November 6, 2016
Scientists of the institute, as part of an international team, have received a new indicator using genetic engineering methods, which allows observing the processes in the neurons of the brain. It is recognized as one of the brightest protein calcium indicators currently created. An analogue of a fluorescent protein isolated from a primitive sea creature, the European lancelet, was used in the experiment. So, the protein of the lower chordate animal turned out to be useful for studying the most complex processes of transmission of nerve impulses in mammals.
As part of an international team, scientists have received a new bright protein indicator for visualizing the processes that occur in the neurons of the brain. This is reported in the International Journal of Molecular Science.
The genetically encoded calcium indicator developed by scientists is a two-part protein. The protein-sensor “senses” how the calcium concentration changes, and the fluorescent protein “interprets” this data into a signal. These indicators make it possible to observe some cellular processes in a living organism, including the transmission of a nerve impulse in the brain.
According to Alena Nikolaeva, the luminous protein mNeonGreen is a derivative of a protein originally isolated from the European lancelet Branchiostoma lanceolatum, a lower chordate animal that lives in the ocean.
The new indicator is called NCaMP. To study the kinetic characteristics of the new protein, the NCaMP gene was introduced into the genome of a model organism, Escherichia coli.
After optimization by several rounds of directed evolution, the version with the best biochemical characteristics was obtained – NCaMP7.
After successfully testing the effectiveness of the artificial protein on nerve cells, animal experiments began. At the final stage, transgenic animals carrying the NCaMP7 gene in their DNA were obtained.
The results of the experiment confirmed that the new indicator allows monitoring the activity of neurons in the cerebral cortex in mice. The researchers noted that the protein of a primitive marine animal turned out to be useful for studying the most complex processes of transmission of nerve impulses in mammals.
“The implementation of such a large-scale multidisciplinary project became possible by virtue of the corporative work of experts from various fields of science. It was attended by molecular biologists, biochemists, cell biologists, specialists in X-ray crystallography and genetic engineering. This is what explains the vastness and depth of the results obtained, ”stressed Alena Nikolaeva.