TY - JOUR
T1 - Differential dynamics of early stages of platelet adhesion and spreading on collagen IV and fibrinogen-coated surfaces [version 2; peer review
T2 - 3 approved]
AU - Horev, Melanie B.
AU - Zabary, Yishaia
AU - Zarka, Revital
AU - Sorrentino, Simona
AU - Medalia, Ohad
AU - Zaritsky, Assaf
AU - Geiger, Benjamin
N1 - Funding Information:
Grant information: AZ is supported by the Israeli Council for Higher Education (CHE) via Data Science Research Center, Ben-Gurion University of the Negev. The research was supported in part from a grant from The Israel Science Foundation (ISF). The work was funded in part by ERC-Syn 810057 and the Swiss National Science Foundation Grant (SNSF 31003A_179418) to O.M.
Publisher Copyright:
© 2020. Horev MB et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
PY - 2021/1/1
Y1 - 2021/1/1
N2 - Background: Upon wound formation, platelets adhere to the neighboring extracellular matrix and spread on it, a process which is critical for physiological wound healing. Multiple external factors, such as the molecular composition of the environment and its mechanical properties, play a key role in this process and direct its speed and outcome. Methods: We combined live cell imaging, quantitative interference reflection microscopy and cryo-electron tomography to characterize, at a single platelet level, the differential spatiotemporal dynamics of the adhesion process to fibrinogen and collagen IV-functionalized surfaces. Results: Initially, platelets sense both substrates by transient rapid extensions of filopodia. On collagen IV, a short-term phase of filopodial extension is followed by lamellipodia-based spreading. This transition is preceded by the extension of a single or couple of microtubules into the platelet’s periphery and their apparent insertion into the core of the filopodia. On fibrinogen surfaces, the filopodia-tolamellipodia transition was partial and microtubule extension was not observed leading to limited spreading, which could be restored by manganese or thrombin. Conclusions: Based on these results, we propose that interaction with collagen IV stimulate platelets to extend microtubules to peripheral filopodia, which in turn, enhances filopodial-to-lamellipodial transition and overall lamellipodia-based spreading. Fibrinogen, on the other hand, fails to induce these early microtubule extensions, leading to full lamellipodia spreading in only a fraction of the seeded platelets. We further suggest that activation of integrin αIIbβ3 is essential for filopodial-to-lamellipodial transition, based on the capacity of integrin activators to enhance lamellipodia spreading on fibrinogen.
AB - Background: Upon wound formation, platelets adhere to the neighboring extracellular matrix and spread on it, a process which is critical for physiological wound healing. Multiple external factors, such as the molecular composition of the environment and its mechanical properties, play a key role in this process and direct its speed and outcome. Methods: We combined live cell imaging, quantitative interference reflection microscopy and cryo-electron tomography to characterize, at a single platelet level, the differential spatiotemporal dynamics of the adhesion process to fibrinogen and collagen IV-functionalized surfaces. Results: Initially, platelets sense both substrates by transient rapid extensions of filopodia. On collagen IV, a short-term phase of filopodial extension is followed by lamellipodia-based spreading. This transition is preceded by the extension of a single or couple of microtubules into the platelet’s periphery and their apparent insertion into the core of the filopodia. On fibrinogen surfaces, the filopodia-tolamellipodia transition was partial and microtubule extension was not observed leading to limited spreading, which could be restored by manganese or thrombin. Conclusions: Based on these results, we propose that interaction with collagen IV stimulate platelets to extend microtubules to peripheral filopodia, which in turn, enhances filopodial-to-lamellipodial transition and overall lamellipodia-based spreading. Fibrinogen, on the other hand, fails to induce these early microtubule extensions, leading to full lamellipodia spreading in only a fraction of the seeded platelets. We further suggest that activation of integrin αIIbβ3 is essential for filopodial-to-lamellipodial transition, based on the capacity of integrin activators to enhance lamellipodia spreading on fibrinogen.
KW - Fibrinogen
KW - Integrin αIIbβ3
KW - Interference Reflection Microscopy (IRM)
KW - Live cell imaging
KW - Microtubules
KW - Platelet spreading
KW - Type IV collagen
UR - http://www.scopus.com/inward/record.url?scp=85117295700&partnerID=8YFLogxK
U2 - 10.12688/F1000RESEARCH.23598.2
DO - 10.12688/F1000RESEARCH.23598.2
M3 - Article
AN - SCOPUS:85117295700
VL - 9
SP - 1
EP - 27
JO - F1000Research
JF - F1000Research
SN - 2046-1402
ER -