Browsing by Author "Kose M."

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Circulating miRNAs in maternal plasma as potential biomarkers of early pregnancy in sheep
(2022-08-17) Hitit M.; Kose M.; Kaya M.S.; Kırbas M.; Dursun S.; Alak I.; Atli M.O.
MicroRNA (miRNA) plays an important role in the control of gene expression and is implied in many biological functions, including embryo implantation and development. The aim was to assess plasma miRNA profiles during the peri-implantation and ascertain potential candidate miRNA markers for early pregnancy diagnosis in ovine plasma. The plasma samples were obtained from a total of 24 ewes on days 12 (pre-implantation; P12, n = 4), 16 (implantation; P16, n = 4) and 22 (post-implantation; P22, n = 4) after mating, and on their corresponding days of 12 (Pre-C; C12, n = 4), 16 (Imp-C; C16, n = 4) and 22 (Post-C; C22, n = 4) of the estrous cycle. The miRNA profiles in plasma were assessed by microarray technology. We detected the presence of 60 ovine-specific miRNAs in plasma samples. Of these miRNAs, 22 demonstrated a differential expression pattern, especially between the estrous cycle and early pregnancy, and targeted 521 genes. Two miRNAs (oar-miR-218a and oar-miR-1185-3p) were confirmed using RT-qPCR in the ovine plasma samples. Protein-protein interaction (PPI) network of target genes established six functional modules, of which modules 1 and 3 were enriched in the common GO terms, such as inflammatory response, defense response, and regulation of immune response. In contrast, module 2 was enriched in the developmental process involved in reproduction, embryo development, embryonic morphogenesis, and regulation of the developmental process. The results indicate that miRNAs profiles of plasma seemed to be modulated during the peri-implantation stage of pregnancy in ewes. Circulating miRNAs could be promising candidates for diagnosis in early ovine pregnancy.
Expression pattern and cellular localization of two critical non-nuclear progesterone receptors in the ovine corpus luteum during the estrous cycle and early pregnancy
(2022-08-01) Atli M.O.; Akbalık M.E.; Kose M.; Alak I.; Atli Z.; Hitit M.
The study aimed to investigate the expression and cellular localization of two critical non-nuclear progesterone receptors, including membrane-associated-progesterone-receptor-component-1 (PGRMC1) and progestin and adipoQ receptor family member 7 (PAQR7) throughout the estrous cycle and early pregnancy in ovine corpus luteum (CL). Ewes were randomly grouped into cyclic (C, n = 4 per group) or pregnant (P, n = 4 per group) groups. Following slaughtering, the CL was obtained from both cyclic and pregnant ewes on days 12 (C12 and P12), 16 (C16 and P16), and 22 (C22 and P22). Western blotting and RT-qPCR were utilized to assess the expression levels of PGRMC1 and PAQR7, whereas immunohistochemistry was performed to determine the localization of PGRMC1 and PAQR7 in CL. Data were evaluated by one-way ANOVA, and the P < 0.05 was considered a significant difference. PGRMC1 was shown to be expressed in both small and large luteal cells and endothelial cells in CL, while PAQR7 expression was only found in small and large luteal cells. Compared to cycle days, pregnancy increased the expression of PGRMC1. PAQR7 did not differ during early pregnancy but reduced during the functional luteolysis stage (C16). mRNA and protein expression patterns for PGRMC1 and PAQR7 were similar on the studied days. This is the first study that demonstrates the expression and cellular localization of PGRMC1 and PAQR7 in ovine CL. We suggest that these receptors could execute a significant role in the ovine CL life span in both cyclic changes and the establishment of pregnancy.
Expression pattern of microRNAs in ovine endometrium during the peri-implantation
(2022-10-01) Kose M.; Hitit M.; Kaya M.S.; Kırbas M.; Dursun S.; Alak I.; Atli M.O.
MicroRNA (miRNA), acting as the transcriptional regulator of gene expression, has been widely demonstrated to be involved in many biological functions, including embryo implantation and development. The objective of the current study was to illuminate the expression pattern of microRNAs (miRNAs) in the endometrium during the peri-implantation in ewes. Intercaruncular endometrial samples was obtained from a total of 24 ewes on days of 12 (pre-implantation, n = 4), 16 (implantation, n = 4) and 22 (post-implantation, n = 4) of pregnancy following mating, and on their corresponding days of 12 (n = 4), 16 (n = 4) and 22 (n = 4) of the estrous cycle. The miRNA profiles were examined in the endometrium by microarray technology. We detected 116 ovine specifics miRNAs in the endometrium. Of these, nineteen were differentially expressed in early pregnancy. Four miRNAs (oar-miR-370-3p, oar-miR-411b-5p, oar-miR-379-3p and oar-miR-411a-3p) that had the most differential fold change were confirmed by RT-qPCR in ovine endometrium. The differentially expressed miRNAs targeted a total of 315 genes, resulting in 39 GO terms in molecular function, 353 in biological process, and 17 in the cellular component. The construction of the PPI network of target genes established two functional modules mostly enriched in the innate immune system, toll receptor cascades in module 1, whereas genes in module 2 were associated with GMCSF-mediated signaling events, insulin pathway, and mTOR signaling pathway. Based on the results, we may imply that miRNAs modulate ovine endometrium during the peri-implantation.
