Fig. 4: Microinjection of sperm RNAs from SARS-CoV-2 infected sires into fertilized oocytes significantly changes anxiety-like behavior in the resultant offspring.
Microinjected-SARS mice show no changes in (A) % time spent in the light zone of the light-dark box (n = 24 MCON M, n = 14 MCON F, n = 22 MSARS M, n = 17 MSARS F), or (B) number of entries into the light zone (n = 24 MCON M, n = 14 MCON F, n = 22 MSARS M, n = 17 MSARS F). There is a significant increase in (C) the latency to enter the light zone in the male microinjected-SARS mice only (n = 19 MCON M, n = 11 MCON F, n = 18 MSARS M, n = 12 MSARS (F)(P = 0.001, general linear model with Bonferroni correction), and significant microinjection x sex interactions in the (D) % time spent in the open arms (n = 24 MCON M, n = 15 MCON F, n = 22 MSARS M, n = 17 MSARS F) and (E) number of entries into the open arms (n = 24 MCON M, n = 15 MCON F, n = 22 MSARS M, n = 17 MSARS (F) of the elevated-plus maze. No differences are seen in the (F) % time spent in the centre of the open-field (n = 24 MCON M, n = 15 MCON F, n = 21 MSARS M, n = 17 MSARS F). There is a significant microinjection x sex interaction for (G) overall anxiety behavioral z-scores (n = 24 MCON M, n = 15 MCON F, n = 22 MSARS M, n = 17 MSARS F)(P = 0.0132, general linear model with Bonferroni correction). No differences seen in H) total distance traveled in the open-field (n = 24 MCON M, n = 15 MCON F, n = 21 MSARS M, n = 17 MSARS F), I the recognition index in trial 2 of the novel object recognition test (n = 22 MCON M, n = 15 MCON F, n = 22 MSARS M, n = 17 MSARS F), (J) overall cognition behavioral z-scores (n = 24 MCON M, n = 15 MCON F, n = 22 MSARS M, n = 17 MSARS F), K % preference for saccharin in the saccharin-preference test (n = 24 MCON M, n = 15 MCON F, n = 22 MSARS M, n = 17 MSARS F), (L) latency to feed in the novelty-suppressed feeding test (n = 24 MCON M, n = 15 MCON F, n = 22 MSARS M, n = 17 MSARS F), (M) overall depression behavioral z-scores (n = 24 MCON M, n = 15 MCON F, n = 22 MSARS M, n = 17 MSARS F), and (N) % bodyweight changed after a 24-h fasting period (n = 24 MCON M, n = 15 MCON F, n = 22 MSARS M, n = 17 MSARS F). There was a trend towards a significant microinjection x sex interaction for O) food consumed in a 5-min period after a 24-h fasting period (n = 24 MCON M, n = 15 MCON F, n = 22 MSARS M, n = 17 MSARS F), but there were no differences in (P) overall feeding behavioral z-scores (n = 24 MCON M, n = 15 MCON F, n = 22 MSARS M, n = 17 MSARS F). Data presented as mean ± SEM. General linear models were used with post-hoc analyses where appropriate (Bonferroni-Holm corrected) except for (L). Cox regression with proportional hazards was used to analyse (L). Each n number refers to the number of individual animals per group. PMicroinjection = main effect of microinjection treatment. PMicroinjection x Sex = interaction effect of microinjection treatment by sex. *P < 0.05, **P < 0.01.
