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Öğe Does Rainbow Trout Seminal Plasma-Cysteine Combination Affect the Cryo-Survivability and Post-Thaw Incubation Resilience of Ram Semen?(Mary Ann Liebert, Inc, 2022) Gökçe, Elif; Üstüner, BurcuThe aim of this study was to evaluate the effect of both pure rainbow trout seminal plasma (RTSP) supplementation and RTSP-cysteine combination on cryopreservation success and post-thaw incubation resilience of ram semen in the nonbreeding season. For this purpose, different doses of RTSP (0%, 1%, 10%, and 15%) with or without cysteine supplementation were used for experiments. Ejaculates chosen for experiments were pooled and then divided into eight equal volumes for grouping (Control-ControlC, RTSP1-RTSP1C, RTSP10-RTSP10C, and RTSP15-RTSP15C). After cryopreservation, frozen-thawed semen samples were incubated for 5 hours at 37 degrees C for determination of post-thaw incubation resistance. Motility, HOST, TUNEL, Rh123-PI, and CTC tests were performed at 0 hour and 3rd and 5th hours of post-thaw incubation to evaluate the efficacy of all experimental groups. The RTSP10 and RTSP10C groups were noted to provide the best protection on motility, plasma membrane integrity, DNA integrity, and mitochondrial function of cryopreserved ram semen. On the other hand, the best protection against cryo-capacitation was observed in RTSP15 and RTSP15C groups. The addition of cysteine was found to be effective when the higher (15%) or lower (1%) doses of RTSP were used, as well as for no use of RTSP.Öğe Effects of triton X-100 pretreatment of lyophilized and frozen-thawed ram sperm on preimplantation embryo developmental competence(Taylor & Francis Inc, 2022) Üstüner, Burcu; Yağcıoğlu, Selin; Nur, Zekariya; Alçay, Selim; Demir, Kamber; Gökçe, Elif; Pabuccuoğlu, SerhatIn this study, it was aimed to determine the effect of destruction of lyophilized and frozen-thawed ram sperm plasma and acrosomal membrane on development of embryos produced by intracytoplasmic sperm injection (ICSI). Semen samples were divided into two groups for lyophilization (L) and freezing (F). For the removal of the plasma membrane, L and F groups were incubated with Triton X-100 (LTX-100 and FTX-100, respectively). Integrities of the plasma membrane, acrosome and chromatin structure were evaluated. Oocytes were injected with these sperm groups. Although no plasma membrane and acrosome integrities of the L (0.0%) group were detected, the plasma membrane integrity of the F group (69.4%) was significantly higher than the FTX-100 group (23.6%) (p < 0.05). The acrosome integrity of the FTX-100 group (3.80%) was significantly lower than the F group (55.6%) (p < 0.05). The chromatin integrities of L and F groups were higher than the Triton X-100 treated groups (p < 0.05). ICSIs with L, LTX-100, F and FTX-100 sperm were produced similar cleavage and blastocyst rates. In conclusion, data presented here confirm that ram spermatozoa can effectively be lyophilized and injected into oocytes for initiation of embryonic development and Triton X-100 pretreatment is not necessary while using lyophilized and frozen semen.Öğe Intracerebroventricular injection of histamine induces the hypothalamic-pituitary-gonadal axis activation in male rats(Elsevier Science Bv, 2018) Niaz, Nasir; Güvenç, Gökçen; Altınbaş, Burçin; Toker, Mehmed Berk; Aydın, Begüm; Udum Küçükşen, Duygu; Yalçın, Murat; Gökçe, ElifBrain histamine holds a key position in the regulation of behavioral states, biological rhythms, body weight, energy metabolism, thermoregulation, fluid balance, stress and reproduction in female animals. However, it is not clear whether central histamine exerts any effect on hypothalamic-pituitary-testicular in male rats and if so, the involvement of type of central histamine receptors. The current study was designed to determine the effect of centrally administrated histamine on plasma gonadotropin hormone-releasing hormone (GnRH), luteinizing hormone (LH), follicle stimulating hormone (FSH) and testosterone level, and sperm parameters, and to show the mediation of the central histaminergic H1, H2 and H3/H4 receptors on histamine-evoked hormonal and sperm parameters' effects. Studies were performed in male Sprague-Dawley rats. A total of 50 or 100 nmol doses of histamine were injected intracerebroventricularly (icy). 