Öznur ASLAN, KORHAN ARSLAN, İhsan KELEŞ, BİLAL AKYÜZ, Mehmet ULUSAN, Esma Gamze İLGAR, AYTAÇ AKÇAY

The Relationship Between von Willebrand Factor Gene and von Willebrand Factor Antigen Levels in Dogs

Bu çalışmada, köpeklerde von Willebrand faktör geninin 42. ekzonunun 2380. pozisyonunda meydana gelen lösin (L) - prolin (P)'nin yer değiştirmesine neden olan mutasyon sonucu oluşan polimorfizm ile plazma vWF antijen seviyeleri arasındaki ilişkinin araştırılması amaçlandı. Çalışmaya, Ocak 2014 ve 2015 yılları arasında, Erciyes Üniversitesi Veteriner Fakültesi Kliniklerine getirilen farklı ırk ve yaşta (95 erkek, 66 dişi) 161 köpek dahil edildi. Köpeklerin klinik muayeneleri yapılarak EDTA ve Na sitratlı tüplere kan örnekleri toplandı. Kan örneklerinde PT, APTT, TT, fibrinojen, D-dimer, trombosit, vWF antigen (vWF: Ag) seviyeleri belirlendi ve VWF elde edildi. Genotipler, yapılan PCR sonucunda elde edilen ürünlerin MspI endonükleaz enzimi ile kesilerek belirlendi. Laboratuar muayenesinde 34 köpek (%21.1) von Willebrand açısından pozitif olarak belirlenirken, 127 köpeğin (%78.9) negatif olduğu belirlendi. Çalışmaya dahil edilen hayvanlarda ırklara göre hastalığın görülme oranları Golden Retreiver (n=35) %34.3, Kangal (n=45) %11.1, German Shepherd (n=11) %36.4, Labrador Retriever (n=7) %42.9, English Cocker Spaniel (n=2) %100, English Pointer (n=8) %25.0, melez (n=8) %33.3, Husky (n=7) %12.5, Malinois (n=1) %100, Dogo Argentino (n=2) %50.0, Samoyed (n=1) %100 ve diğer ırklar %0 olarak belirlendi. Hem pozitif hem de negatif köpeklerde VWF genotipi ile vWF: Ag değerleri arasında istatistiksel olarak bir ilişkili bulunamadı (P=0.675). vWD pozitif ve negatif köpeklerde oransal dağılım açısından karşılaştırıldığında hastalık oranı ve genotip arasında istatistik bir önem gözlenmedi (P=0.969). Sonuç olarak, köpeklerde vWF antigen seviyesi ve VWF arasında ilişki bulunmadı. Bilgilerimize göre bu çalışma, VWF and vWF: Ag arasındaki ilişkinin araştırıldığı ilk çalışma olması nedeniyle önemlidir.

Köpeklerde von Willebrand Faktör Antijen Seviyesi ve von Willebrand Faktör Geni Arasındaki İlişkinin Belirlenmesi

In this study, the relationship between plasma vWF antigen level and polymorphism occurring due to the leucine (L) to proline (P) substitution at position 2380 in exon 42 in von Willebrand factor gene (VWF) in dogs was aimed to investigate. The present study was performed on 161 dogs of various breeds and ages (95 male, 66 female) referred to Faculty of Veterinary Clinics at Erciyes University between January 2014 and 2015. Blood samples were collected in EDTA and Na citrate tubes after clinical examination of the dogs. PT, APTT, TT, fibrinogen, D-dimer, thrombocyte, vWF antigen (vWF: Ag) levels and VWF were determined from the blood samples. Genotypes were examined using PCR and restriction endonucleas enzymes. In the laboratory examination, 34 (21.1%) of the cases were positive and 127 (78.9%) of the cases were negative with concern to von Willebrand. Prevalence of the disease in different dog breeds that included to the present study were Golden Retreiver (n=35) 34.3%, Kangal (n=45) 11.1%, German Shepherd (n=11) 36.4%, Labrador Retriever (n=7) 42.9%, English Cocker Spaniel (n=2) 100%, English Pointer (n=8) 25.0%, mix breed (n=8) 33.3%, Husky (n=7) 12.5%, Malinois (n=1) 100%, Dogo Argentino (n=2) 50.0%, Samoyed (n=1) 100% and some other breeds were 0%. The genotype of VWF was not statistically significant in both positive and negative dogs with concern to vWF: Ag values (P=0.675). When comparison were made in terms of proportional distribution in positive and negative dogs, statistical importance were not observed between genotype and the disease ratio (P=0.969). In conclusion, relationship between VWF and vWF antigen level were not determined in dogs. According to our knowledge; this study is the first repot on VWF and vWF: Ag relationship therefore believed to be important.

