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Öğe Angiogenic and anti-angiogenic factors during the post-hatching growth of the quail (Coturnix coturnix japonica) spleen(Wiley, 2024) Bayram, Bayram; Liman, Narin; Alan, Emel; Sağsöz, Hakan; 0000-0002-5738-918X; 0000-0001-5489-2719; 0000-0002-5456-697X; 0000-0003-4990-3991Vascular endothelial growth factor (VEGF) family members are responsible for endothelial cells' growth, proliferation, migration, angiogenesis, vascular permeability, and differentiation and proliferation of non-endothelial cell types. VEGF and its receptors are found in mammalian lymphoid organs. The present study was conceived to determine (a) the presence and localization of angiogenic VEGF and its receptors (Fms-like tyrosine kinase 1 [Flt1/fms], fetal liver kinase 1 [Flk1]/kinase insert domain receptor [KDR], Fms-like tyrosine kinase 4 [Flt4]) and vascular endothelial growth inhibitor (VEGI) in the quail spleen; and (b) whether their expressions in the spleen components change during the post-hatching growth of the organ, using immunohistochemistry. Immunohistochemical stainings showed that VEGI, VEGF, and VEGF receptors were expressed in many components, including the vascular endothelial and smooth muscle cells, ellipsoid-associated cells (EACs), and immune cells, of quail spleen and that VEGF and its receptors' immunostaining intensity scores (ISs) varied depending on the post-hatching growth period, while VEGI-IS did not change. In addition, ISs of VEGI, VEGF, Flt1/fms, and Flt4 in EACs were weak to moderate, while flk1/KDR-IS in EACs adjacent to the capsule of Schweigger-Seidel sheaths (ellipsoids) was higher than other proteins, supports a more important and specific role of Flk1/KDR in the EAC function. These specific expressions of VEGI, VEGF, flt1/fms, flk1/KDR, and flt4 proteins in splenic cell types suggest their particular roles, in the functional development of splenic components and thus, are critical to post-hatching maturation of quail spleen. These findings indicate that the expression levels of VEGF, Flt1/fms, and Flt4, except Flk1/KDR, are low in the quail spleen, and only a few components of the spleen express VEGF, Flt1/fms, and Flt4 under normal conditions.Öğe Epidermal growth factor receptors and their ligands are expressed in the spleen of the Japanese Quail (Coturnix coturnix japonica) during the post-hatch period(Taylor and Francis Ltd., 2023) Alan, Emel; Liman, Narin; Sağsöz, Hakan1. The epidermal growth factor (EGF) family plays an important role in the development, differentiation, migration and apoptosis of cells, as well as in wound healing, which are all essential to the viability of multicellular organisms. The avian spleen is a principal organ of systemic immunity and its importance in disease resistance is presumably accentuated by the scarcity of avian lymph nodes. 2. The aim of this study was to determine whether EGF receptors (ErbB1-4) and their ligands (EGF, AREG and NRG) are expressed in the structural components of the quail spleen during the post-hatch period. At each selected age, from 1 d to 7, 14, 21 and 60 d, 10 quails were euthanised under ether anaesthesia and their spleens were fixed in a 10% formaldehyde-alcohol solution. Following routine histological processing, the streptavidin-biotin-peroxidase method was used for immunohistochemical examination. 3. Strong cytoplasmic immunoreactions for ErbB2, ErbB4 and NRG were observed in the ellipsoid associated cells (EAC) of the quail spleen throughout the post-hatch period. This immunoreactivity in the EAC increased after the 7th d post-hatch. ErbB1 and ErbB3 immunoreactions were relatively similar and weak in all components of the spleen during the post-hatch period. Some immune cells of the peri-arterial lymphatic sheath (PALS) and peri-ellipsoidal lymphatic sheath (PELS) showed positive immunoreactivity for the ErbB receptors and their ligands. In the vascular smooth muscle cells, immunoreactivity for ErbB2 was stronger than that for the other ErbB receptors and their ligands. 