Vol. 26, No. 4, April1975 Printed in U.S.A.
FERTILITY AND STERILITY Copyright ., 1975 The American Fertility Society
RESPONSE OF LUTEINIZING HORMONE AND FOLLICLE-STIMULATING HORMONE TO DIFFERENT DOSES OF SYNTHETIC LUTEINIZING HORMONE-RELEASING HORMONE BY INTRAMUSCULAR ADMINISTRATION IN NORMAL AND OLIGOSPERMIC MEN: PRELIMINARY REPORT DIEGO TURNER, PH.D., ELDA A. TURNER, PH.D., LUIS SCHWARZSTEIN, M.D.,* AND NESTOR J. APARICIO, M.D. Grupo de Estudio en Fertilidad y Endrocrinologia de Rosario (G.E.F.E.R.), Clinica de las Gltlndulas Endocrinas, Cordoba 1764, Rosario, Argentina
Both naturaP and synthetic2 luteinizing hormone-releasing hormones (LH-RH) stimulate the release of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) from the hypophysis. 3 Similarly, the dose-response correlation of LH and FSH to LH-RH for intravenous application is well established. 4 The present work attempted to establish a similar correlation for intramuscular LH-RH application in normal and oligospermic men, and to determine a suitable dose for chronic treatment. MATERIALS AND METHODS
by venipuncture of a cubital vein immediately before and 60, 120, 180, and 240 minutes after the LH-RH injection. During each test, the subjects remained in a resting position. Serum was extracted from each sample after clot retraction and was stored at -20° C until hormonal determinations were performed. The serum levels of LH and FSH were determined by radioimmunoassay with a double-antibody technique. 5•6 The results were expressed in international milliunits per milliliter of serum (2nd IRP-HMG).
Two healthy fertile men (ages, 26 and 28 years), and three oligospermic patients (ages, 19, 30, and 36 years), volunteered for study. All had normal weights in relation to their heights. Prior to the study, a clinical, endocrinologic examination found each subject normal. Spermogram results were normal for the healthy volunteers. Spermogram results showed counts fewer than 10 million spermatozoids/ml for the oligospermic men. Each subject underwent four tests a week apart. After an all-night fast, at 8 AM the subject was intramuscularly injected with 50, 100, 250, or 500 #Lg of synthetic LH-RH (Hoechst) which had been dissolved in 2 ml of 0.9% saline solution. Blood samples were extracted
RESULTS
No relevant differences in the basal values of LH and FSH were observed between the normal and the oligospermic men (Fig. 1). In the two normal subjects, both hormones responded under stimulation with synthetic LH-RH. Peak values were obtained 60 to 180 minutes after each injection and with doses between 100 and 250 #J-g. The 500-~J.g injection stimulated similar or even lower responses than the lower doses. In one normal subject, the LH levels at the end of the test remained above the basal levels; in the other subject, they returned to their initial levels. The FSH values at 240 minutes showed no relevant differences with the corresponding basal levels. Received May 7, 1974. One of the three oligospermic patients *Reprint requests: Dr. Schwarzstein, Clinics showed congruent responses of LH and de las Glimdulas Endocrinas, Cordoba 1764, Rosario, FSH. Another patient had an evident Argentina. 337
TURNERETAL
338
April1975
NORMAL SUBJECTS
Time (in minut••J OLIGOSPERMIC SUBJECTS
l~ fE ~
~ o.
/
A
-"'--~ ,-...:..,_,.
',,
,/
. 7·- ' ... _
' _.
/
:"'
_ .,
\
-- -" ',,
·1..
• /' ...-~- ''\·,:-"" ___.- - -·--
~~?~~ 0
0
eo
1111
110
2o10
o
eo
o2o
110
240
o
&a
120
,.,
uo
o
eo
o2o
,.,
2•o
Time (in minutes)
FIG. 1. Curves of the individual values of LH and FSH in two healthy volunteers and three oligospermic men before and after an intramuscular injection of 50, 100, 250, or 500 ILg of LH-RH.