100 nmol dose of histamine significantly caused increases in plasma GnRH, LH, FSH and testosterone levels of animals, but not 50 nmol dose of histamine. Moreover, central pretreatment with chlorpheniramine, histaminergic H1 receptor antagonist (100 nmol), ranitidine and histaminergic H2 receptor antagonist (100 nmol) completely prevented histamine evoked increase in plasma GnRH, LH, FSH and testosterone levels, while thioperamide, histaminergic H3/H4 receptor antagonist (100 nmol) pretreatment failed to reverse sex hormones responses to histamine. Both central histamine treatment alone and central histamine treatment after central histaminergic receptors antagonists' pretreatments did not alter any sperm parameters in rats. In conclusion, our findings show that centrally administered histamine increases plasma GnRH, LH, FSH and testosterone levels of conscious male rats without change any sperm parameters. Moreover, according to our findings, central histaminergic H1, and H2 receptors mediate these histamine-induced effects.Öğe Kollajenin Saanen Teke Spermasının Dondurulabilirliği Üzerine Etkisi(Bursa Uludağ Üniversitesi, 2020) Üstüner, Burcu; Aktar, Ahmet; Yılmaz, Mehmet; Mülkpınar, Emine; Gökçe, Elif; Boz, Ezgi; Alçay, SelimBu çalışmada, Sazan Balığı (Cyprinus carpio) pullu derisinden elde edilen kollajen içerikli sıvı özütün teke spermasının dondurulabilirliği üzerine etkisinin belirlenmesi amaçlandı. Çalışmada, toplam 36 ejakülat altı baş tekeden gün aşırı elektro-ejakülatör ile alındı. En az +++ mass aktivite, %70 motilite ve 2x109 spermatozoon/mL özelliğe sahip sperma örnekleri birleştirilerek her grup için 4 eşit kısma bölündü. Sperma iki aşamalı sulandırma methodu ile final konsantrasyonu 1/5 (sperma/sulandırıcı) olacak şekilde; kollajen içermeyen kontrol grubu (K)ve farklı konsantrasyonda kollajen içeren (%1, %5 ve %10; sırasıyla K1, K5 ve K10) Tris-Na sitrat sulandırıcısı ile sulandırıldı. Payetlerprogramlanabilir dondurma makinasında donduruldu ve daha sonra sıvı azot içine aktarıldı. Her gruptan en az 3 payet 37ºC/30sn’de eritilerek eritme sonrası değerlendirmeler yapıldı. Sperma taze ve dondurma sonrası aşamalarda; motilite, plazma membran bütünlüğü Hypo-Osmotic Swelling Test (HOST) ve akrozom hasarı yönünden FITC-Pisum sativum agglutinin (FITC-PSA) boyama ile değerlendirildi. Eritmesonrası deney gruplarının motilite değerleri karşılaştırıldığında, kollajen içeren grupların motilitelerinin kollajen içermeyen kontrol grubuna göre daha yüksek olduğu gözlemlendi (P<0.05). Plazma membran bütünlüğünün, kollajen ilave edilen gruplarda kontrol grubuna göredaha yüksek olduğu tespit edildi (P<0.05). Akrozom hasarının sayısal olarak en yüksek kontrol grubunda olduğu gözlemlenmesine rağmen gruplar arasında istatistiksel bir fark tespit edilmedi (P>0.05). Sonuç olarak; teke spermasının dondurulmasında kullanılan sulandırıcılara%1, %5 ve %10 oranında kollajenin katılmasının motilite ve plazma membran bütünlüğü üzerine olumlu etkisi gözlemlendi.Öğe Long Term Incubation Resilience of Post-Thaw Ram Semen Diluted with Lecithin-Based Extender Supplemented with Bovine Serum Albumin(Kafkas Univ, Veteriner Fakultesi Dergisi, 2019) Alcay, Selim; Toker, Mehmed Berk; Gökçe, Elif; Önder, Nail Tekin; Üstüner, Burcu; Nur, ZekariyaThe objective of the study was to determine the optimum concentration of BSA in lecithin-based extender for post-thawing quality and incubation resilience (0 h, 6 h and 10 h) of ram sperm. Ejaculates were collected from five rams via electro ejaculation. Ejaculates were mixed to obtain pooled semen.Then, pooled semen was diluted with soybean lecithin-based extender without BSA (control) or supplemented with different concentrations of BSA (2.5 mg/mL, 5 mg/mL, 7.5 mg/mL and 10 mg/mL), at a final concentration of 150x106 spermatozoon/mL. Sperm motility, plasma membrane functional integrity (HOST), mitochondrial activity (rhodamine123), capacitation status (CTC), and DNA integrity (TUNEL) were evaluated. At the 10 h incubation, motility, plasma membrane functional integrity and mitochondria! function were better preserved in the BSA5 group compared to the control group. It was determined that high doses of BSA (5 mg/mL, 7.5 mg/mL and 10 mg/mL) affected acrosome reaction.The highest acrosome reaction rates were obtained in BSA10 groups in 6 h and 10 h incubation (P<0.05). TUNEL assay demonstrated that there were no differences among groups for DNA fragmentation at post-thaw and during incubation periods. The study shows that BSA supplemented extenders may have beneficial effect on ram semen parameters at 0 h, 6 h and 10 h of incubation. The results of the present study demonstrated a remarkable advantage of using 5 mg/mL of BSA in 1% lecithin-based extender.