Spatially-Resolved Proteomics: Rapid Quantitative Analysis of Laser Capture Microdissected Alveolar Tissue Samples.

Academic Article

Abstract

  • Laser capture microdissection (LCM)-enabled region-specific tissue analyses are critical to better understand complex multicellular processes. However, current proteomics workflows entail several manual sample preparation steps and are challenged by the microscopic mass-limited samples generated by LCM, impacting measurement robustness, quantification and throughput. Here, we coupled LCM with a proteomics workflow that provides fully automated analysis of proteomes from microdissected tissues. Benchmarking against the current state-of-the-art in ultrasensitive global proteomics (FASP workflow), our approach demonstrated significant improvements in quantification (~2-fold lower variance) and throughput (>5 times faster). Using our approach we for the first time characterized, to a depth of >3,400 proteins, the ontogeny of protein changes during normal lung development in microdissected alveolar tissue containing only 4,000 cells. Our analysis revealed seven defined modules of coordinated transcription factor-signaling molecule expression patterns, suggesting a complex network of temporal regulatory control directs normal lung development with epigenetic regulation fine-tuning pre-natal developmental processes.
  • Published In

  • Scientific Reports  Journal
  • Keywords

  • Animals, Animals, Newborn, Automation, Chromatography, High Pressure Liquid, Laser Capture Microdissection, Lung, Mice, Mice, Inbred C57BL, Proteome, Proteomics, Tandem Mass Spectrometry
  • Digital Object Identifier (doi)

    Pubmed Id

  • 7993522
  • Author List

  • Clair G; Piehowski PD; Nicola T; Kitzmiller JA; Huang EL; Zink EM; Sontag RL; Orton DJ; Moore RJ; Carson JP
  • Start Page

  • 39223
  • Volume

  • 6