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Abstrak :
ABSTRAK
Proses pematangan spermatozoa terjadi karena adanya interaksi antara protein dengan membran plasma spermatozoa. Walaupun proses pematangan spermatozoa ini sangat penting, namun gen yang berperan dalam sekresi protein di epididimis ini masih banyak yang belum dikarakterisasi. Gen-gen yang berperan dalam proses pematangan spermatozoa umumnya merupakan protein sekretorik, terekspresi pada segmen spesifik, diregulasi androgen, faktor testikular dan perkembangan postnatal. Pada penelitian sebelumnya diketahui bahwa b-defensin merupakan gen yang banyak terekspresi di organ reproduksi pria dan memiliki peran dalam pertahanan tubuh dan pematangan spermatozoa. Penelitian ini dilakukan untuk mengkarakterisasi ekspresi gen Defb20 untuk mengetahui perannya dalam proses pematangan spermatozoa. Studi in silico dilakukan untuk prediksi struktur gen, signal peptide dan domain fungsional. Quantitative real-time PCR digunakan untuk mengukur ekspresi gen Defb20 pada analisis sebaran jaringan, regulasi androgen dan faktor testikular serta postnatal developmen. Hasil penelitian mendapatkan bahwa sekuen Defb20 mengandung domain penting seperti N-myristoilation dan beberapa situs phosporilasi protein kinase yang mungkin berperan dalam mekanisme interaksi protein dengan membran plasma. Sekuen asam amino Defb20 mengandung signal peptides, mengindikasikan protein yang disekresikan dan terlibat dalam proses pematangan spermatozoa. -defensins 20 (Defb20) terekspresi spesifik di epididimis dengan ekspresi tertinggi terdapat pada kaput epididimis. Defb20 diregulasi oleh androgen yang ditunjukkan dengan adanya penurunan ekspresi Defb20 paska dilakukan gonadektomi dan kondisi ini dapat diperbaiki dengan pemberian hormon pengganti. Defb20 juga diregulasi oleh faktor testikular, yang dibuktikan dari menurunnya ekspresi Defb20 setelah ligasi pada duktus eferen (efferent duct ligation (EDL)). Defb20 mulai terekspresi pada hari ke-21 setelah lahir yang mengindikasikan gen Defb20 terekspresipada suatu periode perkembangan epididimis. Berdasarkan hasil penelitian disimpulkan bahwa Defb20 memiliki karakteristik ekspresi : mengandung signal peptide yang mengarahkan sintesis protein pada jalur sekretorik, spesifik terekspresi di epididimis, diregulasi androgen dan faktor testikular serta mulai terekspresi pada masa pubertas hingga dewasa

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ABSTRACT
Epididymal sperm maturation occurs via interactions between sperm and proteins secreted by the epididymal epithelium. Although this is an important process, the genes that encode secreted proteins remain largely uncharacterized. The genes that play a role in sperm maturation process has character, among others; is a secretory protein, expressed specifically in the epididymis, regulated by androgen, testicular factor, and postnatal development. Previous studies showed that family of-defensins preferentially eaxpressed in male reproductive tracts and play an important role in both innate immunity and sperm fertility. This study aimed to characterize Defb20 to understand its role in sperm maturation. This study using in silico analyses and quantitative real-time PCR (qRT-PCR). In silico analyses were performed to predict gene structure, signal peptides and functional domains. Defb20 expression in various tissues, after gonadectomy, efferent duct ligation and postnatal development were measured using quantitative real-time RT-PCR. Defb20 sequence contains important domains such as N-myristoilation and kinase binding sites which are putatively involved in the protein activation and protein-plasma membrane interaction. The amino acid sequence of Defb20 contains signal peptides, indicating characteristic of secretory proteins involved in the sperm maturation. β-defensins 20 (Defb20) was expressed exclusively in the epididymis with the highest expression in the caput region. Defb20 was regulated by androgen showing down-regulation after gonadectomy and the expression was recovered after testosterone replacement. However, Defb20 was also regulated by testicular factors in which the expression was down-regulated after efferent duct ligation (EDL). The dependency on the androgen was further confirmed by postnatal expression analysis in which Defb20 begin to express at day21 postnatal indicating specific stage of expression after initial development of the epididymis. In conclusion, Defb20 have a potential to be involved in the epididymal sperm maturation process. Defb20 has characteristic expression; has a signal peptide sequence that directs synthesis in the secretory pathway, specifically expressed in the epididymis, androgen and testicular factors regulated, and expressed in puberty to adulthood

