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Meiotic Indices in the Oligospermic Male
Vol. 28, No.9, September 1977 Printed in U.s.A.
FERTILITY AND STERILITY Copyright' 1977 The American Fertility Society
MEIOTIC INDICES IN THE OLIGOSPERMIC MALE
DONALD J. MEHAN, M.D.* MICHEAL J. CHEHVAL, M.D. SISTER LEO RITA YOLK
Departments of Urology, St. Mary's Hospital and St. Louis University Hospital, St. Louis, Missouri 63117
Thirty-four oligospermic males were evaluated by determining the meiotic index in testicular biopsy specimens. Only 1 patient of 1 0 with a meiotic index of less than 15 achieved pregnancy, whereas 16 of24 patients with a meiotic index of more than 15 had a successful outcome. Corresponding increases in the semenogram were also noted to be related to the meiotic index.
For the past 4 years we have been studying the clinical application of meiotic preparations to the evaluation and treatment of the subfertile male. Our initial studies dealt with the identification of figures of meiosis in testicular biopsy specimens. 1 These studies dealt with the ability to predict the results of therapy purely on the basis of noting the stage of meiosis present in the biopsy specimen. The clinical correlation of prognosis based on the presence of meiotic figures was disappointing and prompted us to count the meiotic figures rather than rely on qualitative analysis. MATERIALS AND METHODS
Thirty-four patients referred for evaluation of subfertility were entered into the study. No significant endocrinologic or physical findings were present with the exception of varicocele (in 25 men). All patients were oligospermic, having a count ofless than 20 x 106 spermatozoa/ml. Treatment consisted ofligation of the internal spermatic vein (13 cases), ligation of the internal spermatic vein plus anterior pituitary-like hormone (Ayerst Laboratories, New York, N. Y.) (APL) (12 cases), or APL only (9 cases). Testicular biopsy specimens were obtained either during the repair of a varicocele or independently as part of the work-up Received April 22, 1977; revised May 16, 1977; accepted May 23,1977. *Reprint requests: Donald J. Mehan, M.D., 1035 Bellevue Avenue, St. Louis, Mo. 63117.
of the oligospermic patient. A part of the biopsy specimen was fixed and stained in our cytogenetic laboratory as previously described. 1 In addition to the usual cellular identification, meiotic counts were carried out in the following fashion. The identifiable meiotic figures per 100 nucleated cells were counted in three separate areas, and an average was calculated, giving us a meiotic index. A quantitation was thus available to us, not realized in previous studies. All patients were followed for at least 9 months after completion of therapy and were evaluated with multiple semenograms. Adequate follow-up was achieved in all patients. The results of a separate study comparing the meiotic "index" of a control group with that of a comparable group of patients treated with estrogen demonstrated the normal range of meiotic figures in a testicular biopsy specimen. 2 Two groups of patients of known fertility were compared. The first group served as a control. The second group was evaluated after receiving estrogen. Significant alteration was noted both microscopically and in the meiotic index. A significant statistical difference existed (p < 0.001). On this basis we estimated that the "normal" meiotic index would be greater than 15.
RESULTS
Ten patients had a meiotic index ofless than 15. Only one of these men subsequently achieved a
Vol. 28, No.9
MEIOTIC INDICES IN THE OLIGOSPERMIC MALE
TABLE 1. Results for Oligospermic Patients with Low Meiotic Indices Meiotic index
Sperm count Patient Pretreatment
953
TABLE 3. Results for Oligospermic Patients with Normal Meiotic Indices Not Resulting in Pregnancy Sperm count
--~----
Result
Post-treatment
Patient
Pretreatment
T.H. P. Z. W.D. D. K. R. E. R. T. J.M. D. K.
