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Abstrak :
Kriopreservasi sperma ikan kerapu kertang Epinephelus lanceolatus (Bloch, 1790) merupakan suatu teknik untuk membantu mengatasi masalah pemijahan yang tidak sinkron antara induk ikan kerapu kertang. Penelitian ini menggunakan konsentrat kurma dan gliserol sebagai krioprotektan. Tujuan dari penelitian yaitu untuk mengetahui pengaruh pemberian konsentrat kurma dan gliserol terhadap kualitas sperma yaitu motilitas, viabilitas, dan abnormalitas 48 jam pascakriopreservasi serta kemampuan fertilisasinya dengan ikan kerapu macan Epinephelus fuscogutatus (Forskal, 1775). Konsentrat kurma yang digunakan dalam penelitian ini dengan berbagai konsentrasi, yaitu 0%; 5%; 10%; 15%; 20%; dan 25%. Rasio antara sperma dengan larutan pengencer yang digunakan yaitu 1:9. Ekstender yang digunakan yaitu larutan marine fish Ringer. Kualitas sperma segar terlebih dahulu dievaluasi secara makroskopis (volume, pH, warna, dan bau) dan secara mikroskopis (motilitas, viabilitas, dan abnormalitas) untuk menguji kelayakan sperma yang akan dikriopreservai dan kemampuan fertilitasnya dengan menghitung persentase fertilisasi. Kriopreservasi dilakukan pada freezer dengan suhu -20⁰C selama 48 jam. Spermatozoa hasil kriopreservasi digunakan untuk membuahi sel telur ikan kerapu macan. Berdasarkan hasil uji statistik ANOVA satu arah dilanjutkan dengan Uji Tukey, diketahui bahwa kombinasi konsentrat kurma dengan berbagai konsentrasi dan gliserol memiliki nilai rata-rata yang berpengaruh nyata terhadap motilitas dan abnormalitas sperma pascakriopreservasi (P<0,05), akan tetapi tidak berpengaruh nyata (P>0,05) terhadap viabilitasnya. Meskipun demikian hasil fertilisasinya dengan ikan kerapu macan menunjukkan hasil yang berpengaruh nyata (P<0,05). Hasil terbaik ditunjukkan pada konsentrasi konsentrat kurma 10% dengan nilai persentase 76,70 ± 1,54% pada motilitas sperma, nilai persentase viabilitas yaitu 77,67 ± 5,78%, serta nilai kemampuan fertilisasi yaitu 66,25 ± 3,23%, dengan nilai persentase rata-rata abnormalitas 21,53 ± 0,84%.

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Cryopreservation of giant grouper sperm Epinephelus lanceolatus (Bloch, 1790) is a technique to overcome the problem of unsynchronized spawning of grouper. Glycerol and palm dates were used as a cryoprotectant. The aim of this study was to determine the effect of palm date concentrate and glycerol on motility, viability, and abnormality 48 hours postcryopreservation and the ability of fertilization using giant grouper sperm postcryopreservation with tiger grouper (Epinephelus fuscogutatus ). The palm date concentrations used in this study were 0%; 5%, 10%, 15%, 20%, and 25%. The ratio between sperm and diluent solution was 1:9. The extender used in this study is a marine fish Ringer solution. Fresh sperm were evaluated macroscopically (volume, pH, color, and odor) and microscopically (motility, viability, and abnormality) to test the sperm feasibility for cryopreservation and fertility ability by calculating the percentage of fertilization. Sperm was stored in a freezer -20⁰C for 48 hours. Spermatozoa postcryopreservation were used to fertilize tiger grouper eggs. Based on the results of statistical tests of one-way ANOVA followed by the Tukey Test, it was shown that palm date concentrate with various concentrations had an average value that significantly affected motility sperm and abnormality after cryopreservation (P<0.05), but had no significant effect (P>0.05) to viability. However, the results of fertilization with tiger grouper showed a significant effect (P<0.05). The best results were shown in the concentration of 10% palm date concentrate with a percentage value of 76.70 ± 1.54% in motility sperm, the value of the viability percentage is 77.67 ± 5.78%, and the value of fertilization ability is 66.25 ± 3.23%, with an average percentage value of abnormality is 21.53 ± 0.84%.

