Table 1.
Names and characteristics of transgenic mouse lines used in the present study
| Name | Definition/Official Name/Source/RRID | Characteristics and use in the present study |
|---|---|---|
| C57Bl/6J | Jackson Laboratories strain #000664; RRID:IMSR_JAX:000664 | Wild type (WT) mice used for breeding, in vitro electrophysiological slice recordings, to assess viral expression and penetrance, and for single-cell RT-PCR |
| nNOS-CreER | B6;129S-Nos1tm1.1(cre/ERT2)Zjh/J; Jackson Laboratories strain #014541; RRID:IMSR_JAX:014541 | Originally described by Taniguchi et al. (2011). When injected with tamoxifen, a ligand for the estrogen receptor (ER), Cre recombinase is induced specifically in neurons that express neuronal nitric oxide synthase (nNOS). In the present study, nNOS-CreER mice were used for breeding |
| Ai14 | B6;129S6-Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J Jackson Laboratories strain #007 914; RRID:IMSR_JAX:007914 | Originally described by Madisen et al. (2010), this reporter strain expresses the red fluorescent protein tdTomato. In the present study, Ai14 mice were used for breeding with nNOS-CreER mice |
| nNOS-CreER;Ai14 | Bigenic mice generated by breeding nNOS-CreER mice with Ai14 mice. In the resultant progeny, nNOS neurons express tdTomato. In the present study, nNOS-CreER;Ai14 mice were used for breeding with Orexin-tTA mice | |
| Orexin-tTA | Orexin-tetracycline-controlled Transcriptional Activator (“ox-tTA” mice) | “Ox-tTA” mice were originally described by Tabuchi et al. (2013). In this strain, tTA, driven by human prepro-orexin promoter, is exclusively expressed in Hcrt/orexin neurons. In the present study, Ox-tTA mice were used for breeding, to assess viral expression and penetrance, to express ChR2 in Hcrt cells for in vitro electrophysiological slice recordings of both Hcrt and nNOS cells, and as monogenic littermate controls in the EEG studies |
| Orexin-tTA; nNOS-CreER;Ai14 | “Ox-tTA;nNOS-CreER;Ai14” mice | Trigenic “ox-tTA;nNOS-CreER;Ai14” mice were generated by breeding Orexin-tTA mice with bigenic nNOS-CreER;Ai14 mice. In the present study, ox-tTA;nNOS-CreER;Ai14 mice were injected with an AAV encoding TetO-ChR2 and then used for in vitro optogenetic stimulation of Hcrt terminals and recording of cortical nNOS cells |
| Orexin-tTA; TetO-DTA | Orexin-tetracycline-controlled Transcriptional Activator; Tetracycline Operator 5; Diphtheria Toxin A fragment (“DTA” mice) | These bigenic “DTA” mice were described by Tabuchi et al. (2014). tTA is exclusively expressed in Hcrt/orexin neurons. In the absence of doxycycline in the chow (“DOX(−)”), tTA binds to TetO and induces production of the toxic DTA protein in the Hcrt neurons which results in degeneration of these cells. In the present study, DTA mice were used for in vitro electrophysiology, immunohistochemistry and EEG/EMG recording |
| Hcrtr1-EGFP |
|
Originally described in Darwinkel et al. (2014), the founder line (KP68Gsat/Mmucd) was from the Mutant Mouse Regional Resource Center. EGFP was inserted upstream of the Hcrtr1 gene. In the present study, Hcrtr1-EGFP mice were used for immunohistochemistry |