Testicular Chromosomal Mosaicism and Infertility THOMAS M. JONES, 1 ANTHONY P. AMAROSE, AND MARTIN LEBOWITZ
,
The University of Chicago Pritzker School of Medicine and The Biomediccd Center for Population Research, Chicago, Illinois 60637 ABSTRACT. A healthy young man with azoospermia and no other endocrinological abnormalities was shown to have chromosomal mosaicism with the cytogenetic errors found only in testicular tissue. Three clones of cells were identified in both meiosis I and meiosis II by cytogenetic analysis of direct testicular smears. Peripheral blood karyotypes and buccal smear preparations revealed no abnormalities.
C
LINICAL studies of male infertility have occasionally employed cytogenetic techniques more extensive than would be required to make a diagnosis of classic genetic problems such as Klinefelter's syndrome. The karyotyping of lymphocytes cultured from peripheral blood has permitted investigators to ascertain the relative frequency of the appearance of genetic errors in infertile men. Indeed, studies reporting populations of men with low sperm counts have shown that such patients are much more likely to have detectable chromosomal abnormalities than are men of proven fertility (1,2). Estimates have ranged as high as twenty per cent. The recent development of techniques for studying cells of the germinal epithelium of the testis in meiotic division has permitted investigators to extend their knowledge of errors discovered in somatic cells in mitotic division. Thus, men with abnormal karyotypes have been shown to have analogous errors in testicular tissue (2,3) and the converse has been found as well— i.e., infertile men with demonstrable persistent errors in testicular preparations have Received July 16, 1975. Presented in part at the 47th Annual Meeting of the Central Society for Clinical Research. Supported by grants no. HD 07110 and no. RR 55. 1 Gerald Sokolek fellow in endocrinology. The authors are indebted to Cecilia Turriff, Barbara Cecil, and Mary Ann Myklebost for their skilled technical assistance.
It is postulated that the gonadal cytogenetic defects account for this patient's azoospermia. In addition, it is hypothesized that this type of incomplete spermatogenesis nonetheless produces suf- * ficient feedback material ("inhibin") so that FSH levels are not affected. (J Clin Endocrinol Me tab A 42: 888, 1976)
been found to have analogous errors when ¥ cells from peripheral blood were karyo- h typed (4). Attempts to contrast the spermatogenic cells of infertile men with those of normal controls have not met with much success. It is true that a high proportion of azoo- r spermic or severely oligospermic men have a germinal epithelium in which either no * meiotic figures are seen or in which nonspecific meiotic disturbances are noted (2,57), but, in general, the meiotic figures encountered are not thought to be distinctive. Skakkebaek and co-workers (8) reported no primary structural abnormalities A in meiotic analyses from a large group of i infertile men. We wish to report here a patient with > replicable primary structural abnormalities in meiotic analyses which were not seen in peripheral blood preparations. * Materials and Methods The patient, a 27-year-old man, came to our attention because his ejaculates were persistently devoid of sperm. The medical history and 4 physical examination were unremarkable. The patient had no history of exposure to radiation * or toxic chemicals. He underwent our standard evaluation for male infertility. Blood samples were drawn for multiple hormonal assays. Plasma estrogen was determined by radioimmunoassay (9). Plasma testosterone was assayed by a com- 1 petitive protein binding technique (10). Serum LH and FSH were estimated by radioimmuno-
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TESTICULAR CHROMOSOMAL MOSAICISM TABLE 1. Hormonal data
assay (11,12). LH and FSH values were obtained every twenty minutes for a three-hour period and results are expressed as a mean of ten samples to minimize the errors of individual variation (13). Peripheral blood cultures and buccal mucosal smears were prepared from the patient and from a normal control. Chromosome profiles of 100 metaphase spreads were counted and 500 epithelial cells were scored for the X-chromatin index (14-16). Testicular tissue was obtained by an open biopsy under local anesthesia. Smears were prepared directly from the testicular tissue and cytogenetic analyses made (17).