12 4 14 0 8 6 4 1
Posttreatment
Meiotic index
Result
28.0 43.3 51.3 21.0 16 49.3 22.6 26.0
No pregnancy" No pregnancy" No pregnancy" No pregnancy" No pregnancy" Significant female factor" Significant female factor" Significant female factor"
x 101;
S. P. P.W. K.M. G.S. J.W. M.B. J.M. R. D. R. L. M.M.
11 3 9 1 8.6 9 0 0 17 18
8 4 10 1 11 12 0 0 12 2
3.6 5.0 3.0 8.6 1.6 10.6 0.6 6.3 2.6 5.3
No pregnancyU No pregnancy" No pregnancy" No pregnancy" No pregnancy" No pregnancy" No pregnancyh No pregnancy" No pregnancy" Pregnancy"
Ligation of the internal spermatic vein + APL. "APL only. ''Ligation of the internal spermatic vein only. U
pregnancy. The minimal follow-up period for patients who did not achieve a pregnancy in this study was 1 year. Seven patients had no improvement in the semenogram, whereas three (including the one achieving pregnancy) experienced a modest increase in the sperm count of 16 million/ml (Table 1). The remaining 24 patients had a meiotic index of more than 15 (16 of these men achieved pregnancy and are considered separately from the 8 who did not). Sixteen patients with a meiotic index greater than 15 were successful in achieving pregnancy. The index ranged from 19.6 to 66.0. Ten patients had a significant increase in their semenogram values (10 to 102 million), three had a slight increase (2, 2, and 8 million), the value for one reTABLE 2. Results for Oligospermic Patients with Normal Meiotic Indices Resulting in Pregnancy Sperm count
Patient
D.M. N.H. T.M. J.M. M.B. J. C. C. S. W.G. G.B. R. W. H.R. R. P. R. S. S. B. J. T. S.M.
------=--------Pretreatment
Post-treatment
5 1 19 1 0 3
20 40 19 10 8 18 69 104 76 32
11
2 8 19 19 0 7 3 3 20
2 77
24 5
Meiotic index
Result
26.3 41.0 57.3 28.3 22.0 26.0 23.6 37.3 24.6 66.0 19.6 20.0 25.3 61.6 40.0 36.3
Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy"
X
101;
41 4 6 0 47 106 41 86
"Ligation of the internal spermatic vein only. bAPL only. "Ligation of the internal spermatic vein + APL.
mained unchanged, and two did not have a followup semenogram (Table 2). Eight men with a meiotic index greater than 15 were unsuccessful in achieving pregnancy. Three of these patients had no improvement in their semenogram values. Five patients in this group had a significant increase in their semenogram values (29, 39, 100, 37, and 85 million). In the latter three cases, a significant female factor (anovulation) was present (Table 3). The average increase in the sperm count was significantly related to the meiotic index. In 10 patients with a meiotic index of less than 15, the average post-therapy increase in the sperm count was 2.4 x 106 • The average increase in sperm count in patients with a meiotic index between 15 and 30 was 33.15 x 106 , which was not significantly different from the average increase of30. 77 x 106 found in patients with meiotic index of more than 30. 35 <0 0
x
"Ligation of the internal spermatic vein + APL. hAPL only. "Ligation of the internal spermatic vein only.
"E
e"
.
E ~
.:
.~
"E >
e
Q.
..
.§
C>
~>
<
5
FIG. 1. Meiotic index related to average increase in sperm count (x 106 ).
954
MEHAN ET AL.
A significant statistical difference existed between those patients whose meiotic index was less than 15 and those whose index was greater than 15. Because of the wide variance in sperm counts, a Cochran's modified test was used (j> < 0.02) (Fig. 1). By applying the X2 test to these results, a significant statistical difference was noted (j> < 0.01). In addition, if the three patients with a known serious female factor are excluded, the statistical difference becomes more marked (j> < 0.005). DISCUSSION
Prior unsuccessful experience with qualitative evaluation of meiotic figures, that is, identification of stages only (diplotene, pachytene, zygotene, diakenesis, and others), prompted us to conduct this quantitative study wherein actual counts of the meiotic figures per 100 nucleated cells were obtained. In our hands this determination has been found to be easily within the capability of
September 1977 any cytogenetic laboratory. Its use should be restricted to the oligospermic male, since little useful information is gained by applying it to the normospermic male. The meiotic index enables us to prognosticate with a reliability not currently available with other means. A reliable prognosis is particularly important in counseling patients with regard to adoption or other alternate means of achieving children. The cryptorchid testis of the teenage patient and the postvasectomy testis pose intriguing possibilities for additional applications of the meiotic index. REFERENCES 1. Chehval MJ, Mehan DJ, Yolk Sr LR: Meiotic figures in
the testicular biopsy of subfertile males. Fertil Steril 28:253, 1977 2. Mehan DJ, Chehval MJ, Yolk Sr LR: Effect of estrogen on the meiotic index of males. Presented at the Annual Meeting of the American Urological Association, Chicago, April,1977
FERTILITY AND STERILITY Copyright' 1977 The American Fertility Society
MEIOTIC INDICES IN THE OLIGOSPERMIC MALE
DONALD J. MEHAN, M.D.* MICHEAL J. CHEHVAL, M.D. SISTER LEO RITA YOLK
Departments of Urology, St. Mary's Hospital and St. Louis University Hospital, St. Louis, Missouri 63117
Thirty-four oligospermic males were evaluated by determining the meiotic index in testicular biopsy specimens. Only 1 patient of 1 0 with a meiotic index of less than 15 achieved pregnancy, whereas 16 of24 patients with a meiotic index of more than 15 had a successful outcome. Corresponding increases in the semenogram were also noted to be related to the meiotic index.
