Hum Genet (1992) 89:590-592
9 Springer-Verlag1992
Original investigations The majority of the marker chromosomes in Japanese patients with stigmata of Turner syndrome are derived from Y chromosomes
Shigeo Nagafuchi 1, Takashi Tamura 1' 2, Yutaka Nakahori 1, Kazue Takano 3, Yoshikazu Nishi 4, Noritaka Iwatani 5, Manabu Kitao 6, Yoshihiro Hori 7, Susumn Konda 8, Tomoko Hasegawa 9, Hironao Numabe 1, Kenji Fujieda 1~ Toshiaki Tanaka 11, ltsuro Hibi ~, and Yasuo Nakagome 1 1Department of Congenital Abnormalities Research, National Children's Medical Research Center, 3-35-31 Taishido, Setagaya-ku, Tokyo 154, Japan 2Department of Clinical Genetics, School of Health Science, Kyorin University, Tokyo, Japan 3Department of Medicine, Tokyo Women's Medical College, Tokyo, Japan 4Department of Pediatrics, Hiroshima Red Cross Hospital, Hiroshima, Japan 5Department of Child Development, Kumamoto University Medical School, Kumamoto, Japan 6Department of Obstetrics and Gynecology, Shimane Medical College, Shimane, Japan 7Department of Pediatrics, Tokyo Medical College, Tokyo, Japan SDepartment of Pediatrics, School of Medicine, Chiba University, Chiba, Japan 9Division of Clinical Genetics and Cytogenetics, Shizuoka Children's Hospital, Shizuoka, Japan ~~ of Pediatrics, Hokkaido University School of Medicine, Sapporo, Japan 11Department of Endocrinology, National Children's Medical Research Center, Tokyo, Japan Received November 22, 1991 / Revised February 7, 1992
Summary. D N A analyses of 41 individuals with stigmata of Turner syndrome and a 45,X/46,X+mar or 46,X+mar karyotype were carried out. Southern-blot analysis employing 17 Y-specific probes was used to determine whether the marker chromosome was Y-chromosomal in origin. Of the 41 D N A samples from these patients, 23 contained detectable Y-chromosomal DNA. Points of chromosome breakage were distributed over the entire length of the Y long arm. Three individuals, who carry different portions of the Y chromosome, had developed gonadoblastoma. GBY (the gonadoblastoma locus on the Y chromosome) is mapped proximal to DYS132, midway between the 13 Yq loci that we have studied. We also used a polymerase chain reaction technique that could detect 7 loci over the length of the Y chromosome. This technique may be useful for the rapid assessment of marker chromosomes, especially for evaluating the risk of gonadoblastoma.
Introduction Turner syndrome is a relatively common disorder occurring in about 1/2500 female births (Nakagome et al. 1963). Short stature and hypergonadotropic hypogonadism are among the most characteristic clinical features. Although 45,X is the most common karyotype, this syndrome may also be caused by the partial absence of one Correspondence to: Y. Nakagome, Department of Human Genetics, School of International Health, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, Japan
of the X chromosomes (Ford et al. 1959; H o o k and Warburton 1982). Features of Turner syndrome may be seen in cases who have 45,X/46,XY mosaicism. Since many of these patients have a testis on one side and a streak gonad on the other, they may well be referred to as showing mixed gonadal dysgenesis (Zah et al. 1975). A marker chromosome is a structurally abnormal chromosome of unknown origin. Patients with clinical features of Turner syndrome often have a 45,X/46,X+mar or, much less frequently, a 4 6 , X + m a r karyotype. It is crucial to know whether their marker chromosomes are of Y-chromosomal origin because gonadoblastoma may develop in patients with dysgenetic gonads, and cells with a Y or a partial Y chromosome (Scully 1970; Schellhas 1974a, b). We have cloned 10 novel Y-specific D N A fragments and mapped them together with 13 known loci on the Y chromosome (Nakahori et al. 1991a). In addition, we have developed a screening system of Y chromosomal aberrations using the polymerase chain reaction (PCR) technique (Nagafuchi et al. 1992). Using these molecular techniques, we have examined 41 Japanese patients with stigmata of Turner syndrome and a marker chromosome to determine whether the marker chromosome contained Y-chromosomal DNA.