Abstrak :
The effect of Dimethyl Formamide (DMF) as a cryoprotectant on spermatozoa quality of carp fish Cyprinus carpio race Majalaya 24 hours post-cryopreservation was studied. This study was conducted in the Environmental Engineering Laboratory. Puspiptek. Serpong. Sperm was collected by the hand stripping method and were immediately diluted with extender [6J and cryoprotectant (DMF). The final concentrations of DMF were 0. 3.75. 7.5. and ! 1.25%. respectively. Sperm were frozen in liquid nitrogen for 24 hours. All sperm were re-examined after thawing for motility. viability, and abnormality. The highest percentage of the molility and the viability of preserved sperm was showed or the DMF 7.5%. On the oiher hand the lowest percentage of the abnormality of preserved sperm was demonstrated by the DMF 7.5%. also. Accordingly. DMF 7.5% is the optimum levels that could preserved spermatozoa still in a good quality 24 hours posl-cryopreservation.

Abstrak :
LATAR BELAKANG: Infertilitas laki-laki salah satunya diakibatkan ketidakseimbangan transpor ion pada spermatozoa, yang dapat menyebabkan gangguan motilitas spermatozoa. Keseimbangan transpor ion untuk memelihara homeostasis spermatozoa yang dimediasi oleh ATPase, diantaranya adalah Na+, K+-ATPase dan Ca2+-ATPase. Beberapa penelitian telah membuktikan bahwa aktivitas spesifik Na+, K+-ATPase dan Ca2+-ATPase menurun pada kelompok astenozoospermia. Akan tetapi belum diketahui bagaimana aktivitas spesifik Na+, K+-ATPase dan Ca2+-ATPase pada kelompok infertilitas lain, yaitu oligoastenoteratozoospermia (OAT) dan nekrozoospermia. Oleh karena itu, pada penelitian ini diteliti aktivitas spesifik Na+, K+-ATPase dan Ca2+-ATPase beserta ekspresi protein Na+, K+-ATPase isoform α4 dan PMCA4 pada kelompok OAT dan nekrozoospermia. METODE: Pada sampel dilakukan analisis semen, isolasi sel, protein dan fraksi membran. Analisis semen dilakukan secara mikroskopik dengan makler, disertai uji HOS dan viabilitas. Aktivitas enzim diukur berdasarkan kemampuan ATPase melepaskan fosfat organik dari ATP dan ditetapkan sebagai aktivitas spesifik. Ekspresi protein Na+, K+-ATPase isoform α4 dan PMCA4 dilakukan dengan teksnik western blot, sedangkan distribusi proteinnya digunakan teknik imunositokimia. HASIL: Pada penelitian ini diperoleh hasil bahwa hampir pada seluruh parameter baik motilitas, viabilitas, integritas membran, aktivitas Na+,K+-ATPase dan Ca2+- ATPase, serta ekspresi protein Na+,K+-ATPase Isoform α4 dan PMCA4 pada kelompok OAT dan nekrozoospermia cenderung lebih rendah dibandingkan dengan normozoospermia. Penurunan ekspresi PMCA4 pada kelompok OAT dan nekrozoospermia berbeda bermakna (p<0,05) dibandingkan dengan normozoospermia. Hasil uji korelasi ekspresi Na+,K+-ATPase Isoform α4 dengan aktivitas spesifik Na+,K+-ATPase menunjukkan korelasi negatif yang lemah namun tidak bermakna. Demikian pula halnya dengan hasil uji ekspresi PMCA4 dan aktifitas spesifik Ca2+-ATPase. KESIMPULAN: Aktivitas spesifik Na+,K+-ATPase dan Ca2+-ATPase serta ekspresi Na+,K+-ATPase isoform α4 dan PMCA4 yang tinggi diperlukan untuk mendukung homeostasis sperma sehingga sperma mempunyai motilitas yang baik.