For the past 4 years we have been studying the clinical application of meiotic preparations to the evaluation and treatment of the subfertile male. Our initial studies dealt with the identification of figures of meiosis in testicular biopsy specimens. 1 These studies dealt with the ability to predict the results of therapy purely on the basis of noting the stage of meiosis present in the biopsy specimen. The clinical correlation of prognosis based on the presence of meiotic figures was disappointing and prompted us to count the meiotic figures rather than rely on qualitative analysis. MATERIALS AND METHODS
Thirty-four patients referred for evaluation of subfertility were entered into the study. No significant endocrinologic or physical findings were present with the exception of varicocele (in 25 men). All patients were oligospermic, having a count ofless than 20 x 106 spermatozoa/ml. Treatment consisted ofligation of the internal spermatic vein (13 cases), ligation of the internal spermatic vein plus anterior pituitary-like hormone (Ayerst Laboratories, New York, N. Y.) (APL) (12 cases), or APL only (9 cases). Testicular biopsy specimens were obtained either during the repair of a varicocele or independently as part of the work-up Received April 22, 1977; revised May 16, 1977; accepted May 23,1977. *Reprint requests: Donald J. Mehan, M.D., 1035 Bellevue Avenue, St. Louis, Mo. 63117.
of the oligospermic patient. A part of the biopsy specimen was fixed and stained in our cytogenetic laboratory as previously described. 1 In addition to the usual cellular identification, meiotic counts were carried out in the following fashion. The identifiable meiotic figures per 100 nucleated cells were counted in three separate areas, and an average was calculated, giving us a meiotic index. A quantitation was thus available to us, not realized in previous studies. All patients were followed for at least 9 months after completion of therapy and were evaluated with multiple semenograms. Adequate follow-up was achieved in all patients. The results of a separate study comparing the meiotic "index" of a control group with that of a comparable group of patients treated with estrogen demonstrated the normal range of meiotic figures in a testicular biopsy specimen. 2 Two groups of patients of known fertility were compared. The first group served as a control. The second group was evaluated after receiving estrogen. Significant alteration was noted both microscopically and in the meiotic index. A significant statistical difference existed (p < 0.001). On this basis we estimated that the "normal" meiotic index would be greater than 15.
RESULTS
Ten patients had a meiotic index ofless than 15. Only one of these men subsequently achieved a
Vol. 28, No.9
MEIOTIC INDICES IN THE OLIGOSPERMIC MALE
TABLE 1. Results for Oligospermic Patients with Low Meiotic Indices Meiotic index
Sperm count Patient Pretreatment
953
TABLE 3. Results for Oligospermic Patients with Normal Meiotic Indices Not Resulting in Pregnancy Sperm count
--~----
Result
Post-treatment
Patient
Pretreatment
T.H. P. Z. W.D. D. K. R. E. R. T. J.M. D. K.
12 4 14 0 8 6 4 1
Posttreatment
Meiotic index
Result
28.0 43.3 51.3 21.0 16 49.3 22.6 26.0
No pregnancy" No pregnancy" No pregnancy" No pregnancy" No pregnancy" Significant female factor" Significant female factor" Significant female factor"
x 101;
