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. 2022 Apr 15;11(8):1350. doi: 10.3390/cells11081350

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Body weight and behavior of adolescent WT and Fmr1^KO male mice in response to cow milk- and soy-based infant formula diets after 2 months treatment. Fmr1^HET females were bred and maintained on Teklad2019 (grain-based but low phytoestrogen levels) diet throughout gestation and lactation. Female Fmr1^HET and Fmr1^KO and male WT and Fmr1^KO pups were weaned at P20 and randomized to diets (Teklad 2019, CIF, SIF). (A) The mice were weighed once per week for 2 months. Female cohorts included Fmr1^HET/Teklad 2019 (n = 4), Fmr1^HET/CIF (n = 5), Fmr1^HET/SIF (n = 6), Fmr1^KO/Teklad 2019 (n = 7), Fmr1^KO/CIF (n = 7) and Fmr1^KO/SIF (n = 6). Statistics were determined by 2-way mixed effects model ANOVA: time F(8230) = 1413, p < 0.0001; genotype/diet F(5,29) = 5.122, p = 0.0017; time x genotype/diet F(40,230) = 8.714, p < 0.0001. Male cohorts included the following: WT/Teklad 2019 (n = 6), WT/CIF (n = 6), WT//SIF (n = 4), Fmr1^KO/Teklad 2019 (n = 5), Fmr1^KO/CIF (n = 5) and Fmr1^KO/SIF (n = 7). Statistics were determined by 2-way mixed-effects model ANOVA: time F(8216) = 2550, p < 0.0001; genotype/diet F(5,27) = 16.78, p < 0.0001; time x genotype/diet F(40,216) = 13.34, p < 0.0001. Tukey’s multiple comparison tests are denoted by p < 0.05 (^x), p < 0.01 (^xx), p < 0.001 (^xxx), and p < 0.0001 (^xxxx) where females x = a for Fmr1^HET Teklad 2019 versus Fmr1^HET CIF, x = b for Fmr1^HET Teklad 2019 versus Fmr1^HET SIF, x = c for Fmr1^HET Teklad 2019 versus Fmr1^KO 2019, x = d for Fmr1^HET Teklad 2019 versus Fmr1^KO CIF, x = e for Fmr1^HET Teklad 2019 versus Fmr1^KO SIF, x = f for Fmr1^HET CIF versus Fmr1^HET SIF, x = g for Fmr1^HET CIF versus Fmr1^KO 2019, x = h for Fmr1^HET CIF versus Fmr1^KO CIF, x = i for Fmr1^HET CIF versus Fmr1^KO SIF, x = j for Fmr1^HET SIF versus Fmr1^KO 2019, x = k for Fmr1^HET SIF versus Fmr1^KO CIF, x = l for Fmr1^HET SIF versus Fmr1^KO SIF, x = m for Fmr1^KO Teklad 2019 versus Fmr1^KO CIF, x = n for Fmr1^KO Teklad 2019 versus Fmr1^KO SIF, and x = o for Fmr1^KO CIF versus Fmr1^KO SIF, and males x = a for WT Teklad 2019 versus WT CIF, x = b for WT Teklad 2019 versus WT SIF, x = c for WT Teklad 2019 versus Fmr1^KO 2019, x = d for WT Teklad 2019 versus Fmr1^KO CIF, x = e for WT Teklad 2019 versus Fmr1^KO SIF, x = f for WT CIF versus WT SIF, x = g for WT CIF versus Fmr1^KO 2019, x = h for WT CIF versus Fmr1^KO CIF, x = i for WT CIF versus Fmr1^KO SIF, x = j for WT SIF versus Fmr1^KO 2019, x = k for WT SIF versus Fmr1^KO CIF, x = l for WT SIF versus Fmr1^KO SIF, x = m for Fmr1^KO Teklad 2019 versus Fmr1^KO CIF, x = n for Fmr1^KO Teklad 2019 versus Fmr1^KO SIF, and x = o for Fmr1^KO CIF versus Fmr1^KO SIF. (B) EchoMRI measurement of lean body mass. Female cohorts include Fmr1^HET/Teklad 2019 (n = 4), Fmr1^HET/CIF (n = 5), Fmr1^HET/SIF (n = 6), Fmr1^KO/Teklad 2019 (n = 6), Fmr1^KO/CIF (n = 7) and Fmr1^KO/SIF (n = 6). Statistics were determined by two-way ANOVA: interaction F(2,28) = 2.702, p = 0.085; genotype F(1,28) = 4.410, p = 0.045; diet F(2,28) = 7.438, p = 0.0026. Male cohorts include WT/Teklad 2019 (n = 6), WT/CIF (n = 6), WT/SIF (n = 4), Fmr1^KO/Teklad 2019 (n = 6), Fmr1^KO/CIF (n = 5) and Fmr1^KO/SIF (n = 7). Statistics were determined by two-way ANOVA: interaction F(2,28) = 0.1425, p = 0.87; genotype F(1,28) = 0.3857, p = 0.54; diet F(2,28) = 46.03, p < 0.0001. Tukey’s multiple comparison tests denoted by p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), and p < 0.0001 (****). Total body weight, fat mass, total water mass, and free water mass measurements assessed by EchoMRI are provided in the Supplementary Data. (C) Nest building female cohorts include Fmr1^HET/Teklad 2019 (n = 3), Fmr1^HET/CIF (n = 5), Fmr1^HET/SIF (n = 5), Fmr1^KO/Teklad 2019 (n = 6), Fmr1^KO/CIF (n = 7) and Fmr1^KO/SIF (n = 6). Statistics were determined by two-way ANOVA: interaction F(2,26) = 0.008736, p = 0.92; genotype F(1,26) = 0.1821, p = 0.67; diet F(2,26) = 1.86, p = 0.18. Male cohorts include WT/Teklad 2019 (n = 6), WT/CIF (n = 6), WT/SIF (n = 4), Fmr1^KO/Teklad 2019 (n = 6), Fmr1^KO/CIF (n = 5) and Fmr1^KO/SIF (n = 7). Statistics were determined by two-way ANOVA: interaction F(2,28) = 2.94, p = 0.069; genotype F(1,28) = 4.376, p = 0.046; diet F(2,28) = 2.845, p = 0.075. Tukey’s multiple comparison tests denoted by p < 0.05 (*) and p < 0.01 (**). (D) Passive avoidance female cohorts include Fmr1^HET/Teklad 2019 (n = 4), Fmr1^HET/CIF (n = 5), Fmr1^HET/SIF (n = 6), Fmr1^KO/Teklad 2019 (n = 6), Fmr1^KO/CIF (n = 7) and Fmr1^KO/SIF (n = 6). Statistics were determined by two-way ANOVA: interaction F(2,28) = 2.093, p = 0.14; genotype F(1,28) = 2.530, p = 0.12; diet F(2,28) = 1.642, p = 0.21. Male cohorts include WT/Teklad 2019 (n = 6), WT/CIF (n = 6), WT/SIF (n = 4), Fmr1^KO/Teklad 2019 (n = 6), Fmr1^KO/CIF (n = 5) and Fmr1^KO/SIF (n = 7). Statistics were determined by two-way ANOVA: interaction F(2,28) = 2.998, p = 0.066; genotype F(1,28) = 6.844, p = 0.014; diet F(2,28) = 3.229, p = 0.055. Tukey’s multiple comparison tests denoted by p < 0.05 (*). Data shown for 6-hours post-foot shock. Results were not statistically different for 24- and 48-h post-foot shock.