Abstrak :
Spag11a diketahui terekspresi secara spesifik pada kaput epididimis sehingga dimungkinkan protein tersebut memiliki fungsi yang spesifik untuk maturasi spermatozoa. Studi peran SPAG11A dalam maturasi spermatozoa di epididimis memerlukan produksi protein SPAG11A untuk dikarakterisasi. Tujuan dari penelitian ini adalah untuk mengklon, mengekspreesikan, dan mengkarakterisasi sifat antimikroba dari protein rekombinan SPAG11A. Insert cDNA Spag11a yang dihasilkan melalui PCR diklon ke dalam vektor pET100/D-TOPO. Plasmid rekombinan kemudian diekspresikan ke Escherichia coli BL21 DE3 star. Deteksi dari fusi protein rekombinan dilakukan dengan SDS-PAGE dan Western Blotting. IMAC Immobilized Metal Affinity Chromatography digunakan untuk mempurifikasi protein rekombinan. Uji antimikroba protein rekombinan dianalisis melalui pengukuran Optical density.PCR amplifikasi dari cDNA kaput epididimis mencit menghasilkan insert Spag11a berukuran 210bp. Insert tersebut kemudian dikloning ke dalam pET100/D-TOPO menghasilkan 1 rekombinan plasmid dari 10 koloni yang diskrining. Ekspresi rekombinan klon ke dalam E.coli BL21 menghasilkan fusi protein setelah diinduksi IPTG selama 4 jam. Fusi protein dikonfirmasi menggunakan Western Blotting menggunakan antibodi yang mengenali N-terminal His-Tag 21kDa dan protein SPAG11A. Uji antimikroba protein rekombinan SPAG11A mununjukkan tidak ada inhibisi yang signifikan terhadap laju pertumbuhan E.coli dan Bacillus subtilis. Insert Spag11a yang berukuran 210bp berhasil diklon ke dalam vektor pET100/D-TOPO. Ekspresi rekombinan Spag11a menghasilkan fusi protein berukuran 21kDa. Protein rekombinan SPAG11A tidak membawa sifat antimikroba terhadap E.coli dan B. subtilis. ...... Spag11a is known to be specifically expressed in the caput region of the epididymis suggesting a specific function for sperm maturation. Study of SPAG11A role in the epididymal sperm maturation requires generating SPAG11A protein for characterization. The objective of this study was to clone, express and characterize antimicrobial property of the recombinant SPAG11A. Spag11a cDNA insert was generated by PCR and cloned in TOPO vector. Recombinant DNA plasmid was subsequently expressed in E coli BL 21 star. Detection of recombinant fusion protein was carried out using SDS PAGE and western immunobloting. IMAC Immobilized Metal Affinity Chromatography was used to purify recombinant protein. Optical density measurement was used to analyse antimicrobial property of the recombinant protein. PCR amplification of mouse caput epididymis cDNA produced a 210 bp insert of Spag11a. Cloning of the insert into TOPO pET100 resulted in 2 recombinants out of 10 colonies that were screened. Expression of recombinant clones in the E coli BL21 produced a fusion protein after being induced IPTG for 4 hours. Fusion protein was confirmed by western immunobloting using two antibodies recognizing N terminal His Tag 21 kDa and SPAG11A protein. Antimicrobial assay for SPAG11A recombinant showed no significant inhibition towards growth rates of E coli and Bacillus subtilis. A 210 bp Spag11a insert was successfully cloned into TOPO pET100 vector. Expression of recombinant spag11a produced a fusion protein of 21 kDa. SPAG11A recombinant protein does not have antimicrobial property towards E coli and B subtilis.