4. The data showed that ErbB receptors and their ligands (EGF, AREG and NRG) are expressed by different structural components of the quail spleen during the post-hatch period.Öğe Expression and localisation of epidermal growth factor receptors and their ligands in the lower genital tract of cycling cows(Csiro Publishing, 2019) Sagsoz, Hakan; Liman, Narin; Saruhan, Berna Guney; Akbalik, Mehmet E.; Ketani, Muzaffer A.; Topaloglu, UgurThe epidermal growth factor receptor (ErbB) family and its ligands are essential for the regulation of multiple cellular processes required for mammalian reproduction. The objectives of this study were to investigate the expression and localisation of ErbB subtypes (ErbB1-4) and selected ligands, namely epidermal growth factor (EGF), amphiregulin (AREG) and neuregulin (NRG), in the cervix and vagina of cycling cows and to determine possible steroid hormone-dependence of their expression using immunohistochemistry. All four ErbBs and EGF, AREG and NRG proteins were found to be localised in the nucleus and cytoplasm of different cells in the cervix and vagina, and their expression differed during the oestrous cycle. During the follicular phase, in both the cervix and vagina, ErbB1, ErbB2, ErbB3, ErbB4 and EGF expression was higher in the luminal epithelium (LE) than in stromal and smooth muscle (SM) cells (P<0.05). During the luteal phase, the expression of ErbB1, ErbB3 and EGF in the LE was significantly different from that in stromal and SM cells in the cervix, whereas the expression of EGF and AREG differed in the vagina compared to the cervix (P<0.05). Throughout the oestrous cycle, in both the cervix and vagina, although ErbB2/human epidermal growth factor receptor 2 expression in the LE and SM cells was significantly higher than in the stromal cells (P<0.05), NRG expression was similar in the LE, stromal and SM cells (P>0.05). Overall, these results suggest that all four ErbBs and the EGF, AREG and NRG proteins may collectively contribute to several cellular processes in the bovine cervix and vagina during the oestrous cycle.Öğe Expression of cadherins and some connective tissue components in cow uterus and placenta during pregnancy(Elsevier Sci Ltd, 2022) Sagsoz, Hakan; Liman, Narin; Akbalik, M. Erdem; Alan, Emel; Saruhan, Berna Guney; Ketani, M. Aydin; Erdog, SerkanThe implantation and placental development processes are regulated with cell adhesion molecules and remod-eling of the maternal endometrium's extracellular matrices (ECM) and fetal chorion. This study aimed to investigate the distribution and localization of some classical cadherins (E-, N-, and P-cadherins) and extracel-lular matrix components collagen type 5 alpha 1, fibronectin, and laminin in the cow placentomes during pregnancy using immunohistochemical and Western blotting analyses. The study results confirmed the expression of E- and P-cadherins, collagen type V alpha 1 (COLV alpha 1), fibronectin, and laminin in the cow placentomes, but not N-cadherin. Throughout the pregnancy, E-and P-cadherins, COLV alpha 1, and laminin were localized in the luminal and glan-dular epithelium of the inter-caruncular endometrium, caruncular epithelium, and the uninucleate (UNCs) and binucleate trophoblast giant cells (BNCs/TGCs). E- cadherin immunoreactivity in the first pregnancy period was strong in the UNCs while moderate in the BNCs/TGCs. However, it was weak in both trophoblast in the second and third pregnancy periods. In the fetal trophoblasts, P-cadherin and laminin immunostainings were more intense in the BNCs/TGCs than UNCs. The fetal and maternal stromal cells were also positive for P-cadherin, COLV alpha 1, fibronectin, and laminin. The immunostaining intensity of COLV alpha 1 and fibronectin in the stromal extracellular matrix of the placentomes decreased as the pregnancy progressed. The endothelia of fetal and maternal vessels were positive for all proteins. The presence and distinct localization of cadherins and ECM proteins in the cow placentome components support the role of these molecules in regulating placental cell growth, migration, and matrix production during pregnancy.