Fig. 1.

Experimental timelines. Experiment 2—establish dose-responsiveness of NPS in female mice (aCSF, NPS – 1, 0.1, 0.01 nmol). Experiment 3—test the efficacy of the biased NPSR agonist RTI-263 (aCSF, NPS 1 nmol, RTI-263 1 nmol). Experiment 4—a larger cohort of female mice were used to test the effects of free estrous cycling on NPS administration (aCSF, NPS 1 nmol). After the completion of the ASR paradigm, 20 mice were removed from the main study to verify the voltage necessary to produce reliable effects in the inhibitory avoidance paradigm. The voltage–response relationship, where we test groups of animals with different intensities of electric shock, is completed to guard against cohort-to-cohort variation in the responsiveness to the shock. The remainder of the mice proceeded into the inhibitory avoidance using the optimal voltage as determined by the voltage–response test (see below). Those mice that are used as part of the voltage–response testing are not used for any subsequent studies.