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. 2023 Jun 21;17:1202099. doi: 10.3389/fnbeh.2023.1202099

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Copyright © 2023 Dunn, Robinson, Nwokafor, Estill, Ferrante, Shen, Lemchi, Creus-Muncunill, Ramirez, Mengaziol, Brynildsen, Leggas, Horn, Ehrlich and Blendy.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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3-Trimester NOWS model recapitulates delayed development and weight gain deficits. (A) Schematic of 3-trimester NOWS model. Weights of male (B) and female (C) pups through the first 14 days of life. For male mice, there was a modest main effect of NOWS treatment [F(1,9.8) = 4.5, p < 0.1], and a significant time x treatment interaction [F(2,70) = 9.7, p < 0.001]. Post hoc testing revealed a significant difference between saline and NOWS groups at PND 7 [χ2(1,N = 37) = 8.24, p < 0.05] and PND 14 [χ2(1,N = 37) = 8.03, p < 0.05]. For females, there was no main effect of NOWS treatment, but there was a significant time x treatment interaction [F(2,86.4) = 3.3, p < 0.05]. Time to reach developmental milestones for male (D) and female (E) pups during the first 2 weeks of life. For male mice, there was a significant effect of NOWS treatment on the time to extinguish pivoting behavior [F(1,9.6) = 13.8, p < 0.01] and time to surface righting [F(1,11.5) = 4.9, p < 0.05]. For female mice, there was a significant effect of NOWS treatment on the time to extinguish pivoting behavior [F(1,8.9) = 7.1, p < 0.05] and time to forelimb grasping [F(1,6.5) = 10.8, p < 0.05]. (F) USV emission, measured by the number of 5 s bins containing a USV during the 5-min test period. There was a significant NOWS treatment x time interaction [χ2(2,N = 39) = 32.08, p < 0.001]. Post hoc testing revealed a significant difference between saline and NOWS groups at 8 h [χ2(1,N = 9) = 47.23, p < 0.0001], 24 h [χ2(1,N = 16) = 19.27, p < 0.0001] and 48 h [χ2(1,N = 14) = 60.27, p < 0.0001]. (G)Total score of somatic signs of withdrawal during 10-min test period. There was a significant NOWS treatment x time interaction [χ2(2,N = 40) = 52.10, p < 0.001]. Post hoc testing revealed a significant difference between saline and NOWS groups at 24 h [χ2(1,N = 16) = 18.87, p < 0.0001]. *p < 0.05, **p < 0.01, ****p < 0.0001. All data was generated from 7 total litters (3 saline, 4 NOWS). (H) Latency to lift hind paw or jump on a 55^oC hotplate assay, at select timepoints after last injection. There was a main effect of NOWS treatment [F(2,35) = 18.0, p < 0.001]. Post hoc testing revealed a significant difference between saline and NOWS groups at 8 h [χ2(1,N = 10) = 11.32, p < 0.01] and 24 h [χ2(1,N = 19) = 6.14, p < 0.05].