Association of Factor V Secretion with Protein Kinase B Signaling in Platelets from Horses with Atypical Equine Thrombasthenia.
Abstract: Two congenital bleeding diatheses have been identified in Thoroughbred horses: Glanzmann thrombasthenia (GT) and a second, novel diathesis associated with abnormal platelet function in response to collagen and thrombin stimulation. Objective: Platelet dysfunction in horses with this second thrombasthenia results from a secretory defect. Methods: Two affected and 6 clinically normal horses. Methods: Ex vivo study. Washed platelets were examined for (1) expression of the αIIb-β3 integrin; (2) fibrinogen binding capacity in response to ADP and thrombin; (3) secretion of dense and α-granules; (4) activation of the mammalian target of rapamycin (mTOR)-protein kinase B (AKT) signaling pathway; and (5) cellular distribution of phosphatidylinositol-4-phosphate-3-kinase, class 2B (PIK3C2B) and SH2 containing inositol-5'-phosphatase 1 (SHIP1). Results: Platelets from affected horses expressed normal amounts of αIIb-β3 integrin and bound fibrinogen normally in response to ADP, but bound 80% less fibrinogen in response to thrombin. α-granules only released 50% as much Factor V as control platelets, but dense granules released their contents normally. Protein kinase B (AKT) phosphorylation was reduced after thrombin activation, but mTOR Complex 2 (mTORC2) and phosphoinositide-dependent kinase 1 (PDK1) signaling were normal. SH2-containing inositol-5'-phosphatase 1 (SHIP1) did not localize to the cytoskeleton of affected platelets and was decreased overall consistent with reduced AKT phosphorylation. Conclusions: Defects in fibrinogen binding, granule secretion, and signal transduction are unique to this thrombasthenia, which we designate as atypical equine thrombasthenia.
Copyright © The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of American College of Veterinary Internal Medicine.
Publication Date: 2015-08-19 PubMed ID: 26290457PubMed Central: PMC4858037DOI: 10.1111/jvim.13595Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research investigates a unique blood-clotting disorder in Thoroughbred horses, referred to as atypical equine thrombasthenia. The study scrutinized the function of platelets, vibrantly analyzing the related defects with fibrinogen binding, secretion of granules, and signal transduction.
Study Design and Methods
- The research was an ex vivo study encompassing two affected horses and six clinically normal horses for comparative analysis.
- Platelets from these horses were washed and evaluated for a range of parameters. These included the expression of the αIIb-β3 integrin, fibrinogen binding capacity in response to ADP and thrombin, secretion of dense and alpha-granules, and the activation of the mammalian target of rapamycin (mTOR)-protein kinase B (AKT) signaling pathway. Additionally, the cellular distribution of phosphatidylinositol-4-phosphate-3-kinase, class 2B (PIK3C2B), and SH2-containing inositol-5′-phosphatase 1 (SHIP1) was also evaluated.
Key Findings
- Platelets from affected horses expressed normal quantities of αIIb-β3 integrin and exhibited an average ability to bind fibrinogen in response to ADP. However, they exhibited an 80% decrease in their capacity to bind fibrinogen in response to thrombin.
- In relation to Factor V secretion, alpha-granules from affected platelets released only 50% of the control level, although the dense granules released their contents normally.
- A marked reduction in Protein Kinase B (AKT) phosphorylation was observed post-thrombin activation. In contrast, signals from mTOR Complex 2 (mTORC2) and phosphoinositide-dependent kinase 1 (PDK1) remained normal.
- SHIP1, an important signaling protein, failed to localize to the cytoskeleton of the affected platelets and was found to be generally decreased. This observation is coherent with the reduction in AKT phosphorylation.
Conclusions
- The study unveils a unique clotting disorder, atypical equine thrombasthenia, in Thoroughbred horses characterized by specific defects in fibrinogen binding, granule secretion, and signal transduction.
- The findings could pave the way for further investigations to improve our understanding of the disorder and potentially inform the development of effective treatments or preventative measures.
Cite This Article
APA
Norris JW, Pombo M, Shirley E, Blevins G, Tablin F.
(2015).
Association of Factor V Secretion with Protein Kinase B Signaling in Platelets from Horses with Atypical Equine Thrombasthenia.
J Vet Intern Med, 29(5), 1387-1394.
https://doi.org/10.1111/jvim.13595 Publication
Researcher Affiliations
- 5A60 Johns Hopkins Asthma and Allergy Center, Johns Hopkins Medical Institute - Sidney Kimmel Comprehensive Cancer Center, Baltimore, MD.
- Department of Anatomy, Physiology and Cell Biology, School of Veterinary Medicine, University of California, Davis, CA.
- Hunters Glen Veterinary Hospital, Inc., Veterinary Acupuncture Services of Tulsa LLC, Tulsa, OK.
- 19463 James Monroe HWY, Leesburg, VA.
- Department of Anatomy, Physiology and Cell Biology, School of Veterinary Medicine, University of California, Davis, CA.
