已婚夫婦中約有15﹪的夫妻有不孕症的困擾, 其中約有一半是男性因素所造成。 於男性不孕症患者中有22.7﹪其致病原因仍不清楚, 但證據顯示遺傳因素扮演重要角色。 近10年來發現有15個基因位Y染色體第5及第六區間上並表現於睪丸組織中。 這15個基因是DFFRY, DBY, UTY, TB4Y, E1F1AY, RBMY1A1, DAZ, BPY1, BPY2, CDY1, CDY2, XKRY, PRY, SMCY, 及 TTY2。 本研究分成兩個部分, 在第一部分我們採用多重引子聚合酶連鎖反應, 以基因特定引子進行這15個基因缺失的篩檢。 我們發現在180位無精蟲或寡精蟲症患者之中, 有19位(10.6%)有一個或一個以上的基因缺失, 而每位基因缺失患者至少有DFFRY, DBY, RBMY1A1及 DAZ四個基因之中, 其中一個基因缺失。 我們推測這四個基因於精子發育過程扮演相當重要的角色。 在第二部份我們採用QC-RT-PCR定量DFFRY, DBY, RBMY1A1及DAZ基因與DAZ的同源基因DAZL及BOULE mRNA在睪丸組織的表現量。 發現於Sertoli cell -only syndrome 病患的睪丸組織中 RBMY1A1、 DAZ、 DAZL及BOULE的mRNA表現量較正常人及hypospermatogenesis + maturation arrest低, 但於正常人及hypospermatogenesis + maturation arrest之間沒有顯著差異。 DFFRY及DBY基因表現量在各組之間則沒有顯著差異。 基因缺失篩檢可以幫助我們了解DFFRY、 DBY、RBMY1A1及 DAZ基因在造精功能上的重要程度。 定量DFFRY、 DBY、RBMY1A1、 DAZ、 DAZL及BOULE基因, 使我們了解這些基因在不同睪丸組織表現型患者之間的差異, 並且利用RBMY1A1、 DAZ、 DAZL及BOULE基因表現量於不同睪丸組織表現型患者之間的差異, 來提供新的分類及診斷睪丸表現型方式。

　　It is estimated that 15- 20% of couples are infertile and male factors account for about half of the cases. The causes of male infertility are unknown in about 22.7﹪of all cases. There are ample scientific evidences showing that genetic factors are involved in most cases of male-factor infertility. Recently, 15 genes located in interval 5 and 6 of human Y chromosome have been shown to be expressed in the testis. These genes (DFFRY, DBY, UTY, TB4Y, E1F1AY, RBMY1A1, DAZ, BPY1, BPY2, CDY1, CDY2, XKRY, PRY, SMCY, and TTY2) potential AZoospermia Factors (AZFs). We designed a novel multiplex PCR protocol to detect deletion status of the 15 Y-chromosomal genes in Taiwanese patients presenting with azoospermia or severe oligospermia. Of 180 patients screened, 19 (10.6%) had deletion of at least one of four genes: DFFRY, DBY, RBMY1A1 and DAZ. Other genes, SMCY, EIF1AY, PRY, TTY2,CDY1, BPY2, CDY2, and XKRY were also deleted in some cases. It was concluded that DFFRY, DBY, RBMY1A1 and DAZ genes play important roles in human spermatogenesis. To explore the roles of these four genes, we measured the amounts of transcripts of these genes and two DAZ-homologous genes (DAZL and BOULE) by quantitative competitive (QC)–RT–PCR in the testicular tissues of patients. The concentrations of RBMY1A1, DAZ, DAZL and BOULE transcripts were significant lower in man with Sertoli cell-only syndrome (SCO). No significant difference was found for the DBY and DFFRY transcripts between different groups of patients and control subjects. Decreased concentrations of RBMY1A1, DAZ, DAZL and BOULE transcripts in patients with SCO can be ascribed to the secondary effect caused by loss of germ cells in these patients because these four genes are germ-cell specific. In contrast, DBY and DFFRY genes are ubiquitous and the concentrations of their transcripts are no different between patients and controls. Expression profiles of genes expressed in the testicular tissue can help to elucidate the roles of these genes in spermatogenic defects and to provide information regarding categorization of patients.

