Fig. 1: MSEW increases avoidance behavior only in diestrus.
a, b Timeline and schematics of MSEW paradigm. c Schematic of the modified EPM, with a fine mist of water sprayed on the open arms to increase avoidance and prevent habituation with repeated trials. d Percent time in the open arms of the modified EPM is not significantly different between control and MSEW mice when stage of estrous during testing is not taken into consideration (Control n = 11, MSEW n = 11). e Number of entries into the open arm between control and MSEW mice is not significantly different when stage of estrous during testing is not taken into consideration (Control n = 11, MSEW n = 11). f Schematic of vaginal cytology used to determine whether mice were in different stages of the estrous cycle (proestrus, estrus, metestrus, and diestrus). g Testing on the modified EPM at specific stages of the estrous cycle shows that during diestrus, MSEW mice display increased avoidance behavior, i.e., reduced percent time in the open arms, compared to control mice (F3,55 = 3.393, p = 0.0242; Control-MSEW Diestrus p = 0.0464; Control n = 11, MSEW n = 11). h MSEW mice also have significantly fewer entries into the open arm while in diestrus (F3,55 = 4.153, p = 0.0101; Control-MSEW Diestrus p = 0.0350; Control n = 11, MSEW n = 11). No significant differences between control and MSEW mice were observed during proestrus, estrus, or metestrus. MSEW = maternal separation early weaning. *p < 0.05 two-sided unpaired t-tests (d, e); mixed-effects model repeated measures followed by Šidák post hoc tests (g, h). Data are presented as mean values + SEM (d, e, g, h). Images in b, c, f were created using BioRender.com. Source data are provided as a Source Data file.