Expression patterns of genes in steroidogenic, cholesterol uptake, and liver x receptor-mediated cholesterol efflux pathway regulating cholesterol homeostasis in natural and PGF2α induced luteolysis as well as early pregnancy in ovine corpus luteum
(2022-05-01) Hitit M.; Kose M.; Kocak N.; Atli M.O.
The aim was to evaluate the expression of genes of steroidogenic, cholesterol uptake, and liver X receptor (LXR) mediated cholesterol efflux pathway in ovine corpus luteum (CL) during natural and prostaglandin F2α (PGF2α) induced luteolysis and early pregnancy. For this study, two experiments were carried out 1); ewes were grouped into two sub-groups as cyclic 12 (C12, n = 4) and 16 (C16, n = 4) and pregnant 12 (P12, n = 4), 16 (P16, n = 4), and 22 (P22, n = 4). Additionally, 2) ewes were grouped into four groups following treatment of PGF2α, the duration of PGF2α challenge at 1 (PG1, n = 4), 4 (PG4, n = 4), and 16 (PG16, n = 4) hours on day 12 of the cycle was compared with 0 h. The corpus luteum tissue samples were collected on the corresponding estrus cycle and pregnancy days, and RNA was extracted using Trizol. mRNA expression levels of the steroidogenic (StAR, CYP11A1, and HSD3B1) and cholesterol uptake receptors (SCARB1 and LDLR) and LXR pathway (NR1H3, NR1H2, ABCA1, and ABCG1) were assessed using quantitative PCR (qPCR), and protein of LXR pathway was investigated using western blot. In-situ hybridization was used to detect mRNA localization. Steroidogenic and cholesterol uptake mRNAs were decreased in C16, while NR1H2 and ABCG1 were increased in C16, compared to C12. Steroidogenic and cholesterol uptake mRNA was greater in P16 than in C16. NR1H2 and ABCA1 protein expression were higher in P16 than in C16. LDLR mRNA was higher in P22 than in P12, while SCARB1 was higher in P16 than in P12. NR1H2 mRNA was greater in P22 than in P12. Steroidogenic and cholesterol uptake mRNA were decreased in PGF2α-induced luteolysis groups against C12. ABCG1 mRNA was higher in PG16 than in PG4 and PG1. The reduction of lipoprotein receptors rather than LXR-mediated reverse transport may contribute to the decline in progesterone (P4) in natural and functional luteolysis.
Expression patterns of Toll-like receptors in the ovine corpus luteum during the early pregnancy and prostaglandin F2α-induced luteolysis
(2018-04-15) Atli M.; Kose M.; Hitit M.; Kaya M.; Bozkaya F.
The aim of this study was to elucidate the expression profiles of Toll-like receptors (TLRs) in the ovine corpus luteum (CL) during early pregnancy and prostaglandin F2α (PGF2α)-induced luteolysis. For this purpose, multiparous Anatolian Merino ewes were selected and randomly allotted into cyclic (including those in the induced luteolysis group, n = 20) and pregnant (n = 12) groups. All of the ewes were scheduled to be slaughtered for predetermined days/hour during the estrous cycle, early pregnancy, and PGF2α induced luteolysis. The CLs were collected from both cyclic and pregnant ewes on days 12 (C12 and P12; n = 8) and 16 (C16 and P16; n = 8) and pregnant ewes on day 22 (P22; n = 4). For the induced luteolysis model, ewes were injected with PGF2α on day 12 of the estrous cycle and CLs were collected at 1 h (PG1h; n = 4), 4 h (PG4h; n = 4), and 16 h (PG16h, n = 4) after injection. Quantitative polymerase chain reaction (qPCR) was used to evaluate the expression profiles of TLR2, TLR4, TLR6, TLR8, and TLR10, while free-floating in situ hybridization and immunohistochemistry were used to define the spatial localization of TLR2, TLR4, and TLR7 in the CL. Data were then analyzed by one-way ANOVA and were considered statistically significant when P values were lower than 0.05. Expression of TLR2 was upregulated in both early and late stages of luteolysis (P <.05). An upregulation of TLR4 was detected at PG16h, while TLR6 was decreased at PG4h (P <.05). Expression of TLR7 and TLR8 was significantly increased during early pregnancy, at both PG16h and regressed groups (C16, P <.05). In contrast, TLR10 was downregulated during PGF2α-induced luteolysis and on P16 (P <.05). TLR4 and TLR7 proteins were particularly localized in endothelial cells on C12/PG0h, but prominent signals corresponding to TLR4 and TLR7 were detected in luteal cells at PG16h. The results suggest an involvement of TLRs in the luteolytic mechanism in ovine CL, as indicated by differential expression levels of TLRs during PGF2α-induced luteolysis. Moreover, the present study indicates that early pregnancy-mediated changes in TLR expression in the CL may contribute to the establishment and maintenance of ovine pregnancy.
