Investigation on porcine aromatase (cyp19) as a specific target gene for boar testis

Sitokrom P450 aromataz östrojen üretiminde önemli bir enzim olup, CYP19 geni tarafından ifade edilir. CYP19 geninin yokluğunda farelerde sperm üretiminin zarar gördüğü ve ilişkili olarak spermlerin yüzme ve yumurtaları dölleme kabiliyetinin azaldığı gözlenmiştir. Buna karşılık CYP19’un domuz sperm üretiminde ve yumurtaların döllenmesinde nasıl bir rol oynadığı bilinmemektedir. Bu nedenle, bu calışmanın amacı CYP19 mRNA ve protein ifadesini farklı sperm kalite özelliklerine sahip olan erkek domuzların üreme organı dokularında araştırmaktır. mRNA ve protein ifadesi çalısması için altı erkek domuz, grup 1 (G-I) ve grup 2 (G-II) olarak iki gruba ayrılmıştır. G-I hayvanlar göreceli olarak G-II hayvanlara göre daha iyi sperm özelliklerine sahiptir. Sonuçlar CYP19 mRNA ifadesinin tüm erkek üreme organlarında ifade edilmediğini göstermiştir. CYP19’un mRNA ifadesi en çok testis dokularında gözlenmiştir ve ayrıca G-I ve G-II domuzlardan alınan testis örneklerinde aynı düzeyde tespit edilmiştir. CYP19’un Western-Blot protein ifadesi sonuçları qRT-PCR sonuçlarından farklı olarak gözlenmiştir. CYP19 protein ifadesi G-II hayvanlarda G-I hayvanlara göre daha yüksek düzeyde gözlenmiştir. Testiste CYP19 protein lokalizasyonu sitoplazma Leydig hücrelerinde güçlü bir sinyal göstermiştir. Sonuçlar, CYP19 geninin domuzlarda sperm üretimi için testislerde spesifik bir hedef gen olduğuna dair yeni bulgular ortaya koymuştur.

Anahtar Kelimeler:

transkripsiyon, leydig hücreleri, sitokrom P-450, gen ifadesi, genler, elçi RNA, domuz, sperma oluşması, testis

Erkek domuzlarda testisler için hedef bir gen olan aromataz (cyp19) üzerine araştırma

Cytochrome P450 aromatase is the key enzyme in estrogen biosynthesis, encoded by CYP19 gene. Impairment of spermatogenesis associated with a decrease in sperm motility and inability to fertilize oocytes in mice due to the lacking of CYP19 was observed. However, it is little known about the CYP19 roles in boar spermatogenesis and fertility. Therefore, the aim of this research was to investigate the mRNA and protein expression of CYP19 in boar reproductive tissues from boars with different sperm quality. For mRNA and protein expression study, a total of six boars were divided into two groups with Group 1 (G-I) and Group 2 (G-II) where G-I was characterized for relatively a better sperm quality. The result showed that the CYP19 transcript was not expressed throughout the male reproductive system. mRNA expression of CYP19 was higher only in testis. CYP19 expression was similar in testis collected from G-I and G-II boars. The CYP19 protein expression results from western blot were different with the results of qRT-PCR. The CYP19 protein was higher in testis collected from G-II than G-I boars. The CYP19 protein localization in testis showed a strong staining only in the cytoplasm Leydig cell. These results shed new light on the roles of porcine CYP19 in spermatogenesis as a specific target gene for testis.

Keywords:

transcription, Leydig cells, cytochrome P-450, gene expression, genes, messenger RNA, pigs, spermatogenesis, testes,

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