Poster Presentation HUPO 2019 - 18th Human Proteome Organization World Congress

MS-based single-cell proteomic screening for oocyte quality during in vitro production a reality? (#824)

Gry Boe Hansen 1 , Rajesh Gupta 2 , Pawel Sadowski 2 , Nana Satake 3
  1. School of Veterinary Sciences, University of Queensland, Gatton, QLD, Australia
  2. Central Analytical Research Facility (CARF), Queensland University of Technology, Brisbane, QLD, Australia
  3. School of Agriculture and Food Science, The University of Queensland, St Lucia, QLD, Australia

Remarkable developments in resolution and sensitivity of MS-based proteomics techniques are continuously revolutionising the way we study cellular communication. Single cell proteome analysis is fast becoming a reality with a variety of sample input methods, such as imaging or flow cytometry interfaces, in addition to traditional chromatography giving a new dimension to study physiological mechanisms at the individual cellular level. The mammalian oocyte is one of the largest single cell with the ability to ultimately produce a multicellular being. Thus, the implications of application and integration of advanced MS-based single cell proteomics is vast. This is particularly true as a powerful tool in in vitro production (IVP) systems where individual oocyte qualities may be verified or embryo health screened prior to implantation without destruction of the main oocyte/embryo. One of the most important cellular collaborations for the successful maturation of an oocyte is the cumulous-oocyte complex (COC). This complex is absolute paramount in oocyte maturational health, with the presumptive changing combined proteome. Within the past few years, our group was given the opportunity to trial a technique developed by Bruker, Parallel Accumulation Serial Fragmentation (PASEF) coupled with Trapped Ion Mobility Spectrometry (timsTOF) on samples consisting of a single digested in vitro matured cattle COCs. The initial preliminary results of this trial showed the identification of approximately 4000 peptides and 2000 proteins, a ten-fold improvement compared to DDA technique trialled within our group on a pooled sample of 10 COCs. To expand and explore the limitations and applications of new MS techniques, the oogenesis to embryogenesis may be an ideal platform to assess changeable proteomes dependant on maturational stage. Furthermore, development of practical screening methods to assess these changes in the live individual oocyte and embryo now appears to be a genuine prospect that would improve production efficiency.