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    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, Selim
    The 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.
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    PLS-DA approach for simultaneous evaluation of genes involved in luteolysis in equine endometrium during estrous cycle and early pregnancy
    (CURRENT BIOLOGY LTD, 2011) Guzeloglu, Aydin; Kayis, Seyit A.; Atli, Mehmet O.; Kurar, Ercan; Semacan, Ahmet; Aslan, Selim
    [Abstract not Available]
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    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, Aydin
    The 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.