Relative abundance and localization of interferon-stimulated gene 15 mRNA transcript in intra- and extra-uterine tissues during the early stages of pregnancy in sheep
(2020-05-01) Alak I.; Hitit M.; Kose M.; Kaya M.S.; Ucar E.H.; Atli Z.; Atli M.O.
The aim of this study was to investigate relative abundance and localization of ISG15 mRNA transcript in intra-uterine (trophoblast, endometrium) and extra-uterine (hypothalamus, anterior pituitary, corpus luteum) tissues before and during the period of conceptus implantation. Multiparous ewes (n = 16) were randomly allotted into four groups: pregnant or estrous cyclic on days of 12 and 16 (n = 4 per group) following estrus. Relative abundances of ISG15 mRNA transcript were determined in the endometrium, corpus luteum, hypothalamus, and anterior-pituitary using real time quantitative PCR. Localization of ISG15 mRNA transcript was evaluated using in situ hybridization. The presence of ISG15 mRNA transcript was only visualized in intra-uterine tissues including the endometrium and trophoblast on day 12 of pregnancy. The ISG15 mRNA transcript was detected in all tissues evaluated on day 16 of pregnancy. The abundance of ISG15 mRNA transcript was greater in the endometrium on day 12 of pregnancy than at other days when evaluations occurred while in all other tissues except the hypothalamus there were large abundances of ISG15 mRNA on day 16 of pregnancy. It is concluded that the ISG15 mRNA transcript is only present in intra-uterine tissues before conceptus implantation. The ISG15 mRNA transcript, however, is present in extra-uterine tissues of ewes during implantation probably due to an increased amount of interferon-tau in blood circulation that is produced by the developing embryo. Results also indicate, for the first time, that pregnancy is associated with an intra-hypothalamus and anterior pituitary increased abundance of ISG15 mRNA transcript in ewes.
The assessment of the thermal behavior of an urban park surface in a dense urban area for planning decisions
(2022-07-01) Adiguzel F.; Cetin M.; Dogan M.; Gungor S.; Kose M.; Bozdogan Sert E.; Kaya E.
The new summer index (SSI) which was established to determine climate comfort was classified according to the index. Climatic comfort areas will increase and thus raising the quality of living in areas. The temperature values felt in the study were calculated with the SSI formula, and then the maps of the study area were classified. The study area was the Hayal Park and its surroundings in the Toros Quarter of Cukurova District, which is surrounded by multi-story and dense buildings in the north of Adana. Although the vertical construction is very high, it is richer in terms of green areas compared to other large central districts. Cukurova, which is one of the largest districts of Adana (according to population), has Seyhan dam lake in the north and Seyhan district in the south. The study area is on the border of Seyhan district and is adjacent to Yüreğir and Sarıçam districts. With the portable Smart SENSOR AS 847 measuring device, temperature and relative humidity were measured at 1.5 m above the ground on Tuesday, August 20 at 07.00, at 14.00, and 21.00. Bioclimatic comfort areas were determined by measuring temperature and relative humidity at 25 points with different textures in the study area. In this study, measured instantaneous temperature (°C) and relative humidity (%) data were transferred to GIS medium by ArcGIS 10.6 program and then modeled with the Kriging method, which is one of the interpolation methods, and temperature and relative humidity maps were created. The temperature map created in degrees Celsius (°C) by the Kriging method and was converted to Fahrenheit degrees (°F) in the Raster Calculator. The reason for this change is that SSI is calculated with the formula Fahrenheit (°F). These maps were then calculated on the Raster Calculator using the SSI formula, and sensed temperature values were obtained The SSI, which normally has 8 classes, has 3 classes in the study area. These classes are 83 ≤ SSI < 91 slightly hot, 91 ≤ SSI < 100 hot, and 100 ≤ SSI < 112 very hot. When we look at the maps, the places outside of Hayal Park are in the hot class at 07.00 in the morning, and we can see that the green spaces have a positive effect on the bioclimatic comfort areas. The small area is in a very hot class. When we look at 14.00 h, it is a totally hot class. Normally, the temperature is the highest of these times and also coincides with the multi-story buildings in the study area; asphalt and concrete areas increased the temperature. As a result, there is a very hot air in the study area. It was determined that the maps identified non-comfortable areas. Since urban form and settlement affect climate comfort values, the urban plan should be revised, and the area should be brought to the bioclimatic comfort value range. For reducing the effect of temperature in the working area and creating comfortable areas is increasing the density of green areas compared to asphalt and concrete areas. Considering that the Cukurova district will grow further, urban planning should be done very well to reach the comfort range in the following settlements.