Plasma testosterone and dihydrotestosterone in ovulatory and anovulatory cycles M. BRIJ Yew
YUSOFF B. York,
D.4WOOD, SAXENA,
M.D.,
CH.B.,
M.ME.D.,
M.K.C.O.G.,
F.A.C.0.G.
PH.D.
Lveu' York
IS WELL ESTABLISHED that the ovary secretes androgens during the menstrual cycle,’ and disruptions in the metabolism and/or production of androgens are associated with polycystic ovarian disease,‘, ” hirsutism,’ and the testicular feminization syndrome.’ In the human subject, estradiol (Et) can be produced by aromatization of testosterone (T), but 5wreduced androgens such as dihydrotestosterone (DHT) cannot be aromatized to estrogens. There is peripheral conversion of T to DHT. Investigations in rats have emphasized the control of luteinizing-hormone (LH) secretion at physiologic levels by androgens that are
not aromatized to estrogens. DHT inhibits estradiol binding by engaging in low-affinity interactions which decreases the rate of formation of the estradiolreceptor complex.’ Recent investigations to establish the pattern of androgen fluctuation have focused attention mainly on T and/or androstenedione (A) and dehydroepiandrosterone (DHEA).‘-’ With the exception of the results of’ ‘4uletta and associates,’ T as measured in these studies usually included DHT as there was no atEempt to separate these taco androgens. In view of the relationship of T and DHT to estrogen levels and estrogenreceptor binding and the possible effect of feedback on gonadotropin release, measurements of T and DHT levels throughout the menstrual cycle in both ovulatoq and anovulaEory cycles would be important to permit more meaningful interpretation of plasma T and DHT levels. Androgen levels throughout anovulatory cycles have not been previously reported. This paper reports our study of circulating T and DHT during ovulatory and anovulatory cycles. in relation to other gonadal steroids and LH.
IT
430
Plasma
l
testosterone
and
dihydrotestosterone
431
0
0
.t 199
zl! 0
0
1
0
l
0
I-
0 0 l
0
0 00
00 0
ti
0 0
0
o 0 0
I
0 0 0 0 00
1
8 l
s*
0
0
0
:
.:
Oo .O
1
8
.
-12
-10
-8
0
,
,
,
s
,
,
1
-6
-4
-2
0
+2
+l
+6
DAYS Fig.
1. Distribution
of’ plasma
T ad
z DHT
Material and methods Ten cycles in seven healthy adult women, 20 to 40 wars old, Ivho \vere not using any form of hormonal corm-aception or receiving hormonal treatment were studied. All the subjects had normal working hours and regular menstrual cycles. Three of the cycles studied in two subjects lasted 35 to 38 days, while the other seven cycles had a duration between 22 and 28 days. Daily basal body temperature was taken and rewrded in each subject in each cycle studied. Blood was drawn daily around the midcycle or every other day when it was at the beginning or end of a cycle, at the same time each day, betlveen 0900 and 1000 hours. The plasma was separated and stored at -20’ C. until ready for hormonal assay. All the plasma samples from a sueject were assayed for each hormone in a single Subjects.
run. Hormonal assays. Plasma T and DHT were measured 1~~ radioimmunoassay as described by Auletta and associates.’ Details of the modification as performed in our laboratory have been described previously.’ The antibody used was raised in rabbits against T-17p hemisuccinate con.jugated to bovine serum albumin. .4fter 1 ml. of plasma was extracted with
o
1
0 l .o
1 +8
0. 0
0
-14
0 0
L 0
1
0.0
+I0
, + l2
,
,
+ 14
l
J
16
l lE
OVULATION throughout
seven
ovulatory
menstrual
chides
da~~.vl
anhydrous ether. the estrac t 13as dried, and chromatography w-as performed on Sephadex l,H-20 with the use of isooctzme : benzene: methCmol (9.5: .5 : .j, v : v : v)‘as eluant to separate T and DH’T ‘r and DHT were then measured by ~adi~)inirl~iirl~~~~s~av. All assays w’ere carried out in duplicate, and procedural losses were monitored by addiCon of T 1.2-H’(K).*’ All results were corrected for procedural losses. The intra-assay coefficient of variaGon was 9 per cent, and the interassay coefficient of’variation was 13.S per cent. Plasma E? was measured by radioi~ll~~~~~~~oassa~ wit11 the use of antiserum raised in sheep agatnst es~radiol17/M?-(0carboxymethyl) oxime coupled to bovine serum albumin. Details of the procedure LS perfortned in our laboratorv have been described bcfi)re.“’ Procedural losses were monitored by additiolt ol l&2. 4, 6, 7-H”(N),+ Recovery of Ez \~as 79./k 2 I. I per
YUSOFF B. York,
D.4WOOD, SAXENA,
M.D.,
CH.B.,
M.ME.D.,
M.K.C.O.G.,
F.A.C.0.G.
