MEHMET KÖSE, MEHMET OSMAN ATLI, Murat GÖRGÜLÜ, MEHMET SALİH KAYA, NURETTİN AYDİLEK, Faruk BOZKAYA, TAHİR BAYRIL, ERCAN KURAR, Aydın GÜZELOĞLU, ZEKERİYA KIYMA

Expression profiles of interferon-tau stimulated genes (ISGs) in peripheral blood leucocytes (PBLs) and milk cells in pregnant dairy cows

Önceki çalışmalarda sütçü ineklerde periferal kan lökositlerindeki interferon-tau stimulated genlerindeki (ISGs) aktivite ölçümünün alternatif erken gebelik teşhis yöntemi olarak kullanılabileceği gösterilmiştir. Bununla birlikte süt gibi lökosit içeren diğer vücut sıvılarında ISGslerin ekspresyon profilindeki değişimlerinceleyen bir veri bulunmamaktadır. Sunulan çalışmada hipotez olarak süt örneklerindeki lökositlerde de, PBLslerde gösterildiği gibi ISGslerin ekspresyon profilinarttırabileceği düşünüldü. Bu amaç için dokuz (n=9) sağmal Holştayn ırkı gebe inek kullanıldı. Tohumlama günü sıfırıncı gün (0. gün) olarak kabul edildi. Kan ve süt örnekleri hücre izolasyonu için 0. ve 18. gün alındı. İzole edilen hücrelerden total RNA elde edildi ve cDNAya dönüştürüldü. Interferon-tau Stimulated Gene 15 (ISG15), Myxovirus (influenza virus) resistance 1 (MX1) and 2 (MX2) genlerinin mRNA transkriptlerinin seviyeleri real-time reverse transcriptase PCR tekniği ile ölçüldü. İstatistiksel analizler için Relative Expression Software Tool (REST2009) programı kullanıldı. Süt örneklerinin 0 ve 18. günleri arasında ISG15, MX1 ve MX2 mRNAlarının ekspresyon seviyelerindeki farklılık istatistiki olarak önemli olmadı. Bununla birlikte, gebe ineklerin 18. gün PBLsdeki ISG15 ve MX2nin transkript seviyeleri 0. gün ile karşılaştırıldığında sırasıyla 6.97±0.68 ve 5.84±1.27 kat arttı (P

Sütçü Gebe İneklerde Periferal Kan Lökositleri (PBLs) ve Süt Hücrelerindeki Interferon-Tau Stimulated Genlerinin (ISGs) Ekspresyon Profili

In previous reports, it was indicated that measurement of activity of Interferon-tau Stimulated Genes (ISGs) in Peripheral Blood Leucocytes (PBLs) may be used as an alternative early pregnancy detection method in dairy cows. However, there are no data showing the expression profiles of ISGs in other body fluids containing leucocytes such as milk. In the present study, it was hypothesized that leucocytes in milk samples may reflect the increases in expression profiles of ISGs as shown in PBLs. For this purpose, nine pregnant lactating Holstein cows were used. Insemination day was accepted as day zero (day 0). Blood and milk samples were collected on day 0 and 18 after insemination for cell isolation. Total RNA was extracted from isolated cells and converted to cDNA. Steady state levels of Interferon-tau Stimulated Gene 15 (ISG15), Myxovirus (influenza virus) resistance 1 (MX1) and 2 (MX2) mRNA transcripts were assayed by using real- time reverse transcriptase PCR. Relative Expression Software Tool (REST2009) was used for statistical analyses. There was no statistical significant difference for expression levels of ISG15, MX1 and MX2 mRNAs between days 0 and 18 in milk samples. However, when compared to day 0, levels of ISG15 and MX2 transcripts were increased 6.97±0.68 fold and 5.84±1.27 fold on day 18 in PBLs in pregnant cows, respectively (P<0.05). According to this result, it may be suggested that milk cells are not suitable measurement of expression profiles of ISGs to detect early pregnancy in lactating dairy cows.

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