PMC

Effects of TPPU on ASD-like behavioral abnormalities in juvenile offspring after maternal glyphosate exposure. (A) Schedule of treatment and behavioral tests. Water or glyphosate (0.098%) was given to pregnant mice. Vehicle (5 mL/kg/d) or TPPU (3 mg/kg/d) was administered orally to pregnant mice from E5 to P21. Subsequently, all mice received normal water. Grooming test and three-chamber social interaction test were performed from P28 to P35. (B) Change of body weight of mothers (n = 5 or 6). Two-way ANOVA (glyphosate, F_1,17 = 7.66, P = 0.013; TPPU, F_1,17 = 9.14, P = 0.008; interaction, F_1,17 = 1.59, P = 0.225). (C) Grooming test. Treatment with TPPU significantly attenuated the increased grooming time in juvenile offspring after maternal glyphosate exposure. Two-way ANOVA (glyphosate, F_1,36 = 14.19, P = 0.001; TPPU, F_1,36 = 25.34, P < 0.001; interaction, F_1,36 = 11.31, P = 0.002). Data are shown as mean ± SEM (n = 10; ***P < 0.01 compared to glyphosate + vehicle group). (D) Three-chamber social interaction test. (Left) Three-way ANOVA (glyphosate, F_1,56 = 9.948, P = 0.003; TPPU, F_1,56 = 0.301, P = 0.585; stranger, F_1,56 = 135.27, P < 0.001; interaction [glyphosate × TPPU], F_1,56 = 0.845, P = 0.362; interaction [glyphosate × stranger], F_1,56 = 38.61, P < 0.001; interaction [TPPU × stranger], F_1,56 = 3.593, P = 0.063; interaction [glyphosate × TPPU × stranger], F_1,56 = 11.06, P = 0.002). (Right) Three-way ANOVA (glyphosate, F_1,56 = 4.168, P = 0.046; TPPU, F_1,56 = 21.96, P < 0.001; stranger, F_1,56 = 104.28, P < 0.001; interaction [glyphosate × TPPU], F_1,56 = 1.902, P = 0.173; interaction [TPPU × stranger], F_1,56 = 3.870, P = 0.054; interaction [glyphosate × stranger], F_1,56 = 13.10, P = 0.001; interaction [glyphosate × TPPU × stranger], F_1,56 = 19.69, P < 0.001). Data are shown as mean ± SEM (n = 8; ***P < 0.01). N.S., not significant.

Abundance of cells of selected taxa in control solution: C_0—the start of the experiment; C_3—after three days of the test; C_7—after seven days of the test; and in glyphosate concentrations of glyphosate tested of: 0.042 g·dm⁻³ glyphosate (0.042_3) after three and (0.042_7) after seven days of tests; 0.85 g·dm⁻³ glyphosate (0.85_3) after three and (0.85_7) after seven days of tests; 8.5 g·dm⁻³ glyphosate (8.5_3) after three and (8.5_7) after seven days of tests. Statistically significant differences were denoted with the asterisk. (A)—dominant species on which glyphosate had a negative effect on growth, (B)—cyanobacteria in which glyphosate stimulated growth selectively, (C)—resistant species on which glyphosate had a positive effect on abundance, (D)—sensitive species on which glyphosate had a negative effect on abundance.

Analysis of chase-phase glyphosate compartmentalization in horseweed leaf tissue exposed during the 10-h pulse phase either to buffer containing 10 mm glyphosate alone or simultaneously to buffer containing 10 mm glyphosate in addition to 10 mm or . Standard in-magnet perfusion conditions were employed: 10-h pulse phase, 2-h wash phase, 12-h chase phase, 20°C, 100% oxygen equilibration, and dark. A, ³¹P-NMR spectra, for the case when and glyphosate are delivered simultaneously, acquired over 2-h signal-averaging periods that occur during hours 14 to 24. B, ³¹P-NMR spectra, for the case when and glyphosate are delivered simultaneously, acquired over 2-h signal-averaging periods that occur during hours 14 to 24. Abbreviations regarding resonance assignments are as follows: CAMPA, cytosolic ; CG, cytosolic glyphosate; CNMG, cytosolic ; CPi, cytosolic ; Sugar-P, sugar phosphates; VAMPA, vacuolar ; VG, vacuolar glyphosate; VNMG, vacuolar ; and VPi, vacuolar . , an external chemical-shift and resonance-amplitude reference, is assigned a ³¹P chemical shift of 18.2 ppm relative to the ³¹P chemical shift of 85% phosphoric acid, which is taken as 0.00 ppm. C, Compartmentalization of cytosolic and vacuolar glyphosate during exposure to glyphosate alone (black bars) or simultaneously to glyphosate and (dark gray bars) or (light gray bars). For each pair of bars, the left bar represents glyphosate occupancy in the cytosol, and the right bar represents glyphosate occupancy in the vacuole. (Glyphosate compartmentalization data obtained when glyphosate is delivered alone were reported previously in bar graph format []). D, The plot shows the time (t) dependence of the glyphosate vacuolar sequestration ratio [VG/(VG + CG)] during the chase phase of the protocol. For leaf tissue exposed to glyphosate alone, fn(t) = 0.024t + 0.19 and r² = 0.99. For leaf tissue simultaneously exposed to glyphosate and , fn(t) = 0.019t + 0.41 and r² = 0.95. For leaf tissue simultaneously exposed to glyphosate and , fn(t) = 0.014t + 0.44 and r² = 0.88. Data in C and D represent means ± se (n = 3) and are normalized (ratio) to the resonance amplitude of the external reference (). Time labels/coordinates indicate the end of each 2-h signal-averaging increment.

FT-IR spectra of glyphosate, montmorillonite, and glyphosate adsorbed onto montmorillonite. M = montmorillonite; GPS = glyphosate, M-GPS = glyphosate adsorbed onto montmorillonite. Glyphosate (800 mg L⁻¹) was dissolved in distilled water and adsorbed onto montmorillonite.