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Öğe Effects of long-term heat stress and dietary restriction on the expression of genes of steroidogenic pathway and small heat-shock proteins in rat testicular tissue(Blackwell Publishing Ltd, 2017) Bozkaya, Faruk; Atlı, Mehmet Osman; Güzeloǧlu, Aydın; Kayış, Seyit Ali; Yıldırım, Mehmet Erol; Kurar, Ercan; Yılmaz, Rahşan; Aydilek, NurettinThe aim was to investigate the effects of long-term heat stress and dietary restriction on the expression of certain genes involving in steroidogenic pathway and small heat-shock proteins (sHSPs) in rat testis. Sprague Dawley rats (n = 24) were equally divided into four groups. Group I and II were kept at an ambient temperature of 22°C, while Groups III and IV were reared at 38°C for 9 weeks. Feed was freely available for Group I and Group III, while Group II and Group IV were fed 60% of the diet consumed by their ad libitum counterparts. At the end of 9 weeks, testicles were collected under euthanasia. Total RNA was isolated from testis tissue samples. Expression profiles of the genes encoding androgen-binding protein, follicle-stimulating hormone receptor, androgen receptor, luteinising hormone receptor, steroidogenic acute regulatory protein (StAR), cyclooxygenase-2 and sHSP genes were assessed at mRNA levels using qPCR. Long-term heat stress decreased the expression of StAR and HspB10 genes while dietary restriction upregulated StAR gene expression. The results suggested that long-term heat stress negatively affected the expression of StAR and HspB10 genes and the dietary restriction was able to reverse negative effect of heat stress on the expression of StAR gene in rat testis.Öğe Evaluation of genes involved in prostaglandin action in equine endometrium during estrous cycle and early pregnancy(Elsevier, 2010) Atli, Mehmet O.; Kurar, Ercan; Kayis, Seyit A.; Aslan, Selim; Semacan, Ahmet; Celik, Sefa; Guzeloglu, AydinThe aim was to evaluate expression of genes involved in the biosynthesis of prostaglandins (PTG), Prostaglandin H Synthase-1 (PTGS1) and PTGS2, PGF synthase (PTGFS), and PGE synthase (PTGES), PGF receptor (PTGFR). PGE receptors (PTGER2 and FTGER4), prostaglandin transporter (SLCO2A1) and hydroxyprostaglandin dehydrogenase-15 (HPGD). Endometrial biopsies were obtained from mares on day of ovulation (d0, n = 4), late diestrus (LD, n = 4), early luteolysis (EL n = 4) and after luteolysis (AL n = 4) during the cycle. Stages of the cycle were confirmed by plasma progesterone concentrations measured daily and ultrasound examinations. Biopsies were also taken on days 14 (P14; n = 4), 15 (P15. n = 4), 18 (P18, n = 4) and 22 (P22; n = 4) of pregnancy. Relative mRNA expressions were quantified using real-time RT-PCR. A mixed model was fitted on the normalized data and least significant difference test (alpha = 0.05) was employed. Expression of PTGS1 mRNA was low throughout the estrous cycle and early days of pregnancy, but upregulated on P18 and P22. PTGS2 expression was increased on EL but it was suppressed by pregnancy on P15, P18, and P22. PTGFS expression was upregulated in both cyclic and pregnant mares compared to d0 and its level was the highest on LD. PTGFR expression was transiently increased on LD and EL and was suppressed during early pregnancy. Both PTGES and PTGER2 expressions were increased on LD, El, and early pregnancy, but were decreased after the luteolysis in cyclic mares as they remained high on P18 and P22. PTGER4 expression did not change throughout the cycle and early pregnancy. Levels of HPGD and SLCO2A1 were significantly increased only on P22. In conclusion, PTGS2 expression increases around the time of luteolysis and concurrent upregulation of PTGFS and PTGES indicates that equine endometrium has increased capability of PTG production around the time of luteolysis. However, pregnancy reduces PTGS2 expression, but maintains the high levels of PTGES during early pregnancy along with PTGER2 while PTGFR expression was suppressed. These findings suggest that possible luteotrophic action of PGE(2) is required in early equine pregnancy. PTGS1 is only upregulated later in the early pregnancy suggesting that it is not involved in luteolysis, but could be the main PTGS enzyme at this time during early pregnancy. An increase in HPGD and SLCO2A1 levels on P22 indicates a tight regulation of PTG action by pregnancy. (C) 2010 Elsevier B.V. All rights reserved.