陳喬維(2000)﹔Y 染色體顯微缺失與精蟲發育缺陷之相關研究。台南市: 國立成功大學碩士論文。

Blagosklonova O, Fellmann F, Clavequin M, Roux C, Bresson J l. AZFa deletions in sertioli cell-only syndrome :a retrospective study. Mol Hum Reprod 2000 ; 6:795-9

Carlo Foresta, Alberto Ferlin, Enrico Moro. Deletion and expression analysis of AZFa genes on the human Y chromosome revealed a major role for DBY in male infertility. Hum Mol Genet 2000; 9 :1161-9

Celi FS, Zenilman ME, Shuldiner AR. A rapid and versatile method to synthesize internal standards for competitive PCR. Nucleic Acids Res 1993; 21: 1047.

Chai NN, Zhou H, Hernandez J, Najmabadi H, Bhasin S, Yen PH. Structure and organization of the RBMY genes on the human Y chromosome: transposition and amplification of an ancestral autosomal hnRNPG gene. Genomics 1998; 49: 283-9.

Chandley AC. The chromosomal basis of human infertility. Br Med Bull 1979; 35:181-6

Cooek Hj, Lee M, Kerr S, Ruggiu M. A murine homologue of the human DAZ gene is autosomal and expressed only in male and female gonads. Hum Mol Genet 1996; 5:513-6.

Eberhart CG, Maines JZ, Wasserman SA. Meiotic cell cycle requirement for a fly homologue of human deleted in azoospermia. Nature 1996; 381: 783-5.

Elliott DJ, Millar MR, Oghene K, Ross A, Kiesewetter F, Pryor J, McIntyre M, Escalier, D : Impact of genetic engineering on the understanding of spermatogenesis. Hum Reprod 2001; 7:191-210.

Ｅ ugene Yujun Xu, Frederick L. Moore, Renee A. Reijo Pera. A gene family required for human germ cell development evolved from an ancient meiotic gene conserved I metazoans. Proc. Natl. Acad. Sci. USA 2001;98:7414-9.

Ferlin A, Boro, E, Garolla A, Foresta C. Human male infertility and Y chromosome deletions: role of the AZF-candidate genes DAZ, RBM and DFFRY. Hum Reprod 1999; 14: 1710-6.

Fernandes S, Huellen K, Goncalves J, Dukal H, Zeisler J, Rajpert E, Meyts De, Skakkebaek N E, Habermann B, Krause W, Sousa M, Barros A, Vogt P H. Hum Reprod 2002;8:286-8.

Foote S, Vollrath D, Hilton A, Page DC. The human Y chromosome: overlapping DNA clones spanning the euchromatic region. Science 1992; 258: 60-6.

Foresta C, Ferlin A, Garolla A, Moro E, Pistorello M, Barbaux S, Paz G, Yavetz H. High frequency of well-defined Y-chromosome deletions in idiopathic Sertoli cell-only syndrome. Hum Reprod 1998;13:302-7.

Foresta C, Ferlin A, Moro E. Deletion and expression analysis of AZFa genes on the human Y chromosome revealed a major role for DBY in male infertility. Hum Mol Genet 2000;9:1161-9.

Graeme M. Brown, Robert A. Furlong, Carole A. Sargent, Robert P. Erickson, Guy Longepied, Michael Mitchell, Michael H. Jones, Timothy B. Hargreave, Howard J. Cooke and Nabeel A. Affara. Characterisation of the coding sequence and fine mapping of the human DFFRY gene and comparative expression analysis and mapping to the Sxrb interval of the mouse Y chromosome of the Dffry gene. Hum Mol Genet 1998;7:97-107.

Hargreave TB, Saunders PTK, Vogt PH, Chandley AC, Cooke H. Expression of RBM in the nuclei of human germ cells is dependent on a critical region of the Y chromosome long arm. Proc. Natl. Acad. Sci. USA 1997; 94: 3848-53.

Houston DW, Zhang J, Maines JZ, Wasserman SA, King ML. A Xenopus DAZ-like gene encodes an RNA component of germ plasm and is a functional homologue of Drosophila boule. Development 1998; 125: 171-80.