LHx FSHx Testosterone Estrogen
Patient
Control
49 ng/ml 51.4 ng/ml 540 ng% 29 pg/ml
2.5-74.8 ng/ml 18.8-197.2 ng/ml 350-870 ng% 0-151.5 pg/ml
for patient and control are consistent for normal males (0%). Cytogenetic studies of the testis showed three clones of cells with different, but consistent, karyotypes, i.e., multiple cells with the same karyotype for each clone in the cells of the first and second meiotic divisions. Forty-five spermatogonial metaphases were scored with no discernible clones upon karyotyping. Figure 1 shows the karyotype of a spermatogonial cell with its coiled chromosomes; the chromosome number is 46 with an X and Y chromosome. In the diakinetic stage of meiosis I, 208 cells were scored of which 162 were in the three clones with remaining cells showing random chromosome loss. In meiosis II, a total of 43 cells were counted of which 35 were in the three clones with the remaining cells showing random loss. Of the 162 cells in the three clones in meiosis I, 89 were in clone 1 (54.9%). Figure 2 shows a cell from the first clone (n = 23,XY) as seen in this stage of meiosis
Results
Table 1 compares the hormonal data of the patient with that of men in the same age range with normal ejaculates. None of the patient's values are outside the normal range. One hundred cells were counted from the peripheral blood cultures of both the patient and the control. The chromosome profiles of the patient were: 92 cells with 46 chromosomes, 6 with 45, 1 with 48, and 1 polyploid. The control showed 94 cells with 46 chromosomes, 1 with 44, 3 with 45, 1 with 47, and 1 with 48. The distribution is within normal limits and in no instance was a standard abnormal karyotype found in the aneuploid cells of the patient or the control. The X-chromatin indices
//
889
ti
%, . \
it
1-3 FIG. 1. Karyotype of meiotic spermatogonial metaphase, 2n = 46,XY. Note the coiling of the chromosomes.
4-5
6-I2 andX
I3-I5
pi
I
19-20
16-18
I . G.»
21-22
The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 10 August 2016. at 13:47 For personal use only. No other uses without permission. . All rights reserved.
JONES, AMAROSE AND LEBOWITZ
890
1-3
JCE & M • 1976 Vol 42 • No 5
4-5
0 $ A
FIG. 2. Clone 1 in diakinesis of meiosis I. n = 23.XY.
6-12
0A
13-15
16-18 XY«-
21-22
19-20
which is in contrast to recent reports that Xbearing spermatozoa are normally more prevalent (18). An example of the second clone detected in the patient (n = 22, XY, -F) is seen in Fig. 4 and appeared in 48 cells (29.6%) in meiosis I. The karyotype is taken from a cell in diakinesis of meiosis I. This second clone showed a persistent absence of a member of Group F, presumably number 20. There is some separation of the XY bivalent. An example of this clone from a secondary spermatocyte in meiosis II is seen in Fig. 5. This appeared in 8 cells (22.9%) scored
I. There is a suggestion of fragility of the XY bivalent but the appearance of near separation of the X and Y chromosomes at this stage is not necessarily abnormal (2,8). However, the continued impression of fragility of the X chromosome is abnormal, even though clone one showed the normal chromosome profile. Figure 3, a karyotype of clone one from meiosis II, shows the flared chromosomes at this stage of meiosis and also shows its Y-bearing status. Twentyfive cells (71.4%) were in clone one in meiosis II and most of the cells at this stage of meiosis from this patient were Y-bearing
A
A
)\ 1-3
A
4-5
%
*
•
*
FIG. 3. Clone 1 in metaphase of meiosis II. n = 23,Y.
•
6-12
_ _
19-20
A
K 16-18
21-22
The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 10 August 2016. at 13:47 For personal use only. No other uses without permission. . All rights reserved.
TESTICULAR CHROMOSOMAL MOSAICISM
A A •%
.1 §
u
1-3
FIG. 4. Clone 2 in diakinesis of meiosis I. Note the absence of a chromosome in Group F. n = 22,XY,-F.
JtJm p i \»sJ
891
4-5
^^ ^ $ »• 6-12
13-15
16-18
19-20
21-22
7t A 1-3
"
4-5
FIG. 5. Clone 2 in metaphase of meiosis II. n = 22,Y,-F. 6-12
13-15
16-18
YL
r
19-20
21-22
&
16-18 2JT22"
I.
mar univ
mar univ
The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 10 August 2016. at 13:47 For personal use only. No other uses without permission. . All rights reserved.
JCE & M • 1976 Vol 42 • No 5
JONES, AMAROSE AND LEBOWITZ
892
1-3
4-5 FIG. 7. Clone 3 in metaphase of meiosis II. n = 24,-X or -Y, +2 marker univalents.