Materials and methods Clinical samples
Some 302 patients were diagnosed as having Turner syndrome by pediatric endocrinologists according to clinical findings, including
591 female external genitalia, short stature and other stigmata, and a karyotype compatible with Turner syndrome (Tanaka et al. 1991). All the patients had at least one of the following cell lines: 45,X; 45,X plus a structurally abnormal X chromosome; or 45,X+mar. Of the 302, 27 had a marker chromosome(s), and of these 27, 22 were available for the present study. An additional 19 patients who had both clinical stigmata compatible with Turner syndrome and a marker chromosome were included. The latter were among the patients who were referred to us for the identification of marker chromosomes. Of 41 patients analyzed by Southern blotting, 37 had 45,X/ 46,X+mar mosaicism and four cases had a 46,X+mar karyotype. Of the 41 cases, individuals (cases 877,878,966, 1017 and 1031 to 1035) had been included in the mapping panel described by Nakahori et al. (1991a). To date, 3 cases (877, 1539 and 1514), each with a 45,X/46,X+mar karyotype, have developed gonadoblastoma. DNA
analysis
Genomic DNAs were prepared from peripheral blood leukocytes by standard methods (Maniatis et al. 1982) and analyzed by Southern blotting using 17 Y-specific fragments as probes. These probes detect 23 loci on the Y chromosome, and their order has been defined by deletion mapping (Nakahori et al. 1991a). Restriction digestion, electrophoresis, transfer, and hybridization of DNA were performed as previously described (Tamura et al. 1991). Southern-hybridization analysis has its limitation in the detection of a low proportion of cells with a Y chromosome. To assess this limitation, a male DNA sample was serially diluted with female control DNA. As little as 1 : 16 (about 6%) of male DNA in female DNA could be detected by Y-specific single copy probes. In cases of mosaic individuals exhibiting less than 20% of cells with a marker chromosome, a dual PCR method amplifying the amelogenin locus (AMGL) was used. The AMGL locus is on the short arm of the Y adjacent to the centromere (Nakahori et al. 1991a). This method could detect one part of male DNA in 25000 parts of female DNA (Nakagome et al. 1991a). PCR detection of 7 loci over the length of the Y chromosome was also carried out in some of the patients for comparison. The oligonucleotide primers used have been described elsewhere: PABY (Ellis et al. 1990), SRY (Nakagome et al. 1991c), AMGL (Nakahori et al. 1991b), DYS139, DYS132, DYS1 (Nagafuehi et al. 1992) and the B fragment of DYZ1 (Nakagome et al. 1991b). For dual amplification of the AMGL locus, a second set of primers was used (Nakagome et al. 1991a).
Results Of 41 D N A samples analyzed by Southern blotting, 23 contained Y-specific D N A sequences. The 23 marker chromosomes consisted of various segments of the Y chromosome including the centromere. Their break points were distributed evenly over the long arm (Fig. 1). Three cases, 925, 1519 and 1523, showed a weaker than normal (but distinctly positive) signal for the DYZ1 locus, although they were quinacrine-negative on chromosome analysis. The dual P C R method (Nakagome et al. 1991b) did not detect Y-specific D N A sequences in any of the 9 individuals out of the 18 Y-negative cases. Five samples were subjected to P C R analysis for 7 loci in addition to Southern-blot analysis. They were cases 1578, 1579, 1580, 1582 and 1583. Segments of Y chromosomes were detected in two cases (1580 and 1583), confirming the results of the Southern blot analysis.