LH response, but no relevant FSH variation after the 250-~tg injection. The third oligospermic patient showed practically no LH response and only a slight FSH response. When responses were obtained in these patients, the maximal peaks occurred between 60 and 180 minutes after the corresponding LH-RH intramuscular injection, and with doses between 100 and 250 l£g. DISCUSSION
The results of this study confirm those of other au'thors 7 ; no differences in the basal LH and FSH values between the normal and oligospermic subjects were found. Since the maximal responses of LH and FSH to intramuscularly injected
LH-RH were obtained with doses ranging between 100 and 250 JLg, we can assume that these would be the doses for chronic treatment; the 50-JLg dose stimulated very low or irrelevant responses, and the 500-JLg dose stimulated, in general, a decrease in LH and FSH levels. Despite random variations and the small number of subjects studied, the congruency of the results is still noteworthy. Maximal peaks in the hormone levels were obtained between 60 and 180 minutes after the injection and decreases occurred, in almost every case, at 240 minutes. Therefore, it would seem necessary to administer LH-RH several times a day if a prolonged gonadotropin stimulation were desired.
339
LH AND FSH RESPONSE
Vol. 26, No.4
Two of the three oligospermic patients had discordant responses of LH and FSH to LH-RH. This fact should be viewed cautiously because of the small number of patients studied; however, a possible difference in the pituitary response of oligospermic patients and normal men should be considered when analyzing LHRH test results.
From the results, we can assume that LH-RH doses between 100 and 250 11-g should be used as a basis for chronic treatment.
SUMMARY
REFERENCES
The response of LH and FSH levels to intramuscularly administered synthetic LH-RH was studied in two healthy volunteers and three oligospermic patients. Four tests with 50, 100, 250, and 500 11-g of LH-RH, respectively, were carried out on each subject at 8 AM; the interval between tests was one week. The serum levels of LH and FSH were determined by radioimmunoassay (double-antibody method) before each injection, and 60, 120, 180, and 240 minutes after each injection. No differences in the basal values of either hormone were observed. In both oligospermic and normal men, maximal responses were obtained with doses between 100 and 250 JLg. With 500 11-g, levels decreased rather than increased. Maximal peaks occurred between 60 and 180 minutes after injection. In the two normal subjects, the responses of LH and FSH were similar. Two of the three oligospermic patients showed discordant responses.
Acknowledgments. Acknowledgment is due to lnstituto Serono, Immunochemical Division (Rome, Italy), for providing material for LH and FSH radioimmunoassays and to Hoechst AG, West Germany, for providing synthetic LH-RH.
1. Baba Y, Matsuo H, Schally AV: Structure of porcine LH and FSH releasing hormone. II. Confirmation of the proposed structure by conventional sequential analysis. Biochem Biophys Res Commun 44:459, 1971 2. Geiger R, Konig W, Wissman H, et al: Synthesis and characterisation of a decapeptide having LH-RH/FSH-RH activity. Biochem Biophys Res Commun 45:767, 1971 3. Schally AV, Kastin AJ, Arimura A: The hypothalamus and reproduction. Am J Obstet Gynecol114:423, 1972 4. Haug E, T01jesen P: Effect of synthetic luteinizing hormone and follicle stimulating hormone-releasing hormone (LH-FSHIRH) on serum levels of LH, FSH, thyrotrophin (TSH) and growth hormone (HGH) in normal male subjects. Acta Endocrinol (Kbh) 73:465, 1973 5. Midgley AR Jr: Radioimmunoassay: a method for human chorionic gonadotropin and human luteinizing hormone. Endocrinology 79:10, 1966 6, Midgley AR Jr: Radioimmunoassay for folliclestimulating hormone. J Clin Endocrinol Metab 27:295, 1967 7. Franchimont P, Millet D, Vendrely E, et al: Relationship between spermatogenesis and serum gonadotropin levels in azoospermia and oligospermia. J Clin Endocrinol Metab 34:1003, 1972
