Fig 1.
Maternal obesity impairs energy balance and glucose homeostasis in the offspring and neonatal TUDCA treatment improves this metabolic malprogramming.
(A) Body weight curves of adult female mice fed a chow or an HFHS diet before and during pregnancy (n = 5 per group). (B) Body composition (n = 3 per group) and (C) GTT (n = 4–5 per group) of pregnant female mice fed a chow or HFHS diet at gestational day 16. (D) Body weight curves of mice born to chow-fed dams, HFHS-fed dams, or HFHS-fed dams and treated with neoTUDCA (n = 5–10 per group). (E) Average body composition (n = 5–8 per group) and (F) representative images and quantification of adipocyte size (immunostained for perilipin, green fluorescence) of 10-week-old mice born to chow-fed dams, HFHS-fed dams, or HFHS-fed dams and treated with TUDCA neonatally (n = 4–5 per group). (G) Food intake, (H) oxygen consumption, (I) energy expenditure, (J) RER, and (K) locomotor activity of 10-week-old mice born to chow-fed dams, HFHS-fed dams, or HFHS-fed dams and treated with TUDCA neonatally (n = 3–8 per group). (L) GTT and (M) ITT of 7- to 8-week-old mice born to chow-fed dams, HFHS-fed dams, or HFHS-fed dams and treated with TUDCA neonatally (n = 4–8 per group). Data are presented as mean ± SEM (panels A, C, D, J, L, M) or mean + SEM (panels B, E–I, K). *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, and ****P ≤ 0.0001 versus chow groups. Statistical significance between groups was determined by one-way ANOVA (panels E–G, I–K), and two-way ANOVA (A–D, H, L, M) followed by Tukey’s Multiple Comparison test. Scale bar, 100 μm. The underlying data are provided in S1 Data. GTT, glucose tolerance test; HFHS, high-fat high-sucrose; ITT, insulin tolerance test; neoTUDCA, tauroursodeoxycholic acid given neonatally; RER, respiratory exchange ratio; TUDCA, tauroursodeoxycholic acid.
Fig 2.
Neonatal TUDCA treatment reverses the elevated expression of ER stress markers in the offspring of obese dams.
Relative expression of Atf4, Atf6, Xbp1, Bip, and Chop mRNA in panels A and D the ARH, (E, F) PVH, (G, H) pancreas, (I, J) liver, and (K, L) fat depot of (A, E, G, I, and K) P10 and (D, F, H, J, and L) adult 10-week-old mice born to chow-fed dams, HFHS-fed dams, or HFHS-fed dams and treated with TUDCA neonatally (n = 4–6 per group). (B) Representative images and quantification of Atf4 and Bip mRNA in arcuate Pomc- and Agrp mRNA-expressing cells of P10 mice born to chow-fed dams, HFHS-fed dams, or HFHS-fed dams and treated with TUDCA neonatally (n = 4–9 per group). (C) Spliced form of Xbp1 in the ARH of P10 and mice born to chow-fed dams, HFHS-fed dams, or HFHS-fed dams and treated with TUDCA neonatally (n = 4–6 per group). Data are presented as mean + SEM. *P ≤ 0.05, **P < 0.01, ***P ≤ 0.001, and ****P ≤ 0.0001 versus other groups. Statistical significance between groups was determined by two-way ANOVA (panels A, B, D–L), and one-way ANOVA (panel C) followed by Tukey’s Multiple Comparison test. Scale bar, 5 μm. The underlying data are provided in S1 Data. ARH, arcuate nucleus; ER, endoplasmic reticulum; neoTUDCA, tauroursodeoxycholic acid given neonatally; PVH, paraventricular nucleus; TUDCA, tauroursodeoxycholic acid.
Fig 3.
Maternal obesity causes neonatal hyperleptinemia and attenuated response to leptin that can be reversed by neonatal TUDCA treatment.
(A) Serum leptin levels in dams at gestational day 16 and E16.5 fetuses of dams fed a chow or HFHS diet and in P10 and 10-week-old mice born to chow-fed dams, HFHS-fed dams, or HFHS-fed dams and treated with TUDCA neonatally (n = 4–8 per group). (B) Confocal images and quantification of the number of leptin-induced pSTAT3-immunoreactive cells in the ARH and DMH of P14 pups born to chow-fed dams, HFHS-fed dams, or HFHS-fed dams and treated with TUDCA neonatally (n = 5 per group). Data are presented as mean + SEM. *P ≤ 0.05 and **P < 0.01 versus chow groups. Statistical significance was determined by unpaired two-tailed Student t test (A), and one-way ANOVA followed by Tukey’s Multiple Comparison test (B). Scale bar, 100 μm. The underlying data are provided in S1 Data. ARH, arcuate nucleus; DMH, dorsomedial nucleus; HFHS, high-fat high-sucrose; neoTUDCA, tauroursodeoxycholic acid given neonatally; pSTAT3, phosphorylated signal transducer and activator of transcription 3; TUDCA, tauroursodeoxycholic acid.