Fig. 1. Data from 23 patients with a portion of the Y chromosome. Each half-tone bar indicates the presence of loci in a particular patient. The loci are indicated as numbers at the top: 1 ZFY; 2 DXYS5; 3 DXYS73Y; 4 DYF27B; 5 DYF27C; 6 DYS131; 7 DYS7A/B; 8 DYS130; 9 AMGL; 10 DYS7D; 11 DYS11; 12 DYS140; 13 DYS139; 14 DYS133; 15 DYS134; 16 DYS132; 17 DYS135; 18 DYF27; 19 DYS131A; 20 DYS7E; 21 DYS7C; 22 DYS1; 23 DYZ1. Numbers 1-9 correspond to loci on the Y short arm, 10 roughly corresponds to the centromere, and 11-23 to Yq. DYZl(23) is a repeated DNA family and its distribution roughly corresponds to the fluorescent part of Yq
Discussion Patients with Turner syndrome often have a structurally abnormal X or Y chromosome in addition to a normal X chromosome. This syndrome might be the result of monosomy for a gene c o m m o n to the X and Y chromosome. Recently, it has been claimed that a gene responsible for Turner syndrome has been cloned (Fisher et al. 1990). This gene has a homologous locus in each of Yp and Xq. Features of Turner syndrome in cases with a 46,XY karyotype (Magenis et al. 1984; Blagowidow et al. 1989) might be caused by the deletion of the gene on Yp. The reason for 2 of our cases (1510 and 1527) with a 46,X+mar karyotype showing features of Turner syndrome is not known, since the marker has a deletion of Yq instead of Yp. These patients might have a very low proportion of the 45,X cell line, or an additional deletion or a mutation of the Turner gene on either the X or the Y chromosome. A gonadoblastoma often develops in patients with both dysgenetic gonads and a Y chromosome (Scully 1970; Schellhas 1974a, b). It has been postulated that there is a gonadoblastoma gene (GBY) on the Y chromosome. By deletion mapping of a 46,XY female with gonadoblastoma, G B Y has been mapped to the Y long arm (Page 1987). In our series, three patients (877, 1539 and 1514), who carry different portions of the Y chromosomes, have developed gonadoblastoma. G B Y can be mapped proximal to DYS132, but not within the distal half of the non-fluorescent part and fluorescent part of Yq (represented by numbers 17-23 in Fig. 1). The P C R technique described by Saiki et al. (1988) has enabled the rapid analysis of many samples. Out of the 7 sets of primers used in this study, 2 sets detecting
592 DYS139 a n d DYS132 cover the p r o x i m a l Y q region, which is p o t e n t i a l l y r e l e v a n t to the d e v e l o p m e n t of g o n a d o b l a s t o m a . W e hope that these p r i m e r s will be useful for screening m a r k e r c h r o m o s o m e s in p a t i e n t s with 4 5 , X / 4 6 , X + m a r karyotypes.
Acknowledgements. We are grateful to Professor S. R. Young, South Carolina University for helpful discussions, and to Mrs. Satoko Seki for technical assistance. This work was supported in part by grants from the Ministry of Health and Welfare, the Ministry of Education, Science and Culture, and the Science and Technology Agency, Japan.
References Blagowidow N, Page DC, Huff D, Mennuti MT (1989) UlrichTurner syndrome in an XY female fetus with deletion of the sex-determining portion of the Y chromosome. Am J Med Genet 34 : 159-162 Ellis N, Taylor A, Bengtsson BO, Kidd J, Rogers J, Goodfellow PN (1990) Population structure of the human pseudoautosomal boundary. Nature 344: 663-665 Fisher EMC, Beer-Romero P, Brown LG, Ridley A, McNeil JA, Lawrence JB, Willard HF, Bieber FR, Page DC (1990) Homologous ribosomal protein genes on the human X and Y chromosomes: escape from X inactivation and possible implications for Turner syndrome. Cell 63 : 1205-1218 Ford CE, Jones KW, Polani PE, Almeida JC de, Briggs JH (1959) A sex chromosome anomaly in a case of gonadal dysgenesis (Turner's syndrome). Lancet I:711-713 Hook EB, Warburton D (1982) The distribution of chromosomal genotypes associated with Turner's syndrome: livebirth prevalence rates and evidence for diminished fetal mortality and severity in genotypes with structural X abnormalities or mosaicism. Hum Genet 64 : 24-27 Magenis RE, Tochen ML, Holahan KP, Carey T, Allen L, Brown MG (1984) Turner syndrome resulting from partial deletion of Y chromosome short arm: localization of male determinants. J Pediatr 105 : 916-919 Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory. Cold Spring Harbor, NY Nagafuchi S, Seki S, Nakahori Y, Tamura T, Numabe H, Nakagome Y (1992) PCR detection of structurally abnormal Y chromosomes. Jpn J Hum Genet 37 (in press)
