Preparation and Immunostaining of the Myenteric Plexus of Prion-Infected Mice

Methods Mol Biol. 2017:1658:285-292. doi: 10.1007/978-1-4939-7244-9_19.

Abstract

Intracerebral inoculation of mice with the M1000 strain of mouse-adapted human prions results in the consistent accumulation of PrPSc in the ileum of the gastrointestinal tract (GIT) of mice with clinical signs of prion disease. The accumulation of PrPSc in the ileum is accompanied by caspase activation and loss of immunoreactivity in subpopulations of neurons in the enteric nervous system. This suggests that like neurons in the central nervous system, cells in the enteric nervous system are also susceptible to prion-induced toxicity. In this chapter we describe the immunostaining of cells in myenteric plexus preparations of whole mounts prepared from the gastrointestinal tract of prion-infected mice.

Keywords: Fluorescence microscopy; Gastrointestinal tract; Ileum; Microdissection; Myenteric plexus; Neuron; Prion infection; Wholemount.

MeSH terms

  • Animals
  • Biomarkers / metabolism
  • Disease Models, Animal
  • Gene Expression
  • Glial Fibrillary Acidic Protein / genetics
  • Glial Fibrillary Acidic Protein / metabolism
  • Humans
  • Ileum / metabolism
  • Ileum / pathology*
  • Immunohistochemistry / methods*
  • Injections, Intraventricular
  • Mice
  • Mice, Transgenic
  • Microdissection / methods
  • Myenteric Plexus / metabolism
  • Myenteric Plexus / pathology*
  • Neurofilament Proteins / genetics
  • Neurofilament Proteins / metabolism
  • Neurons / metabolism
  • Neurons / pathology*
  • PrPSc Proteins / genetics
  • PrPSc Proteins / metabolism
  • Prion Diseases / genetics
  • Prion Diseases / metabolism
  • Prion Diseases / pathology*
  • Tissue Fixation / methods

Substances

  • Biomarkers
  • Glial Fibrillary Acidic Protein
  • Neurofilament Proteins
  • PrPSc Proteins
  • glial fibrillary astrocytic protein, mouse
  • neurofilament protein M