Öğe Expression of enzymes and receptors of leukotriene pathway genes in equine endometrium during the estrous cycle and early pregnancy(Elsevier Science Inc, 2013) Guzeloglu, Aydin; Atli, Mehmet O.; Kurar, Ercan; Kayis, Seyit A.; Handler, Johannes; Semacan, Ahmet; Aslan, SelimThe aims of the present study were to elucidate the expression profiles of leukotriene (LT) pathway mRNA transcription and to determine the possible interaction of LT and prostaglandin (PTG) pathways genes in equine endometrium during the estrous cycle and early pregnancy. Endometrial biopsies were obtained from mares on the day of ovulation (d0), at late diestrous (LD, n = 4), and after luteolysis in the estrus phase (AL n = 4) of the cycle. Biopsies were also taken on Days 14 (P14; n = 4), 18 (P18, n = 4), and 22 (P22, n = 4) during early pregnancy that were comparable days to cyclic sampling days. A mixed model was fitted on the normalized relative mRNA levels, quantified by qPCR in duplicate, and least significant difference test was employed to detect significantly different group(s). In addition, to determine the degree of contribution of each gene to separation of treatment groups, the multivariate projection method partial least square regression discriminant analysis was used. The expression of 5-lipoxygenase mRNA was greater on d0 and LD, declined at AL, and was suppressed by early pregnancy. Leukotriene A(4) hydrolase mRNA expression increased at LD and during early pregnancy, but Was significantly greater at LD compared with P14. The expression of LT C-4 synthase mRNA was only induced at LD. Cysteinyl leukotriene receptors (CysLT1 and CysLT2) mRNA expressions were decreased by both cyclic changes and early pregnancy, whereas 5-lipoxygenase-activating protein and B leukotriene receptor mRNA expressions were not affected by early pregnancy or stages of the estrous cycle. Partial least square discriminant analysis suggests that LT and PTG pathway enzymes and receptors appear to behave similarly in terms of mRNA expression. In conclusion, the expression profiles of LT pathway genes are demonstrated in equine endometrium for the first time by the present study, and the present data suggest that LT pathway mRNA transcriptions are tightly regulated during early pregnancy in mares. (C) 2013 Elsevier Inc. All rights reserved.Öğe Expression of Epidermal Growth Factor (EGF) and Heparin-Binding EGF (HB-EGF) mRNA in Mare Endometrium(Kafkas Univ, Veteriner Fakultesi Dergisi, 2012) Atli, Mehmet Osman; Guzeloglu, Aydin; Kurar, Ercan; Kayis, Seyit Ali; Aslan, Selim; Semacan, Ahmet; Celik, SefaEGF and HB-EGF play crucial roles in embryonic development and peri-implantation. In this study, aim was to characterize expression profiles of EGF and HB-EGF in mare endometrium during estrous cycle and early pregnancy. Endometrium biopsies were obtained from mares on day of ovulation (d0), late diestrus (LD) and after luteolysis (AL) in the estrus phase. In pregnant groups, biopsies were taken on days 14 (P14), 18 (P18), 22 (P22) and 60 (P60). Relative expression levels of genes were quantified using real-time RT-PCR. A mixed model was fitted on the normalized data and Least Significant Difference (LSD) test was employed to determine significantly different group(s). EGF mRNA expression was up-regulated at LD compared to d0 while HB-EGF expression was not changed throughout the cycle. EGF expression was also increased during early pregnancy with the highest expression level observed on P60. Similarly, HB-EGF mRNA level was increased on P60. Pregnancy induced EGF expression on P14 and P18 compared to LD and AL whereas expression of HB-EGF was only significantly higher on P18 than that of AL. These results indicate that EGF expression is up-regulated during the cycle at late diestrus when P4 is high and is increased by pregnancy. HB-EGF expression is induced later in the pregnancy. In conclusion, EGF and HB-EGF appear to involve in the events that happen in the mare endometrium during peri-implantation period.