Abstrak :
[ABSTRAK
Latar belakang: Pematangan sperma di epididimis terjadi melalui interaksi antara protein yang disekresikan oleh epitel dengan spermatozoa. Proses tersebut diregulasi oleh androgen dan lingkungan spesifik di region epididimis. Androgendependent gene yang hanya terekspresi di region tertentu namun tidak terekspresi di region lain menimbulkan dugaan peran androgent reseptor (AR) koregulator. Gelsolin (Gsn) adalah AR koregulator ditemukan predominan di epididimis Holstein, namun perannya masih belum diketahui. Penelitian ini bertujuan untuk mengkarakterisasi Gsn pada epididimis mencit. Metode: In silico untuk memprediksi struktur gen dan domain fungional. Quantitative Real Time RT-PCR untuk menganalisa sebaran jaringan, ketergantungan terhadap faktor endokrin dan faktor testikular, dan regulasi postnatal. Hasil: Gsn merupakan protein yang mengandung signal peptide. Ekspresi Gsn tidak spesifik di epididimis. Gsn dipengaruhi oleh androgen dan faktor testikular. Pasca gonadektomi, ekspresi Gsn menurun setelah 3 hari dan injeksi T eksogen meningkatkan ekspresi Gsn. Hasil ini diperkuat dengan pemberian flutamide yang menurunkan ekspresi Gsn. Ekspresi Gsn pada perkembangan individu konstan postnatal 5 hari. Kesimpulan: Gsn adalah protein yang disekresikan oleh epitel epididimis, diregulasi oleh androgen dan faktor testikular. Ekspresi Gsn yang tidak spesifik pada region tertentu di epididimis, diperlukan penelitian lanjut untuk mengetahui peran Gsn dalam menentukan ekspresi gen-gen yang terlibat dalam pematangan sperma.

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ABSTRACT
Background: Sperm maturation in epididymis occurs through interaction between proteins secreted by epithels with spermatozoa. The process is regulated by androgen and specific environment in epididimal region. The androgendependent gene that is only expressed in a particular region, but not expressed in other regions led to allegations of androgen receptor (AR) coregulator action. Gelsolin (Gsn) is AR coregulator found predominant in epididimal Holstein, but its role still unknown. The aim is to characterize Gsn in mouse epididymis. Methods: In silico analyses to predict gene strucure and functional domain. Quantitative Real Time RT-PCR to analyse tissue distribution, androgen dependent, testicular factor and postnatal regulation. Results: Gsn is protein that contains signal peptide. Gsn is not spesific expressed in epididymis. It is regulated by androgen and testicular factor. Post gonadectomy, Gsn expression decrease in 3 days while injected by T exogen increasing Gsn expression. This expression confirmed by flutamide that decreasing Gsn expression. Gsn expression was constant at day 5 in postnatal development. Conclusions: Gsn is protein that secreted by epididymal epitels and regulated by androgen and testicular factor. Gsn expression was not spesific in epididymal region. It is needed to do future research to know the role of Gsn in determining genes expression that related to sperm maturation, Background: Sperm maturation in epididymis occurs through interaction between proteins secreted by epithels with spermatozoa. The process is regulated by androgen and specific environment in epididimal region. The androgendependent gene that is only expressed in a particular region, but not expressed in other regions led to allegations of androgen receptor (AR) coregulator action. Gelsolin (Gsn) is AR coregulator found predominant in epididimal Holstein, but its role still unknown. The aim is to characterize Gsn in mouse epididymis. Methods: In silico analyses to predict gene strucure and functional domain. Quantitative Real Time RT-PCR to analyse tissue distribution, androgen dependent, testicular factor and postnatal regulation. Results: Gsn is protein that contains signal peptide. Gsn is not spesific expressed in epididymis. It is regulated by androgen and testicular factor. Post gonadectomy, Gsn expression decrease in 3 days while injected by T exogen increasing Gsn expression. This expression confirmed by flutamide that decreasing Gsn expression. Gsn expression was constant at day 5 in postnatal development. Conclusions: Gsn is protein that secreted by epididymal epitels and regulated by androgen and testicular factor. Gsn expression was not spesific in epididymal region. It is needed to do future research to know the role of Gsn in determining genes expression that related to sperm maturation]