Öğe Lyophilized Extender Supplemented with Rainbow Trout (Oncorhynchus mykiss) Seminal Plasma Improves Cryopreservation of Ram Sperm(Kafkas Univ, Veteriner Fakultesi Dergisi, 2022) Üstüner, Burcu; Alçay, Selim; Gökçe, Elif; Yılmaz, M. Melih; Aktar, Ahmet; Hüraydın, Oğuzhan; Nur, ZekariyaThis study aimed to investigate the effect of the Rainbow trout seminal plasma (RTSP) supplemented (control, 10% or 15%) lyophilized extender on freezability of ram semen. Collected semen was pooled and split into two aliquots, and each of the ejaculates was diluted with fresh or lyophilized extenders with RTSP (0%, FC and LC; 10% F10 and L10or 15% F15 and L15) using two-step dilution method. Semen was frozen using the programmable freezing machine. Semen samples were examined for sperm motility, defective acrosomes, plasma membrane integrity and DNA fragmentation at native and post-thaw stage. The highest percentages of post-thaw motility and plasma integrity were observed in the F10, F15, and L15 (P 0.05). There was no significant difference in the rate of post-thaw defective acrosomes when the other extender groups were compared with the control (P 0.05). The highest percentage of post-thaw DNA fragmentation values were observed in the FC group, while the lowest DNA fragmentation was obtained in the F15 but only significant different from FC and LC groups. In conclusion, the findings of this study show that the lyophilized extender with 15% RTSP added can be used successfully for freezing ram semen.Öğe Successful cryopreservation of honey bee drone spermatozoa with royal jelly supplemented extenders(Academic Press Inc Elsevier Science, 2019) Alcay, Selim; Çakmak, Selvinar; Çakmak, İbrahim; Mulkpinar, Emine; Gökçe, Elif; Üstüner, Burcu; Nur, ZekariyaThe aim of the present study was to evaluate different concentrations of royal jelly (RJ) supplemented extenders for post-thawing quality of drone sperm. Semen samples were collected from sexually mature drones. Pooled semen was diluted with extender without RJ (control) or supplemented with different concentrations of RJ (1, 2, 4 or 8%). Sperm motility, plasma membrane functional integrity, and acrosomal integrity were evaluated. At post thaw, the highest sperm motility and acrosomal integrity rates were obtained in the RJ1 group. Functional integrity of sperm membrane was better preserved in the RJ1 and RJ2 groups compare to the other groups. The study shows that RJ supplemented extenders have beneficial effects on drone semen parameters. The results of the present study demonstrated advantage of using 1% RJ supplemented extender.Öğe The Combined Effect of Melatonin Implant and Removal of Buck Seminal Plasma on Cryopreservation During the Nonbreeding Season(Mary Ann Liebert, Inc, 2022) Üstüner, Burcu; Üstüner, Hakan; Gökçe, Elif; Önder, N. Tekin; Yılmaz, M. Melih; Huraydin, Oğuzhan; Toker, M. BerkThis study aimed to determine how melatonin (MT) and seminal plasma affected the freezability of buck sperm during the nonbreeding season. Semen was collected from eight bucks before (pre-MT) and after (post-MT) MT application in the nonbreeding season. Individual ejaculates were collected from the bucks, split into two equal groups according to the removal of seminal plasma (SP) (-) or nonremoval of SP (+). For washing, the groups of ejaculates were centrifuged, and the supernatant was separated, SP (-) and SP (+) ejaculates were diluted, then frozen. Semen samples were examined for sperm motility, plasma membrane integrity, defective acrosomes, DNA fragmentation, and mitochondrial membrane function at the native and post-thaw stages. When the general average post-thaw motility (p < 0.01), plasma membrane (p < 0.05), acrosome (p < 0.05), and DNA integrity rates (p < 0.05) and mitochondrial membrane potential (MMP) (p < 0.01) were evaluated, it was seen that MT administration caused a statistically significant improvement. The dramatic effect of nonremoval of seminal plasma on motility and plasma membrane integrity is more clearly observed in individual semen samples frozen in the pre-MT group (p < 0.05). Also, it was observed that removing seminal plasma in the post-MT group caused even milder post-thaw acrosome damage compared with the SP (+) group (p < 0.05). The effect of removing seminal plasma was not observed in terms of DNA integrity and MMP rates in pre- and post-MT groups. As a result, it was concluded that MT application and removal of seminal plasma in the nonbreeding season result in improvement in the freezability of buck semen.
