Details

Autor(en) / Beteiligte

• Clair, Geremy
• Piehowski, Paul D
• Nicola, Teodora
• Kitzmiller, Joseph A
• Huang, Eric L
• Zink, Erika M
• Sontag, Ryan L
• Orton, Daniel J
• Moore, Ronald J
• Carson, James P
• Smith, Richard D
• Whitsett, Jeffrey A
• Corley, Richard A
• Ambalavanan, Namasivayam
• Ansong, Charles

Titel

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

Ist Teil von

• Scientific reports, 2016-12, Vol.6 (1), p.39223-39223, Article 39223

Ort / Verlag

England: Nature Publishing Group

Erscheinungsjahr

2016

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• 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.