PDF

___

Brooks MB, Castillo-Juarez H, Oltenacu P: Heritability of plasma von Willebrand factor antigen concentration in German Wirehaired pointers. Vet Q, 23, 126-128, 2001. DOI: 10.1080/01652176.2001.9695098
Burgess HJ, Woods JP, Abrams-Ogg AC, Wood RD: Evaluation of laboratory methods to improve characterization of dogs with von Willebrand disease. Can J Vet Res, 73 (4): 252-259, 2009.
Kramer JW, Venta PJ, Klein SR, Cao Y, Schall WD, Yuzbasiyan-Gurkan VA: von Willebrand's factor genomic nucleotide variant and polymerase chain reaction diagnostic test associated with inheritable type-2 von Willebrand's disease in a line of German Shorthaired Pointer dogs. Vet Pathol, 41, 221-228, 2004. DOI: 10.1354/vp.41-3-221
DeWilde L: Canine inherited bleeding disorders. The North American Veterinary Conference, 48-51, 2007.
Burgess H, Wood D: Validation of a von Willebrand factor antigen enzyme-linked immunosorbent assay and newly developed collagen- binding assay. Can J Vet Res, 72 (5): 420-427, 2008.
Sadler JE: Biochemistry and genetics of von Willebrand factor. Annu Rev Biochem, 67, 395-424, 1998. DOI: 10.1146/annurev.biochem.67.1.395
Venta PJ, Li J, Yuzbasiyan-Gurkan V, Brewer GJ, Schall WD: Mutation causing von Willebrand's disease in Scottish Terriers. J Vet Intern Med, , 10-19, 2000. DOI: 10.1111/j.1939-1676.2000.tb01493.x
Venta PJ, Vidal M: MspI RSP in exon 42 of the canine von Willebrand factor (VWF) gene. Animal Genetics, 30, 229, 1999. DOI: 10.1046/j.1365- 1999.00404-5.x
Lillicrap D: The molecular genetics of von Willebrand disease. Haematol Rep, 1, 3-8, 2005.
Ginsburg D, Bowie WEJ: Molecular genetics of von Willebrand Disease. Blood, 79 (10): 2507-2519, 1992.
Brooks M, Raymond S, Catalfamo J: Plasma von Willebrand factor antigen concentration as a predictor of von Willebrand's disease status in German Wirehaired Pointers. J Am Vet Med Assoc, 209 (5): 930-933, 1996.
Kayar A, Or ME, Gönül R, Akdenizli E, Yaramiş Ç, Bakirel U, Dodurka HT: von Willebrand's disease in German Shepherd Dogs in the Marmara Region (Turkey). Revue Méd Vét, 157 (1): 11-15, 2006.
Raymond SL, Jones DW, Brooks MB, Dodds J: Clinical and laboratory features of a severe form of von Willebrand disease in Shetland Sheepdogs. J Am Vet Med Assoc, 197, 1342-1346, 1990.
Pathak EJ: Type 3 von Willebrand's disease in a Shetland sheepdog. Can Vet J, 45 (8): 685-687, 2004.
Thomas JS: von Willebrand's disease in the dog and cat. Vet Clin North Am: Small Anim Pract, 26, 1089-1110, 1996. DOI: 10.1016/S0195- (96)50057-4
Desch KC, Ozel AB, Siemieniak D, Kalish Y, Shavit JA, Thornburg CD, Sharathkumar AA, McHugh CP, Laurie CC, Crenshaw A, Mirel DB, Kim Y, Cropp CD,Molloy AM, Kirke PN, Bailey-Wilson JE, Wilson AF, Mills JL, Scott JM, Brody LC, Li JZ, Ginsburg D: Linkage analysis identifies a locus for plasma von Willebrand factor undetected by genome-wide association. Proc Natl Acad Sci U S A, 110, 588-593, 2013. DOI: 10.1073/pnas.1219885110
Castaman G, Federici AB, Rodeghiero F, Mannucci PM: von Willebrand's disease in the year 2003: Towards the complete identification of gene defects for correct diagnosis and treatment. Haematologica, 88 (1): 94-108, 2003.
Venta PJ, Li J, Yuzbasiyan-Gurkan V, Brewer GJ, Schall WD: Mutation causing von Willebrand's disease in Scottish Terriers. J Vet Intern Med, 14, 10-19, 2000. DOI: 10.1892/0891-6640(2000)014_____0010:MCVWDI> 3.CO;2
Brooks M, Dodds WJ, Raymond SL: Epidemiologic features of von Willebrand's disease in Doberman Pinschers, Scottish Terriers, and Shetland Sheepdogs: 260 cases (1984-1988). J Am Vet Med Assoc, 200, 1127, 1992.
Stokol T, Parry BW, Mansell PD: Von Willebrand's disease in Scottish Terriers in Australia. Aust Vet J, 72, 404-407, 1995. DOI: 10.1111/j.1751- 1995.tb06188.x
Green RA, Thomas JS: Hemostatic disorders: Coagulopathies and thrombosis. In, Ettinger SJ, Feldman EC (Eds): Textbook of Veterinary Internal Medicine. 4th ed., 1946-1963, WB Sanders, Philadelphia, 1995.
Slappendel RJ, Beijer EG, van Leeuwen M: Type III von Willebrand's disease in Dutch Kooiker dogs. Vet Q, 20 (3): 93-97, 1998.
Wannamethee SG, Whincup PH, Lennon L, Rumley A, Lowe GD: Fibrin D-dimer, tissue-type plasminogen activator, von Willebrand factor, and risk of incident stroke in older men. Stroke, 43, 1206-1211, 2012. DOI: 10.1161/STROKEAHA.111.636373
Clark P, Parry BW: Survey of Irish Wolfhound in Australia for von Willebrands disease. Aust Vet J, 72, 393, 1995. DOI: 10.1111/j.1751- 1995.tb06179.x
Moser J,Meyers KM, Russon RH: Inheritance of von Willebrand factor deficiency in Doberman Pinschers. J Am Vet Med Assoc, 209, 1103- , 1996.
Sadler JE, Ginsburg D: A database of polymorphisms in the von Willebrand factor gene and pseudogene. For the consortium on von Willebrand factor mutations and polymorphisms and the subcommittee on von Willebrand factor of the scientific and standardization committee of the international society on thrombosis and haemostasis. Thromb Haemost, 69 (2): 185-191, 1993.