MeSH Terms
- Animals
- Blood Platelets / physiology
- Blotting, Western
- Case-Control Studies
- Factor V / analysis
- Fibrinogen / physiology
- Horse Diseases / blood
- Horse Diseases / physiopathology
- Horses
- Proto-Oncogene Proteins c-akt / blood
- Proto-Oncogene Proteins c-akt / physiology
- Signal Transduction / physiology
- Thrombasthenia / blood
- Thrombasthenia / physiopathology
- Thrombasthenia / veterinary
References
This article includes 35 references
- Livesey L, Christopherson P, Hammond A, Perkins J, Toivio-Kinnucan M, Insalaco T, Boudreaux MK. Platelet dysfunction (Glanzmann's thrombasthenia) in horses.. J Vet Intern Med 2005 Nov-Dec;19(6):917-9.
- Christopherson PW, van Santen VL, Livesey L, Boudreaux MK. A 10-base-pair deletion in the gene encoding platelet glycoprotein IIb associated with Glanzmann thrombasthenia in a horse.. J Vet Intern Med 2007 Jan-Feb;21(1):196-8.
- Christopherson PW, Insalaco TA, van Santen VL, Livesey L, Bourne C, Boudreaux MK. Characterization of the cDNA Encoding alphaIIb and beta3 in normal horses and two horses with Glanzmann thrombasthenia.. Vet Pathol 2006 Jan;43(1):78-82.
- Sanz MG, Wills TB, Christopherson P, Hines MT. Glanzmann thrombasthenia in a 17-year-old Peruvian Paso mare.. Vet Clin Pathol 2011 Mar;40(1):48-51.
- Macieira S, Lussier J, Bédard C. Characterization of the cDNA and genomic DNA sequence encoding for the platelet integrin alpha IIB and beta III in a horse with Glanzmann thrombasthenia.. Can J Vet Res 2011 Jul;75(3):222-7.
- Macieira S, Rivard GE, Champagne J, Lavoie JP, Bédard C. Glanzmann thrombasthenia in an Oldenbourg filly.. Vet Clin Pathol 2007 Jun;36(2):204-8.
- Bevers EM, Comfurius P, Nieuwenhuis HK, Levy-Toledano S, Enouf J, Belluci S, Caen JP, Zwaal RF. Platelet prothrombin converting activity in hereditary disorders of platelet function.. Br J Haematol 1986 Jun;63(2):335-45.
- Fry MM, Walker NJ, Blevins GM, Magdesian KG, Tablin F. Platelet function defect in a thoroughbred filly.. J Vet Intern Med 2005 May-Jun;19(3):359-62.
- Norris JW, Pratt SM, Auh JH, Wilson SJ, Clutter D, Magdesian KG, Ferraro GL, Tablin F. Investigation of a novel, heritable bleeding diathesis of Thoroughbred horses and development of a screening assay.. J Vet Intern Med 2006 Nov-Dec;20(6):1450-6.
- Norris JW, Pratt SM, Hunter JF, Gardner IA, Tablin F. Prevalence of reduced fibrinogen binding to platelets in a population of Thoroughbreds.. Am J Vet Res 2007 Jul;68(7):716-21.
- Woulfe D, Jiang H, Morgans A, Monks R, Birnbaum M, Brass LF. Defects in secretion, aggregation, and thrombus formation in platelets from mice lacking Akt2.. J Clin Invest 2004 Feb;113(3):441-50.
- Faraday N, Goldschmidt-Clermont P, Dise K, Bray PF. Quantitation of soluble fibrinogen binding to platelets by fluorescence-activated flow cytometry.. J Lab Clin Med 1994 May;123(5):728-40.
- Skaer RJ, Flemans RJ, McQuilkan S. Mepacrine stains the dense bodies of human platelets and not platelet lysosomes.. Br J Haematol 1981 Nov;49(3):435-8.
- Sarbassov DD, Guertin DA, Ali SM, Sabatini DM. Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex.. Science 2005 Feb 18;307(5712):1098-101.
- Sarbassov DD, Ali SM, Kim DH, Guertin DA, Latek RR, Erdjument-Bromage H, Tempst P, Sabatini DM. Rictor, a novel binding partner of mTOR, defines a rapamycin-insensitive and raptor-independent pathway that regulates the cytoskeleton.. Curr Biol 2004 Jul 27;14(14):1296-302.
- Guertin DA, Stevens DM, Thoreen CC, Burds AA, Kalaany NY, Moffat J, Brown M, Fitzgerald KJ, Sabatini DM. Ablation in mice of the mTORC components raptor, rictor, or mLST8 reveals that mTORC2 is required for signaling to Akt-FOXO and PKCalpha, but not S6K1.. Dev Cell 2006 Dec;11(6):859-71.
- Segura D, Monreal L. Poor reproducibility of template bleeding time in horses.. J Vet Intern Med 2008 Jan-Feb;22(1):238-41.
- Brass LF. Thrombin and platelet activation.. Chest 2003 Sep;124(3 Suppl):18S-25S.