Öğe Expression of epidermal growth factor (EGF) and heparin-binding EGF (HB-EGF) mrna in mare endometrium(2012) Kurar, Ercan; Güzeloğlu, Aydın; Kayış, Seyit Ali; Çelik, Sefa; Aslan, Selim; Atlı, Mehmet Osman; Semacan, AhmetEGF ve HB-EGF implantasyon sırasında ve embriyonik gelişim esnasında çok önemli roller oynamaktadır. Bu çalışmanın amacı, erken gebelik ve östrus siklusunda at endometriumunda EGF ve HB-EGF ekspresyon profilini karakterize etmektir. Endometrium biyopsileri östrustaki kısraklardan ovulasyon gününde (d0), geç diöstrusta (LD) ve luteolizis sonrası (AL) dönemlerde toplandı. Gebe kısraklardan ise gebeliğin 14. (P14), 18. (P18), 22. (P22) ve 60. (P60) günlerinde biyopsiler toplandı. Nispi mRNA ekspresyon seviyeleri real-time PZR kullanılarak tespit edildi. Normalize edilmiş veriler karışık model kullanılarak analiz edildi ve istatistiki olarak farklı olan gruplar Asgari Önemli Fark (AÖF) testi ile farklı gruplar tespit edildi. EGF mRNA ekspresyonu LD’de d0 göre artmasına rağmen, HB-EGF ekspresyonu siklus boyunca değişmedi. EGF ayrıca erken gebelik boyunca arttı ve en yüksek seviyeye 60. günde ulaştı. Benzer şekilde HB-EGF mRNA seviyesi 60. günde arttı. Gebelik, EGF ekspresyonunu 14. ve 18. günlerde LD ve AL göre arttırırken, HB-EGF sadece gebeliğin 18. gününde AL’e göre önemli oranda arttı. Bu sonuçlar, EGF’nin P4 seviyesinin yüksek olduğu geç diöstrusta ve gebeliğe bağlı olarak arttığını göstermektedir. HB-EGF ise gebeliğin daha ileri dönemlerinde uyarılmaktadır. Sonuç olarak, kısrak endometriumunda EGF ve HB-EGF’nin peri-implantasyon dönemindeki görülen olaylara katıldığı gözlenmiştir.Öğe Expression of platelet activating factors and its receptor in cat uterus during early pregnancy(Ankara Univ Press, 2013) Agaoglu, Ali Reha; Agaoglu, Ozgecan Korkmaz; Kurar, Ercan; Guzeloglu, Aydin; Kayis, Seyit Ali; Atli, Mehmet Osman; Schafer-Somi, SabinePlatelet activating factors (PAF1 beta, PAF1 gamma, PAF:ah, PAF-R) have a significant role during mammalian pregnancy process. This study aimed to investigate expression profiles of PAF gene family in cat uterus during pregnancy. Tissue samples were collected via ovariohysterectomy at days 7 (Gr1; n=3), 18-20 (Gr2; n=7), 24-25 (Gr3; n=5) after mating from the different regions of the uterus. Relative expression levels of genes were quantified using real-time qRT-PCR. A mixed model was fitted on the normalized data and Least Significant Difference (LSD) test was employed to determine significant differences between groups. Expressions of PAP genes were assessed in the cat uterus during pregnancy around implantation period; however, no significant differences were detected between the preimplantation and the implantation stage. Similarly, no significant difference concerning mRNA expression was assessed between different parts of the uterus during pregnancy In conclusion, PAP genes are expressed in the cat uterus during early pregnancy. However, this expression is not associated with changes in the uterus during early pregnancy.Öğe Expression of Toll-like receptors (TLRs) in the equine peripheral blood mononuclear cells (PBMCs) during the early pregnancy and estrous cycle(2012) Atlı, Mehmet Osman; Kayış, Seyit Ali; Kurar, Ercan; Semacan, Ahmet; Güzeloğlu, AydınAmaç: Günümüzde kısraklarda birçok gebelik teşhis metodu olmasına rağmen, mümkün olabilecek en erken zamanda gebeliğinin teşhis edilmesi için pratik, güvenilir ve ekonomik bir yönteme halen ihtiyaç bulunmaktadır. Allojenik fötusa karşı immun tolerans erken gebelik esnasında kritik bir öneme sahiptir. Toll-like reseptörlerinin (TLRs) doğal immun sistemin önemli bir parçası olduğu bilinmektedir. Sunulan çalışmada, kısraklarda erken gebelikte periferal kan mononükler hücrelerinde (PKMH) TLRs ekspresyonunun incelenmesi amaçlandı. Gereç ve Yöntem: Kan numuneleri aynı üç