Öğe The expression of epidermal growth factor receptors and their ligands (epidermal growth factor, neuregulin, amphiregulin) in the bitch uterus during the estrus cycle(Elsevier Science Bv, 2014) Sagsoz, Hakan; Liman, Narin; Saruhan, Berna Guney; Kucukaslan, IbrahimIn order to study the possible role of EGFR receptors in the bitch reproductive process, we have analyzed the expression pattern and localization of EGFR receptors and some of their ligands epidermal growth factor (EGF), neuregulin (NRG), amphiregulin (AREG), in the uterus during the estrus cycle using immunohistochemistry. The immunostaining for receptors and ligands of EGFR/ligand system was confined to membrane and cytoplasm of the target cells. Variations were observed, not only at the different stages of the estrous cycle, but also in the different tissue compartments of the uterus. However, it was detected that the immunostainings for NRG and AREG in the different cells do not show important differences at stages of the estrus cycle. In the luminal epithelium, strong immunostaining for ErbB1/HER1, ErbB2/HER2, ErbB4/HER4 and EGF was found at estrus. In the glandular epithelium, strong immunostaining for ErbB4/HER4 was observed at diestrus, while strong immunostaining for EGF was detected in both of estrus and diestrus. ErbB3/HER3 immunoreactivity in the stromal cells was higher at diestrus and anestrus, while ErbB4/HER4 immunoreactivity was lower at anestrus. In the myometrium, the highest levels of immunoreactivity of ErbB2/HER2 were found at estrus, while ErbB3/HER3 immunoreactivity was higher at anestrus. EGF immunoreactivity was lower at anestrus compared to other stage of cycle. Altered EGFR/ligand system expression during the estrus cycle suggests this growth factor system is a potent regulator of proliferation and differentiation events during preparation for implantation of bitch uterus. (C) 2014 Elsevier B.V. All rights reserved.Öğe Gebelik Süresince İnek Uterus ve Plasentasında Kaderinler ve Bazı Bağdoku Elemanlarının Dağılımı(2018) Sağsöz, Hakan; Erdoğan, Serkan; Saruhan, Berna Güney; Akbalık, Mehmet Erdem; Liman, Narin; Ketani, Muzaffer Aydın; Alan, EmelHücre adezyon molekülleri (Cell adhesion molecules-CAM), bir hücrenin baska bir hücre ile ya da ekstrasellüler matriksle iliskisini saglayan membran bagımlı (transmembranöz) proteinlerdir. Ekstrasellüler matriks ise fibröz proteinler ve glikozaminoglikanlardan olusur ve farklı kompozisyonu nedeniyle, destekleme, hücreler arası iletisim ve hücre dinamiginin düzenlenmesi gibi pek çok isleve sahiptir. Plasenta ekstrasellüler matriks proteinlerinden zengin bir doku olup, normal plasental gelisim ve bariyer fonksiyonu için hücre adezyon molekülleri ve ekstrasellüler matrix proteinleri gerekli moleküllerdir. Bu çalısma, ineklerde plasentada gebelik süresince hücre adezyon moleküllerinden biri olan kaderinler (E-, P-, V-,Nkaderinler) ile bazı ekstrasellüler matriks komponentlerinin (laminin, fibronektin, tip V kollagen) lokalizasyonları ile ekspresyonlarındaki olası degisimleri ortaya koymak için yapıldı. Bu çalısmada toplam 40 adet gebe Holstein inek uterusu kullanıldı. Uteruslar Diyarbakır?da bulunan özel kesimhanelerden temin edildi. Uterusların gebeligin hangi dönemine ait oldugunu belirlemek amacıyla tasımıs oldugu fötusların yasları hesaplandı. Doku örnekleri gebe kornunun plasentomal ve interplasentomal olmak üzere iki farklı bölgesinden alındı ve %10 nötral formalin solüsyonunda 24 saat tespit edildi. Rutin histolojik islemleri takiben de parafinde bloklandı. Hazırlanan parafin bloklarından, 5 mikrometre kalınlıgında seri kesitler alındı ve çalısmada belirtilen faktörlere iliskin uygun antikorlar kullanılarak immunohistokimyasal yöntemle boyandı. Western Blot için ise dokular alındı ve -84ºC?de islem yapılana