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Growth Arrest-Specific 6 (Gas6) and TAM Receptors in Mouse Platelets
Fikriye Uras1, Burhanettin Küçük, Özlem Bingöl Özakpınar
1Marmara University Faculty of Pharmacy, Department of Biochemistry, İstanbul, Turkey.
Objective:
Growth arrest-specific 6 (Gas6) is a newly discovered vitamin K-dependent protein, which is a ligand for TAM receptors [Tyro3 (Sky), Axl, and Mer] from the tyrosine kinase family. Gas6 knockout mice were resistant to venous and arterial thrombosis. There are contradictory reports on the presence of Gas6 and its receptors in mouse platelets. The objective of this study was to investigate whether Gas6 and its receptors were present in mouse platelets or not.
Materials And Methods:
Specific pathogen-free BALB/c male and female mice of 8-10 weeks old and 25-30 g in weight were anesthetized under light ether anesthesia and blood samples were taken from their hearts. RNAs were isolated from isolated platelets, and then mRNAs encoding Gas6 and TAM receptors were detected by reverse transcription-polymerase chain reaction (RT-PCR). Protein concentrations of Gas6 and TAM receptors in platelets were measured by ELISA, but not those of Mer, because of the absence of any commercial ELISA kit for mouse specimens.
Results:
RT-PCR results indicated the presence of mRNAs encoding Gas6 and Mer in mouse platelets. However, although RT-PCR reactions were performed at various temperatures and cycles, we could not detect the presence of mRNAs encoding Axl and Tyro3 (Sky). Receptor protein levels of Axl and Tyro3 were below the detection limits of the ELISA method.
Conclusion:
We found the presence of mRNAs encoding Gas6 and the receptor Mer in mouse platelets, but not Axl and Tyro3. Gas6, Axl, and Tyro3 protein levels were below the detection limits of the ELISA. The presence of mRNA is not obvious evidence of protein expression in platelets that have no nucleus or DNA. Further studies are required to clarify the presence of Gas6/TAM receptors in platelets using real-time PCR and more sensitive immunological methods, and future studies on mechanisms will indicate whether the Gas6/TAM pathway is a strategy for treatment of disorders.
Insights
This study investigated Growth arrest-specific 6 (Gas6) and its TAM receptors in mouse platelets. Researchers detected Gas6 and Mer mRNA, but not Axl or Tyro3, suggesting a potential role in platelet function.
Area of Science:
- Hematology
- Molecular Biology
- Tyrosine Kinase Signaling
Background:
- Growth arrest-specific 6 (Gas6) is a vitamin K-dependent protein and a ligand for TAM receptors (Tyro3, Axl, Mer).
- Gas6 knockout mice exhibit resistance to thrombosis, highlighting its role in hemostasis.
- Conflicting reports exist regarding Gas6 and TAM receptor presence in mouse platelets.
Purpose of the Study:
- To determine the presence of Gas6 and its TAM receptors (Tyro3, Axl, Mer) in mouse platelets.
- To clarify the expression of Gas6 and TAM receptor genes and proteins at the platelet level.
Main Methods:
- RNA isolation from mouse platelets followed by reverse transcription-polymerase chain reaction (RT-PCR) to detect specific mRNAs.
- Enzyme-linked immunosorbent assay (ELISA) to quantify protein levels of Gas6 and TAM receptors in platelets.
Main Results:
- RT-PCR confirmed the presence of mRNA for Gas6 and the Mer receptor in mouse platelets.
- mRNA for Axl and Tyro3 receptors was not detected in mouse platelets.
- Protein levels for Gas6, Axl, and Tyro3 were below ELISA detection limits.
Conclusions:
- Mouse platelets express mRNA for Gas6 and the Mer receptor, but not Axl or Tyro3.
- The absence of detectable protein levels necessitates further investigation using more sensitive methods.
- Clarifying Gas6/TAM receptor presence in platelets is crucial for understanding their role in thrombosis and potential therapeutic strategies.
