberg","tag-methods","tag-microscopy","tag-monterotondo","tag-neurobiology","tag-optogenetics","tag-rome"],"acf":{"article_intro":"

Flick the switch, and illumination follows. The comical image of a light bulb pinging on when a brainwave occurs became remarkably prescient just a few years ago when suddenly, out of the dark, a brand new technique for controlling brain activity lit up the scene.<\/p>\n","related_links":[{"link_description":"Cornelius Gross from EMBL Monterotondo discussed optogenetics' potential in the 2014 ELLS Insight Lecture \u2013 video now available on demand","link_url":"http:\/\/news.embl.de\/events\/1502-insight-lecture\/"}],"article_sources":false,"vf_locked":false,"featured":false,"color":"#007B53"},"embl_taxonomy_terms":[],"yoast_head":"\nCell control in a flash | EMBL<\/title>\n<meta name=\"description\" content=\"Optogenetics: a technique that uses light to control brain activity is being adapted to illuminate fruit fly development and mice\u2019s sense of touch.\" \/>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" 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