Joep H A, M Tuerlings, Ron JT, Van Golde, Astrid R, Oudakker, Helger G Yntema, Jan A M Kremer. Familial oligoasthenoteratozoospermia : evidence of autosomal dominant inheritance with sex-limited expression. Fertil Steril 2002; 77 : 415-8.

Lahn BT, Page DC. Functional coherence of the human Y chromosome. Science 1997; 278: 675-80.

Lerry P, Alzari P, Sassoon D, Wolgemuth D, Fellous M. The protein encoded by a murine male germ cell-specific transcript is a putative ATP-dependent RNA helicas. Cell 1989; 57: 549-59.

Liford R, Jones AM, Bishop DT, Thornton J, Mueller R. Case-control study of whether subfertility in men is familial. Br Med J 1994; 309: 570-3.

Margaret T. Fuller. Genetic control of cell proliferation and differentiation in Drosophila spermatogenesis. Cell Dev Bio. 1998; 9 : 433-44

Johannes H, P Hackstein, Ron Hochstenbach, Peter L Pearson. Towards an understanding of the genetics of human male infertility. E S 2000;12 565-572.

Kamp C, Hirschmann P, Voss H, Huellen K, Vogt PH. Two long homologous retroviral sequence blocks in proximal Yq11 cause AZFa microdeletions as a result of intrachromosomal recombination events. Hum Mol Genet 2000;9:2563-72.

Karashima T, Sugimoto A, Yamamoto M. Caenorhabditis elegans homologue of the human azoospermia factor DAZ is required for oogenesis but not for spermatogenesis. Development 2000; 127: 1069-79.

Kent-First M, Muallem A, Shultz J, Pryor J, Roberts K, Nolten W, et al. Defining regions of the Y-chromosome responsible for male infertility and identification of a fourth AZF region (AZFd) by Y-chromosome microdeletion detection. Mol Reprod Dev 1999;53:27-41.

Kirsch S, Keil R, Edelmann A, Henegariu O, Hirschmann P, LePaslier D, Vogt PH. Molecular analysis of the genomic structure of the human Y chromosome in the euchromatic part of its long arm (Yq11). Cytogenet Cell Genet 1996; 75: 197-206.

Kleiman SE, Yogev L, Gamzu R, Hauser R, Botchan A, Lessing JB, Paz G, Yavezt H. Genetic evaluation of infertile men. Hum Reprod 1999;14:33-8.

Krausz C, Bussani-Mastellone C, Granchi S, McElreavey K, Scarselli G, Forti G. Screening for microdeletions of Y chromosome genes in patients undergoing intracytoplasmic sperm injection. Hum Reprod 1999;14:1717-21.

Krausz C, Quintana-Murci L, McElreavey K. Prognostic value of Y deletion analysis: what is the clinical prognostic value of Y chromosome microdeletion analysis? Hum Reprod 2000;15:1431-4.

Larry I. Lipshultz, Stuart S. Infertility in the male. Third edition. Mosby-Year Book, Inc., St. Louis, 1997; 173-174.McElreavey K, Krausz C. Sex chromosoem genetics `99 male infertlity and the Y chromosome. Am J Hum Genet 1999; 64: 928-33.

Leroy P, Alzari P, Saaaoon D, Wolgemuth D, Fellous M. The protein encoded by a murine male germ cell-specific transcript is a putative ATP-dependent RNA helicase.Cell 1989;57:549-59.

Margaret L. Delbridge, Harry JL, Toder R, Waugh O`Neill RJ, Ma K, Chandley AC, Marshall Graves JA. A human candidate spermatogenesis gene, RBM1, is conserved and amplified on the marsupial Y chromosome. Nat Genet 1997; 15: 131-6.

Pryor JL, Kent-First M, Muallen A, Van Bergen AH, Nolten WE, Meisner L, Roberts KP. Micordeletions in the Y chromosome of infertile man. N Engl J Med 1997; 336: 534-9.

Reijo R, Lee TY, Salo P, Alagappan R, Brown LG, Rosenberg M, Rozen S, Jaffe T, Straus D, Hovatta O, Chapelle A, Silber S, Page DC. Diverse spermatogenic defects in humans caused by Y chromosome deletions encompassing a novel RNA-binding protein gene. Nat Genet 1995; 10: 383-93.

