Planning and Analysis of Axon Degeneration Screening Experiments

Methods Mol Biol. 2020:2143:63-82. doi: 10.1007/978-1-0716-0585-1_6.

Abstract

A network of intersecting molecular pathways interacts to initiate and execute axon destruction. Maximum protection against axon degeneration likely requires more than manipulation of a single target. Here, we describe the process of designing a high-throughput arrayed screening assay for the identification of key factors responsible for axon destruction and/or protection. First, we go over some existing screens in the literature, then discuss the planning, tracking, analysis, and statistics around such a screening experiment. Prioritization of perturbations may allow laboratories to cost-effectively explore the process of screening. We also present the pairing of a combinatorial drug screen with a machine learning algorithm, predicting how to best modulate neurodegenerative and neuroprotective components.

Keywords: Analysis; Antitargets; Axon degeneration; Discovery; High-throughput; Hits; Nonhits; Screening; Support vector machine learning; Targets.

MeSH terms

  • Animals
  • Axons / physiology*
  • Axotomy
  • CRISPR-Cas Systems
  • Combinatorial Chemistry Techniques
  • Computer Simulation
  • Drug Evaluation, Preclinical / methods
  • Finches / embryology
  • High-Throughput Screening Assays / instrumentation
  • High-Throughput Screening Assays / methods*
  • Image Processing, Computer-Assisted
  • Nerve Degeneration / physiopathology*
  • Phenotype
  • Quality Control
  • RNA Interference
  • Retinal Ganglion Cells / cytology
  • Sensitivity and Specificity
  • Support Vector Machine