In situ seeding assay: A novel technique for direct tissue localization of bioactive tau

J Neuropathol Exp Neurol. 2024 Oct 1;83(10):870-881. doi: 10.1093/jnen/nlae059.

Abstract

Proteins exhibiting prion-like properties are implicated in tauopathies. The prion-like traits of tau influence disease progression and correlate with severity. Techniques to measure tau bioactivity such as RT-QuIC and biosensor cells lack spatial specificity. Therefore, we developed a histological probe aimed at detecting and localizing bioactive tau in situ. We first induced the recruitment of a tagged probe by bioactive Tau in human brain tissue slices using biosensor cell lysates containing a fluorescent probe. We then enhanced sensitivity and flexibility by designing a recombinant probe with a myc tag. The probe design aimed to replicate the recruitment process seen in prion-like mechanisms based on the cryo-EM structure of tau aggregates in Alzheimer disease (AD). Using this novel probe, we observed selective staining of misfolded tau in pre- and post-synaptic structures within neurofibrillary tangles and neurites, whether or not associated with neuritic plaques. The probe specifically targeted AD-associated bioactive tau and did not recognize bioactive tau from other neurodegenerative diseases. Electron microscopy and immunolabeling further confirmed the identification of fibrillar and non-fibrillar tau. Finally, we established a correlation between quantifying bioactive tau using this technique and gold standard biosensor cells. This technique presents a robust approach for detecting bioactive tau in AD tissues and has potential applications for deciphering mechanisms of tau propagation and degradation pathways.

Keywords: Alzheimer disease; astrocyte; histopathology; prion-like; synapse; tau; tauopathies.

MeSH terms

  • Alzheimer Disease* / metabolism
  • Alzheimer Disease* / pathology
  • Biosensing Techniques / methods
  • Brain* / metabolism
  • Brain* / pathology
  • Humans
  • Neurofibrillary Tangles / metabolism
  • Neurofibrillary Tangles / pathology
  • Tauopathies / metabolism
  • Tauopathies / pathology
  • tau Proteins* / metabolism

Substances

  • tau Proteins