Cell and Tissue Research

, Volume 376, Issue 3, pp 325–340 | Cite as

Intestinal nerve cell injury occurs prior to insulin resistance in female mice ingesting a high-fat diet

  • Yvonne Nyavor
  • Rachel Estill
  • Hannah Edwards
  • Hailey Ogden
  • Kaila Heideman
  • Kiefer Starks
  • Christopher Miller
  • George May
  • Lance Flesch
  • John McMillan
  • Martin Gericke
  • Larry Forney
  • Onesmo BalembaEmail author
Regular Article


Diabetic patients suffer from gastrointestinal disorders associated with dysmotility, enteric neuropathy and dysbiosis of gut microbiota; however, gender differences are not fully known. Previous studies have shown that a high-fat diet (HFD) causes type two diabetes (T2D) in male mice after 4–8 weeks but only does so in female mice after 16 weeks. This study seeks to determine whether sex influences the development of intestinal dysmotility, enteric neuropathy and dysbiosis in mice fed HFD. We fed 8-week-old C57BL6 male and female mice a standard chow diet (SCD) or a 72% kcal HFD for 8 weeks. We analyzed the associations between sex and intestinal dysmotility, neuropathy and dysbiosis using motility assays, immunohistochemistry and next-generation sequencing. HFD ingestion caused obesity, glucose intolerance and insulin resistance in male but not female mice. However, HFD ingestion slowed intestinal propulsive motility in both male and female mice. This was associated with decreased inhibitory neuromuscular transmission, loss of myenteric inhibitory motor neurons and axonal swelling and loss of cytoskeletal filaments. HFD induced dysbiosis and changed the abundance of specific bacteria, especially Allobaculum, Bifidobacterium and Lactobacillus, which correlated with dysmotility and neuropathy. Female mice had higher immunoreactivity and numbers of myenteric inhibitory motor neurons, matching larger amplitudes of inhibitory junction potentials. This study suggests that sex influences the development of HFD-induced metabolic syndrome but dysmotility, neuropathy and dysbiosis occur independent of sex and prior to T2D conditions. Gastrointestinal dysmotility, neuropathy and dysbiosis might play a crucial role in the pathophysiology of T2D in humans irrespective of sex.


Gut microbiota Diabetes mellitus Gastrointestinal motility Enteric nervous system Diabetic enteric neuropathy 



We would like to thank Catherine Brands and the UI Laboratory Animal Research Facility staff for their assistance during animal handling and Forrest Potter and Ann Norton (IBEST Optical Imaging Core) for their assistance with imaging and analysis. We thank Dan New, Dr. Matt Settles, Dr. Celeste Brown, Dr. Ben Ridenhour and the IBEST Genomics Resources Core for their help with microbial community analysis. We also thank Dr. Vanda A. Lennon of the Mayo Clinic for the gift of ANNA1 positive human serum.

Author contributions

Conception and design: YN, LF and OB. Development of methodology: YN, MG and OB. Acquisition of data: YN, RE, KS, CM, HE, KH, HO, JM, GM, MG and OB. Analysis and interpretation of data: YN, MG and OB. Writing, review and/or revision of the manuscript: YN, LF, MG and OB. Study supervision: OB.

Funding information

The research reported in this publication was supported by the University of Idaho—Dyess Faculty Fellowship and Institutional Development Awards (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant numbers P30GM103324 and P20 GM103408.

Compliance with ethical standards

Competing interests

The authors declare that they have no conflict of interest.


All authors have no competing interests financial or otherwise and have nothing to disclose.

Ethical approval

All applicable international, national and/or institutional guidelines for the care and use of animals were followed and all procedures performed in studies involving animals were in accordance with the ethical standards of the University of Idaho.

Ethical statement

All authors declare that this research was done by strictly adhering to the rules of good scientific practice and are responsible for its content. All experiments were performed in a manner that maximized rigor and reproducibility and without bias.

Supplementary material

441_2019_3002_MOESM1_ESM.docx (108 kb)
ESM 1 (DOCX 107 kb)
441_2019_3002_Fig7_ESM.png (232 kb)
Supplementary Fig. 1

HFD has different effects on the metabolism of male and female mice. Serum concentrations of leptin (a), resistin (b) and plasmogen activator inhibitor 1 (c, PAI-1) were measured by Milliplex assay. HFD increased the leptin concentration in both male and female mice but male mice had a significantly higher leptin concentration than female mice overall (a). Conversely, resistin was not affected by HFD in female mice even though it was increased in male mice (b). PAI-1 concentrations were increased by HFD ingestion in male mice but unaffected (reduced) in female mice (c). Data are expressed as mean ± SEM and analyzed by one-way analysis of variance followed by Tukey’s post-tests. (PNG 231 kb)

High Resolution Image (TIF 793 kb)
441_2019_3002_Fig8_ESM.png (1.4 mb)
Supplementary Fig. 2

HFD effects on duodenal motility. Pictures of duodenal segments from SCD-M (a), HFD-M (b), SCD-F (e) and HFD-F (f) mice during motility assays. Corresponding spatiotemporal maps (c-d, g-h) were generated in the GIMM program. Pictures and corresponding spatiotemporal maps show distinct contractions in SCD mice of both sexes (a, c, e, g). In contrast, HFD mice duodenums lack distinct uniform contractions (b, d, f, h) and in some cases, oral parts were severely distended and unable to contract (red asterisks) or relax properly. (PNG 1463 kb)

High Resolution Image (TIF 2679 kb)
441_2019_3002_Fig9_ESM.png (305 kb)
Supplementary Fig. 3

HFD determines microbial community composition. PCoA plot (a) shows gut microbial samples from both male and female mice fed SCD and HFD (PNG 305 kb)

High Resolution Image (TIF 877 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Yvonne Nyavor
    • 1
  • Rachel Estill
    • 1
  • Hannah Edwards
    • 1
  • Hailey Ogden
    • 1
  • Kaila Heideman
    • 1
  • Kiefer Starks
    • 1
  • Christopher Miller
    • 1
  • George May
    • 1
  • Lance Flesch
    • 1
  • John McMillan
    • 1
  • Martin Gericke
    • 2
  • Larry Forney
    • 1
  • Onesmo Balemba
    • 1
    Email author
  1. 1.Department of Biological SciencesUniversity of IdahoMoscowUSA
  2. 2.Institute for AnatomyUniversity of LeipzigLeipzigGermany

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