- Jonnalagadda D, Izu LT, Whiteheart SW. Platelet secretion is kinetically heterogeneous in an agonist-responsive manner.. Blood 2012 Dec 20;120(26):5209-16.
- Wolberg AS. Thrombin generation and fibrin clot structure.. Blood Rev 2007 May;21(3):131-42.
- Duvernay M, Young S, Gailani D, Schoenecker J, Hamm HE. Protease-activated receptor (PAR) 1 and PAR4 differentially regulate factor V expression from human platelets.. Mol Pharmacol 2013 Apr;83(4):781-92.
- Chen J, De S, Damron DS, Chen WS, Hay N, Byzova TV. Impaired platelet responses to thrombin and collagen in AKT-1-deficient mice.. Blood 2004 Sep 15;104(6):1703-10.
- O'Brien KA, Stojanovic-Terpo A, Hay N, Du X. An important role for Akt3 in platelet activation and thrombosis.. Blood 2011 Oct 13;118(15):4215-23.
- Moore SF, Hunter RW, Hers I. mTORC2 protein complex-mediated Akt (Protein Kinase B) Serine 473 Phosphorylation is not required for Akt1 activity in human platelets [corrected].. J Biol Chem 2011 Jul 15;286(28):24553-60.
- Moore SF, van den Bosch MT, Hunter RW, Sakamoto K, Poole AW, Hers I. Dual regulation of glycogen synthase kinase 3 (GSK3)α/β by protein kinase C (PKC)α and Akt promotes thrombin-mediated integrin αIIbβ3 activation and granule secretion in platelets.. J Biol Chem 2013 Feb 8;288(6):3918-28.
- Li D, August S, Woulfe DS. GSK3beta is a negative regulator of platelet function and thrombosis.. Blood 2008 Apr 1;111(7):3522-30.
- Dangelmaier C, Manne BK, Liverani E, Jin J, Bray P, Kunapuli SP. PDK1 selectively phosphorylates Thr(308) on Akt and contributes to human platelet functional responses.. Thromb Haemost 2014 Mar 3;111(3):508-17.
- Chen X, Zhang Y, Wang Y, Li D, Zhang L, Wang K, Luo X, Yang Z, Wu Y, Liu J. PDK1 regulates platelet activation and arterial thrombosis.. Blood 2013 May 2;121(18):3718-26.
- Manna D, Albanese A, Park WS, Cho W. Mechanistic basis of differential cellular responses of phosphatidylinositol 3,4-bisphosphate- and phosphatidylinositol 3,4,5-trisphosphate-binding pleckstrin homology domains.. J Biol Chem 2007 Nov 2;282(44):32093-105.
- Banfić H, Downes CP, Rittenhouse SE. Biphasic activation of PKBalpha/Akt in platelets. Evidence for stimulation both by phosphatidylinositol 3,4-bisphosphate, produced via a novel pathway, and by phosphatidylinositol 3,4,5-trisphosphate.. J Biol Chem 1998 May 8;273(19):11630-7.
- Zhang J, Banfić H, Straforini F, Tosi L, Volinia S, Rittenhouse SE. A type II phosphoinositide 3-kinase is stimulated via activated integrin in platelets. A source of phosphatidylinositol 3-phosphate.. J Biol Chem 1998 Jun 5;273(23):14081-4.
- Giuriato S, Payrastre B, Drayer AL, Plantavid M, Woscholski R, Parker P, Erneux C, Chap H. Tyrosine phosphorylation and relocation of SHIP are integrin-mediated in thrombin-stimulated human blood platelets.. J Biol Chem 1997 Oct 24;272(43):26857-63.
- Giuriato S, Pesesse X, Bodin S, Sasaki T, Viala C, Marion E, Penninger J, Schurmans S, Erneux C, Payrastre B. SH2-containing inositol 5-phosphatases 1 and 2 in blood platelets: their interactions and roles in the control of phosphatidylinositol 3,4,5-trisphosphate levels.. Biochem J 2003 Nov 15;376(Pt 1):199-207.
- Damen JE, Liu L, Ware MD, Ermolaeva M, Majerus PW, Krystal G. Multiple forms of the SH2-containing inositol phosphatase, SHIP, are generated by C-terminal truncation.. Blood 1998 Aug 15;92(4):1199-205.
- Séverin S, Gratacap MP, Lenain N, Alvarez L, Hollande E, Penninger JM, Gachet C, Plantavid M, Payrastre B. Deficiency of Src homology 2 domain-containing inositol 5-phosphatase 1 affects platelet responses and thrombus growth.. J Clin Invest 2007 Apr;117(4):944-52.
Citations
This article has been cited 2 times.- Dahlgren AR, Tablin F, Finno CJ. Genetics of equine bleeding disorders. Equine Vet J 2021 Jan;53(1):30-37.
- Satué K, Gardon JC, Muñoz A. Clinical and laboratorial description of the differential diagnoses of hemostatic disorders in the horse. Iran J Vet Res 2020 Winter;21(1):1-8.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