kadar muhafaza edildi. Gebelik süresince, plasentanın yapısına katılan maternal epitel ve fötal trofoblastlarda E- ve P-kaderinin güçlü bir sekilde ekspresse oldugu, N-kaderinin fötal mezensimal hücrelerde, VE-kaderinin ise plasental damarların endotel ve düz kas hücrelerinde lokalize oldugu belirlendi. Fibronektinin, özellikle maternal stromal hücreler ve fötal mezensimal hücrelerde bulundugu, laminin ve kollajen tip V?in ise ekstraselüler matriks, maternal epitel ve fötal trofoblastlarda daha baskın oldugu ortaya konulmustur. Sonuç olarak, ineklerde gebeligin farklı dönemlerinde plasentanın föto-maternal bölümü ile interplasentomal bölgedeki uterus mukozasından kaderinlerin (E-, P-, N- ve VEkaderin) ve bazı bagdoku komponentlerinin (laminin, fibronektin ve kollajen tip V) degisik oranlarda lokalize olması, diger memeli türlerinde oldugu gibi plasental gelisimde ve gebeligin devamlılıgında bu faktörlerin ineklerde de önemli oldugunu göstermistir.Öğe Immunohistochemical localization of epidermal growth factor system in the lung of the Japanese quail (Coturnix coturnix japonica) during the post-hatching period(Wiley, 2015) Alan, Emel; Liman, Narin; Sağsöz, HakanThe purpose of this study is to determine the possible changes in the localization of the four Epidermal Growth Factor Receptors and three ligands in quail lungs from the first day of hatching until the 125th after hatching using immunohistochemical methods. Immunohistochemical results demonstrated that four EGFRs and their ligands are chiefly located in the cytoplasm of cells. Additionally, ErbB4, AREG, and NRG1 are localized to the nucleus and nucleolus, but EGF is present in the nucleolus. ErbB2 was also found in the cell membrane. In the epithelium of secondary bronchi, the goblet cells only exhibited ErbB1 and ErbB2, whereas the basal and ciliated cells exhibited EGFRs and ligands immunoreactivity. The atrial granular cells displayed moderate levels of ErbB1-ErbB3 and EGF and strong levels of ErbB4, AREG, and NRG1 immunoreactivity. While the squamous atrial cells and squamous respiratory cells of air capillaries and endothelial cells of blood capillaries exhibited moderate to strong ErbB2, ErbB4, AREG, and NRG1 immunoreactivity, they had negative or weak ErbB1, ErbB3, and EGF immunoreactivity. The expression levels of ErbB2-ErbB4, EGF, AREG, and NRG1 were also detected in fibroblasts. Although ErbB2 was highly expressed in the bronchial and vascular smooth muscle cells, weak expression of ErbB1, ErbB3, AREG and EGF and moderate expression of ErbB4 and NRG1 were observed. Macrophages were only negative for ErbB1. In conclusion, these data indicate that the EGFR-system is functionally active at hatching, which supports the hypothesis that the members of EGFR-system play several cell-specific roles in quail lung growth after hatching.Öğe The immunolocalization of cadherins and beta-catenin in the cervix and vagina of cycling cows(Springer Science and Business Media, 2023) Liman, Narin; Sağsöz, HakanThe adherens junctions (AJs) maintain the epithelial cell layers' structural integrity and barrier function. AJs also play a vital role in various biological and pathological processes. AJs perform these functions through the cadherin-catenin adhesion complex. This study investigated the presence, cell-specific localization, and temporal distribution of AJ components such as classical type I cadherins and beta-catenin in the cow cervix and vagina during the estrous cycle. Immunohistochemistry and Western blot analysis results demonstrated that beta-catenin and epithelial (E)-, neural (N)-, and placental (P)-cadherins are expressed in the cow cervix and vagina during the estrous cycle. These adhesion molecules were localized in the membrane and cytoplasm of the ciliated and non-ciliated cervical cells and the stratified vaginal epithelial cells. Positive immunostaining for P-, N-cadherin, and beta-catenin was also observed in the vascular endothelial cells of the cervical and vaginal