Roberts KP. Y-chromosome deletions and male infertility: state of the art and clinical implications. Int J Androl 1998; 19: 255-9.

Ruggiu M, Speed R, Taggart M, McKay SJ, Kilanowski F, Saunders P, Dorin J, Cooke HJ. The mouse Dazla gene encodes a cytoplasmic protein essential for gametogenesis. Nature 1997; 389: 73-77.

Ruggiu M, Cooke H. Y bind RNA for spermatogenesis? Inl J Androl 1999; 22: 19 -27.

Schlegel PN, Palermo GD, and Alikani M. Micropucture retieval of epididy male sperm with in vitro micromanipulation techniques. Urology 1995; 46: 238-241.

Sherry L Gee, John G Conboy. Mouse erythroid cells express multiple putative RNA helicase genes exhibiting high sequence conservative from yeast to mammals.E S 1994;140:171-7.

Silber SJ, Alagappan R, Brown LG, Page DC. Y chromosome deletions in azoospermic and severely oligozoospermic men undergoing intracytoplasmic sperm injection after testicular sperm extraction. Hum Reprod 1998;13:3332-7.

Simoni M, Kamischke A, Nieschlag E. Current status of the molecular diagnosis of Y-chromosomal microdeletions in the work-up of male infertility. Initiative for international quality control. Hum Reprod 1998;13:1764-8.

Slee R, Grimes B, Speed RM, Taggart M, Maguire SM, Ross A, McGill NI,Saunders PTK, Cooke HJ. A human DAZ transgene confers partial rescue of the mouse Dazl null phenotype. Proc. Natl. Acad. Sci. USA 1999; 96: 8040-5.

Stuppia L, Gatta V, Calabrese G, Franchi PG, Morizio E, Bombieri C, Mingarelli R, Sforza V, Frajese G, Tenaglia R, Palka G. A quarter of men with idiopathic oligo-azoospermia display chromosomal abnormalities and microdeletions of different types in interval 6 of Yq11. Hum Genet 1998; 102: 566-570.

Sun C, Skaletsky H, Rozen S, Gromoll J, Nieschlag E, Oates R, Page D C. Deletion of azoospermia factor a (AZFa) region of human Y chromosome caused by recombination between HERV15 proviruses. Hum Mol Genet 2000;9:2291-6.

Tsai SJ, Wiltbank MC. Quantification of mRNA using competitive RT-PCR with standardcurve methodology. Biotechniques 1996; 21: 862-6.

Tiepolo L, Zuffardi O. Localization of factors controlling spermatogenesis in the nonfluorescent portion of the human Y chromosome long arm. Hum Genet 1976; 34:119-24.

Tilford Charles A, Tomoko Kuroda-Kawaguchi, Helen Skaletsky, Steve Rozen, Laura G. Brown, Michaei Rosenberg, John D. Mc Pherson, Kristine Wylie, Mandeep Sekhon, Tomara A.Kucaba, Robert H. Waterston & David C.Page. A physical map of the human chromosome. Science 2001;409:943-5.

Tomoko Kuroda-Kawaguchi, Helen Skaletsky, Laura G. Brown, Patrick J. Minx Holland S. Cordum, Robert H. Waterston, Richard K. Wilson, Sherman Silber,Robert Oates, Steve Rozen ＆ David C. Page. The AZFc region of the Ychromosome features massive palindromes and uniform recurrent Deletions in infertile men. Nat genet 2001:29:279-86

Vogt PH, Edelmann A, Kirsch S, Henegariu O, Hirschmann P, Kiesewetter F,Kohn FM, Schill WB, Farah S, Ramos C, Hartmann M, Hartschuh W, Meschede D, Behre HM, Castel A, Nieschlag E, Weidner W, Grone H-J, Jung A, Engel W, Haidl G. Human Y chromosome azoospermia factors (AZF) mapped to different subregions in Yq11. Hum Mol Genet 1996; 5: 933-43.

Word Health Organization (WHO). Towards more objectivity in diagnosis and management of male infertility. Int J Androl 1987 Suppl 7.

Wylie C. Germ cells. Cell 1999; 96, 165-74.

Yen PH. Advances in Y chromosome mapping. Curr Opinn Obstet Gynecol 1999; 11:275-81.