Nakagome Y, Hibi I, Konoshita K, Nagao T, Aikawa M (1963) Incidence of Turner's syndromes in Japanese dwarfed girls. Lancet II : 412 Nakagome Y, Seki S, Nagafuchi S, Nakahori Y, Sato K (1991a) Absence of fetal cells in maternal circulation at a level of 1 in 25 000. Am J Med Genet 40 : 506-508 Nakagome Y, Nagafuchi S, Seki S, Nakahori Y, Tamura T, Yamada M, Iwaya M (1991b) A repeating unit of the DYZ1 family on the human Y chromosome consists of segments with partial male-specificity. Cytogenet Cell Genet 56:74-77 Nakagome Y, Seki S, Fukutani K, Nagafuchi S, Nakahori Y, Tamura T (1991c) PCR detection of distal Yp sequences in an XX true hermaphrodite. Am J Med Genet 41:112-114 Nakahori Y, Tamura T, Nagafuchi S, Fujieda K, Minowada S, Fukutani K, Fuse H, Hayashi K, Kuroki Y, Fukushima Y, Agematsu K, Kuno T, Kaneko S, Yamada K, Kitagawa T, Nonomura M, Fukuda S, Kusano M, Onigata S, Hibi I, Nakagome Y (1991a) Molecular cloning and mapping of 10 new probes on the human Y chromosome Genomics 9:765769 Nakahori Y, Hamano K, Iwaya M, Nakagome Y (1991b) Sex identification by polymerase chain reaction using X-Y homologous primer. Am J Med Genet 39 : 472-473 Page DC (1987) Hypothesis: a Y-chromosomal gene causes gonadoblastoma in dysgenetic gonads. Development 101 [Suppl]: 151-155 Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239 : 487-491 Schellhas HF (1974a) Malignant potential of the dysgenetic gonads. I. Obstet Gynecol 44 : 298-309 Schellhas HF (1974b) Malignant potential of the dysgenetic gonads. II. Obstet Gynecol 44 : 455-462 Scully RE (1970) Gonadoblastoma: a review of 74 cases. Cancer 25 : 1340-1356 Tamura T, Kuroki Y, Nagafuchi S, Suwa S, Nakahori Y, Terashima K, Furusho T, Nakagome Y (1991) DNA analysis of a patient with two different marker chromosome using Y-specific DNA probes. Jpn J Hum Genet 36:195-199 Tanaka T, Hibi I, Shizume K (1991) GH-secretion capacity in Turner syndrome and its relation to clinical characteristics and effect of GH treatment (in Japanese). Folia Endocrinol 67: 597-610 Zah W, Kalderon HE, Tucci JR (1975) Mixed gonadal dysgenesis. Acta Endocrinol 79 : 3-39
9 Springer-Verlag1992
Original investigations The majority of the marker chromosomes in Japanese patients with stigmata of Turner syndrome are derived from Y chromosomes
Shigeo Nagafuchi 1, Takashi Tamura 1' 2, Yutaka Nakahori 1, Kazue Takano 3, Yoshikazu Nishi 4, Noritaka Iwatani 5, Manabu Kitao 6, Yoshihiro Hori 7, Susumn Konda 8, Tomoko Hasegawa 9, Hironao Numabe 1, Kenji Fujieda 1~ Toshiaki Tanaka 11, ltsuro Hibi ~, and Yasuo Nakagome 1 1Department of Congenital Abnormalities Research, National Children's Medical Research Center, 3-35-31 Taishido, Setagaya-ku, Tokyo 154, Japan 2Department of Clinical Genetics, School of Health Science, Kyorin University, Tokyo, Japan 3Department of Medicine, Tokyo Women's Medical College, Tokyo, Japan 4Department of Pediatrics, Hiroshima Red Cross Hospital, Hiroshima, Japan 5Department of Child Development, Kumamoto University Medical School, Kumamoto, Japan 6Department of Obstetrics and Gynecology, Shimane Medical College, Shimane, Japan 7Department of Pediatrics, Tokyo Medical College, Tokyo, Japan SDepartment of Pediatrics, School of Medicine, Chiba University, Chiba, Japan 9Division of Clinical Genetics and Cytogenetics, Shizuoka Children's Hospital, Shizuoka, Japan ~~ of Pediatrics, Hokkaido University School of Medicine, Sapporo, Japan 11Department of Endocrinology, National Children's Medical Research Center, Tokyo, Japan Received November 22, 1991 / Revised February 7, 1992
Summary. D N A analyses of 41 individuals with stigmata of Turner syndrome and a 45,X/46,X+mar or 46,X+mar karyotype were carried out. Southern-blot analysis employing 17 Y-specific probes was used to determine whether the marker chromosome was Y-chromosomal in origin. Of the 41 D N A samples from these patients, 23 contained detectable Y-chromosomal DNA. Points of chromosome breakage were distributed over the entire length of the Y long arm. Three individuals, who carry different portions of the Y chromosome, had developed gonadoblastoma. GBY (the gonadoblastoma locus on the Y chromosome) is mapped proximal to DYS132, midway between the 13 Yq loci that we have studied. We also used a polymerase chain reaction technique that could detect 7 loci over the length of the Y chromosome. This technique may be useful for the rapid assessment of marker chromosomes, especially for evaluating the risk of gonadoblastoma.