stroma. Quantitative immunohistochemistry examinations revealed that in the cervical and vaginal epithelia, P-cadherin's optical density values (ODv) were the highest; in contrast, the N-cadherin ODv were the lowest. The ODv of P-cadherin and beta-catenin in the cervical epithelium and E-cadherin in the vagina were significantly higher in the luteal phase versus the follicular phase of the estrous cycle. Furthermore, the ODv of P-cadherin, N-cadherin, and beta-catenin in the cervix's central and peripheral epithelial regions were different during the estrous cycle. These findings indicate that classical cadherins and beta-catenin in the cervix and vagina exhibit cell- and tissue-specific expression patterns under the influence of estrogen and progesterone hormones during the estrous cycle.Öğe Immunolocalization of vascular endothelial growth factor, its receptors (flt1/fms, flk1/KDR, flt4) and vascular endothelial growth inhibitor in the bitch uterus during the sexual cycle(Elsevier Science Bv, 2013) Sagsoz, Hakan; Liman, Narin; Kucukaslan, Ibrahim; Saruhan, Berna GuneyAngiogenesis is regulated by proangiogenic and antiangiogenic factors. Vascular endothelial growth factor (VEGF) is a prime proangiogenic regulator, whereas vascular endothelial growth inhibitor (VEGI) is a specific antiangiogenic cytokine. To clarify temporal changes in the localization of pro-angiogenic and anti-angiogenic factors in the uterus of normal bitches during the proestrus, estrus, diestrus and anestrus phases of the estrous cycle, the expressions of VEGF and its receptors (flt1/fms, flk1/KDR and flt4) and their correlation with VEGI were analyzed using immunohistochemistry. Uteruses were collected after ovariohysterectomy. Immunohistochemical staining was evaluated semi-quantitatively by an immunohistochemical total score consisting of the sum of the intensity and proportional scores. The results in the bitch uterus demonstrated that positive immunohistochemical staining was found exclusively in the cytoplasm and apical membrane of luminal and glandular epithelial, stromal and smooth muscle cells and nuclear staining was observed in the fit1/fms, flk4 and VEGI during proestrous and estrous. Semi-quantitative analyses revealed that the total score for VEGF in the glandular epithelial cells was significantly higher than that of luminal, endometrial stromal and myometrial smooth muscle cells during proestrous (p < 0.05). The total score for flk1/KDR and flt4 in the glandular epithelium was also significantly higher than that of endometrial stromal cells during proestrous, whilst the total score for flt1/fms in the glandular epithelium was significantly higher than that of endometrial stromal cells during anestrus (p < 0.05). We conclude that, in the bitch uterus, cyclic changes may be precisely regulated by the combined functions of VEGF family members, angiogenic VEGF and VEGF receptors, and the angiogenesis inhibitor VEGI. (C) 2013 Elsevier B.V. All rights reserved.Öğe Physiological roles of the angiogenic factors during posthatching development period and adults in the quail lung(Wiley, 2016) Sagsoz, Hakan; Liman, Narin; Alan, EmelThe bronchus and vasculature form an intrinsic functional component of the avian lung, and its growth must be tightly regulated and coordinated by lung epithelial and endothelial development. Vascular endothelial growth inhibitor (VEGI), vascular endothelial growth factor (VEGF) and its receptors (flk1/KDR, flt1/fms, flt4) are required for epithelial and endothelial cell survival and apoptosis. Especially, VEGF and its receptors are critical for the development of the lung and serve as a maintenance factor during adult life. To determine the function of VEGI, VEGF and its receptors in the posthatching lung development, we revealed its expression and localization using by immunohistochemical procedure. VEGI, VEGF and its receptors were observed in the structural components of the bronchi, atria and air capillaries, as well as in the pulmonary blood vessels throughout the posthatching development period. On the other hand, immunostaining for VEGI, VEGF and its receptors was faintly detected in the glands of the secondary bronchi. Furthermore, it was determined that the secondary bronchial and atrial muscles did not display VEGF immunoreactions. Our results showed that VEGF and its receptors (flt1/fms, flk1/KDR and flt4) and VEGI were expressed at varying intensity by different cell groups. Therefore, they are also required for the development of the lung component during posthatching period.