PH.D.
Lveu' York
IS WELL ESTABLISHED that the ovary secretes androgens during the menstrual cycle,’ and disruptions in the metabolism and/or production of androgens are associated with polycystic ovarian disease,‘, ” hirsutism,’ and the testicular feminization syndrome.’ In the human subject, estradiol (Et) can be produced by aromatization of testosterone (T), but 5wreduced androgens such as dihydrotestosterone (DHT) cannot be aromatized to estrogens. There is peripheral conversion of T to DHT. Investigations in rats have emphasized the control of luteinizing-hormone (LH) secretion at physiologic levels by androgens that are
not aromatized to estrogens. DHT inhibits estradiol binding by engaging in low-affinity interactions which decreases the rate of formation of the estradiolreceptor complex.’ Recent investigations to establish the pattern of androgen fluctuation have focused attention mainly on T and/or androstenedione (A) and dehydroepiandrosterone (DHEA).‘-’ With the exception of the results of’ ‘4uletta and associates,’ T as measured in these studies usually included DHT as there was no atEempt to separate these taco androgens. In view of the relationship of T and DHT to estrogen levels and estrogenreceptor binding and the possible effect of feedback on gonadotropin release, measurements of T and DHT levels throughout the menstrual cycle in both ovulatoq and anovulaEory cycles would be important to permit more meaningful interpretation of plasma T and DHT levels. Androgen levels throughout anovulatory cycles have not been previously reported. This paper reports our study of circulating T and DHT during ovulatory and anovulatory cycles. in relation to other gonadal steroids and LH.
IT
430
Plasma
l
testosterone
and
dihydrotestosterone
431
0
0
.t 199
zl! 0
0
1
0
l
0
I-
0 0 l
0
0 00
00 0
ti
0 0
0
o 0 0
I
0 0 0 0 00
1
8 l
s*
0
0
0
:
.:
Oo .O
1
8
.
-12
-10
-8
0
,
,
,
s
,
,
1
-6
-4
-2
0
+2
+l
+6
DAYS Fig.
1. Distribution
of’ plasma
T ad
z DHT
Material and methods Ten cycles in seven healthy adult women, 20 to 40 wars old, Ivho \vere not using any form of hormonal corm-aception or receiving hormonal treatment were studied. All the subjects had normal working hours and regular menstrual cycles. Three of the cycles studied in two subjects lasted 35 to 38 days, while the other seven cycles had a duration between 22 and 28 days. Daily basal body temperature was taken and rewrded in each subject in each cycle studied. Blood was drawn daily around the midcycle or every other day when it was at the beginning or end of a cycle, at the same time each day, betlveen 0900 and 1000 hours. The plasma was separated and stored at -20’ C. until ready for hormonal assay. All the plasma samples from a sueject were assayed for each hormone in a single Subjects.
run. Hormonal assays. Plasma T and DHT were measured 1~~ radioimmunoassay as described by Auletta and associates.’ Details of the modification as performed in our laboratory have been described previously.’ The antibody used was raised in rabbits against T-17p hemisuccinate con.jugated to bovine serum albumin. .4fter 1 ml. of plasma was extracted with
o
1
0 l .o
1 +8
0. 0
0
-14
0 0
L 0
1
0.0
+I0
, + l2
,
,
+ 14
l
J
16
l lE
OVULATION throughout
seven
ovulatory
menstrual
chides
da~~.vl
anhydrous ether. the estrac t 13as dried, and chromatography w-as performed on Sephadex l,H-20 with the use of isooctzme : benzene: methCmol (9.5: .5 : .j, v : v : v)‘as eluant to separate T and DH’T ‘r and DHT were then measured by ~adi~)inirl~iirl~~~~s~av. All assays w’ere carried out in duplicate, and procedural losses were monitored by addiCon of T 1.2-H’(K).*’ All results were corrected for procedural losses. The intra-assay coefficient of variaGon was 9 per cent, and the interassay coefficient of’variation was 13.S per cent. Plasma E? was measured by radioi~ll~~~~~~~oassa~ wit11 the use of antiserum raised in sheep agatnst es~radiol17/M?-(0carboxymethyl) oxime coupled to bovine serum albumin. Details of the procedure LS perfortned in our laboratorv have been described bcfi)re.“’ Procedural losses were monitored by additiolt ol l&2. 4, 6, 7-H”(N),+ Recovery of Ez \~as 79./k 2 I. I per
