Release date: 2017-02-22
Under the support of the National Natural Science Foundation of China National Science Fund for Distinguished Young Scholars (project number), the Tang Shiming team of the School of Life Sciences of Peking University has made important progress in the long-time two-photon imaging technology of awake macaques. The research results were published online on February 16th, 2017 in Neuron, under the title "Long-Term Two-Photon Imaging in Awake Macaque Monkey". Dr. Li Ming, Ph.D., Life Science Center of Peking University, is the first author. Researcher Tang Shiming of the School of Life Sciences of Peking University and McGovern Institute of Brain Science is the author of the communication. The paper link is: http://(17)30051-X.pdf.
Rhesus macaques are very close to humans in terms of cognitive behavior, brain structure and function. They are ideal model animals for human brain diseases, visual cognition and other advanced cognitive functions. Two-photon imaging technology has been widely used in the field of neuroscience in recent years because it can observe a large number of neuronal activities in the cortex with single cell or even single synaptic resolution. However, due to the difficulty of large brain tissue hyperplasia, large brain beating, and low efficiency of gene-encoded probes in large animals, long-term two-photon imaging of brain activity in awake macaques has not been realized, and it is one of the researches in this field. An internationally recognized technology bottleneck.
Tang Shiming's group passed microinjection of adeno-associated virus (AAV) into the visual cortex of macaques, and transferred into the calcium-encoding GCaMP5 and GCaMP6s genes, and obtained long-stable and high-efficiency expression in the macaque visual cortex. Combined with the newly designed, it can effectively prevent intracranial infection. And the imaging window of dural hyperplasia, obtaining a stable optical quality that can last for several months; at the same time, by improving the head fixture and image movement correction, a stable image and high-quality neural activity signals are obtained, and finally the awake macaque recognizes Two-photon imaging with a stabilization time of more than 6 months was achieved under known behavioral conditions. For the first time, the research team realized the electrophysiological operation and research of neuronal intracellular recording and single cell electroporation under the two-photon imaging of awake macaques, and proved that the calcium probe signal has a large linear relationship with neuron activity (10Hz~ 150Hz), and achieve sparse transfection and dendritic imaging of macaque cortical neurons.
The long-time two-photon imaging technology of awake macaques will bring great convenience to the study of human brain diseases, visual cognition and other advanced cognitive functions.
Two-photon imaging of macaque brain. A: imaging window of visual cortex V1; B: high quality nerve cell image; C: imaging of different levels/depth.
Source: National Natural Science Foundation
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