Introduction Turner syndrome is a relatively common disorder occurring in about 1/2500 female births (Nakagome et al. 1963). Short stature and hypergonadotropic hypogonadism are among the most characteristic clinical features. Although 45,X is the most common karyotype, this syndrome may also be caused by the partial absence of one Correspondence to: Y. Nakagome, Department of Human Genetics, School of International Health, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, Japan
of the X chromosomes (Ford et al. 1959; H o o k and Warburton 1982). Features of Turner syndrome may be seen in cases who have 45,X/46,XY mosaicism. Since many of these patients have a testis on one side and a streak gonad on the other, they may well be referred to as showing mixed gonadal dysgenesis (Zah et al. 1975). A marker chromosome is a structurally abnormal chromosome of unknown origin. Patients with clinical features of Turner syndrome often have a 45,X/46,X+mar or, much less frequently, a 4 6 , X + m a r karyotype. It is crucial to know whether their marker chromosomes are of Y-chromosomal origin because gonadoblastoma may develop in patients with dysgenetic gonads, and cells with a Y or a partial Y chromosome (Scully 1970; Schellhas 1974a, b). We have cloned 10 novel Y-specific D N A fragments and mapped them together with 13 known loci on the Y chromosome (Nakahori et al. 1991a). In addition, we have developed a screening system of Y chromosomal aberrations using the polymerase chain reaction (PCR) technique (Nagafuchi et al. 1992). Using these molecular techniques, we have examined 41 Japanese patients with stigmata of Turner syndrome and a marker chromosome to determine whether the marker chromosome contained Y-chromosomal DNA.
Materials and methods Clinical samples
Some 302 patients were diagnosed as having Turner syndrome by pediatric endocrinologists according to clinical findings, including
591 female external genitalia, short stature and other stigmata, and a karyotype compatible with Turner syndrome (Tanaka et al. 1991). All the patients had at least one of the following cell lines: 45,X; 45,X plus a structurally abnormal X chromosome; or 45,X+mar. Of the 302, 27 had a marker chromosome(s), and of these 27, 22 were available for the present study. An additional 19 patients who had both clinical stigmata compatible with Turner syndrome and a marker chromosome were included. The latter were among the patients who were referred to us for the identification of marker chromosomes. Of 41 patients analyzed by Southern blotting, 37 had 45,X/ 46,X+mar mosaicism and four cases had a 46,X+mar karyotype. Of the 41 cases, individuals (cases 877,878,966, 1017 and 1031 to 1035) had been included in the mapping panel described by Nakahori et al. (1991a). To date, 3 cases (877, 1539 and 1514), each with a 45,X/46,X+mar karyotype, have developed gonadoblastoma. DNA
analysis
Genomic DNAs were prepared from peripheral blood leukocytes by standard methods (Maniatis et al. 1982) and analyzed by Southern blotting using 17 Y-specific fragments as probes. These probes detect 23 loci on the Y chromosome, and their order has been defined by deletion mapping (Nakahori et al. 1991a). Restriction digestion, electrophoresis, transfer, and hybridization of DNA were performed as previously described (Tamura et al. 1991). Southern-hybridization analysis has its limitation in the detection of a low proportion of cells with a Y chromosome. To assess this limitation, a male DNA sample was serially diluted with female control DNA. As little as 1 : 16 (about 6%) of male DNA in female DNA could be detected by Y-specific single copy probes. In cases of mosaic individuals exhibiting less than 20% of cells with a marker chromosome, a dual PCR method amplifying the amelogenin locus (AMGL) was used. The AMGL locus is on the short arm of the Y adjacent to the centromere (Nakahori et al. 1991a). This method could detect one part of male DNA in 25000 parts of female DNA (Nakagome et al. 1991a). PCR detection of 7 loci over the length of the Y chromosome was also carried out in some of the patients for comparison. The oligonucleotide primers used have been described elsewhere: PABY (Ellis et al. 1990), SRY (Nakagome et al. 1991c), AMGL (Nakahori et al. 1991b), DYS139, DYS132, DYS1 (Nagafuehi et al. 1992) and the B fragment of DYZ1 (Nakagome et al. 1991b). For dual amplification of the AMGL locus, a second set of primers was used (Nakagome et al. 1991a).
