İnsan sperminde kisspeptin ve sperm katyon kanallarının (CatSper) birbirleriyle olan ilişkisi
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Dosyalar
Tarih
2024
Yazarlar
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Cilt Başlığı
Yayıncı
Selçuk Üniversitesi, Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
KISS1R, G proteinin alt biriminden Gq/11α'yla bağlantılı G proteiniyle bağlı reseptördür. KISS1'in KISS1R'ye bağlanmasıyla Fosfotidilinositol (PI)'ü uyarıp fosfolipaz C'nin aktivasyonunu sağlar. Aktivasyonu sonucu fosfatidil inositol bifosfatın üzerinden inositol-trifosfat (IP3) ve diaçilgliserol (DAG) oluşumunu sağlamaktadır. IP3 Ca2+ salınımını sağlamaktadır. Bu döngü ile KISS1'in sperm hareketliliğinde etkisi kabul edilmekle birlikte yeterince bilinmemektedir. Benzer şekilde spermdeki kalsiyum kanallarından CatSper3'ün etkinliği CatSper1 ve CatSper2'ye nazaran yeterince araştırılmamıştır. Çalışmanın, insan sperminde CatSper3 ve KISS1 ekspresyonlarının, sperm motilitesi ile ilişkilerinin değerlendirilerek, erkek infertilitesindeki rollerinin belirlenmesine katkıda bulunması amaçlanmaktadır. Çalışma kapsamında Selçuk Üniversitesi Tıp Fakültesi Tüp Bebek Merkezine rutin spermiyogram testi için başvuran 18-45 yaş aralığındaki 52 erkek birey dahil edildi. WHO'nun belirttiği kriterlere göre semen parametrelerinin; semen hacmi, sperm konsantrasyonu, toplam sperm sayısı, sperm motilitesi, progresif motilite (PM), hızlı progressif motilite, yavaş progresif motilite, değerlendirildi. Çalışmada 2 grup oluşturuldu. Bunlar; birinci grup Normozoospermi Grubu (n=26) sperm konsantrasyonu 15 milyon/ml'den büyük, sperm motilitesi >39 veya PM >30, ikinci grup olarak da Astenozoospermi Grubu (n=26) sperm konsantrasyonu 15 milyon/ml'den büyük, sperm motilitesi <39 veya PM <30 koşullarını sağlayanlar olarak belirlenmiştir. Semen örnekleri immünfloresan işaretleme ile Anti-CatSper3 ve Anti-Kiss1R primer antikorları ile işaretlendi. Kesitler, flüoresan mikroskop altında değerlendirilerek ekspresyon yüzdeleri belirlendi. Semen örneklerinin genetik analizi yapılarak CatSper3 ve KISS1R gen ekspresyonlarına bakıldı. KISS1R ve CatSper3 ekspresyonları ve gen ifadeleri gruplar arasında karşılaştırıldı. Sonuçlarımıza göre CatSper3 ekspresyonlarının, normozoospermi grubunda astenozoospermi grubuna göre total boyanma yüzdesinde anlamlı bir fark gözlenmedi. Ancak boyanma bölgeleri değerlendirildiğinde: normozoospermik bireylerde sperm membranı ve boyun bölgesindeki Catsper3 işaretlenme oranının astenozoospermik bireylerden daha yüksek olduğu görüldü. Kiss-Peptin işaretlenme düzeyleri incelendiğinde, normozoospermik bireyler ve astenozoospermi bireylerdeki ekspresyonlar arasında anlamlı bir istatistiksel fark saptandı. Membran ve baş-kuyruk bölgesinde Kiss-Peptin işaretlenme yüzdelerinin normozoospermik bireylerde astenozoospermik bireylere kıyasla anlamlı şekilde yüksek olduğu görüldü. Öte yandan baş bölgesinde Kiss-Peptin işaretlenme yüzdelerinin normozoospermik bireylerde astenozoospermik bireylerden anlamlı bir şekilde düşük olduğu tespit edildi. Yapılan genetik analizler sonucunda ise CatSper3, KISS1R gen ifadelerinde gruplar arası anlamlı fark saptanmadı. Sperm hareketliliği için değerli olan bu yolakların molekülleri erkek infertilitesinin belirteçleri olarak kullanılabilir. Tanı ve tedavide yeni hedef ve stratejilerin geliştirilmesine katkı sağlayabilir.