6-12 ; \
13-15
19-20
16-18
21-22
in metaphase II. Again, chromosome 20 is missing. No fragments were present in any of the cells to account for its absence. Finally, the patient had a third clone of cells in which the abnormality was in the sex chromosome bivalent (n = 24, -XY, +2 mar). This clone appeared in 25 cells in meiosis I (15.4%). Figure 6 shows a kary-
otype from diakinesis of meiosis I. No abnormalities were seen in the 22 autosomes but the XY bivalent could not be located. Instead, two marker univalents, i.e., unidentifiable, non-paired chromosome fragments were seen. In meiosis II, the same held true (Fig. 7), with the autosomes intact, but, in place of either an X or Y chromosome, we find fragments which are probably the broken portions of the X chromosomes. Two cells were counted in clone 3 (5.7%) in metaphase II. In addition to cytogenetic studies, the testicular biopsy was processed for routine light microscopy. Figure 8 shows a typical section of seminiferous tubules demonstrating that the spermatogenesis, though by no means normal, is not typical of testicular biopsies from azoospermic men. Some degree of hypospermatogenesis was present, though all stages of spermatogenesis could be observed. The lumina tended to be occluded by germ cells, as if the cells were being shed prematurely. No tubules were hyalinized. The interstitial cells were normal in size and number. Discussion
FiG. 8. Cross section of seminiferous tubule showing hypospermatogenesis.
In this patient the cytogenetic errors were found in a non-random fashion so that three distinct clones of cells were seen in the primary and secondary spermatocytes but no
The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 10 August 2016. at 13:47 For personal use only. No other uses without permission. . All rights reserved.
TESTICULAR CHROMOSOMAL MOSAICISM discernible clones were detected in the chromosome profiles of spermatogonial or peripheral blood cells. Such highly specific chromosomal mosaicism has not, to our knowledge, been previously reported. Unlike many azoospermic patients (1), the patient did not have an FSH elevation. In addition, the histological picture was not typical of many of the syndromes associated with azoospermia. It may be that the production even of genetically imperfect spermatozoa which presumably fail to survive into the ejaculatory ducts is associated with the production of sufficient quantities of the frequently postulated, but never isolated, feed-back product, inhibin (19,20), so that the hypothalamic-pituitary axis is not alerted to the absence of complete spermatogenesis. Clearly the degree of cytogenetic disruption found in the testis of the patient could account for his persistent infertility and although this is the first such case we have found in the over 80 cases of male infertility which we have studied under our current protocol, we anticipate that similar cases may be found in the future. References 1. de Kretser, D. M., H. G. Burger, D. Fortune, B.
2. 3.
4.
5.
Hudson, A. R. Long, C. A. Paulsen, and H. D. Taft, Hormonal, histological, and chromosomal studies in adult males with testicular disorders, J Clin Endocrinol Metab 35: 392, 1972. Hulten, M., and J. Lindsten, Cytogenetic aspects of human male meiosis, Adv Hum Genet 4: 327, 1973. Tettenborn, U., A. Gropp, J.-D. Murken, W. Tinnefeld, W. Fuhrman, and E. Schwinger, Meiosis and testicular histology in XYY males, Lancet 2: 267, 1970. Mcllree, M. E., W. S. Tulloch, and J. E. Newsam, Studies on human meiotic chromosomes from testicular tissue, Lancet 1: 679, 1966. Hulten, M., R. Eliasson, and K. G. Tillinger, Low chiasma count and other meiotic irregularities in two infertile 46, XY men with spermatogenic arrest, Hereditas 65: 285, 1970.