Results Of 41 D N A samples analyzed by Southern blotting, 23 contained Y-specific D N A sequences. The 23 marker chromosomes consisted of various segments of the Y chromosome including the centromere. Their break points were distributed evenly over the long arm (Fig. 1). Three cases, 925, 1519 and 1523, showed a weaker than normal (but distinctly positive) signal for the DYZ1 locus, although they were quinacrine-negative on chromosome analysis. The dual P C R method (Nakagome et al. 1991b) did not detect Y-specific D N A sequences in any of the 9 individuals out of the 18 Y-negative cases. Five samples were subjected to P C R analysis for 7 loci in addition to Southern-blot analysis. They were cases 1578, 1579, 1580, 1582 and 1583. Segments of Y chromosomes were detected in two cases (1580 and 1583), confirming the results of the Southern blot analysis.
Fig. 1. Data from 23 patients with a portion of the Y chromosome. Each half-tone bar indicates the presence of loci in a particular patient. The loci are indicated as numbers at the top: 1 ZFY; 2 DXYS5; 3 DXYS73Y; 4 DYF27B; 5 DYF27C; 6 DYS131; 7 DYS7A/B; 8 DYS130; 9 AMGL; 10 DYS7D; 11 DYS11; 12 DYS140; 13 DYS139; 14 DYS133; 15 DYS134; 16 DYS132; 17 DYS135; 18 DYF27; 19 DYS131A; 20 DYS7E; 21 DYS7C; 22 DYS1; 23 DYZ1. Numbers 1-9 correspond to loci on the Y short arm, 10 roughly corresponds to the centromere, and 11-23 to Yq. DYZl(23) is a repeated DNA family and its distribution roughly corresponds to the fluorescent part of Yq
Discussion Patients with Turner syndrome often have a structurally abnormal X or Y chromosome in addition to a normal X chromosome. This syndrome might be the result of monosomy for a gene c o m m o n to the X and Y chromosome. Recently, it has been claimed that a gene responsible for Turner syndrome has been cloned (Fisher et al. 1990). This gene has a homologous locus in each of Yp and Xq. Features of Turner syndrome in cases with a 46,XY karyotype (Magenis et al. 1984; Blagowidow et al. 1989) might be caused by the deletion of the gene on Yp. The reason for 2 of our cases (1510 and 1527) with a 46,X+mar karyotype showing features of Turner syndrome is not known, since the marker has a deletion of Yq instead of Yp. These patients might have a very low proportion of the 45,X cell line, or an additional deletion or a mutation of the Turner gene on either the X or the Y chromosome. A gonadoblastoma often develops in patients with both dysgenetic gonads and a Y chromosome (Scully 1970; Schellhas 1974a, b). It has been postulated that there is a gonadoblastoma gene (GBY) on the Y chromosome. By deletion mapping of a 46,XY female with gonadoblastoma, G B Y has been mapped to the Y long arm (Page 1987). In our series, three patients (877, 1539 and 1514), who carry different portions of the Y chromosomes, have developed gonadoblastoma. G B Y can be mapped proximal to DYS132, but not within the distal half of the non-fluorescent part and fluorescent part of Yq (represented by numbers 17-23 in Fig. 1). The P C R technique described by Saiki et al. (1988) has enabled the rapid analysis of many samples. Out of the 7 sets of primers used in this study, 2 sets detecting
592 DYS139 a n d DYS132 cover the p r o x i m a l Y q region, which is p o t e n t i a l l y r e l e v a n t to the d e v e l o p m e n t of g o n a d o b l a s t o m a . W e hope that these p r i m e r s will be useful for screening m a r k e r c h r o m o s o m e s in p a t i e n t s with 4 5 , X / 4 6 , X + m a r karyotypes.
Acknowledgements. We are grateful to Professor S. R. Young, South Carolina University for helpful discussions, and to Mrs. Satoko Seki for technical assistance. This work was supported in part by grants from the Ministry of Health and Welfare, the Ministry of Education, Science and Culture, and the Science and Technology Agency, Japan.
