Impact of the Maternal Microbiome on ENS Development and GI Motility of Offspring Público
Smoller, Charlie (Spring 2022)
Published
Abstract
The gut microbiome has become a growing field of research in biology due to its connection with multiple facets of human health including juvenile development, homeostasis, and metabolic function. Recently, research has found that the maternal gut microbiome plays a role in the development of the nervous system. However, studies have neglected to focus on the enteric nervous system (ENS), which is an autonomous component of the nervous system that regulates GI function and physiology. Here we show that the depletion of the maternal mouse microbiome through antibiotic administration during pregnancy causes the offspring to have structural deficits in the ENS and motility issues in the GI tract. By utilizing functional motility assays, we saw abnormalities in gut motility such as reduced transit through the gastrointestinal (GI) tract and signs of constipation in the 3-week-old offspring of antibiotic-treated mothers. Moreover, by utilizing immunohistochemistry techniques such as CLARITY to image the gut, we found deficits in the number of developing neuronal cells as well as lower myenteric plexus intensity and epithelial innervation in the ENS in both postnatal day 2 and 3-week-old mice. These data suggest that prescribing antibiotics to a mother during pregnancy could cause possible harm to offspring. It is also the first to find that the depletion of the maternal gut microbiome can cause deficits in the development of the ENS of offspring.
Table of Contents
Introduction …………………………………………………………………………………..….1
Methods…………………………………………………………………………………………....6
Ethical approval…………………………………………………………………………………..6
Animal Studies…………………………………………………………………………………….6
Antibiotic-Treated (Abx) Mice Experimental Plan………………………………………..6
DNA Extraction and qPCR to Validate the Maternal Antibiotic Model……………….7
GI Motility Assays…………………………………………………………………………………7
CLARITY Immunostaining………………………………………………………………………9
Confocal Imaging for CLARITY………………………………………………………………..10
Immunohistochemistry on Frozen Ileal Sections………………………………………….10
Arbitrary Fluorescence Quantification……………………………………………………….12
Statistical Analysis ……………………………………………………………………………….12
Results…………………………………………………………………………………………........12
Discussion…………………………………………………………………………………………...20
Overall Findings and Importance……………………………………………………………....20
Validation of the Antibiotic Model……………………………………………………………….21
Functional GI Issues of Offspring Born from Pregnant Mothers Treated with Antibiotics...21
Structural Issues of Offspring Born from Pregnant Mothers Treated with Antibiotics……..22
Limitations…………………………………………………………………………………………. 24
Conclusion…………………………………………………………………………………………. 25
References…………………………………………………………………………………………. 27
Tables and Figures
Table 1…………………………………………………………………………………………...11
Figure 1………………………………………………………………………………………….13
Figure 2………………………………………………………………………………………….16
Figure 3………………………………………………………………………………………….18
Figure 4………………………………………………………………………………………….20
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