To generate dynorphin reporter mice, we crossed Pdyn-IRES-Cre mice with R26-lsl-L10-GFP mice, two lines of mice that were produced and characterized in the Lowell lab and were published previously (Krashes et al., 2014 (link)). The Pdyn-IRES-Cre mice are available from The Jackson Laboratory (stock no. 027958). Pdyn-IRES-Cre mice selectively express Cre recombinase in cells that have expressed prodynorphin, and L10-GFP mice have a loxP-flanked STOP cassette which prevents the transcription of downstream green fluorescent protein (GFP). We crossed these mice with Pdyn-IRES-Cre mice because in the presence of Cre recombinase, the STOP cassette is deleted, resulting in offspring that express GFP selectively in Cre-expressing dynorphin cells, creating Pdyn-GFP reporter mice.
We housed all mice on a 12:12 light:dark cycle with lights on at 0600 at 22°C ambient temperature with ad libitum access to food and water. All protocols and care of the mice followed National Institute of Health guidelines and were approved by the University of Iowa Institutional Animal Care and Use Committee.
We housed all mice on a 12:12 light:dark cycle with lights on at 0600 at 22°C ambient temperature with ad libitum access to food and water. All protocols and care of the mice followed National Institute of Health guidelines and were approved by the University of Iowa Institutional Animal Care and Use Committee.