893
6. McDermott, A., Meiotic studies on azoospermic men, Fertil Steril 25: 79, 1974. 7. Mcllree, M. E., W. M. Price, W. M. Court Borwn, W. S. Tuloch, J. E. Newsam, and N. MacLean, Chromosome studies on testicular cells from 50 subfertile men, Lancet 2: 64, 1966. 8. Skakkebaek, N. E., J. I. Bryant, and J. Philip, Studies on meiotic chromosomes in infertile men and controls with normal karyotypes, J Reprod Fertil 35: 23, 1973. 9. Chung-Hsiu, W., L. E. Lundy, and S. G. Lee, A rapid radioimmunoassay for plasma estrogen, AmJ Obstet Gynecol 115: 169, 1973. 10. Rosenfield, R. L., Plasma testosterone-binding globulin and indexes of the concentration of unbound plasma androgens in normal and hirsute subjects,/ Clin Endocrinol Metab 32: 717, 1971. 11. Midgley, A. R., Radioimmunoassay: A method for human chorionic gonadotropin and human luteinizing hormone, Endocrinology 79: 10, 1966. 12. Faiman, C , and R. J. Ryan, Radioimmunoassay for human follicle-stimulating hormone, J Clin Endocrinol Metab 27: 444, 1967. 13. Santen, R. J., and C. W. Bardin, Episodic luteinizing hormone secretion in man, J Clin Invest 52: 2617, 1973. 14. Amarose, A. P., E. J. Plotz, and K. E. Markin, Congenital rudimentary gonadal syndrome (XO/ XX) (Cytogenetic and hormonal studies), Obstet Gynecol 27: 673, 1966. 15. Amarose, A. P., E. J. Plotz, and A. A. Stein, Residual chromosomal aberrations in female cancer patients following irradiation therapy, Exp Mol Pathol 7: 58, 1967. 16. Amarose, A. P., and C. R. Schuster, Chromosome analyses of bone marrow and peripheral blood in subjects with a history of illicit drug use, Arch Gen Psychiatry 25: 181, 1971. 17. Evans, E. P., G. Breckon, and C. E. Ford, An air-drying method for meiotic preparations from mammalian testes, Cytogenetics 3: 289, 1964. 18. Quinlivan, W. L. G., and H. Sullivan, The ratios and separation of X and Y spermatozoa in human semen, Fertil Steril 25: 315, 1974. 19. Rosen, S. W., and D. D. Weintraub, Monotropic increase of serum FSH correlated with low sperm count in young men with idiopathic oligospermia and aspermia, J Clin Endocrinol Metab 32: 410, 1971. 20. Vanthiel, D. H., R. J. Sherins, G. H. Myers, and V. T. DeVita, Evidence for a specific seminiferous tubular factor affecting follicle-stimulating hormone secretion in man, J Clin Invest 51: 1009, 1972.
The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 10 August 2016. at 13:47 For personal use only. No other uses without permission. . All rights reserved.
,
The University of Chicago Pritzker School of Medicine and The Biomediccd Center for Population Research, Chicago, Illinois 60637 ABSTRACT. A healthy young man with azoospermia and no other endocrinological abnormalities was shown to have chromosomal mosaicism with the cytogenetic errors found only in testicular tissue. Three clones of cells were identified in both meiosis I and meiosis II by cytogenetic analysis of direct testicular smears. Peripheral blood karyotypes and buccal smear preparations revealed no abnormalities.
C
LINICAL studies of male infertility have occasionally employed cytogenetic techniques more extensive than would be required to make a diagnosis of classic genetic problems such as Klinefelter's syndrome. The karyotyping of lymphocytes cultured from peripheral blood has permitted investigators to ascertain the relative frequency of the appearance of genetic errors in infertile men. Indeed, studies reporting populations of men with low sperm counts have shown that such patients are much more likely to have detectable chromosomal abnormalities than are men of proven fertility (1,2). Estimates have ranged as high as twenty per cent. The recent development of techniques for studying cells of the germinal epithelium of the testis in meiotic division has permitted investigators to extend their knowledge of errors discovered in somatic cells in mitotic division. Thus, men with abnormal karyotypes have been shown to have analogous errors in testicular tissue (2,3) and the converse has been found as well— i.e., infertile men with demonstrable persistent errors in testicular preparations have Received July 16, 1975. Presented in part at the 47th Annual Meeting of the Central Society for Clinical Research. Supported by grants no. HD 07110 and no. RR 55. 1 Gerald Sokolek fellow in endocrinology. The authors are indebted to Cecilia Turriff, Barbara Cecil, and Mary Ann Myklebost for their skilled technical assistance.