Abstrak :
ABSTRAK
Latar Belakang: Pematangan spermatozoa di epididimis terjadi melalui interaksi antara spermatozoa dengan protein yang disekresikan oleh sel epitel yang melapisi duktus epididimis. Sekresi protein tersebut menciptakan microenvironment yang diregulasi oleh gen-gen tertentu. Studi sebelumnya menunjukkan bahwa gen yang terlibat dalam pematangan spermatozoa pada umumnya terekspresi secara spesifik di epididimis dan dipengaruhi oleh androgen. Spink2 merupakan salah satu gen yang terekspresi di epididimis, namun regulasi ekspresinya masih belum diketahui. Tujuan penelitian ini adalah untuk mengkarakterisasi ekspresi dan regulasi gen Spink2 pada epididimis mencit jantan.Metode: Analisis secara in silico digunakan untuk mengetahui struktur gen dan prediksi sinyal peptida, serta domain fungsional gen Spink2. Quantitative real-time RT-PCR digunakan dalam mengukur ekspresi relatif gen Spink2 pada analisis spesifisitas jaringan, ketergantungan terhadap androgen dan faktor testikuler, serta post-natal development.Hasil: Spink2 termasuk dalam famili serine protease inhibitor yang ditandai dengan adanya domain Kazal type 2. Analisis signal peptide menunjukkan bahwa Spink2 merupakan protein sekretori. Spink2 terekspresi di testis dan epididimis, dengan ekspresi tertinggi berada di kaput epididimis. Ekspresi Spink2 pada mencit yang digonadektomi mengalami peningkatan setelah 6 jam, kemudian menurun mulai dari hari ke-1 hingga hari ke-5. Pemberian testosteron mampu mempertahankan ekspresi Spink2 pada 3 dan 5 hari setelah gonadektomi. Selain itu, pada analisis pengaruh faktor testikuler, ekspresi Spink2 menunjukkan adanya regulasi dari faktor testikuler pada semua kelompok setelah dilakukan efferent duct ligation EDL . Spink2 menujukkan regulasi post-natal yakni mulai terekspresi pada umur mendekati 22 hari.Kesimpulan: SPINK2 merupakan protein sekretori yang terekspresi pada kaput epididimis, serta diregulasi oleh androgen dan faktor testikuler. Spink2 tidak terekspresi secara konstitutif. Berdasarkan data tersebut Spink2 sangat berpotensi terlibat dalam proses pematangan spermatozoa di epididimis. Penelitian lebih lanjut diperlukan untuk mengkonfirmasi potensi tersebut.

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ABSTRACT
Background Sperm maturation in the epididymis occurs through interactions between sperm and proteins secreted by epithelium cells lining the epididymal duct. The secretion of these proteins creates a microenvironment that is regulated by certain genes. Previous studies showed that genes which are involved in sperm maturation process are expressed specifically in the epididymis and regulated by androgen. Spink2 is one of the epididymal genes, but the regulation of its expression is still unknown. Therefore, this study was aimed to characterize Spink2 expression and its regulation in the mouse epididymis.Method s In silico analysis was performed to determine the gene structure and identify the signal peptide, as well as the functional domain of Spink2. Quantitative real time RT PCR was performed to measure relative expression of Spink2 in the analyses of the tissue specificity, androgen dependency, testicular factor and post natal development.Result s Spink2 belongs to the serine protease inhibitor family which is characterized by the presence of Kazal type 2 domain. Signal peptide analysis showed that Spink2 amino acid sequence contains a signal peptide, indicating Spink2 is a secretory protein. Spink2 was expressed specifically in the testis and epididymis, with the highest level of its expression was in the epididymal caput. Spink2 expression increased after six hours and started to decrease on day 1 throughout day 5. Interestingly, administration of exogenous testosterone was able to maintain expression at the physiological level. In addition, Spink2 was slightly affected by testicular factors. During post natal development, Spink2 start to be expressed at day 22 before increased dramatically throughout day 60.Conclusion s Spink2 is a secretory protein that is expressed in caput region of the mouse epididymis and regulated by androgen. Spink2 is not constitutively expressed throughout development. Based on our data, may be involved in epididiymal sperm maturation process. Further studies are required to confirm its role.