Öğe The profile of the epidermal growth factor system in rat endometrium during postpartum involution period(Springer, 2015) Alan, Emel; Liman, Narin; Sagsoz, HakanThe epidermal growth factor (EGF) plays a crucial role in the control of uterine cell proliferation, growth and differentiation. This study was designed to investigate the spatiotemporal expression pattern and localization of the EGF receptor/ligand system during the process of uterine involution using immunohistochemistry. Our results indicated that the expression of the ErbB/HER receptors and their ligands varied with structural changes in the uterus at different days of involution. Supranuclear punctate ErbB1 immunostaining was observed in the luminal and glandular epithelial cells and endometrial fibroblasts. Moderate ErbB2/HER2 immunoreactivity was observed in the lateral membrane and cytoplasm of the epithelial cells on the 1st, 3rd and 5th days and was decreased on the other days of involution. The amount of nuclear and cytoplasmic ErbB3/HER3 and ErbB4/HER4 immunostaining remained constant throughout the postpartum period. The EGF immunoreaction was weak in the luminal and glandular epithelium throughout the involution period. Although the cytoplasmic AREG immunoreactivity in the glandular epithelium was stronger on the 1st and 3rd days compared with the other days of involution, NRG1 immunostaining was weak on the 1st and 3rd days and was moderate in the apical cytoplasm on the 10th and 15th days of involution. The macrophages displayed strong cytoplasmic immunoreactivity for ErbB3/HER3, ErbB4/HER4, EGF, AREG and NRG. Strong, moderate and weak immunostaining for ErbB2/HER2, ErbB4/HER4 and other proteins (ErbB1, ErbB3, AREG and NRG), respectively, was present in the myometrial smooth muscle cells. These findings support the hypothesis that the EGFsystem plays a role in the development of various physiological changes associated with uterine involution.Öğe Tissue distribution of some immune cells in bovine reproductive tract during follicular and luteal phase(Wiley, 2018) Akbalık, Mehmet Erdem; Liman, Narin; Sağsöz, Hakan; Saruhan, Berna Güney; 0000-0001-9898-0593; 0000-0002-5456-697X; 0000-0002-5111-5524More recent studies indicate that immune cells which secrete their secretory products or cytokines play an important role in reproductive system. In our study, immune cell populations (CD8(+) T lymphocytes, CD68(+) macrophages, plasma cells, siderophages, eosinophils) and expression of major histocompatibility complex (MHC) class I and class II were examined in female reproductive tract during follicular (n=13) and luteal phase (n=10). Plasma cells and eosinophil granulocytes are present in few numbers in luminal epithelium, but abundant in longitudinal muscle layer of uterus, whereas siderophages are the dominant cell type in stroma. Moreover, MHC-I and -II+ cells are expressed by individual cells in organ layers, while CD8(+) T cells and CD68(+) macrophages are dominant in epithelium and muscle layer, respectively. In conclusion, we did not found significant changes in immune cells according to follicular and luteal phases, but localization and numbers in each organ have changed according to both organ and layers. These results indicate that these factors may play a crucial role not only to generate an immune response but also to have a role in regulation of physiological functions in female reproductive organs.
