Long-term all-optical interrogation of cortical neurons in awake-behaving nonhuman primates

• Published in: PLoS biology 2018-08, Vol.16 (8), p.e2005839-e2005839
• Main Authors: Ju, Niansheng, Jiang, Rundong, Macknik, Stephen L, Martinez-Conde, Susana, Tang, Shiming
• Format: Article
• Language: English
• Subjects: Activation; Animals; Behavior; Biology and Life Sciences; Brain - physiology; Brain research; Calcium; Calcium - metabolism; Calcium imaging; Calcium-binding protein; Calmodulin; Coding; Cognition; Cortex (temporal); Dependovirus; Education; Engineering and Technology; Eye movements; Feasibility studies; Genetic code; Genetics; Information processing; Insects; Interrogation; Laboratories; Lasers; Life sciences; Macaca mulatta; Medical research; Medicine and Health Sciences; Methods; Methods and Resources; Microscopy; Neuroimaging; Neurons; Neurons - physiology; Optics; Optogenetics - methods; Photic Stimulation; Physiological aspects; Populations; Primates; Prostheses; Questioning; Recording; Recording sessions; Research and Analysis Methods; Rhesus monkey; Rhodopsin; Rodents; Social Sciences; Stimulation; Viruses; Visual cortex; Visual Cortex - physiology; Wakefulness
• ISSN: 1545-7885; 1544-9173; 1545-7885
• DOI: 10.1371/journal.pbio.2005839

Whereas optogenetic techniques have proven successful in their ability to manipulate neuronal populations-with high spatial and temporal fidelity-in species ranging from insects to rodents, significant obstacles remain in their application to nonhuman primates (NHPs). Robust optogenetics-activated behavior and long-term monitoring of target neurons have been challenging in NHPs. Here, we present a method for all-optical interrogation (AOI), integrating optical stimulation and simultaneous two-photon (2P) imaging of neuronal populations in the primary visual cortex (V1) of awake rhesus macaques. A red-shifted channel-rhodopsin transgene (ChR1/VChR1 [C1V1]) and genetically encoded calcium indicators (genetically encoded calmodulin protein [GCaMP]5 or GCaMP6s) were delivered by adeno-associated viruses (AAVs) and subsequently expressed in V1 neuronal populations for months. We achieved optogenetic stimulation using both single-photon (1P) activation of neuronal populations and 2P activation of single cells, while simultaneously recording 2P calcium imaging in awake NHPs. Optogenetic manipulations of V1 neuronal populations produced reliable artificial visual percepts. Together, our advances show the feasibility of precise and stable AOI of cortical neurons in awake NHPs, which may lead to broad applications in high-level cognition and preclinical testing studies.