Abstrak :

Latar Belakang: Beberapa gen yang terekspresi spesfik di epididimis diduga
terlibat dalam proses pematangan sperma. Karakteristik gen yang terlibat dalam
pematangan sperma selain ekspresinya spesifik di epididimis juga dipengaruhi
oleh androgen, faktor testikuler, dan terekspresi pada saat masa pubertas. Salah
satu famili gen yang cukup banyak ditemukan terekspresi di epididimis adalah
Beta Defensin. Gen Beta Defensin diketahui memiliki peran sebagai pertahanan
terhadap mikroba, namun diduga memiliki keterlibatan dalam proses pematangan
sperma karena ekspresinya banyak ditemukan di epididimis. Oleh karena itu,
penelitian pada gen Beta Defensin terhadap perannya dalam proses pematangan
sperma perlu dilakukan. Berdasarkan studi sebelumnya diketahui bahwa salah
satu gen Beta Defensin yang terekspresi di epididimis yaitu Beta Defensin 2
(Defb2), namun karakterisasi terhadap gen ini belum dilakukan. Dengan
demikian, pada penelitian ini bertujuan untuk mengkarakterisasi gen Defb2 terkait
dengan perannya pada proses pematangan sperma.
Metode: Analisis bioinformatika digunakan untuk mendapatkan informasi
mengenai struktur gen, signal peptide, dan domain fungsional pada gen Defb2.
Analisis qRT-PCR untuk mengetahui ekspresi relatif gen Defb2 pada berbagai
jaringan, regulasinya oleh androgen, pengaruh dari faktor testikular dan
ekspresinya pada perkembangan postnatal.
Hasil: Defb2 merupakan protein sekretori karena memiliki signal peptide. Defb2
memiliki domain fungsional berupa N-myristoylation dan protein kinase-C. Gen
Defb2 terekspresi spesifik di epididimis khususnya pada bagian caput epididimis.
Defb2 ekspresinya dipengaruhi oleh androgen terbukti setelah perlakuan
gonadektomi, ekspresi Defb2 menjadi menurun dan kembali mengalami kenaikan
ketika diberikan testosteron eksogen. Defb2 juga ekspresinya dipengaruhi oleh
faktor testikuler terbukti setelah diberi perlakuan Efferent Duct Ligation (EDL)
maka ekspresi Defb2 langsung menurun bahkan terjadi apoptosis sel sehingga
pola ekspresi gen Defb2 sudah tidak bisa diamati. Begitu juga pada analisis
postnatal development terlihat ekspresi gen Defb2 mulai terdeteksi jelas pada hari
ke-15 yang merupakan masa pubertas mencit jantan.
Kesimpulan: Defb2 merupakan gen yang terlibat dalam proses pematangan
sperma di epididimis yang dibuktikan dengan ekspresi spesifik di epididimis,
diregulasi oleh androgen dan faktor testikuler, serta mulai terekspresi pada masa
pubertas.

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Background: Some of the specific genes expression in the epididymis are
suspected to be involved in the process of sperm maturation. Characteristics of the
genes involved in sperm maturation in the epididymis-specific expression in
addition also influenced by androgens, testicular factors, and expressed at the time
of puberty. One of a family of genes that is pretty much found expressed in the
epididymis is a Beta Defensins. Beta Defensin genes known to have a role as a
defence against microbes, but suspected to have involvement in the process of
sperm maturation because the expression is found in the epididymis. Therefore,
research on Beta Defensin genes against its role in sperm maturation process
needs to be done. Based on previous studies it is known that one of the Beta
Defensin genes which expressed in the epididymis that is Beta Defensins 2
(Defb2), but the characterization of this gene has not been made against. Thus,
this research aims to characterize genes associated with the Defb2 role in the
process of sperm maturation.
Methods: Bioinformatics analysis was used to obtain information about the
structure of genes, signal peptides, and functional domains of the Defb2 gene.
qRT-PCR analysis to find out the relative gene expression of Defb2 on various
tissue, regulation by androgens, the effect of testicular factors and its expression
in postnatal development.
Results: Defb2 is a secreted protein because it has signal peptides. Defb2 has a
functional domain in the form of N-myristoylation and kinase-C protein. Specific
genes expression of Defb2 in the epididymis is especially in the caput epididymis.
Defb2 expression influenced by androgens is proven after the gonadectomy, the
expression of Defb2 to be decreased and start increase again when exogenous
testosterone is given. Defb2 also its expression influenced by testicular factors
that proven after being given the treatment by Efferent Duct Ligation (EDL), then
the Defb2 expressions directly decreased and the cell apoptosis occurs even so
that the pattern of gene expression Defb2 already could not be observed. So also
on analysis of postnatal development seen gene expression Defb2 begins to be
detected clearly at day 15 which is a male mice puberty.
Conclusions: Defb2 is a gene which is involved in the process of maturation of
sperm in the epididymis that is evidenced by specific expression in the
epididymis, be regulated by androgens and testicular factors, and as well as start
expressed at puberty.