kısraktan östrus siklusunun ve gebeliğin 0 ve 8. günlerinde toplandı. Total RNA örnekleri PKMH’lerinden izole edildi ve cDNA sentezlendi. mRNA düzeyindeki TLRs ekspresyonları iki defa realtime PZR kullanılarak kuantifiye edildi. İstatistiksel analiz için Relative Expression Software Tool (REST2009) kullanıldı. Bulgular: RT-PZR ile PKMH’lerde TLRs (1-10) ekspresyonları tespit edildi. Bununla beraber, real-time PZR ölçümleri siklik veya gebelik durumunun TLRs ekspresyonu üzerine etkisini göstermekte başarısız kaldı. Öneri: Bu sonuçlara bakıldığında, atlarda gebeliğin erken döneminde PKMH’lerinde TLR genlerinin ekspresyonlarının gebeliğin erken anlaşılması için iyi bir markör olmadığı söylenebilir.Öğe Expression of Toll-like Receptors (TLRs) Is Regulated by Early Pregnancy in the Mare Endometrium.(Soc Study Reproduction, 2010) Atli, Mehmet Osman; Kurar, Ercan; Kayis, Seyit A.; Aslan, Selim; Semacan, Ahmet; Celik, Sefa; Guzeloglu, Aydin[Abstract Not Available]Öğe Expression profiles of interferon-tau stimulated genes (ISGs) in peripheral blood leucocytes (PBLs) and milk cells in pregnant dairy cows(Kafkas Üniversitesi Veteriner Fakültesi, 2014) Köse, Mehmet; Görgülü, Mehmet; Kaya, Mehmet Salih; Aydilek, Nurettin; Bozkaya, Faruk; Bayrıl, Tahir; Kurar, Ercan; Kıyma, Zekeriya; Güzeloğlu, Aydın; Atlı, Mehmet OsmanIn 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.Öğe Expressions of Insulin-Like Growth Factor-Binding Proteins (IGFBP) in Mare Endometrium during Estrous Cycle and Early Pregnancy.(Soc Study Reproduction, 2010) Kurar, Ercan; Atli, Mehmet O.; Kayis, Seyit A.; Arslan, Selim; Semacan, Ahmet; Celik, Sefa; Guzeloglu, Aydin[Abstract Not Available]Öğe Kısrak endometriyumunda Phosphatase and Tensin Homolog (PTEN) geni mRNA ekspresyonunun araştırılması(2015) Kayış, Seyit Ali; Güzeloğlu, Aydın; Şen, Gonca; Hitit, Mustafa; Atlı, Mehmet Osman; Kurar, Ercan; Özel, ÇağlayanAmaç: Bu çalışmanın amacı, siklüs ve erken gebelik dönemlerinde dinamik bir yapıya sahip olan kısrak endometriyumunda phosphatase and tensin homolog (PTEN) gen ekspresyonunun mRNA düzeyinde belirlenmesidir.Gereç ve Yöntem: Araştırmada her güne 4 farklı kısrak olacak şekilde (n=4/gün) siklik kısraklardan ovulasyon gününde (d0), geç diöstrusta (LD) ve luteolizis sonrası östrusta (AL), gebe kısraklardan ise gebeliğin 14. (P14), 18. (P18) ve 22. (P22) günlerinde endometriyum biyopsi örnekleri toplandı. Doku örneklerinden total RNA elde edildi ve cDNA'ya dönüştürüldü. PTEN ekspresyonlarında siklus ve gebeliğe bağlı muhtemel değişiklikler mRNA seviyesinde kantitatif polimeraz zincir reaksiyonu (qPZR) kullanılarak araştırıldı. Referans gen olarak GAPDH ekspresyonu ile normalize edilen veriler karışık model kullanılarak analiz edildi. Farklı olan grup(lar) Asgari Önemli Fark (LSD) testi ile tespit edildi.Bulgular: PTEN ekspresyonu çalışmaya konu olan tüm siklüs ve erken gebelik dönemlerinde at endometriyumunda mRNA düzeyinde tespit edildi. LD'e göre AL'de ekspresyon seviyesinde anlamlı olmayan bir düşüş gözlendi. Benzer şekilde d0'a göre araştırılan P14, P18 ve P22 erken gebelik günlerinde PTEN ekspresyonunun baskılanmış olduğu tespit edildi. Ancak fark istatistiksel olarak anlamlı bulunmadı (P>0.10).Öneri: PTEN aktivitesinin ve protein düzeyinde ekspresyonun araştırılmasının anlamlı olacağı kanaatine varılmıştır.Öğe Kısraklarda erken gebelik tespiti(2012) Semacan, Ahmet; Kurar, Ercan; Köse, Mehmet; Atlı, Mehmet Osman; Güzeloğlu, AydınAt yetiştiriciliğinde kısa olan üreme sezonunun iyi değerlendirilip, sezon boşa geçirilmeden kısraklarda gebeliğinoluşturulması öncelikli hedefler arasında yer almaktadır. Sınırlı üreme sezonuna sahip kısrakların ülkemizin de içindebulunduğu coğrafyada ilkbahar aylarında başlayan ovaryum aktiviteleri sonbahara kadar devam etmektedir. Bu süreç içindereprodüktif herhangi bir sorunu olmayan kısraklarda düzenli östrus siklusu sayısıda oldukça azdır. Dolayısıyla gebeliğinmümkün olan en kısa zamanda tespitinin kritik önemi bulunmaktadır. Bu amaçla farklı tanı yöntemleri kullanılmasınarağmen, gebeliğin mümkün olan en kısa zamanda teşhisine olanak sağlayacak etkin ve pratik bir yönteme halen ihtiyaçduyulmaktadır. Bu derleme kapsamında öncelikle kısrakların erken gebelik fizyolojisi hakkında kısaca bilgi verilecektir.Mevcut gebelik tanı yöntemleri (östrusların izlenmesi, rektal palpasyon, ultrasonografi, progesteron, östrojen, eCG, erkengebelik faktörünün ölçümü ve gebeliğe bağlı gen ekspresyonları ile gebelik tespiti) avantajları ve dezavantajları açısındantartışılacaktır. Ayrıca, gebeliğin maternal kabul mekanizmasının moleküler düzeyde anlaşılması amacıyla yapılan güncelçalışmalar ve gebeliğin tanısında alternatif yaklaşımlar hakkında bilgi verilecektir.