Hasil Pencarian  ::  Simpan CSV :: Kembali

Hasil Pencarian

Ditemukan 201823 dokumen yang sesuai dengan query
cover
Andika Setyoadi
"

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.


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.

"
Depok: Fakultas Kedokteran Universitas Indonesia, 2019
T-pdf
UI - Tesis Membership  Universitas Indonesia Library
cover
Syaifiyatul H.
"[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.

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]"
2015
T-Pdf
UI - Tesis Membership  Universitas Indonesia Library
cover
Silvani Permatasari
"[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.

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.
]"
2015
T-Pdf
UI - Tesis Membership  Universitas Indonesia Library
cover
Dewi Muliawati
"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

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"
Depok: Fakultas Kedokteran Universitas Indonesia, 2019
T-Pdf
UI - Tesis Membership  Universitas Indonesia Library
cover
Seruni Tyas Khairunissa
"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.
"
Jakarta: Fakultas Kedokteran Universitas Indonesia, 2017
T-Pdf
UI - Tesis Membership  Universitas Indonesia Library
cover
Purba, Stefanus Raditya
"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.

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."
Jakarta: Fakultas Kedokteran Universitas Indonesia, 2013
S70359
UI - Skripsi Membership  Universitas Indonesia Library
cover
Cut Yasmin
"Latar Belakang : Proses pematangan sperma terjadi karena adanya interaksi antara sel sperma dan protein yang disekresikan oleh sel epitel yang melapisi saluran epididimis, sehingga menyebabkan perubahan morfologi dan biokimia pada membran spermatozoa. Sekresi protein pada saluran epididimis ditentukan oleh gen-gen yang diekspresikan secara spesifik di region tertentu misalnya pada region initial segmen, caput, corpus atau cauda sehingga pada masing-masing segmen terbentuk lingkungan spesifik (microenvironment) yang diperlukan untuk proses pematangan sperma. Gen yang menyandi protein yang terlibat dalam proses pematangan sperma diekspresikan secara spesifik pada epididimis dan ekspresinya diregulasi oleh androgen. Salah satu gen yang diduga berperan dalam proses pematangan sperma adalah gen Serpina1f. Berdasarkan urutan asam amino, Serpina1f dianggap sebagai anggota baru dari keluarga serpin dengan fungsi putatif sebagai serin protease inhibitor. Karakterisasi Serpina1f pada organ reproduksi pria khususnya epididimis dapat memberikan petunjuk mengenai perannya dalam proses pematangan sperma.
Tujuan : Mengkarakterisasi gen Serpina1f di epididimis mencit.
Desain : Penelitian ini menggunakan analisis bioinformatika dan eksperimental
Metode : Struktur gen, batas ekson-intron dan domain fungsional serta deteksi signal peptide pada Serpina1f dilakukan analisis bioinformatika. Untuk menganalisis ekspresi SERPINA1F tingkat protein dilakukan western imunoblotting, sedangkan untuk mengetahui lokasi protein SERPINA1F dilakukan imunohistokimia dan imunositokimia.
Hasil : SERPINA1F merupakan anggota family SERPIN. Berdasarkan analisis Signal peptide, SERPINA1F merupakan protein sekretori. Hasil imunohistokimia pada jaringan epididimis mencit menunjukkan adanya reaksi antibodi dengan terwarnainya nukleus dan sitoplasma pada sel epitel initial segment dan caput. Sedangkan pada corpus dan cauda SERPINA1F terdeteksi hanya pada sitoplasma. Hasil analisis western imunoblotting dan imunositokimia pada protein sperma menunjukkan adanya asosiasi SERPINA1F dengan spermatozoa dan terdapat di seluruh bagian spermatozoa.
Kesimpulan : SERPINA1F memiliki signal peptide pada asam amino 1 - 27 dan termasuk protein sekretori. Berdasarkan urutan asam aminonya, SERPINA1F yang termasuk famili SERPIN. Protein SERPINA1F diekspresikan di sel epitel initial segment, caput, corpus dan cauda. SERPINA1F berasosiasi dengan spermatozoa dan terdapat pada seluruh bagian spermatozoa.

