Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Nov 28;15(23):4944.
doi: 10.3390/nu15234944.

Analysis of Gut Characteristics and Microbiota Changes with Maternal Supplementation in a Neural Tube Defect Mouse Model

Juan Antonio Cordero-Varela  1 , Marta Reyes-Corral  1 , Miguel Lao-Pérez  1 , Beatriz Fernández-Santos  1 , Fernando Montenegro-Elvira  1 , Lluis Sempere  1 , Patricia Ybot-González  1   2
Affiliations

Analysis of Gut Characteristics and Microbiota Changes with Maternal Supplementation in a Neural Tube Defect Mouse Model

Juan Antonio Cordero-Varela et al. Nutrients. .

Abstract

Adequate nutrient supply is crucial for the proper development of the embryo. Although nutrient supply is determined by maternal diet, the gut microbiota also influences nutrient availability. While currently there is no cure for neural tube defects (NTDs), their prevention is largely amenable to maternal folic acid and inositol supplementation. The gut microbiota also contributes to the production of these nutrients, which are absorbed by the host, but its role in this context remains largely unexplored. In this study, we performed a functional and morphological analysis of the intestinal tract of loop-tail mice (Vangl2 mutants), a mouse model of folate/inositol-resistant NTDs. In addition, we investigated the changes in gut microbiota using 16S rRNA gene sequencing regarding (1) the host genotype; (2) the sample source for metagenomics analysis; (3) the pregnancy status in the gestational window of neural tube closure; (4) folic acid and (5) D-chiro-inositol supplementation. We observed that Vangl2+/Lp mice showed no apparent changes in gastrointestinal transit time or fecal output, yet exhibited increased intestinal length and cecal weight and gut dysbiosis. Moreover, our results showed that the mice supplemented with folic acid and D-chiro-inositol had significant changes in their microbiota composition, which are changes that could have implications for nutrient absorption.

Keywords: Wnt/PCP; folic acid; inositol; loop-tail; metagenomics; microbiota; neural tube defects; pregnancy; prevention.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Experimental design for the study of gut microbiota in loop-tail mice. We investigated the changes in gut microbiota in relation to the host genotype (Vangl2+/+ vs Vangl2+/Lp), the sample source (feces vs cecum), the pregnancy status, and folic acid (FA) or D-chiro-inositol (CI) supplementation. To this end, feces and/or cecum samples were collected from five different groups of n = 9 female mice each: wild type Vangl2+/+ non-supplemented (NS) and non-pregnant mice (group 1); heterozygous Vangl2+/Lp non-supplemented and non-pregnant mice (group 2); and heterozygous Vangl2+/Lp pregnant mice non-supplemented (group 3); supplemented with FA (group 4); and supplemented with CI (group 5).
Figure 2
Figure 2
Functional and macroscopic morphological analysis of the intestinal tract of Vangl2+/+ (purple) and Vangl2+/Lp (dark yellow) female mice. (A) Total gastrointestinal transit time measured after oral gavage of dye. (B) Stool production in 1 h. (C) Average stool weight. (D) Stool water content. (E) Total gut length. (F) Small intestine length. (G) Colon length. (H) Cecum weight. Measurements in (E–H) were normalized to the weight of each animal (in grams). Unpaired two-sided t-test was applied to study differences between Vangl2+/+ and Vangl2+/Lp mice: ns, non-significant; ** p < 0.01; *** p < 0.001 (n = 8–10 mice per group).
Figure 3
Figure 3
Effect of the host genotype on gut microbiota. Richness and diversity measures of the microbiota from Vangl2+/+ (purple) and Vangl2+/Lp (dark yellow) female mice stool samples, according to the phylum (A–D) and the genus (E–G). (A,E) Alpha diversity described by the Hill numbers for each group: 0, species richness; 1, modified version of Shannon index; 2, equivalent to Simpson index. The higher the number, the stronger focus on dominant taxa. A Mann–Whitney test was applied to alpha diversity values to determine differences among groups of interest. (B,F) Non-metric multidimensional scaling (NMDS) analysis for beta microbiome diversity. (C,G) Significant difference (p < 0.05) between Vangl2+/+ and Vangl2+/Lp female mice stool samples at phylum and genera level. (D) Firmicutes/Bacteroidetes ratio.
Figure 4
Figure 4
Effect of the pregnancy status on gut microbiota. Richness and diversity measures of the microbiota from Vangl2+/Lp non-pregnant (green) and Vangl2+/Lp pregnant (orange) female mice cecum samples, according to the phylum (A,B) and the genera (C–E). (A,D) Alpha diversity described by the Hill numbers for each group: 0, species richness; 1, modified version of Shannon index; 2, equivalent to Simpson index. Higher number means a stronger focus on dominant taxa. A Mann–Whitney test was applied to alpha diversity values to determine differences among groups of interest. (B,E) NMDS analysis for beta microbiome diversity. (C) Firmicutes/Bacteroidetes ratio. (F) Significant differences (p < 0.05) in the relative abundance between Vangl2+/Lp non-pregnant and Vangl2+/Lp pregnant female mice cecum samples at genera level.
Figure 5
Figure 5
Effect of the dietary supplementation on gut microbiota. Richness and diversity measures of the microbiota from Vangl2+/Lp pregnant female mice cecum samples under different dietary conditions: control diet non-supplemented (NS, orange), supplemented with folic acid (FA, pink) and supplemented with D-chiro-inositol (CI, yellow) according to the phylum (A–D) and the genera (E–G). (A,E), Alpha diversity described by the Hill numbers for each group: 0, species richness; 1, modified version of Shannon index; 2, equivalent to Simpson index. Higher number means a stronger focus on dominant taxa. A Mann–Whitney test was applied to alpha diversity values to determine differences among groups of interest. (B,F) NMDS analysis for beta microbiome diversity. (C,G) Significant difference (p < 0.05) between Vangl2+/Lp pregnant female mice cecum samples from NS, FA and CI phylum and genus in cecum samples. (D) Firmicutes/Bacteroidetes ratio.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Cetin I., Bühling K., Demir C., Kortam A., Prescott S.L., Yamashiro Y., Yarmolinskaya M., Koletzko B. Impact of Micronutrient Status during Pregnancy on Early Nutrition Programming. Ann. Nutr. Metab. 2019;74:269–278. doi: 10.1159/000499698. - DOI - PubMed
    1. Tunçalp Ӧ., Pena-Rosas J.P., Lawrie T., Bucagu M., Oladapo O.T., Portela A., Metin Gülmezoglu A. WHO recommendations on antenatal care for a positive pregnancy experience-going beyond survival. BJOG. 2017;124:860–862. doi: 10.1111/1471-0528.14599. - DOI - PubMed
    1. Czeizel A.E., Dudas I. Prevention of the first occurrence of neural-tube defects by periconceptional vitamin supplementation. N. Engl. J. Med. 1992;327:1832–1835. doi: 10.1056/NEJM199212243272602. - DOI - PubMed
    1. Berry R.J., Li Z., Erickson J.D., Li S., Moore C.A., Wang H., Mulinare J., Zhao P., Wong L.Y., Gindler J., et al. Prevention of neural-tube defects with folic acid in China. China-U.S. Collaborative Project for Neural Tube Defect Prevention. N. Engl. J. Med. 1999;341:1485–1490. doi: 10.1056/NEJM199911113412001. - DOI - PubMed
    1. Berry R.J., Li Z. Folic acid alone prevents neural tube defects: Evidence from the China study. Epidemiology. 2002;13:114–116. doi: 10.1097/00001648-200201000-00021. - DOI - PubMed