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BACKGROUND: Male infertility could be caused by sperm motility disorders. This condition is caused by imbalance ion transport. The balance of ion transport to maintain homeostasis of spermatazoa, is mediated by the ATPase, including the Na+,K+-ATPase and Ca2+-ATPase. Several studies have shown that the specific activity of Na+,K+-ATPase and Ca2+-ATPase decreased in group astenozoospermia. There is no data about the specific activity of Na+,K+-ATPase and Ca2+-ATPase in the other infertility cases, i.e, oligoasthenoteratozoospermia (OAT) dan necrozoospermia group. Therefore, this study investigated the specific activity of Na+,K+-ATPase and Ca2+-ATPase and the expression of Na+,K+-ATPase isoform α4 and PMCA4 protein in both cases. METHODS: Samples were examined for semen analysis, cell isolation, proteins and membrane fractionation. Semen analysis performed microscopically by Makler, accompanied by HOS and viability test. Enzyme activity is measured by the ability ATPase to release organic phosphate from ATP and defined as a specific activity. The protein expression of Na+,K+-ATPase isoform α4 and PMCA4 done with western blot technique, while the distribution of the protein used immunocytochemistry techniques. RESULT: This study showed that almost all the parameters such as motility, viability, membrane integrity, the activity of Na+,K+-ATPase and Ca2+-ATPase, and protein expression of Na+,K+-ATPase isoforms α4 and PMCA4 in the OAT and necrozoospermia were tendency lower than the normozoospermia. Decrease expression of PMCA4 on OAT and nekrozoospermia group were significantly difference compared to normozoospermia. The correlation of Na+,K+-ATPase isoform α4 intensity with specific activity of Na+,K+-ATPase as well as PMCA4 was weak negatife but not significantly. CONCLUSSION: The specific activity of Na+,K+-ATPase and Ca2+-ATPase and high expression of Na+,K+-ATPase isoform α4 and PMCA4 support homeostasis condition in sperm leading achievement of good motility.

Abstrak :
LATAR BELAKANG : Salah satu hambatan dalam program reproduksi berbantu adalah rendahnya viabilitas dan motilitas sel spermatozoa. Analisis proteomik menunjukkan adanya banyak protein yang diduga berperan dalam regulasi motilitas dan viabilitas spermatozoa antara lain progesteron. Penelitian ini bertujuan untuk menganalisis efek prosurvival progesteron terhadap spermatozoa melalui penekanan apoptosis.BAHAN DAN CARA KERJA : Sampel spermatozoa dicuci dengan sentrifugasi gradient. Sampel spermatozoa di tambahkan progesteron dengan konsentrasi 0 kontrol , 250, 500, 750 dan 1000 ng/mL. Setelah perlakuan, sampel dilakukan pemeriksaan integritas membran dengan metode HOS dan pemeriksaan motilitas dengan Computer Assisted Sperm Analyzer CASA . Deteksi protein fosforilasi tirosin dan Akt serta aktivitas kaspase dilakukan dengan metode western blot.HASIL : Efek penambahan progesteron meningkatkan rerata motilitas spermatozoa namun berbeda tidak bermakna p>0.05 . Integritas membran spermatozoa tidak berpengaruh pada pemberian progesteron. Analisis western blot menunjukkan peningkatan fosforilasi protein tirosin antara kelompok kontrol dan setelah diberikan progesteron p>0.05 . Demikian halnya dengan hasil fosforilasi protein Akt juga mengalami peningkatan pada kelompok kontrol dan setelah diberikan progesteron berbagai dosis. Aktivitas kaspase-3 mengalami penurunan bila dibandingkan antara kelompok kontrol dan setelah diberikan progesteron p

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BACKGROUND One of the obstacles in assisted reproduction programs is the low viability and motility of spermatozoa cells. Proteomic analysis indicates that many proteins are thought to play a role in the regulation of motility and viability of spermatozoa, among others, progesterone. This study aims to analyze the prosurvival effect of progesterone against spermatozoa through apoptosis suppression.METHODS Spermatozoa was washed with gradient centrifugation. Progesterone is added to each sample with a final concentration 0 control , 250, 500, 750 and 1000 ng mL. After the sample treatment was done, membrane integrity checking with hypoosmotic swelling test and motility examination with Computer Assisted Sperm Analyzer CASA . Detection of protein in the western blot will be done that recognizes the phosphorylation of tyrosine residues and Akt and caspase activity.RESULT The effect of addition of progesterone increases sperm motility but not signicantly different p 0.05 . The integrity of the spermatozoa membrane is no effect in progesterone. Western blot analysis revealed an increase of tyrosine phosphorylation protein levels between control and after progesterone group p 0.05 . Similarly, the results of Akt protein phosphorylation also increased in control and after progesterone group. Caspase 3 activity decreased when compared between control and after progesterone group p