Background: Sperm maturation occur due to the interaction between the sperm cell and the proteins secreted by the epithelial cells lining the epididymis duct, causing morphological and biochemical changes in the sperm membrane. The secretion of proteins in epididymis duct is determined by genes that are expressed specifically in certain regions, for example in the region of the initial segment, caput, corpus or cauda that each segment forms microenvironment that is required for sperm maturation process. Genes that encode proteins involved in the process of sperm maturation in the epididymis specifically expressed and its expression is regulated by androgens. One of the genes thought to play a role in sperm maturation process is Serpina1f. Based on the amino acid sequence, Serpina1f considered as a new member of the serpin family with putative functions as a serine protease inhibitor. Characterization Serpina1f on male reproductive organs, especially the epididymis may provide a clue as to its role in sperm maturation process.
Objective: To characterize Serpina1f in the epididymis of mice.
Design: This study used a bioinformatics analysis and experimental.
Methods: Bioinformatics analysis were used to know gene structure, exon-intron boundaries and functional domains as well as the detection of signal peptide on Serpina1f. To analyze protein expression levels SERPINA1F performed western immunoblotting, whereas for the location of the protein immunohistochemistry and immunocytochemistry SERPINA1F done.
Results: SERPINA1F a member of the serpin family. Based on the analysis of signal peptide, a protein secretory SERPINA1F. The results of immunohistochemistry on epididymis tissue of mice showed an antibody reaction with colored nucleus and cytoplasm in epithelial cells of the initial segment and caput. While SERPINA1F detected only in the cytoplasm corpus and cauda. The results of western imunoblotting analysis and immunocytochemistry showed SERPINA1F associated with spermatozoa and found in all parts of the spermatozoa.
Conclusion: SERPINA1F has a signal peptide at amino acids 1-27 and include secretory proteins. Based on their amino acid sequence, which includes SERPINA1F serpin family. The protein is expressed in epithelial cells SERPINA1F initial segment, caput, corpus and cauda. SERPINA1F associated with spermatozoa and found on all parts."
Jakarta: Fakultas Kedokteran Universitas Indonesia, 2014
T-Pdf
UI - Tesis Membership  Universitas Indonesia Library
cover
Indri Aderni
"Latar belakang: Beta defensin diekspresikan terutama oleh sel epitel pada permukaan mukosa berbagai organ seperti kulit, usus, mulut dan saluran genital. Studi sebelumnya menunjukkan bahwa beta defensin 30 (Defb30) terekspresi spesifik di epididimis. Defb30 merupakan peptida kationik berukuran kecil yang diduga berperan penting pada proses pematangan spermatozoa di epididimis dan juga berperan sebagai pertahanan host terhadap infeksi mikroba. Untuk mempelajari aktivitas antimikroba Defb30 ini diperlukan analisis pada tingkat protein dan hal tersebut memerlukan protein dalam jumlah yang cukup. Karena itu perlu dilakukan suatu rekayasa genetika untuk pembuatan protein rekombinan DEFB30.
Metode: Gen sintetik penyandi protein DEFB30 yang telah dioptimasi kodonnya diklona ke dalam vektor pQE-80L, suatu plasmid yang mengandung sistem ekspresi untuk prokariota. Plasmid rekombinan yang mengandung sisipan gen target dikonfirmasi dengan analisis enzim restriksi dan sekuensing. Selanjutnya plasmid rekombinan di ekpresikan ke dalam E. coli BL21 dan diinduksi menggunakan IPTG (Isopropyl-1-Thio-d-Galactopyranoside) dengan berbagai waktu inkubasi. Deteksi protein rekombinan dilakukan dengan SDS-PAGE dan westernblotting. IMAC (Immobilized Metal Affinity Chromatography) digunakan untuk mempurifikasi protein rekombinan. Uji antimikroba protein rekombinan dilakukan dengan cara pengukuran nilai optical density (OD) dan dianalisis hasilnya menggunakan uji one way anova.
Hasil: Gen sintetik penyandi protein rekombinan DEFB30 berhasil dikonstruksi pada plasmid pQE-80L. Ekspresi ke dalam E. coli BL21 menghasilkan suatu protein fusi setelah diinduksi menggunakan IPTG selama 4 jam. Hasil analisis protein rekombinan dengan westernblotting menggunakan antibodi Anti-His G-HRP menunjukkan terbentuk pita tebal yang berukuran diatas 10 kDa (±12 kDa). Uji antimikroba protein rekombinan DEFB30 menunjukkan bahwa DEFB30 dapat menghambat pertumbuhan bakteri Eschericia coli dan Bacillus subtilis.
Kesimpulan: Gen sintetik penyandi beta defensin 30 berhasil diklona ke dalam plasmid pQE-80L. Ekspresikan protein rekombinan DEFB30 menghasilkan suatu protein fusi berukuran ±12kDa. Protein rekombinan DEFB30 terbukti memiliki sifat antimikroba terhadap Eschericia coli dan Bacillus subtilis.

ackground: Beta defensins are primarily expressed by epithelial cells at mucosal surfaces, such as those in skin, gut, mouth and genital tracts. Previous studies have demonstrated that beta defensin 30 (Defb30) is exclusively expressed in the epididymis. Defb30 is known as a small cationic antimicrobial peptide which plays an important role in epididymal sperm maturation and also acts as a host defence against microbial infection. Study of Defb30 role in the antimicrobial activity requires generating DEFB30 protein for characterization. For the purpose of this study, genetic engineering was done for the manufacture of the DEFB30 recombinant protein.
Methods: In this study, according to the preferred codon in E. coli, the Defb30 gene was optimized and synthesized. The gene was cloned into pQE-80L vector and subsequently expressed in E. coli BL21; using IPTG (Isopropyl-1-Thio-d-Galactopyranoside) as an inducer. Detection of recombinant protein was carried out by using SDS-PAGE and westernblotting. IMAC (Immobilized Metal Affinity Chromatography) was used to purify recombinant protein. Optical density measurement was used to analyze antimicrobial property of the DEFB30 recombinant protein.
Results: The synthetic gene was successfully constructed into pQE-80L plasmid and expression of the recombinant protein in E. coli BL21 produced a fusion protein after being induced by IPTG for 4 hours. Westernblotting analysis using Anti-His G-HRP antibody showed band above 10kDa (±12kDa). Antimicrobial assay for DEFB30 recombinant protein showed inhibition towards growth rates of Eschericia coli and Bacillus subtilis.
Conclusion: Defb30 synthetic gene was succesfully cloned into pQE-80L plasmid. Expression of recombinant DEFB30 produced a fusion protein of ±12kDa. This recombinant protein has antimicrobial property towards Eschericia coli and Bacillus subtilis.
"
Depok: Fakultas Kedokteran Universitas Indonesia, 2020
T-pdf
UI - Tesis Membership  Universitas Indonesia Library
cover
Meidika Dara Rizki
"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.

