Open AccessFunctional Control of Transplantable Human ESC‐Derived Neurons Via Optogenetic Targeting
Author(s) -
Weick Jason P.,
Johnson M. Austin,
Skroch Steven P.,
Williams Justin C.,
Deisseroth Karl,
Zhang SuChun
Publication year - 2010
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1002/stem.514
Subject(s) - biology , optogenetics , mcherry , neuroscience , microbiology and biotechnology , channelrhodopsin , excitatory postsynaptic potential , induced pluripotent stem cell , transgene , inhibitory postsynaptic potential , embryonic stem cell , green fluorescent protein , genetics , gene
Current methods to examine and regulate the functional integration and plasticity of human ESC (hESC)‐derived neurons are cumbersome and technically challenging. Here, we engineered hESCs and their derivatives to express the light‐gated channelrhodopsin‐2 (ChR2) protein to overcome these deficiencies. Optogenetic targeting of hESC‐derived neurons with ChR2 linked to the mCherry fluorophore allowed reliable cell tracking as well as light‐induced spiking at physiological frequencies. Optically induced excitatory and inhibitory postsynaptic currents could be elicited in either ChR2 + or ChR2 − neurons in vitro and in acute brain slices taken from transplanted severe combined immunodeficient (SCID) mice. Furthermore, we created a clonal hESC line that expresses ChR2‐mCherry under the control of the synapsin‐1 promoter. On neuronal differentiation, ChR2‐mCherry expression was restricted to neurons and was stably expressed for at least 6 months, providing more predictable light‐induced currents than transient infections. This pluripotent cell line will allow both in vitro and in vivo analysis of functional development as well as the integration capacity of neuronal populations for cell‐replacement strategies. STEM CELLS 2010;28:2008–2016