It is postulated that the gonadal cytogenetic defects account for this patient's azoospermia. In addition, it is hypothesized that this type of incomplete spermatogenesis nonetheless produces suf- * ficient feedback material ("inhibin") so that FSH levels are not affected. (J Clin Endocrinol Me tab A 42: 888, 1976)
been found to have analogous errors when ¥ cells from peripheral blood were karyo- h typed (4). Attempts to contrast the spermatogenic cells of infertile men with those of normal controls have not met with much success. It is true that a high proportion of azoo- r spermic or severely oligospermic men have a germinal epithelium in which either no * meiotic figures are seen or in which nonspecific meiotic disturbances are noted (2,57), but, in general, the meiotic figures encountered are not thought to be distinctive. Skakkebaek and co-workers (8) reported no primary structural abnormalities A in meiotic analyses from a large group of i infertile men. We wish to report here a patient with > replicable primary structural abnormalities in meiotic analyses which were not seen in peripheral blood preparations. * Materials and Methods The patient, a 27-year-old man, came to our attention because his ejaculates were persistently devoid of sperm. The medical history and 4 physical examination were unremarkable. The patient had no history of exposure to radiation * or toxic chemicals. He underwent our standard evaluation for male infertility. Blood samples were drawn for multiple hormonal assays. Plasma estrogen was determined by radioimmunoassay (9). Plasma testosterone was assayed by a com- 1 petitive protein binding technique (10). Serum LH and FSH were estimated by radioimmuno-
The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 10 August 2016. at 13:47 For personal use only. No other uses without permission. . All rights reserved.
TESTICULAR CHROMOSOMAL MOSAICISM TABLE 1. Hormonal data
assay (11,12). LH and FSH values were obtained every twenty minutes for a three-hour period and results are expressed as a mean of ten samples to minimize the errors of individual variation (13). Peripheral blood cultures and buccal mucosal smears were prepared from the patient and from a normal control. Chromosome profiles of 100 metaphase spreads were counted and 500 epithelial cells were scored for the X-chromatin index (14-16). Testicular tissue was obtained by an open biopsy under local anesthesia. Smears were prepared directly from the testicular tissue and cytogenetic analyses made (17).
LHx FSHx Testosterone Estrogen
Patient
Control
49 ng/ml 51.4 ng/ml 540 ng% 29 pg/ml
2.5-74.8 ng/ml 18.8-197.2 ng/ml 350-870 ng% 0-151.5 pg/ml
for patient and control are consistent for normal males (0%). Cytogenetic studies of the testis showed three clones of cells with different, but consistent, karyotypes, i.e., multiple cells with the same karyotype for each clone in the cells of the first and second meiotic divisions. Forty-five spermatogonial metaphases were scored with no discernible clones upon karyotyping. Figure 1 shows the karyotype of a spermatogonial cell with its coiled chromosomes; the chromosome number is 46 with an X and Y chromosome. In the diakinetic stage of meiosis I, 208 cells were scored of which 162 were in the three clones with remaining cells showing random chromosome loss. In meiosis II, a total of 43 cells were counted of which 35 were in the three clones with the remaining cells showing random loss. Of the 162 cells in the three clones in meiosis I, 89 were in clone 1 (54.9%). Figure 2 shows a cell from the first clone (n = 23,XY) as seen in this stage of meiosis
Results
Table 1 compares the hormonal data of the patient with that of men in the same age range with normal ejaculates. None of the patient's values are outside the normal range. One hundred cells were counted from the peripheral blood cultures of both the patient and the control. The chromosome profiles of the patient were: 92 cells with 46 chromosomes, 6 with 45, 1 with 48, and 1 polyploid. The control showed 94 cells with 46 chromosomes, 1 with 44, 3 with 45, 1 with 47, and 1 with 48. The distribution is within normal limits and in no instance was a standard abnormal karyotype found in the aneuploid cells of the patient or the control. The X-chromatin indices
//
889
ti
%, . \
it
1-3 FIG. 1. Karyotype of meiotic spermatogonial metaphase, 2n = 46,XY. Note the coiling of the chromosomes.
4-5
6-I2 andX
I3-I5
pi
I
19-20
16-18
I . G.»
21-22
The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 10 August 2016. at 13:47 For personal use only. No other uses without permission. . All rights reserved.
JONES, AMAROSE AND LEBOWITZ
890
1-3
JCE & M • 1976 Vol 42 • No 5
4-5
0 $ A
FIG. 2. Clone 1 in diakinesis of meiosis I. n = 23.XY.