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. "
2017
T-Pdf
UI - Tesis Membership  Universitas Indonesia Library
cover
Annisa Parisudha
"ABSTRAK
Latar Belakang : Proses pematangan spermatozoa terjadi melalui interaksi spermatozoa dengan protein yang disekresikan ke lumen oleh sel-sel epitel epididimis. Sekresi protein akan menciptakan lingkungan yang mendukung proses pematangan spermatozoa. Namun gen penyandi protein yang terlibat dalam proses pematangan spermatozoa di epididmis masih belum banyak diketahui. Berdasarkan penelitian sebelumnya, gen-gen yang terlibat dalam proses pematangan spermatozoa ini memiliki kriteria antara lain protein sekretori, terekspresi spesifik di epididimis, dan menunjukkan eskpresi regional, diregulasi oleh faktor androgen dan faktor testikular. Defb30 merupakan salah satu gen yang perlu dilakukan karakterisasi lebih lanjut untuk mengetahui apakah gen tersebut memenuhi kriteria sebagai gen yang terlibat dalam proses pematangan spermatozoa. Tujuan dari penelitian ini adalah untuk melakukan karakterisasi gen Defb30 pada epididimis mencit.Desain : Penelitian ini menggunakan analisis bioinformatika dan Quantitative real-time PCR qRT-PCR .Metode : Analisis bioinformatika digunakan untuk memprediksi struktur gen, sinyal peptida dan domain fungsional. Analisis qRT-PCR digunakan untuk mengukur ekspresi relatif gen Defb30 terhadap analisis sebaran jaringan, regulasi terhadap androgen dan faktor testikular serta postnatal development.Hasil : Analisis sinyal peptida menggunakan signalP 4.1 menunjukkan bahwa Defb30 merupakan protein sekretori. Defb30 terekspresi secara spesifik di epididimis dan memiliki nilai spesifitas tinggi di bagian kaput epididimis. Ekspresi relatif gen Defb30 diregulasi oleh faktor endokrin berupa androgen, penurunan ekspresi relatif gen Defb30 terlihat pada hari pertama hingga hari ketiga gonadektomi dan testosteron diketahui mampu mencegah penurunan ekspresi Defb30 pada mencit yang telah digonadektomi. Analisis eksperimen efferent duct ligation menunjukkan gen Defb30 diregulasi oleh faktor testikular. Analisis postnatal development menunjukkan bahwa gen Defb30 mulai terekspresi pada hari ke-15 postnatal dan meningkat hingga usia dewasa.Kesimpulan : Defb30 merupakan protein sekretori yang terekspresi spesifik pada kaput epididimis dan diregulasi oleh androgen dan faktor testikluar.

ABSTRACT
Background The process of sperm maturation occurs through interaction between sperm and proteins secreted by epididymal epithelial cells. The secretion of proteins will create micro environment suitable for spermmaturation. However, the role of protein encoding genes involved in the maturation process are not widely known. Based on previous studies the genes that are involved in spermmaturation process have characteristics such as secretory protein, specific expression in the epididymis and shows region specific expression, regulated by androgen and testicular factors. Defb30 is one of the genes that need further characterization to determine the putative function. Therefore, this study was aimed to characterize expression and regulation of Defb30 in the mouse epididymis.Methods Bioinformatics analysis was used to predict the structure of genes, peptide signals and functional domains. qRT PCR analysis was performed to measure the level of Defb30 expressionin the tissue distribution, regulation of andorgen and testicular factors and postnatal development.Result Peptide signal analysis using signalP 4.1 indicated that Defb30 was a secretory protein. Defb30 was expressed exclusively in the epididymis and had a high specificity in the caput. The expression of the Defb30 gene was regulated by androgen in which decreased of Defb30 expression was observed at the first day to the third day of gonadectomy and exogenous T was able to maintain Defb30 expression at 3d and 5d gonadectomized mice. efferent duct ligation showed that Defb30 was slightly regulated by testicular factors. Defb30 was developmentally regulated being expressed start at day 15 postnataly.Conclusions Defb30 is a secretory protein which is expressed specifically in the caput epididymis and it is regulated by androgen and testicular factors. "
2017
T-Pdf
UI - Tesis Membership  Universitas Indonesia Library
<<   1 2 3 4 5 6 7 8 9 10   >>