Whole-Mount Adult Ear Skin Imaging Reveals Defective Neuro-Vascular Branching Morphogenesis in Obese and Type 2 Diabetic Mouse Models

Sci Rep. 2018 Jan 11;8(1):430. doi: 10.1038/s41598-017-18581-7.

Abstract

Obesity and type 2 diabetes are frequently associated with peripheral neuropathy. Though there are multiple methods for diagnosis and analysis of morphological changes of peripheral nerves and blood vessels, three-dimensional high-resolution imaging is necessary to appreciate the pathogenesis with an anatomically recognizable branching morphogenesis and patterning. Here we established a novel technique for whole-mount imaging of adult mouse ear skin to visualize branching morphogenesis and patterning of peripheral nerves and blood vessels. Whole-mount immunostaining of adult mouse ear skin showed that peripheral sensory and sympathetic nerves align with large-diameter blood vessels. Diet-induced obesity (DIO) mice exhibit defective vascular smooth muscle cells (VSMCs) coverage, while there is no significant change in the amount of peripheral nerves. The leptin receptor-deficient db/db mice, a severe obese and type 2 diabetic mouse model, exhibit defective VSMC coverage and a large increase in the amount of smaller-diameter nerve bundles with myelin sheath and unmyelinated nerve fibers. Interestingly, an increase in the amount of myeloid immune cells was observed in the DIO but not db/db mouse skin. These data suggest that our whole-mount imaging method enables us to investigate the neuro-vascular and neuro-immune phenotypes in the animal models of obesity and diabetes.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blood Vessels / diagnostic imaging*
  • Diabetes Mellitus, Type 2 / complications*
  • Disease Models, Animal
  • Ear / blood supply
  • Ear / diagnostic imaging*
  • Ear / innervation
  • Imaging, Three-Dimensional
  • Mice
  • Microscopy, Confocal / methods
  • Morphogenesis
  • Obesity / complications*
  • Peripheral Nerves / diagnostic imaging*
  • Peripheral Nervous System Diseases / diagnostic imaging*
  • Receptors, Leptin / genetics
  • Skin / blood supply
  • Skin / innervation

Substances

  • Receptors, Leptin
  • leptin receptor, mouse