KISS1R is a G protein-coupled receptor associated with the Gq/11α subunit of G protein. By binding of KISS1 to KISS1R, it stimulates phosphotidylinositol (PI) and activates phospholipase C. As a result of its activation, it provides the form of inositol-triphosphate (IP3) and diacylglycerol (DAG) from phosphatidyl inositol biphosphate. IP3 provides Ca2+ release. Although the effect of KISS1 on sperm motility with this cycle is accepted, it is not known enough. Similarly, the effectiveness of CatSper3, one of the calcium channels in sperm, has not been adequately investigated compared to CatSper1 and CatSper2. The main aim of this thesis is to contribute to determining their roles at male infertility by evaluating the expressions of CatSper3 and KISS1 in human sperm and their relationship with sperm motility. Within the scope of the study, 52 male individuals who are between 18-42 years old and applied to Selçuk University Faculty of Medicine Hospital, In Vitro Fertilization Center for routine spermiogram test were included. According to the criteria specified by WHO, main parameters of semen; concentration of sperma, volume, motility of semen, total count of sperma, progressive motility (PM), fast progressive motility, slow progressive motility were evaluated. Two groups were created in the study; normozoospermia (n=26) sperm concentration >15 million/ml sperm motility >39 or PM >30 is the first group. Asthenozoospermia (n=26) sperm concentration >15 million/ml, sperm motility <39 or PM <30 are the second group, was determined as. Semen samples were labeled with CatSper3 and Anti-Kıss1R primary antibodies by immunofluorescence staining. The fraction were evaluated under a fluorescent microscope and expression percentages were determined. Genetic analysis of semen samples was performed and CatSper3 and KISS1R gene expressions were examined. KISS1R and CatSper3 expressions and gene expressions of them were compared by groups. Our results showed that no significant difference was observed in the total labeling percentage of CatSper3 expressions in the normozoospermia group compared to the asthenozoospermia group. However, when the staining areas were evaluated, it was seen that the percentage of Catsper3 staining in the membrane and neck in normozoospermia individuals was higher than in asthenozoospermia individuals. When Kiss-Peptin staining levels were examined, a significant statistical difference was detected in their expression between normozoospermia individuals and asthenozoospermia individuals. It was observed that Kiss-Peptin labeling percentages in the membrane and head-tail region were significantly higher in normozoospermia individuals compared to asthenozoospermia individuals. On the other hand, it was determined that Kiss-Peptin staining percentages in the Head region were significantly lower in normozoospermia individuals than in asthenozoospermia individuals. As a result of the genetic analysis, no significant difference was detected between the groups in CatSper3 and KISS1R gene expressions. Molecules of these pathways, which are valuable for sperm motility, can be used as markers of male infertility. It may contribute to the development of new targets and strategies in diagnosis and treatment.
KISS1R is a G protein-coupled receptor associated with the Gq/11α subunit of G protein. By binding of KISS1 to KISS1R, it stimulates phosphotidylinositol (PI) and activates phospholipase C. As a result of its activation, it provides the form of inositol-triphosphate (IP3) and diacylglycerol (DAG) from phosphatidyl inositol biphosphate. IP3 provides Ca2+ release. Although the effect of KISS1 on sperm motility with this cycle is accepted, it is not known enough. Similarly, the effectiveness of CatSper3, one of the calcium channels in sperm, has not been adequately investigated compared to CatSper1 and CatSper2. The main aim of this thesis is to contribute to determining their roles at male infertility by evaluating the expressions of CatSper3 and KISS1 in human sperm and their relationship with sperm motility. Within the scope of the study, 52 male individuals who are between 18-42 years old and applied to Selçuk University Faculty of Medicine Hospital, In Vitro Fertilization Center for routine spermiogram test were included. According to the criteria specified by WHO, main parameters of semen; concentration of sperma, volume, motility of semen, total count of sperma, progressive motility (PM), fast progressive motility, slow progressive motility were evaluated. Two groups were created in the study; normozoospermia (n=26) sperm concentration >15 million/ml sperm motility >39 or PM >30 is the first group. Asthenozoospermia (n=26) sperm concentration >15 million/ml, sperm motility <39 or PM <30 are the second group, was determined as. Semen samples were labeled with CatSper3 and Anti-Kıss1R primary antibodies by immunofluorescence staining. The fraction were evaluated under a fluorescent microscope and expression percentages were determined. Genetic analysis of semen samples was performed and CatSper3 and KISS1R gene expressions were examined. KISS1R and CatSper3 expressions and gene expressions of them were compared by groups. Our results showed that no significant difference was observed in the total labeling percentage of CatSper3 expressions in the normozoospermia group compared to the asthenozoospermia group. However, when the staining areas were evaluated, it was seen that the percentage of Catsper3 staining in the membrane and neck in normozoospermia individuals was higher than in asthenozoospermia individuals. When Kiss-Peptin staining levels were examined, a significant statistical difference was detected in their expression between normozoospermia individuals and asthenozoospermia individuals. It was observed that Kiss-Peptin labeling percentages in the membrane and head-tail region were significantly higher in normozoospermia individuals compared to asthenozoospermia individuals. On the other hand, it was determined that Kiss-Peptin staining percentages in the Head region were significantly lower in normozoospermia individuals than in asthenozoospermia individuals. As a result of the genetic analysis, no significant difference was detected between the groups in CatSper3 and KISS1R gene expressions. Molecules of these pathways, which are valuable for sperm motility, can be used as markers of male infertility. It may contribute to the development of new targets and strategies in diagnosis and treatment.
Açıklama
Anahtar Kelimeler
Erkek İnfertilitesi, Kisspeptin, KISS1/KISS1R, Sperm Motilitesi, CatSper3, Male Infertility, Sperm Motility
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Aniç S., (2024). İnsan Sperminde Kisspeptin ve Sperm Katyon Kanallarının (Catsper) Birbirleriyle Olan İlişkisi. (Yüksek Lisans Tezi). Selçuk Üniversitesi, Fen Bilimleri Enstitüsü, Konya.