Abstrak :
ABSTRAK
Salah satu kendala yang dihadapi manusia saat ini adalah pertambahan penduduk yang tidak terkendali yang menimbulkan banyak masalah baru di berbagai aspek kehidupan. Oleh sebab itu, pengendalian pertumbuhan penduduk harus dilakukan dengan berbagai metode kontrasepsi. Pengembangan kontrasepsi pria non-hormonal dengan menghambat proses pematangan sperma di epididimis menjadi hal yang menjanjikan. Sayangnya, gen yang berperan di dalam proses pematangan sperma di epididimis masih belum banyak dipelajari. Kami menganalisis beberapa kandidat gen yang diekspresikan di epididimis, salah satunya adalah Defensin Beta-42 (Defb42). Beberapa langkah metode yang kami lakukan adalah isolasi epididimis dan ekstraksi RNA, analisis bioinformatika, dan real-time Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) untuk menganalisa ketergantungan ekspresi terhadap androgen dari gen Defb42 karena pematangan sperma bergantung pada androgen. Selain itu, proses pematangan sperma juga terjadi akibat interaksi antara sperma dan protein yang disekresikan oleh epitel epididimis. Oleh sebab itu, peptida sinyal harus dianalisis juga untuk mengecek apabila gen ini adalah protein sekretori. Hasilnya adalah gen ini diregulasi oleh androgen dan memiliki peptida sinyal. Hal ini membuat gen Defb42 menjadi kandidat gen yang menjanjikan untuk diteliti lebih lanjut dalam upaya pengembangan kontrasepsi non-hormonal pada pria.

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ABSTRACT
One of the problem nowadays is the uncontrolled population growth. This raises many other problems in every aspect of life. Therefore, the population growth must be controlled with many types of contraceptive agents. The development of male non-hormonal contraceptive agent by inhibiting the sperm maturation process in epididymis seems to be promising. We analyzed several candidates of gene which are expressed in epididymis; one of them is Defensin Beta-42 (Defb42) gene. Several method we conducted in this research are epididymis isolation and RNA extraction, bioinformatics analysis, and real time Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) to analyze the expression dependency towards androgen of Defb42 gene because the sperm maturation is androgen-dependent process. Besides, the sperm maturation process occurs due to the interaction between sperm and protein secreted by epididymal epithelium. Therefore, the signal peptide has to be analyzed to confirm whether the candidate gene is a secretory protein. The results are this gene is regulated by androgen and has signal peptide properties. Therefore we can conclude that the gene is promising to be studied further in effort to develop male non-hormonal contraceptive agent.

Abstrak :
[ABSTRAK
Latar belakang: Proses pematangan sperma terjadi melalui interaksi spermatozoa dengan protein yang disekresikan ke lumen oleh sel epitel epididimis. Sekresi protein pada epididimis ditentukan oleh gen-gen yang terekspresi spesifik di epididimis. Ekspresi gen di epididimis dapat dipengaruhi oleh androgen atau faktor testikular. CD52 telah diketahui terekspresi di epididimis, namun regulasi yang mempengaruhi ekspresi gen CD52 di epididimis belum diketahui. Tujuan dari penelitian ini adalah untuk menganalisis ekspresi dan regulasi gen CD52 agar dapat memprediksi perannya di epididimis mencit. Metode: Analisis bioinformatika dilakukan untuk memprediksi sinyal peptida dan domain fungsional dari CD52. Quantitative real time RT-PCR digunakan untuk mengukur ekspresi relatif gen CD52 pada analisis spesifisitas jaringan, ketergantungan terhadap androgen dan faktor testikular, serta postnatal development. Hasil: CD52 memiliki sinyal peptida yang menunjukkan ciri protein sekretori dan terekspresi secara spesifik di epididimis. Ekspresi CD52 yang tertinggi terdapat di bagian cauda. Ekspresi CD52 pada mencit diregulasi oleh androgen yang ditandai dengan penurunan pada hari pertama dan ketiga setelah digonadektomi dan pemberian testosteron eksogen setelah gonadektomi dapat menjaga ekspresi CD52 50% dari kadar normalnya. Eksperimen dengan memberikan reseptor androgen antagonis (flutamide) juga mendukung bahwa ekspresi CD52 sangat tergantung terhadap androgen. Ekspresi CD52 menurun sangat bermakna hingga mencapai 93% dibandingkan dengan kontrol. Selain androgen, ekspresi CD52 juga dipengaruhi oleh faktor testikular. Ekspresi CD52 mengalami penurunan bermakna dari hari pertama hingga kelima setelah perlakuan efferent duct ligation (EDL) hingga mencapai 75% dari kontrol. Selain itu ekspresi CD52 juga dipengaruhi oleh perkembangan pasca lahir. Ekspresi CD52 meningkat di hari ke-15 hingga hari ke-60 pasca lahir. Kesimpulan: CD52 merupakan gen penyandi protein sekretori yang terekspresi spesifik di epididimis pada region cauda dan regulasinya dipengaruhi oleh androgen, faktor testikular, dan perkembangan pasca lahir.

