Abstract
Optogenetic methods are now widespread in neuroscience research. Here we present a detailed surgical procedure to inject adeno-associated viruses and implant optic fiber cannulas in the dorsal raphe nucleus (DRN) of living mice. Combined with transgenic mouse lines, this protocol allows specific targeting of serotonin-producing neurons in the brain. It includes fixing a mouse in a stereotaxic frame, performing a craniotomy, virus injection and fiber implantation. Animals can be later used in behavioral experiments, combined with optogenetic manipulations (Dugué et al., 2014; Correia et al., 2017) or monitoring of neuronal activity (Matias et al., 2017). The described procedure is a fundamental step in both optogenetic and fiber photometry experiments of deep brain areas. It is optimized for serotonin neurons in the DRN, but it can be applied to any other cell type and brain region. When using transgenic mouse lines that express functionally relevant levels of optogenetic tools or reporter lines, the virus injection step can be skipped and the protocol is reduced to the cannula implantation procedure.
Highlights
The described procedure is a fundamental step in both optogenetic and fiber photometry experiments of deep brain areas. It is optimized for serotonin neurons in the dorsal raphe nucleus (DRN), but it can be applied to any other cell type and brain region
When using transgenic mouse lines that express functionally relevant levels of optogenetic tools or reporter lines, the virus injection step can be skipped and the protocol is reduced to the cannula implantation procedure
Virus injections and fiber cannula implantations have been previously described for diverse areas in the brain
Summary
5. Absorbable sponges (Spongostan Dental, Ferrosan Medical Devices) (Ethicon, catalog number: MS0005). Cleaning wipes (Kimwipes, Kimtech) (KCWW, Kimberly-Clark, catalog number: 34120). SERT-Cre C57BL/6 mice (Slc6a4tm1(cre)Xz, THE JACKSON LABORATORY, catalog number: 014554) and WT C57BL/6 mice (littermates control). Virus AAV2.9.EF1a.DIO.hChR2(H134R)-eYFP.WPRE.hGH (1013 GC/ml) for photostimulation or AAV2/1-Syn-Dio-GCaMP6s (1013 GC/ml) for fiber photometry Analgesic (e.g., Dolorex, Butorphanol, http://www.dolorex.info/dolorex/dolorex.asp, 10 mg/ml, injectable solution, Intervet, Schering-Plough Animal Health).
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