FERTILITY AND STERILITY Copyright ., 1975 The American Fertility Society
RESPONSE OF LUTEINIZING HORMONE AND FOLLICLE-STIMULATING HORMONE TO DIFFERENT DOSES OF SYNTHETIC LUTEINIZING HORMONE-RELEASING HORMONE BY INTRAMUSCULAR ADMINISTRATION IN NORMAL AND OLIGOSPERMIC MEN: PRELIMINARY REPORT DIEGO TURNER, PH.D., ELDA A. TURNER, PH.D., LUIS SCHWARZSTEIN, M.D.,* AND NESTOR J. APARICIO, M.D. Grupo de Estudio en Fertilidad y Endrocrinologia de Rosario (G.E.F.E.R.), Clinica de las Gltlndulas Endocrinas, Cordoba 1764, Rosario, Argentina
Both naturaP and synthetic2 luteinizing hormone-releasing hormones (LH-RH) stimulate the release of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) from the hypophysis. 3 Similarly, the dose-response correlation of LH and FSH to LH-RH for intravenous application is well established. 4 The present work attempted to establish a similar correlation for intramuscular LH-RH application in normal and oligospermic men, and to determine a suitable dose for chronic treatment. MATERIALS AND METHODS
by venipuncture of a cubital vein immediately before and 60, 120, 180, and 240 minutes after the LH-RH injection. During each test, the subjects remained in a resting position. Serum was extracted from each sample after clot retraction and was stored at -20° C until hormonal determinations were performed. The serum levels of LH and FSH were determined by radioimmunoassay with a double-antibody technique. 5•6 The results were expressed in international milliunits per milliliter of serum (2nd IRP-HMG).
Two healthy fertile men (ages, 26 and 28 years), and three oligospermic patients (ages, 19, 30, and 36 years), volunteered for study. All had normal weights in relation to their heights. Prior to the study, a clinical, endocrinologic examination found each subject normal. Spermogram results were normal for the healthy volunteers. Spermogram results showed counts fewer than 10 million spermatozoids/ml for the oligospermic men. Each subject underwent four tests a week apart. After an all-night fast, at 8 AM the subject was intramuscularly injected with 50, 100, 250, or 500 #Lg of synthetic LH-RH (Hoechst) which had been dissolved in 2 ml of 0.9% saline solution. Blood samples were extracted
RESULTS
No relevant differences in the basal values of LH and FSH were observed between the normal and the oligospermic men (Fig. 1). In the two normal subjects, both hormones responded under stimulation with synthetic LH-RH. Peak values were obtained 60 to 180 minutes after each injection and with doses between 100 and 250 #J-g. The 500-~J.g injection stimulated similar or even lower responses than the lower doses. In one normal subject, the LH levels at the end of the test remained above the basal levels; in the other subject, they returned to their initial levels. The FSH values at 240 minutes showed no relevant differences with the corresponding basal levels. Received May 7, 1974. One of the three oligospermic patients *Reprint requests: Dr. Schwarzstein, Clinics showed congruent responses of LH and de las Glimdulas Endocrinas, Cordoba 1764, Rosario, FSH. Another patient had an evident Argentina. 337
TURNERETAL
338
April1975
NORMAL SUBJECTS
Time (in minut••J OLIGOSPERMIC SUBJECTS
l~ fE ~
~ o.
/
A
-"'--~ ,-...:..,_,.
',,
,/
. 7·- ' ... _
' _.
/
:"'
_ .,
\
-- -" ',,
·1..
• /' ...-~- ''\·,:-"" ___.- - -·--
~~?~~ 0
0
eo
1111
110
2o10
o
eo
o2o
110
240
o
&a
120
,.,
uo
o
eo
o2o
,.,
2•o
Time (in minutes)
FIG. 1. Curves of the individual values of LH and FSH in two healthy volunteers and three oligospermic men before and after an intramuscular injection of 50, 100, 250, or 500 ILg of LH-RH.