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ABSTRACT
Background. Epididymal sperm maturation is occurs via interactions between sperm and proteins secreted by epididymal epithelium. These proteins are encoded by genes that are specifically expressed in a region-specific manner. Previous studies have demonstrated that epididymal genes are regulated by androgen and testicular factors. CD52 is an epididymal gene putatively involved in sperm maturation. However, the regulation of its expression in the epididymis has not been fully understood and little is known about its role during sperm maturation process. Therefore, this study was aimed to analyze the expression and regulation of CD52 in the mouse epididymis. Method. Bioinfomatic analyses were perfomed to predict signal peptides and functional domains of CD52. Quantitative real-time RT-PCR was used to analyze tissue distribution, androgen, testicular factors dependency and postnatal development. Results. CD52 amino acid sequence contains a signal peptide, indicating it is a secretory protein. CD52 exhibited region-spesific expression in the epididymis with the highest level was in cauda. Mice CD52 expression was regulated by androgen indicated by a decrease started at day 1 following a gonadectomy. Interestingly, testosterone replacement therapy was able to maintain the expression at 50% of normal level. Experiment by given androgen receptor antagonist, flutamide showed decrease of CD52 expression about 93% than control. It?s confirming that CD52 expression depend on androgen. Moreover, testicular factors also influenced CD52 expression. This was revealed by efferent duct ligation in which CD52 expression was reduced at day 1 to day 5 following the ligation. Finally, CD52 expression was developmentally regulated, this was indicated by increase in the level of expression start at day 15 postnatally. Conclusion: CD52 is a secretory protein and exhibited region-spesific expression in the cauda epididymis. It is regulated by androgen, testicular factors, and also affected by development stage. , Background. Epididymal sperm maturation is occurs via interactions between sperm and proteins secreted by epididymal epithelium. These proteins are encoded by genes that are specifically expressed in a region-specific manner. Previous studies have demonstrated that epididymal genes are regulated by androgen and testicular factors. CD52 is an epididymal gene putatively involved in sperm maturation. However, the regulation of its expression in the epididymis has not been fully understood and little is known about its role during sperm maturation process. Therefore, this study was aimed to analyze the expression and regulation of CD52 in the mouse epididymis. Method. Bioinfomatic analyses were perfomed to predict signal peptides and functional domains of CD52. Quantitative real-time RT-PCR was used to analyze tissue distribution, androgen, testicular factors dependency and postnatal development. Results. CD52 amino acid sequence contains a signal peptide, indicating it is a secretory protein. CD52 exhibited region-spesific expression in the epididymis with the highest level was in cauda. Mice CD52 expression was regulated by androgen indicated by a decrease started at day 1 following a gonadectomy. Interestingly, testosterone replacement therapy was able to maintain the expression at 50% of normal level. Experiment by given androgen receptor antagonist, flutamide showed decrease of CD52 expression about 93% than control. It’s confirming that CD52 expression depend on androgen. Moreover, testicular factors also influenced CD52 expression. This was revealed by efferent duct ligation in which CD52 expression was reduced at day 1 to day 5 following the ligation. Finally, CD52 expression was developmentally regulated, this was indicated by increase in the level of expression start at day 15 postnatally. Conclusion: CD52 is a secretory protein and exhibited region-spesific expression in the cauda epididymis. It is regulated by androgen, testicular factors, and also affected by development stage. ]