Life takes shape with the motion of a single cell. In response to signals from certain proteins and enzymes, a cell can start to move and shake, leading to contractions that cause it to squeeze, pinch, and eventually divide. As daughter cells follow suit down the generational line, they grow, differentiate, and ultimately arrange themselves into a fully formed organism.
Now MIT scientists have used light to control how a single cell jiggles and moves during its earliest stage of development. The team studied the motion of egg cells produced by starfish — an organism that scientists have long used as a classic model for understanding cell growth and development.
The researchers focused on a key enzyme that triggers a cascade of motion within a starfish egg cell. They genetically designed a light-sensitive version of the same enzyme, which they injected into egg cells, and then stimulated the cells with different patterns of light.
They found that the light successfully triggered the enzyme, which in turn prompted the cells to jiggle and move in predictable patterns. For instance, the scientists could stimulate cells to exhibit small pinches or sweeping contractions, depending on the pattern of light they induced. They could even shine light at specific points around a cell to stretch its shape from a circle to a square.
Their results, which will appear in the journal Nature Physics, provide scientists with a new optical tool for controlling cell shape in its earliest developmental stages. Such a tool, they envision, could guide the design of synthetic cells, such as therapeutic “patch” cells that contract in response to light signals to help close wounds, or drug-delivering “carrier” cells that release their contents only when illuminated at specific locations in the body. Overall, the researchers see their findings as a new way to probe how life takes shape from a single cell.
“By revealing how a light-activated switch can reshape cells in real time, we’re uncovering basic design principles for how living systems self-organize and evolve shape,” says the study’s senior author, Nikta Fakhri, associate professor of physics at MIT. “The power of these tools is that they are guiding us to decode all these processes of growth and development, to help us understand how nature does it.”
The study’s MIT authors include first author Jinghui Liu, Yu-Chen Chao, and Tzer Han Tan; along with Tom Burkart, Alexander Ziepke, and Erwin Frey of Ludwig Maximilian University of Munich; John Reinhard of Saarland University; and S. Zachary Swartz of the Whitehead Institute for Biomedical Research.
Cell circuitry
Fakhri’s group at MIT studies the physical dynamics that drive cell growth and development. She is particularly interested in symmetry, and the processes that govern how cells follow or break symmetry as they grow and divide. The five-limbed starfish, she says, is an ideal organism for exploring such questions of growth, symmetry, and early development.
“A starfish is a fascinating system because it starts with a symmetrical cell and becomes a bilaterally symmetric larvae at early stages, and then develops into pentameral adult symmetry,” Fakhri says. “So there’s all these signaling processes that happen along the way to tell the cell how it needs to organize.”
Scientists have long studied the starfish and its various stages of development. Among many revelations, researchers have discovered a key “circuitry” within a starfish egg cell that controls its motion and shape. This circuitry involves an enzyme, GEF, that naturally circulates in a cell’s cytoplasm. When this enzyme is activated, it induces a change in a protein, called Rho, that is known to be essential for regulating cell mechanics.
When the GEF enzyme stimulates Rho, it causes the protein to switch from an essentially free-floating state to a state that binds the protein to the cell’s membrane. In this membrane-bound state, the protein then triggers the growth of microscopic, muscle-like fibers that thread out across the membrane and subsequently twitch, enabling the cell to contract and move.
In previous work, Fakhri’s group showed that a cell’s movements can be manipulated by varying the cell’s concentrations of GEF enzyme: The more enzyme they introduced into a cell, the more contractions the cell would exhibit.
“This whole idea made us think whether it’s possible to hack this circuitry, to not just change a cell’s pattern of movements but get a desired mechanical response,” Fakhri says.
Lights and action
To precisely manipulate a cell’s movements, the team looked to optogenetics — an approach that involves genetically engineering cells and cellular components such as proteins and enzymes, such that they activate in response to light.
Using established optogenetic techniques, the researchers developed a light-sensitive version of the GEF enzyme. From this engineered enzyme, they isolated its mRNA — essentially, the genetic blueprint for building the enzyme. They then injected this blueprint into egg cells that the team harvested from a single starfish ovary, which can hold millions of unfertilized cells. The cells, infused with the new mRNA, then began to produce light-sensitive GEF enzymes on their own.
In experiments, the researchers then placed each enzyme-infused egg cell under a microscope and shone light onto the cell in different patterns and from different points along the cell’s periphery. They took videos of the cell’s movements in response.
They found that when they aimed the light in specific points, the GEF enzyme became activated and recruited Rho protein to the light-targeted sites. There, the protein then set off its characteristic cascade of muscle-like fibers that pulled or pinched the cell in the same, light-stimulated spots. Much like pulling the strings of a marionette, they were able to control the cell’s movements, for instance directing it to morph into various shapes, including a square.
Surprisingly, they also found they could stimulate the cell to undergo sweeping contractions by shining a light in a single spot, exceeding a certain threshold of enzyme concentration.
“We realized this Rho-GEF circuitry is an excitable system, where a small, well-timed stimulus can trigger a large, all-or-nothing response,” Fakhri says. “So we can either illuminate the whole cell, or just a tiny place on the cell, such that enough enzyme is recruited to that region so the system gets kickstarted to contract or pinch on its own.”
The researchers compiled their observations and derived a theoretical framework to predict how a cell’s shape will change, given how it is stimulated with light. The framework, Fakhri says, opens a window into “the ‘excitability’ at the heart of cellular remodeling, which is a fundamental process in embryo development and wound healing.”
She adds: “This work provides a blueprint for designing ‘programmable’ synthetic cells, letting researchers orchestrate shape changes at will for future biomedical applications.”
This work was supported, in part, by the Sloan Foundation, and the National Science Foundation.
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Written by Jennifer Chu, MIT News
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