Tesis

Analisis Aktivitas Spesifik Dinein ATPase dan Ekspresi Dinein ATPase-associated with various cellular activities (AAA) Pada Spermatozoa Laki-laki Oligoastenozoospermia dan Nekrozoospermia = Analysis of dynein ATPase specific activity and expression of dynein ATPase-associated with various cellular activities (AAA) on human sperm with oligoasthenoteratozoospermia and nekrozoospermia men.

LATAR BELAKANG: Analisis semen merupakan pemeriksaan yang dapat memberikan informasi tentang jumlah, motilitas dan morfologi spermatozoa pada ejakulat. Oligoastenoteratozoospermia (OAT) dan nekrozoospermia merupakan keadaan infertil yang berkaitan dengan gangguan motilitas, rendahnya jumlah serta abnormalitas morfologi spermatozoa. Motilitas spermatozoa ditentukan oleh bagian ekor yang di dalamnya terdapat struktur kompleks disebut aksonem. Motilitas spermatozoa yang ditentukan oleh komponen dalam aksonem seperti dinein, memerlukan energi hasil hidrolisis ATP melalui aktivasi enzim ATPase pada lengan dinein. Sampai saat ini belum diketahui bagaimana ekspresi domain penting dari dinein yaitu AAA1 dan AAA2 serta aktivitas dinein ATPAse pada spermatozoa pasien Oligoastenoteratozoospermia dan nekrozoospermia. METODE: Penelitian ini merupakan studi kasus kontrol yang menganalisis aktivitas dinein ATPase serta ekspresi dinein AAA1 dan AAA2 pada spermatozoa 14 laki-laki OAT dan 10 nekrozoospermia dibandingkan dengan 14 laki-laki normozoospermia. Analisis aktivitas spesifik dinein ATPase dilakukan dengan metode Fiske Subarrow. Ekspresi dan distribusi protein AAA dinein dilakukan dengan metode ELISA dan imunositokimia. Analisis statistik yang dilakukan adalah uji t tidak berpasangan, uji Mann Whitney, Uji Annova One Way dan Uji Kruskal-wallis dengan kemaknaan p < 0.05. HASIL: Aktivitas spesifik dinein ATPase pada spermatozoa kelompok OAT dan nekrozoospermia lebih rendah dibandingkan dengan spermatozoa kelompok normozoospermia (p = 0,000). Terdapat perbedaan bermakna antara ekspresi domain AAA1 protein dinein spermatozoa kelompok OAT dan nekrozoospermia dibandingkan dengan kelompok normozoospermia (p = 0,003 dan p = 0,000); serta ekspresi AAA2 dinein pada kelompok OAT dan nekrozoospermia dibandingkan dengan normozoospermia (p = 0,048 dan p = 0,008). Distribusi dinein AAA1 dan AAA2 yang kurang padat dan tidak merata pada ekor dan kepala spermatozoa kelompok OAT dan nekrozoospermia, dibandingkan kelompok normozoospermia. KESIMPULAN: Aktivitas dinein ATPase dan ekspresi dinein AAA1 serta AAA2 pada spermatozoa kelompok OAT dan nekrozoospermia lebih rendah dibandingkan dengan kelompok normozoospermia.
Kata Kunci: dinein ATPase, dinein AAA1, AAA2, Oligoastenoteratozoospermia, Nekrozoospermia.


Background: Semen analysis is an examination which provide information about the number, motility and morphology of sperm. Oligoasthenoteratozoospermia (OAT) and necrozoospermia are some of sperm analysis disorders, associated with motility disorder, as well as low number of sperm and morphological abnormalities. An axial complex, known as axoneme, plays a major role in sperm motility. Sperm motility determined by the component in axoneme like dynein, requires the energy of ATP hydrolysis through the activation of ATPase at dynein arm. Recently, there were limited study about dynein ATPase activity on sperm. The expression of main domain of dynein, AAA1 and AAA2, and the activity of dynein ATPase in OAT and necrozoospermia patients is still not fully understood. Methods: This research was a case-control study, comparing as many as 14 men with normozoospermia and OAT and 10 necrozoospermia. Analysis of dynein ATPase activity was conducted using the enzyme activity assay Fiske Subarrow method. Expression of AAA protein was done by ELISA and immunocytochemistry. Statistical analysis was performed by unpaired t test, Mann Whitney test, Annova One Way Test and the Kruskal-Wallis test with significance level (p < 0.05). Result: The activity of dynein ATPase in OAT and necrozoospermia were significantly lower compared to normozoospermia (p=0,000). We also found a significantly different of AAA1 domain of dynein expression between OAT and necrozoospermia as compared to normozoospermia (p=0,003; p=0,000), as well as AAA2 domain (p=0,048; p=0,008). This result was confirmed by immunocytochemistry test that demonstrated the distribution of proteins which are less compact in the head and almost not detected in the tail on sperm of OAT and necrozoospermia, compared to normozoospermia group. Conclusion: Dynein ATPase activity and the expression of AAA1 and AAA2 domain in sperm of OAT and necrozoospermia were lower compared to normozoospermia. Expression of AAA1 and AAA2 dynein experienced a significant reduction in OAT and necrozoospermia compared to normozoospermia group.
Keywords: Dynein ATPase, AAA1 dynein, AAA2 dynein, Oligoasthenoteratozoospermia, Necrozoospermia