Gastroenterology

Gastroenterology

Volume 153, Issue 6, December 2017, Pages 1594-1606.e2
Gastroenterology

Original Research
Full Report: Basic and Translational—Alimentary Tract
Paneth Cell Defects Induce Microbiota Dysbiosis in Mice and Promote Visceral Hypersensitivity

https://doi.org/10.1053/j.gastro.2017.08.044Get rights and content

Background & Aims

Separation of newborn rats from their mothers induces visceral hypersensitivity and impaired epithelial secretory cell lineages when they are adults. Little is known about the mechanisms by which maternal separation causes visceral hypersensitivity or its relationship with defects in epithelial secretory cell lineages.

Methods

We performed studies with C3H/HeN mice separated from their mothers as newborns and mice genetically engineered (Sox9flox/flox-vil-cre on C57BL/6 background) to have deficiencies in Paneth cells. Paneth cell deficiency was assessed by lysozyme staining of ileum tissues and lysozyme activity in fecal samples. When mice were 50 days old, their abdominal response to colorectal distension was assessed by electromyography. Fecal samples were collected and microbiota were analyzed using Gut Low-Density Array quantitative polymerase chain reaction.

Results

Mice with maternal separation developed visceral hypersensitivity and defects in Paneth cells, as reported from rats, compared with mice without maternal separation. Sox9flox/flox-vil-Cre mice also had increased visceral hypersensitivity compared with control littermate Sox9flox/flox mice. Fecal samples from mice with maternal separation and from Sox9flox/flox-vil-cre mice had evidence for intestinal dysbiosis of the microbiota, characterized by expansion of Escherichia coli. Daily gavage of conventional C3H/HeN adult mice with 109 commensal E coli induced visceral hypersensitivity. Conversely, daily oral administration of lysozyme prevented expansion of E coli during maternal separation and visceral hypersensitivity.

Conclusions

Mice with defects in Paneth cells (induced by maternal separation or genetically engineered) have intestinal expansion of E coli leading to visceral hypersensitivity. These findings provide evidence that Paneth cell function and intestinal dysbiosis are involved in visceral sensitivity.

Section snippets

Mouse Models

All experimental protocols described in this study were approved by the local Animal Care Use Committee (Comité d'Ethique de Pharmacologie-Toxicologie de Toulouse - Midi-Pyrénées, France) registered as N°86 at the Ministry of Research and Higher Education (N° 0029/SMVT), and conducted in accordance with the European directive 2010/63/UE. All experiments were performed on Day (D) 50 mice. More details are in Supplementary Materials and Methods.

MS Protocol

To minimize cannibalism induced by perinatal stress,

MS in Mice Induced Visceral Hypersensitivity

As a first step in this study, we established a MS model in C3H/HeN mice.

No weight differences were observed among groups of mice from D2 to D50.

The electromyographic (EMG) response to graded colorectal distension was used as an index of visceral sensitivity. MS led to allodynia at day 50 in response to colorectal distension with low volume (P = .028 at 0.02 mL; P = .007 at 0.04 mL; P = .003 at 0.06 mL compared with control; Figure 1A; representative EMG recording is presented in Supplementary

Discussion

Paneth cells producing enteric antimicrobial peptides are important players in small intestine innate immunity. In mice, Paneth cells appear only 2 weeks after birth31, 32, 33, 34 and they are sensitive to environmental factors including enteric infections35, 36 and early life stress.22 Indeed, MS performed in rats decreases epithelial secretory cell lineages number including Paneth cells.22 Furthermore, neonatal MS induces visceral hypersensitivity in adulthood.23 This observation questioned a

Acknowledgments

The authors gratefully acknowledge Yannick Lippi and Claire Naylies from Transcriptomic impact of Xenobiotics platform for technical assistance for microbiota analysis.

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    Conflicts of interest The authors disclose no conflicts.

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