6-12
0A
13-15
16-18 XY«-
21-22
19-20
which is in contrast to recent reports that Xbearing spermatozoa are normally more prevalent (18). An example of the second clone detected in the patient (n = 22, XY, -F) is seen in Fig. 4 and appeared in 48 cells (29.6%) in meiosis I. The karyotype is taken from a cell in diakinesis of meiosis I. This second clone showed a persistent absence of a member of Group F, presumably number 20. There is some separation of the XY bivalent. An example of this clone from a secondary spermatocyte in meiosis II is seen in Fig. 5. This appeared in 8 cells (22.9%) scored
I. There is a suggestion of fragility of the XY bivalent but the appearance of near separation of the X and Y chromosomes at this stage is not necessarily abnormal (2,8). However, the continued impression of fragility of the X chromosome is abnormal, even though clone one showed the normal chromosome profile. Figure 3, a karyotype of clone one from meiosis II, shows the flared chromosomes at this stage of meiosis and also shows its Y-bearing status. Twentyfive cells (71.4%) were in clone one in meiosis II and most of the cells at this stage of meiosis from this patient were Y-bearing
A
A
)\ 1-3
A
4-5
%
*
•
*
FIG. 3. Clone 1 in metaphase of meiosis II. n = 23,Y.
•
6-12
_ _
19-20
A
K 16-18
21-22
The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 10 August 2016. at 13:47 For personal use only. No other uses without permission. . All rights reserved.
TESTICULAR CHROMOSOMAL MOSAICISM
A A •%
.1 §
u
1-3
FIG. 4. Clone 2 in diakinesis of meiosis I. Note the absence of a chromosome in Group F. n = 22,XY,-F.
JtJm p i \»sJ
891
4-5
^^ ^ $ »• 6-12
13-15
16-18
19-20
21-22
7t A 1-3
"
4-5
FIG. 5. Clone 2 in metaphase of meiosis II. n = 22,Y,-F. 6-12
13-15
16-18
YL
r
19-20
21-22
&
16-18 2JT22"
I.
mar univ
mar univ
The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 10 August 2016. at 13:47 For personal use only. No other uses without permission. . All rights reserved.
JCE & M • 1976 Vol 42 • No 5
JONES, AMAROSE AND LEBOWITZ
892
1-3
4-5 FIG. 7. Clone 3 in metaphase of meiosis II. n = 24,-X or -Y, +2 marker univalents.
6-12 ; \
13-15
19-20
16-18
21-22
in metaphase II. Again, chromosome 20 is missing. No fragments were present in any of the cells to account for its absence. Finally, the patient had a third clone of cells in which the abnormality was in the sex chromosome bivalent (n = 24, -XY, +2 mar). This clone appeared in 25 cells in meiosis I (15.4%). Figure 6 shows a kary-
otype from diakinesis of meiosis I. No abnormalities were seen in the 22 autosomes but the XY bivalent could not be located. Instead, two marker univalents, i.e., unidentifiable, non-paired chromosome fragments were seen. In meiosis II, the same held true (Fig. 7), with the autosomes intact, but, in place of either an X or Y chromosome, we find fragments which are probably the broken portions of the X chromosomes. Two cells were counted in clone 3 (5.7%) in metaphase II. In addition to cytogenetic studies, the testicular biopsy was processed for routine light microscopy. Figure 8 shows a typical section of seminiferous tubules demonstrating that the spermatogenesis, though by no means normal, is not typical of testicular biopsies from azoospermic men. Some degree of hypospermatogenesis was present, though all stages of spermatogenesis could be observed. The lumina tended to be occluded by germ cells, as if the cells were being shed prematurely. No tubules were hyalinized. The interstitial cells were normal in size and number. Discussion
FiG. 8. Cross section of seminiferous tubule showing hypospermatogenesis.
In this patient the cytogenetic errors were found in a non-random fashion so that three distinct clones of cells were seen in the primary and secondary spermatocytes but no
The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 10 August 2016. at 13:47 For personal use only. No other uses without permission. . All rights reserved.