Öğe PLS-DA approach for simultaneous evaluation of genes involved in luteolysis in equine endometrium during estrous cycle and early pregnancy(Elsevier Sci Ltd, 2011) Guzeloglu, Aydin; Kayis, Seyit A.; Atli, Mehmet O.; Kurar, Ercan; Semacan, Ahmet; Aslan, Selim[Abstract Not Available]Öğe POMC, iNOS, PGES, IL-4, IL-5 and IL-10 gene expression in peripheral blood mononuclear cells of cyclic and pregnant mares(Kafkas Üniversitesi Veteriner Fakültesi, 2011) Kurar, Ercan; Atlı, Mehmet Osman; Güzeloğlu, Aydın; Semacan, AhmetAlthough a number of diff erent clinical and laboratory methods are used, there is still need for a practical, reliable and economical diagnosis method for early pregnancy in mares. Objective was to provide a preliminary background for understanding immunological modifications at molecular level during early pregnancy in mare and thereby to allow development of a practical pregnancy test. Blood samples were collected from 10 pregnant and 4 cyclic mares on days of ovulation (d0), 4 and 8. Total RNA samples were isolated from peripheral blood mononuclear cells (PBMC) and cDNA synthesis was performed. Pregnancy dependent gene expression profiles of proopiomelanocortin (POMC), nitric oxide synthase (iNOS), prostaglandin E synthase (PGES), interleukin-4 (IL-4), IL-5 and IL-10 were evaluated at mRNA level using semiquantitative Reverse Transcriptase-Polymerase Chain Reaction. Data were analyzed by General Linear Model and possible diff erences between all mean factors were determined by Tukey’s analysis. No gene expressions were observed for PGES, IL-5 and iNOS in PMBCs. Expressions of POMC, IL-4 and IL-10 were not significant on d0, 4 and 8, which suggested that pregnancy or cyclic status had no eff ect on expression of these genes. Only, IL-10 mRNA expression was lower in pregnant mares than cyclic mares (P<0.05).Öğe Rating of putative housekeeping genes for quantitative gene expression analysis in cyclic and early pregnant equine endometrium(Elsevier Science Bv, 2011) Kayis, Seyit A.; Atli, Mehmet O.; Kurar, Ercan; Bozkaya, Faruk; Semacan, Ahmet; Aslan, Selim; Guzeloglu, AydinThe aim was an evaluation of a set of housekeeping genes (HKGs) to be used in the normalization of gene expression in the equine endometrium. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), hypoxanthine ribosyl transferase 1 (HPRT1), ubiquitin B (UBB), tubulin alpha 1 (TUBA1), ribosomal protein L32 (RPL32), beta-2-microglobulin (B2M), 18S rRNA (18S), and 285 rRNA (28S) HKGs were evaluated using real-time PCR and were compared in different physiological stages of the endometrium. Endometrial biopsies were obtained from mares on day of ovulation (d0, n = 4), at late diestrus (LD, n = 4), after luteolyis (AL, n = 4) of the cycle and on days 14 (P14; n = 3), 18 (P18, n = 3) and 22 (P22; n = 3) of pregnancy. A model based on REML with support of descriptive statistics was proposed in accordance with experimental design and was further confirmed with principal component analysis (PCA). Results were compared with widely used software including geNorm, BestKeeper, and NormFinder. Results indicated that GAPDH was the most stable HKG and RPL32 was ranked as the second best. 18S and 285 were found to be the least stable. The proposed model, PCA, geNorm, and BestKeeper were in agreement in detecting the most stable and the least stable HKGs in the equine endometrium during the estrous cycle and early pregnancy. (C) 2011 Elsevier B.V. All rights reserved.
