Abstract Number: PB0367
Meeting: ISTH 2022 Congress
Theme: Platelets and Megakaryocytes » Platelet Proteomics and Genomics
Background: Platelets inherit mRNA from their parent megakaryocyte. Young newly formed platelets are characterized as those with the highest levels of inherited mRNA, as this is rapidly lost once platelets are released, and age, within the circulation. Previous studies have identified the general decay half-life of platelet mRNA to be approximately 6 hours, but it is unclear if this is decay is similar for all mRNAs.
Aims: To use in vivo temporal labelling to investigate platelet mRNA dynamics.
Methods: Fluorescent anti-CD42c antibodies (Emfret) were injected (i.v) at various timepoints to C57Bl6 mice (12-14wk) to identify age-specific platelet populations: younger ( < 24h), older (>1 days old) and global. Following isolation by cell sorting, RNA was extracted (2.5million per sample) and qRT-PCR performed for a panel of highly expressed platelet mRNAs. Unbiased clustering analysis was performed to compare age and abundance of individual RNAs.
Results: Principal component analysis demonstrated each analyzed population clustered individually but with differential directionality for individual mRNAs; indicating potential for individual patterns of decreased abundance. Focused comparison and grouping of mRNAs revealed the amount of loss varied by specific mRNA (median fold change ranging from -5.3 to -66.4) but was not contingent on starting abundance. In particular, those mRNAs classified as associated with signaling proteins (Gnas, Atp2a3, Alox12, Rgs18, Sh3bgrl3) demonstrated the greatest reduction in abundance, whilst those associated with granule proteins (Srgn, Nrgn, B2m, Pf4, Pbpp) demonstrated the least (mean ±s.e.m fold change: -35.6 ±6.8 versus -9.2 ±1.9 respectively, p < 0.05, n=4).
Conclusion(s): The variable rates of loss observed in this dataset strongly suggest that platelet mRNA is under more refined control than previously appreciated. Further research is required to determine the mechanisms and purpose of such regulation, particularly if retention of granule protein encoding mRNAs correlates with the phenomenon of downstream de novo protein production previously reported by some groups.
To cite this abstract in AMA style:
Armstrong P, Kirkby N, Gutmann C, Allan H, Joshi A, Crescente M, Mitchell J, Mayr M, Warner T. Temporal labelling of ageing platelets in mice indicates differential regulation and retention of granule protein encoding mRNAs [abstract]. https://abstracts.isth.org/abstract/temporal-labelling-of-ageing-platelets-in-mice-indicates-differential-regulation-and-retention-of-granule-protein-encoding-mrnas/. Accessed March 21, 2024.« Back to ISTH 2022 Congress
ISTH Congress Abstracts - https://abstracts.isth.org/abstract/temporal-labelling-of-ageing-platelets-in-mice-indicates-differential-regulation-and-retention-of-granule-protein-encoding-mrnas/