Fig 4.
TUDCA treatment reverses neonatal disruption of POMC axonal projections induced by maternal obesity.
(A) Confocal images and quantification of the density of POMC-, αMSH-, and AgRP-immunoreactive fibers in the PVH of P14 mice born to chow-fed dams, HFHS-fed dams, or HFHS-fed dams and treated with TUDCA neonatally (n = 5–7 per group). (B) Relative expression of Pcsk1, Pcsk2, Cpe, Prcp, and Pam in the ARH of P10 mice born to chow-fed dams, HFHS-fed dams, or HFHS-fed dams and treated with TUDCA neonatally (n = 6 per group). (C) Confocal images and quantification of the density of POMC-, and AgRP-immunoreactive fibers in the PVH of 10- to 12-week-old mice born to chow-fed dams, HFHS-fed dams, or HFHS-fed dams and treated with TUDCA neonatally (n = 5–7 per group). Data are presented as mean + SEM. *P ≤ 0.05, **P < 0.01, ***P ≤ 0.001, and ****P ≤ 0.0001 versus other groups. Statistical significance was determined by one-way ANOVA (A-B) followed by Tukey’s Multiple Comparison test. Scale bar, 50 μm. The underlying data are provided in S1 Data. AgRP, agouti-related peptide; ARH, arcuate nucleus of the hypothalamus; HFHS, high-fat high-sucrose; neoTUDCA, tauroursodeoxycholic acid given neonatally; POMC, pro-opiomelanocortin; PVH, paraventricular nucleus of the hypothalamus; TUDCA, tauroursodeoxycholic acid; αMSH, α-melanocyte-stimulating hormone.
Fig 5.
Saturated fatty acid treatment causes ER stress-induced disruption of axon growth.
(A) Serum fatty acid levels in dams, P10 and 10-week-old mice born to chow-fed dams, HFHS-fed dams, or HFHS-fed dams and treated with TUDCA neonatally (n = 4–7 per group). (B) Relative expression of Atf4, Atf6, Xbp1, Bip, and Chop mRNA in mouse hypothalamic mHypoE-N43/5 cells treated with vehicle (BSA with 0.1% ethanol), or a cocktail of palmitate (250 μM) with lauric (1 mM) and myristic acids (200 μM) (PA+LA+MA), or OA alone for 24 h (n = 4–5 cultures per condition). (C) Representative images and quantification of the density of long-chain fatty acid analog BODIPY (green fluorescence) immunoreactivity in mHypoE-N43/5 cells treated with vehicle (BSA with 0.1% ethanol) or palmitate (250 μM) with lauric (1 mM) and myristic acids (200 μM) (PA+LA+MA) for 24 h (n = 5–7 cultures per condition). Red fluorescence and blue fluorescence depict actin filaments phalloidin and DAPI nuclear staining, respectively. (D) Confocal images and quantification of TUJ1 (neuron-specific class III beta-tubulin) immunoreactive fibers derived from isolated ARH explants incubated with vehicle (0.1% ethanol) or a combination of palmitate (250 μM) with lauric (1 mM) and myristic acids (200 μM) (PA+LA+MA) with or without TUDCA (750 μg/ml, n = 6 cultures per condition). Data are presented as mean + SEM. *P < 0.05, **P ≤ 0.01, ***P < 0.001 versus other groups. Statistical significance was determined by unpaired two-tailed Student t test (A, C, D), two-way ANOVA followed by Tukey’s Multiple Comparison test (B). Scale bars, 20 μm (C), and 50 μm (D). The underlying data are provided in S1 Data. ARH, arcuate nucleus; ER, endoplasmic reticulum; HFHS, high-fat high-sucrose; neoTUDCA, tauroursodeoxycholic acid given neonatally; OA, oleic acid; TUDCA, tauroursodeoxycholic acid; TUJ1, neuron-specific Class III β-tubulin.
Table 1.
Fatty acid composition of D12331 diet.