Fig. 1.

Zygote microinjection of nine sperm miRs recapitulated the HPA axis phenotype. (A) Schematic of experimental design in which single-cell mouse zygotes were injected with nine miRs (multi-miR; 1 ng/μL final concentration of miR-29c, miR-30a, miR-30c, miR-32, miR-193-5p, miR-204, miR-375, miR-532-3p, and miR-698) or with one of two control conditions: an injection control (PBS only) or a control for the total amount of injected miRs (single miR; 1 ng/μL final concentration miR-193-5p alone) before implantation into surrogate females or preparation for later analyses. (B and D) Corticosterone levels in male and female adult mice were significantly reduced in the multi-miR group relative to PBS controls in response to a 15-min restraint from t = 0–15 min. (C and E) Total AUC of corticosterone production over 120 min was significantly decreased in the multi-miR group compared with PBS controls, and expected sex differences were observed. Data are presented as mean ± SEM n = 6–10 mice per group. *P < 0.05 multi-miR vs. PBS. (F) Model demonstrating the similarity in the shape of the male corticosterone response curve in the multi-miR–injected group with the outcome previously reported in offspring following paternal stress (11).