LH response, but no relevant FSH variation after the 250-~tg injection. The third oligospermic patient showed practically no LH response and only a slight FSH response. When responses were obtained in these patients, the maximal peaks occurred between 60 and 180 minutes after the corresponding LH-RH intramuscular injection, and with doses between 100 and 250 l£g. DISCUSSION
The results of this study confirm those of other au'thors 7 ; no differences in the basal LH and FSH values between the normal and oligospermic subjects were found. Since the maximal responses of LH and FSH to intramuscularly injected
LH-RH were obtained with doses ranging between 100 and 250 JLg, we can assume that these would be the doses for chronic treatment; the 50-JLg dose stimulated very low or irrelevant responses, and the 500-JLg dose stimulated, in general, a decrease in LH and FSH levels. Despite random variations and the small number of subjects studied, the congruency of the results is still noteworthy. Maximal peaks in the hormone levels were obtained between 60 and 180 minutes after the injection and decreases occurred, in almost every case, at 240 minutes. Therefore, it would seem necessary to administer LH-RH several times a day if a prolonged gonadotropin stimulation were desired.
339
LH AND FSH RESPONSE
Vol. 26, No.4
Two of the three oligospermic patients had discordant responses of LH and FSH to LH-RH. This fact should be viewed cautiously because of the small number of patients studied; however, a possible difference in the pituitary response of oligospermic patients and normal men should be considered when analyzing LHRH test results.
From the results, we can assume that LH-RH doses between 100 and 250 11-g should be used as a basis for chronic treatment.
SUMMARY
REFERENCES
The response of LH and FSH levels to intramuscularly administered synthetic LH-RH was studied in two healthy volunteers and three oligospermic patients. Four tests with 50, 100, 250, and 500 11-g of LH-RH, respectively, were carried out on each subject at 8 AM; the interval between tests was one week. The serum levels of LH and FSH were determined by radioimmunoassay (double-antibody method) before each injection, and 60, 120, 180, and 240 minutes after each injection. No differences in the basal values of either hormone were observed. In both oligospermic and normal men, maximal responses were obtained with doses between 100 and 250 JLg. With 500 11-g, levels decreased rather than increased. Maximal peaks occurred between 60 and 180 minutes after injection. In the two normal subjects, the responses of LH and FSH were similar. Two of the three oligospermic patients showed discordant responses.
Acknowledgments. Acknowledgment is due to lnstituto Serono, Immunochemical Division (Rome, Italy), for providing material for LH and FSH radioimmunoassays and to Hoechst AG, West Germany, for providing synthetic LH-RH.
1. Baba Y, Matsuo H, Schally AV: Structure of porcine LH and FSH releasing hormone. II. Confirmation of the proposed structure by conventional sequential analysis. Biochem Biophys Res Commun 44:459, 1971 2. Geiger R, Konig W, Wissman H, et al: Synthesis and characterisation of a decapeptide having LH-RH/FSH-RH activity. Biochem Biophys Res Commun 45:767, 1971 3. Schally AV, Kastin AJ, Arimura A: The hypothalamus and reproduction. Am J Obstet Gynecol114:423, 1972 4. Haug E, T01jesen P: Effect of synthetic luteinizing hormone and follicle stimulating hormone-releasing hormone (LH-FSHIRH) on serum levels of LH, FSH, thyrotrophin (TSH) and growth hormone (HGH) in normal male subjects. Acta Endocrinol (Kbh) 73:465, 1973 5. Midgley AR Jr: Radioimmunoassay: a method for human chorionic gonadotropin and human luteinizing hormone. Endocrinology 79:10, 1966 6, Midgley AR Jr: Radioimmunoassay for folliclestimulating hormone. J Clin Endocrinol Metab 27:295, 1967 7. Franchimont P, Millet D, Vendrely E, et al: Relationship between spermatogenesis and serum gonadotropin levels in azoospermia and oligospermia. J Clin Endocrinol Metab 34:1003, 1972