References Blagowidow N, Page DC, Huff D, Mennuti MT (1989) UlrichTurner syndrome in an XY female fetus with deletion of the sex-determining portion of the Y chromosome. Am J Med Genet 34 : 159-162 Ellis N, Taylor A, Bengtsson BO, Kidd J, Rogers J, Goodfellow PN (1990) Population structure of the human pseudoautosomal boundary. Nature 344: 663-665 Fisher EMC, Beer-Romero P, Brown LG, Ridley A, McNeil JA, Lawrence JB, Willard HF, Bieber FR, Page DC (1990) Homologous ribosomal protein genes on the human X and Y chromosomes: escape from X inactivation and possible implications for Turner syndrome. Cell 63 : 1205-1218 Ford CE, Jones KW, Polani PE, Almeida JC de, Briggs JH (1959) A sex chromosome anomaly in a case of gonadal dysgenesis (Turner's syndrome). Lancet I:711-713 Hook EB, Warburton D (1982) The distribution of chromosomal genotypes associated with Turner's syndrome: livebirth prevalence rates and evidence for diminished fetal mortality and severity in genotypes with structural X abnormalities or mosaicism. Hum Genet 64 : 24-27 Magenis RE, Tochen ML, Holahan KP, Carey T, Allen L, Brown MG (1984) Turner syndrome resulting from partial deletion of Y chromosome short arm: localization of male determinants. J Pediatr 105 : 916-919 Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory. Cold Spring Harbor, NY Nagafuchi S, Seki S, Nakahori Y, Tamura T, Numabe H, Nakagome Y (1992) PCR detection of structurally abnormal Y chromosomes. Jpn J Hum Genet 37 (in press)
Nakagome Y, Hibi I, Konoshita K, Nagao T, Aikawa M (1963) Incidence of Turner's syndromes in Japanese dwarfed girls. Lancet II : 412 Nakagome Y, Seki S, Nagafuchi S, Nakahori Y, Sato K (1991a) Absence of fetal cells in maternal circulation at a level of 1 in 25 000. Am J Med Genet 40 : 506-508 Nakagome Y, Nagafuchi S, Seki S, Nakahori Y, Tamura T, Yamada M, Iwaya M (1991b) A repeating unit of the DYZ1 family on the human Y chromosome consists of segments with partial male-specificity. Cytogenet Cell Genet 56:74-77 Nakagome Y, Seki S, Fukutani K, Nagafuchi S, Nakahori Y, Tamura T (1991c) PCR detection of distal Yp sequences in an XX true hermaphrodite. Am J Med Genet 41:112-114 Nakahori Y, Tamura T, Nagafuchi S, Fujieda K, Minowada S, Fukutani K, Fuse H, Hayashi K, Kuroki Y, Fukushima Y, Agematsu K, Kuno T, Kaneko S, Yamada K, Kitagawa T, Nonomura M, Fukuda S, Kusano M, Onigata S, Hibi I, Nakagome Y (1991a) Molecular cloning and mapping of 10 new probes on the human Y chromosome Genomics 9:765769 Nakahori Y, Hamano K, Iwaya M, Nakagome Y (1991b) Sex identification by polymerase chain reaction using X-Y homologous primer. Am J Med Genet 39 : 472-473 Page DC (1987) Hypothesis: a Y-chromosomal gene causes gonadoblastoma in dysgenetic gonads. Development 101 [Suppl]: 151-155 Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239 : 487-491 Schellhas HF (1974a) Malignant potential of the dysgenetic gonads. I. Obstet Gynecol 44 : 298-309 Schellhas HF (1974b) Malignant potential of the dysgenetic gonads. II. Obstet Gynecol 44 : 455-462 Scully RE (1970) Gonadoblastoma: a review of 74 cases. Cancer 25 : 1340-1356 Tamura T, Kuroki Y, Nagafuchi S, Suwa S, Nakahori Y, Terashima K, Furusho T, Nakagome Y (1991) DNA analysis of a patient with two different marker chromosome using Y-specific DNA probes. Jpn J Hum Genet 36:195-199 Tanaka T, Hibi I, Shizume K (1991) GH-secretion capacity in Turner syndrome and its relation to clinical characteristics and effect of GH treatment (in Japanese). Folia Endocrinol 67: 597-610 Zah W, Kalderon HE, Tucci JR (1975) Mixed gonadal dysgenesis. Acta Endocrinol 79 : 3-39