Abstrak :
ABSTRAK Latar Belakang: Untuk meningkatkan kemungkinan konsepsi pada pasangan yang menjalani inseminasi intrauterin (IIU), dilakukan preparasi spematozoa dengan metode pencucian swim-up (SU) yang dapat meningkatkan kualitas spermatozoa. Aktivitas dari dinein ATPase dapat terlibat dalam proses preparasi spermatozoa, namun nilai yang pasti dari aktivitas dinein ATPase pada spermatozoa kelompok astenozoosperma yang menjalani pencucian SU belum diketahui. Tujuan: Studi ini dilakukan untuk melakukan evaluasi terhadap efisiensi dari metode preparasi spermatozoa dengan pencucian SU pada sampel astenozoospermia pada laki-laki infertil. Metode: Sampel semen didapatkan dari 6 laki-laki pasangan infertil (astenozoospermia) yang akan menjalani terapi inseminasi intrauterin. Analisis semen dilakukan sebelum dan sesudah dilakukannya preparasi spermatozoa. Preparasi spermatozoa dilakukan dengan metode swim-up (SU). Kemudian, aktivitas dinein ATPase diuji dengan metode Vivenes setelah fraksi aksonem sperma dikumpulkan dengan metode Olson. Dilakukan uji statistik paired t-test atau uji Wilcoxon Signed Rank untuk melihat derajat kemaknaan, dengan nilai bermakna jika p<0,05. Hasil: Berdasarkan analisis semen, ditemukan peningkatan signifikan terhadap motilitas dan morfologi progresif spermatozoa kelompok astenozoospermia setelah dilakukannya preparasi sperma dengan metode swim-up (p<0,05). Didapatkan pula peningkatan pada aktivitas spesifik dinein ATPase pasca-pencucian (p>0,05). Walaupun begitu, terdapat penurunan pada nilai konsentrasi sperma (p>0,05). Kesimpulan: Terdapat peningkatan kualitas spermatozoa kelompok astenozoospermia yang signifikan disertai peningkatan aktivitas spesifik dinein ATPase setelah pencucian dengan metode swim-up.

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ABSTRACT Background: To increase the likelihood of conception in couples undergoing intrauterine insemination (IIU), spematozoa preparation was carried out with a swim-up (SU) washing method that could improve the quality of spermatozoa. The activity of dinein ATPase can be involved in the preparation process of spermatozoa, but the exact value of dinein ATPase activity in the spermatozoa of the astenozoosperm group undergoing SU washing is unknown. Objective: This study was conducted to evaluate the efficiency of the spermatozoa preparation method by swim-up washing in the asthenozoospermia sample in infertile men. Methods: Semen samples were obtained from 6 men from infertile couples (asthenozoospermia) who would undergo intrauterine insemination therapy. Cement analysis was carried out before and after the preparation of spermatozoa. Preparation of spermatozoa is carried out by the Swim-up (SU) method. Then, the dinein ATPase activity was tested by the Vivenes method after the axoneme fraction of the sperm was collected by the Olson method. Paired t-test statistics or the Wilcoxon were conducted to see the degree of significance, with a significant value if p <0.05. Results: Based on semen analysis, it was found a significant increase in the progressive motility and morphology of the asthenozoospermia spermatozoa after swim-up method of sperm preparation (p <0.05). There was also an increase in post-washing dinein ATPase specific activity (p> 0.05). However, there was a decrease in the value of sperm concentration (p> 0.05). Discussion: There was an increase in the quality of the asthenozoospermia spermatozoa and significant specific dinein ATPase activity after spermatozoa preparation with swim-up method.