S. P. P.W. K.M. G.S. J.W. M.B. J.M. R. D. R. L. M.M.
11 3 9 1 8.6 9 0 0 17 18
8 4 10 1 11 12 0 0 12 2
3.6 5.0 3.0 8.6 1.6 10.6 0.6 6.3 2.6 5.3
No pregnancyU No pregnancy" No pregnancy" No pregnancy" No pregnancy" No pregnancy" No pregnancyh No pregnancy" No pregnancy" Pregnancy"
Ligation of the internal spermatic vein + APL. "APL only. ''Ligation of the internal spermatic vein only. U
pregnancy. The minimal follow-up period for patients who did not achieve a pregnancy in this study was 1 year. Seven patients had no improvement in the semenogram, whereas three (including the one achieving pregnancy) experienced a modest increase in the sperm count of 16 million/ml (Table 1). The remaining 24 patients had a meiotic index of more than 15 (16 of these men achieved pregnancy and are considered separately from the 8 who did not). Sixteen patients with a meiotic index greater than 15 were successful in achieving pregnancy. The index ranged from 19.6 to 66.0. Ten patients had a significant increase in their semenogram values (10 to 102 million), three had a slight increase (2, 2, and 8 million), the value for one reTABLE 2. Results for Oligospermic Patients with Normal Meiotic Indices Resulting in Pregnancy Sperm count
Patient
D.M. N.H. T.M. J.M. M.B. J. C. C. S. W.G. G.B. R. W. H.R. R. P. R. S. S. B. J. T. S.M.
------=--------Pretreatment
Post-treatment
5 1 19 1 0 3
20 40 19 10 8 18 69 104 76 32
11
2 8 19 19 0 7 3 3 20
2 77
24 5
Meiotic index
Result
26.3 41.0 57.3 28.3 22.0 26.0 23.6 37.3 24.6 66.0 19.6 20.0 25.3 61.6 40.0 36.3
Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy" Pregnancy"
X
101;
41 4 6 0 47 106 41 86
"Ligation of the internal spermatic vein only. bAPL only. "Ligation of the internal spermatic vein + APL.
mained unchanged, and two did not have a followup semenogram (Table 2). Eight men with a meiotic index greater than 15 were unsuccessful in achieving pregnancy. Three of these patients had no improvement in their semenogram values. Five patients in this group had a significant increase in their semenogram values (29, 39, 100, 37, and 85 million). In the latter three cases, a significant female factor (anovulation) was present (Table 3). The average increase in the sperm count was significantly related to the meiotic index. In 10 patients with a meiotic index of less than 15, the average post-therapy increase in the sperm count was 2.4 x 106 • The average increase in sperm count in patients with a meiotic index between 15 and 30 was 33.15 x 106 , which was not significantly different from the average increase of30. 77 x 106 found in patients with meiotic index of more than 30. 35 <0 0
x
"Ligation of the internal spermatic vein + APL. hAPL only. "Ligation of the internal spermatic vein only.
"E
e"
.
E ~
.:
.~
"E >
e
Q.
..
.§
C>
~>
<
5
FIG. 1. Meiotic index related to average increase in sperm count (x 106 ).
954
MEHAN ET AL.
A significant statistical difference existed between those patients whose meiotic index was less than 15 and those whose index was greater than 15. Because of the wide variance in sperm counts, a Cochran's modified test was used (j> < 0.02) (Fig. 1). By applying the X2 test to these results, a significant statistical difference was noted (j> < 0.01). In addition, if the three patients with a known serious female factor are excluded, the statistical difference becomes more marked (j> < 0.005). DISCUSSION
Prior unsuccessful experience with qualitative evaluation of meiotic figures, that is, identification of stages only (diplotene, pachytene, zygotene, diakenesis, and others), prompted us to conduct this quantitative study wherein actual counts of the meiotic figures per 100 nucleated cells were obtained. In our hands this determination has been found to be easily within the capability of
September 1977 any cytogenetic laboratory. Its use should be restricted to the oligospermic male, since little useful information is gained by applying it to the normospermic male. The meiotic index enables us to prognosticate with a reliability not currently available with other means. A reliable prognosis is particularly important in counseling patients with regard to adoption or other alternate means of achieving children. The cryptorchid testis of the teenage patient and the postvasectomy testis pose intriguing possibilities for additional applications of the meiotic index. REFERENCES 1. Chehval MJ, Mehan DJ, Yolk Sr LR: Meiotic figures in
the testicular biopsy of subfertile males. Fertil Steril 28:253, 1977 2. Mehan DJ, Chehval MJ, Yolk Sr LR: Effect of estrogen on the meiotic index of males. Presented at the Annual Meeting of the American Urological Association, Chicago, April,1977