Evidence of the p75 pan-neurotrophic receptor in platelets and

comparison to the isoform found in brain

Samuel 1,2
Fleury , Imane 1,2
Boukhatem , Mélanie 1
Welman , Jessica Le 1,2
Blanc , Marie Lordkipanidzé 1,2
1Research center, Montreal Heart Institute, 2Faculty of Pharmacy, Université de Montréal

INTRODUCTION RESULTS DISCUSSION

Platelets 1. A band consistent with p75NTR is seen in human platelet lysates • Antibody-based methods suggest
• Crucial to thrombosis & haemostasis • Full-length receptor p75NTR may be present, but give
• Share many similarities with neurons expected to run between divergent results
Brain-derived neurotrophic factor (BDNF) 72 and 85 kDa as seen in • A more robust method of protein
the human cortex (C) identification (e.g. mass
• Involved in learning and memory through the
spectroscopy) is required to
tropomyosin receptor kinase B (TrkB) and the
• Truncated receptor expected confirm protein identity
p75 pan-neurotrophic receptor (p75NTR)
to run at 62-65 kDa CONCLUSIONS
• 100-1000x more abundant in platelets
• Blood and brain levels dysregulated in many • Differences in molecular • Platelets appear to express the full-
neurological diseases mass are observed in the length p75NTR isoform
p75NTR receptor cortex vs platelets • Antibodies should be carefully
• Two isoforms (full-length and truncated) characterized to study this receptor
• Many types of post-translational modifications • Not all antibodies correctly outside of the central nervous
identify p75NTR system
RESEARCH QUESTIONS
2. Differences in mass suggests different post-translational modifications • Platelets as peripheral biomarkers
of neuronal health: not ready for
• Do platelets express the p75NTR receptor?
prime time
• Does this receptor bear similarities to the
receptor found in the brain? FUTURE WORK
• Could platelets be used as peripheral • Investigate the function and
biomarkers of neuronal health? signaling of p75NTR in human
platelets
METHODS
• Determine the potential role of
Western Blotting
this receptor in platelet biology
• Denaturing and reducing conditions
ACKNOWLEDGMENTS
• Healthy human cortex & platelet lysates
3. Discrepancies are also seen with flow cytometry-based assays
• Antibodies raised against the intracellular and
extracellular domains
Protein Deglycosylation
• All N-glycans and simple O-linked glycans were
removed
Flow Cytometry
• Humanized (REA844) and mouse (M-1818)
antibodies against the extracellular domain
• Fixation in 1% paraformaldehyde
• Permeabilization with 0.1% Triton X-100