TESTICULAR CHROMOSOMAL MOSAICISM discernible clones were detected in the chromosome profiles of spermatogonial or peripheral blood cells. Such highly specific chromosomal mosaicism has not, to our knowledge, been previously reported. Unlike many azoospermic patients (1), the patient did not have an FSH elevation. In addition, the histological picture was not typical of many of the syndromes associated with azoospermia. It may be that the production even of genetically imperfect spermatozoa which presumably fail to survive into the ejaculatory ducts is associated with the production of sufficient quantities of the frequently postulated, but never isolated, feed-back product, inhibin (19,20), so that the hypothalamic-pituitary axis is not alerted to the absence of complete spermatogenesis. Clearly the degree of cytogenetic disruption found in the testis of the patient could account for his persistent infertility and although this is the first such case we have found in the over 80 cases of male infertility which we have studied under our current protocol, we anticipate that similar cases may be found in the future. References 1. de Kretser, D. M., H. G. Burger, D. Fortune, B.
2. 3.
4.
5.
Hudson, A. R. Long, C. A. Paulsen, and H. D. Taft, Hormonal, histological, and chromosomal studies in adult males with testicular disorders, J Clin Endocrinol Metab 35: 392, 1972. Hulten, M., and J. Lindsten, Cytogenetic aspects of human male meiosis, Adv Hum Genet 4: 327, 1973. Tettenborn, U., A. Gropp, J.-D. Murken, W. Tinnefeld, W. Fuhrman, and E. Schwinger, Meiosis and testicular histology in XYY males, Lancet 2: 267, 1970. Mcllree, M. E., W. S. Tulloch, and J. E. Newsam, Studies on human meiotic chromosomes from testicular tissue, Lancet 1: 679, 1966. Hulten, M., R. Eliasson, and K. G. Tillinger, Low chiasma count and other meiotic irregularities in two infertile 46, XY men with spermatogenic arrest, Hereditas 65: 285, 1970.
893
6. McDermott, A., Meiotic studies on azoospermic men, Fertil Steril 25: 79, 1974. 7. Mcllree, M. E., W. M. Price, W. M. Court Borwn, W. S. Tuloch, J. E. Newsam, and N. MacLean, Chromosome studies on testicular cells from 50 subfertile men, Lancet 2: 64, 1966. 8. Skakkebaek, N. E., J. I. Bryant, and J. Philip, Studies on meiotic chromosomes in infertile men and controls with normal karyotypes, J Reprod Fertil 35: 23, 1973. 9. Chung-Hsiu, W., L. E. Lundy, and S. G. Lee, A rapid radioimmunoassay for plasma estrogen, AmJ Obstet Gynecol 115: 169, 1973. 10. Rosenfield, R. L., Plasma testosterone-binding globulin and indexes of the concentration of unbound plasma androgens in normal and hirsute subjects,/ Clin Endocrinol Metab 32: 717, 1971. 11. Midgley, A. R., Radioimmunoassay: A method for human chorionic gonadotropin and human luteinizing hormone, Endocrinology 79: 10, 1966. 12. Faiman, C , and R. J. Ryan, Radioimmunoassay for human follicle-stimulating hormone, J Clin Endocrinol Metab 27: 444, 1967. 13. Santen, R. J., and C. W. Bardin, Episodic luteinizing hormone secretion in man, J Clin Invest 52: 2617, 1973. 14. Amarose, A. P., E. J. Plotz, and K. E. Markin, Congenital rudimentary gonadal syndrome (XO/ XX) (Cytogenetic and hormonal studies), Obstet Gynecol 27: 673, 1966. 15. Amarose, A. P., E. J. Plotz, and A. A. Stein, Residual chromosomal aberrations in female cancer patients following irradiation therapy, Exp Mol Pathol 7: 58, 1967. 16. Amarose, A. P., and C. R. Schuster, Chromosome analyses of bone marrow and peripheral blood in subjects with a history of illicit drug use, Arch Gen Psychiatry 25: 181, 1971. 17. Evans, E. P., G. Breckon, and C. E. Ford, An air-drying method for meiotic preparations from mammalian testes, Cytogenetics 3: 289, 1964. 18. Quinlivan, W. L. G., and H. Sullivan, The ratios and separation of X and Y spermatozoa in human semen, Fertil Steril 25: 315, 1974. 19. Rosen, S. W., and D. D. Weintraub, Monotropic increase of serum FSH correlated with low sperm count in young men with idiopathic oligospermia and aspermia, J Clin Endocrinol Metab 32: 410, 1971. 20. Vanthiel, D. H., R. J. Sherins, G. H. Myers, and V. T. DeVita, Evidence for a specific seminiferous tubular factor affecting follicle-stimulating hormone secretion in man, J Clin Invest 51: 1009, 1972.
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