Selective viral transduction of adult-born olfactory neurons for chronic in vivo optogenetic stimulation.
Local interneurons are continuously regenerated in the olfactory bulb of adult rodents. In this process, called adult neurogenesis, neural stem cells in the walls of the lateral ventricle give rise to neuroblasts that migrate for several millimeters along the rostral migratory stream (RMS) to reach and incorporate into the olfactory bulb. To study the different steps and the impact of adult-born neuron integration into preexisting olfactory circuits, it is necessary to selectively label and manipulate the activity of this specific population of neurons. The recent development of optogenetic technologies offers the opportunity to use light to precisely activate this specific cohort of neurons without affecting surrounding neurons. Here, we present a series of procedures to virally express Channelrhodopsin2(ChR2)-YFP in a temporally restricted cohort of neuroblasts in the RMS before they reach the olfactory bulb and become adult-born neurons. In addition, we show how to implant and calibrate a miniature LED for chronic in vivo stimulation of ChR2-expressing neurons.
Laboratory for Perception and Memory, Institut Pasteur and Centre National de la Recherche Scientifique.
Gabriel
G
Lepousez
Mariana
M
Alonso
Sebastian
S
Wagner
Benjamin W
BW
Gallarda
Pierre-Marie
PM
Lledo
D000818:Animals; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D000075402:Channelrhodopsins; D016086:Lentivirus / Q000235:genetics; D008164:Luminescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D009474:Neurons / Q000502:physiology*; D009830:Olfactory Bulb / Q000166:cytology / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D014161:Transduction, Genetic
D001426:Bacterial Proteins; D000075402:Channelrhodopsins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria