Estradiol-induced follicular atresia in rhesus monkeys is not prevented by exogenous gonadotropins

Estradiol-17β (E₂) induces atresia of the dominant preovulatory follicle (DF) when administered on day 6 of the menstrual cycle. The present study was designed to determine whether the atretogenic effect of E₂ could be averted by the administration of exogenous gonadotropins, in an attempt to determine whether E₂-induced atresia in primates is due to a direct action at the ovarian level or is mediated via pituitary secretion. After identification of the DF during laparoscopy, cyclic monkeys received Silastic capsules containing E₂ placed s.c. for 24 hours, plus one of the following treatments: phosphate-buffered saline, or 25 I.U. of either human urinary menopausal gonadotropin (hMG), FSH-rich hMG, human urinary FSH (uFSH), or human pituitary FSH (pFSH) injected i.m. twice daily for 2 days. The control treatment resulted in atresia of the DF and extended follicular phases (26.3 ± 5.9 days, x? ± S.D.), but in normal luteal phases following ovulation of a substitute DF. Similar results occurred in all animals receiving FSH-rich hMG or pFSH, and in 11 of 16 animals receiving hMG or uFSH (P > 0.05). Since all possible routes and regimens of gonadotropin administration were not attempted, a central action of E₂ cannot be ruled out. However, we believe that the experimental observations support our contention that the atretogenic action of E₂ is exerted in part at the ovary.     

Incidence of atresia or of luteinization without rupture of the dominant ovarian follicle in rhesus monkeys treated with estradiol-17β on day 8 of the menstrual cycle

Previous results have established that 17β-estradiol (E₂) administered in capsules for 24 h on Day 6 of the menstrual cycle results in atresia of the dominant follicle (DF). The present experiment was designed to determine if atresia could be induced similarly as late as Day 8, when the DF is presumably larger, to facilitate biochemical analyses. On the morning of Day 8, laparoscopy was used to confirm the presence of the DF, and 4, 6, or 8 Silastic capsules containing E₂ were placed s.c. for 12 or 24 h. Treatment with E₂ for 12 h resulted in atresia of the dominant follicle: 2/4,4/5, and 2/5 with 4, 6, and 8 capsules, respectively. Increases in luteinizing hormone (LH) usually occurred following removal of E₂ after 12 h; similar increases resulted previously from treatment on Day 6, and are interpreted as the result of removal of E₂-induced negative feedback rather than an effect of positive feedback. In contrast to the efficacy of treatment for 12 h, treatment with E₂ for 24 h resulted in a lower incidence of atresia (2/5,1/6, and 1/4 with 4, 6, and 8 capsules), and premature LH surges were seen during treatment in almost all animals, especially with 6 or 8 capsules, initiating luteinization of the DF without ovulation. Regarding intrafollicular changes, E₂ treatment resulted in a 10-fold reduction in follicular fluid concentrations of estrogen, while progesterone (P) levels were augmented; the output of P by granulosa cells in culture was also enhanced by E₂ treatment. We conclude that the DF in monkeys remains susceptible to the atretogenic effects of E₂ as late as Day 8 of the cycle, but the amount of follicular material recoverable is not increased with the later treatment. Furthermore, the similarity between the luteinized unruptured follicles induced in this study and those occurring spontaneously in women suggests that this model may be valuable in elucidating the factors involved in this source of infertility in women. © 1994 Wiley-Liss, Inc.     

Direct effect of estradiol-17β on progesterone accumulation by ovarian granulosa cells from rhesus monkeys

We have previously demonstrated that estradiol-17β (E₂) administered in vivo induces atresia of the dominant ovarian follicle (DF). Whether this effect is exerted directly at the ovarian level or by central mediation has not been confirmed. The present study was designed to assess whether E₂ in amounts similar to those found in monkey follicular fluid (FF) directly alters in vitro progesterone (P) accumulation by granulosa cells (GC) aspirated from follicles in cycling rhesus monkeys. Follicular contents were aspirated from three to five animals on each of days 8–13 of the cycle. GC were plated at a density of 50,000 viable GC/0.5 ml medium; GC were incubated with 0 or 2–2,000 ng/ml E₂, and cultures were maintained for 72 h. P accumulation by GC collected on day 8 and treated with 2 ng/ml E₂ was augmented 37.5 ± 5.5% (X ± S. E. M.; P<.05) over controls but was diminished significantly at 20 ng/ml ( ?55 ± 18% with respect to controls), 200 ng/ml ( ?73.7 ± 13.2%), and 2,000 ng/ml ( ?77.3 ± 18.4%). A similar dose-response relationship was noted on other cycle days. At a concentration of 2,000 ng/ml, E₂ significantly reduced P 91.5 ± 8.5% (day 10), 81.5 ± 18.5% (day 11), 84.3 ± 4.7% (day 12), and 53.7 ± 15.8% (day 13). We conclude that E₂ at concentrations found in FF can inhibit P output by monkey GC through a direct action.     

Menstrual cycles in rhesus monkeys (Macaca mulatta) are unaffected by a single dose of the anesthetics ketamine and xylazine administered during the midfollicular phase at laparoscopy

To identify an anesthetic regimen that produces more complete relaxation and analgesia than ketamine hydrochloride (Ketaset®) alone, a combination of ketamine (15 mg/kg body weight) and the hypnotic xylazine (Rompun®, 0.33 mg/kg) was evaluated. Since the desired experimental application required that the anesthetic not interfere with normal hormonal events during the menstrual cycle, this combination administered on day 6 of the cycle was tested to determine whether hormonal surges, incidence of ovulation, or cycle length would be altered relative to the use of ketamine alone. In five of six animals, ketamine plus xylazine had no effect on the occurrence of timely surges of estrogen, luteinizing hormone (LH), or follicle-stimulating hormone (FSH), or on ovulation as determined by the presence of a corpus luteum at laparoscopy and normal serum concentrations of progesterone. There were no significant differences between the cycle during treatment and previous cycles in the same animal for length of the menstrual cycle (26.0 ± 2.3 [5] days; X? ± S.D. [n] or luteal phase (13.4 ± 2.4 [5] days). Likewise, these values did not differ from those of ten control monkeys treated with ketumine only on day 5 or 6 of the cycle (incidence of ovulation, 10/10; cycle length, 27.9 ± 1.8 [10]; luteal phase length, 15.1 ± 1.4 [10], P > 0.05). Patterns of circulating progesterone were not altered by the addition of xylazine anesthesia. These findings indicate that xylazine, given in the midfollicular phase, did not alter ovulatory events or menstrual cycle characteristics in rhesus monkeys. Ketamine plus xylazine apparently provides anesthesia appropriate for laparoscopy.     

Atresia of the dominant ovarian follicle in rhesus monkeys is detected within 24 hours of estradiol treatment

Following treatment with estradiol-17β (E₂) on day 6 of the menstrual cycle, degenerative alterations in the microenvironment of the dominant follicle (DF) (follicular fluid [FF], granulosa cells [GC], and oocyte) are readily apparent on day 10, or 96 h after E₂ administration. The present study was designed to determine how early such changes could be detected and which indices of atresia were observed first. The DF was identified during laparoscopy on day 5 or 6 of the cycle, and four capsules containing crystalline E₂ were inserted s.c. for 24 h. Contents of the DF were aspirated at 24, 48, and 72 h following initiation of E₂ treatment. General size and appearance of the DF did not change distinctly with E₂ treatment; however, by 48 h FF viscosity was increased markedly. GC viability was not altered with treatment. FF concentrations of estrogen (E) were dramatically reduced at 24 h. These differences were maintained at 48 h and at 72 h. E accumulation by cultured GC was significantly reduced by > eightfold. There appeared a similar trend for reduced progesterone (P) in FF and decreased P production by GC in vitro. These results demonstrate that degenerative alterations in the DF indicative of atresia can be detected as early as 24 h after initiation of E₂ treatment; the index of atresia appearing earliest is a reduction in FF concentrations of E, and the first morphological changes in the DF can be observed 24 h later. This study indicates that biochemical alterations precede morphologic changes with E₂-induced atresia, and should allow us to begin to determine the earliest events and putative initiation sites of atresia.     

Variables affecting the yield and developmental potential of embryos following superstimulation and in vitro fertilization in rhesus monkeys

Three follicular-stimulation protocols were compared to evaluate the yield and quality of oocytes obtained from rhesus monkeys. Five animals received a high-dose regimen of PMSG (protocol I), three received a lower-dose regimen (protocol II), and two received Clomid-Pergonal (protocol III). Oocytes were recovered at laparoscopy after HCG injection, fertilized in vitro, and cultured up to the blastocyst stage. Yields of mature oocytes were 17.2 ± 13.0 (80% of total recovered), 6.7 ± 6.6 (41%), and 4.5 ± 2.5 (90%) per stimulated cycle for protocols I, II, and III, respectively. Of mature oocytes, 72%, 45%, and 89% were fertilized for protocols I, II and III, respectively. Protocol I produced the most fertilized oocytes per stimulated cycle (11.6 ± 11.6) and the greatest E₂ production (approximately fourfold that maximally expected for an unstimulated cycle). For the combined protocol I and II results, there was a significant correlation (P ? 0.05) between mean embryo development score and E₂ production. Fertilized oocytes from protocol I yielded 7.8 ± 8.0 morulae and 6.8 ± 7.2 early zonal blastocysts per cycle. After transfer of nine singleton embryos to surrogate recipients, one live birth resulted. We conclude that our high-dose PMSG regimen offers the best means at present for obtaining susbstantial numbers of developmentally competent oocytes in rhesus monkeys and that more extensive use of rapid serum E₂ assays for monitoring both stimulated cycles and those of potential surrogate recipients could help to predict the success of in vitro fertilization and embryonic development following embryo transfer in rhesus monkeys.     

Patterns of estrogen and progesterone receptors in monkey endometrium during the normal menstrual cycle

Total concentrations of estradiol-17β (E₂) and progesterone (P) receptors (R) were measured in the endometrium of rhesus monkeys ($\text{Macaca mulatta}$) during the normal menstrual cycle. The endometrium was collected at abdominal fundal hysterotomy on days 8, 12, 15, 18 and 24 of the menstrual cycle. Visual inspection of the ovaries and measurement of E₂, P, follicle stimulating hormone (FSH) and luteinizing hormone (LH) provided assuredness of normal ovarian function. Exchange procedures were used in order to measure the total concentrations of E₂R and PR in nuclear and cytosol fractions. The pattern of estrogen receptor showed a slight increase in the cytosol and nuclear concentrations at the preovulatory interval. Later, the total E₂R concentration was decreased when P increased during the luteal phase. Cytosol PR synthesis was parallel to the serum E₂ increase during the late follicular phase. Secretion of P by the corpus luteum was accompanied by a rapid nuclear translocation and concomitant decrease in cytoplasmic PR. Thereafter the total PR concentration declined during the second half of the luteal phase. These findings in monkey endometrium are similar to those reported for human endometrium during the normal menstrual cycle and further establish the utility of these surrogate primates in investigations indicative of human endometrial function.     

Excessive ovarian follicular development in pregnant pigtailed macaques (Macaca nemestrina)

To evaluate the status and possible control of ovarian follicular development during pregnancy, circulating levels of estrone (E₁), estradiol-17β (E₂), and follicle-stimulating hormone (FSH) were measured throughout gestation in both intact and ovariectomized pregnant pigtailed monkeys (Macaca nemestrina). From an additional group of pregnant monkeys, ovaries were obtained at late gestation (on day 150 or 159 of pregnancy) for histological studies. Circulating concentrations of E₁ and E₂ increased on day 13 and remained elevated for about 10 days; they then declined and reached low levels on day 32 of gestation. After day 60, there were gradual but smaller increases in estrogen levels to day 140, after which both E₁ and E₂ levels increased significantly, reaching maximum levels (E₁ = 832.2 ± 210.8 pg/ ml; E₂ = 1.66 ± 0.32 ng/ml) at the end of pregnancy. Removal of ovaries on day 35 of gestation did not affect pregnancy or the pattern of estrogen secretion. Serum concentrations of FSH demonstrated only minor fluctuations during pregnancy but were similar to those found during the early follicular phase of cycling pigtailed monkeys investigated in this study. Ovarian histology revealed extensive follicular growth; in addition to the corpus luteum of pregnancy, ovaries were packed with pre-antral, small antral, and medium-sized Graafian follicles. Some of these follicles appeared to be cystic and showed various degree of atresia; their general appearance was similar to the follicles of human females with polycystic ovary syndrome. Our data suggest that FSH may initiate ovarian follicular growth during gestation. High levels of estrogens were incapable of suppressing FSH secretion but may be responsible for the induction of atresia in a large number of follicles in pregnant pigtailed monkeys.     

Laparoscopic observations of ovaries before and after ovulation in the Japanese monkey (Macaca fuscata)

Laparoscopic observations of ovaries before and after ovulation were made in 74 cycles in 39 Japanese monkeys (Macaca fuscata). Lengths of menstrual cycles, the follicular phase and the luteal phase in these animals were 25.3±2.8 days, 13.2±1.4 days, and 12.8±2.3 days, respectively. Ovarian morphology during the menstrual cycle in the Japanese monkey agreed in some features and disagreed in others, with findings inMacaca mulatta andMacaca fascicularis reported in the previous papers using laparotomy or laparoscopy. Follicular appearance before and after ovulation varied so considerably from ovary to ovary that it was hard to predict the accurate ovulation time or to estimate the precise age of corpus luteum by means of observations on the morphological appearance only. However, diagnosis of whether ovulation had already occurred or not, was possible when careful observations were made, although some ovaries without haemorrhagic and luteinized appearance after ovulation were very similar to pre-ovulatory follicle. Clear luteinization was detected without fail in many cases three days after ovulation. This study was supported by the Scientific Research Fund of the Ministry of Education of Japan (No. 856165).     

Absence of direct effects of prostaglandins on rabbit blastocysts in vitro

A total of 27 monkeys (M. Fascicularis) whose control cycle lengths ranged from 28 to 32 days were used in this study. All the treatments described below started either on day 17 or 18 of the cycle. Six monkeys received daily injections of 20 μg estradiol-17β (E₂) for 5 consecutive days. Although a drop in blood progesterone (P) did occur due to this treatment, no shortening of the luteal phase of the cycle was recorded. Seven monkeys received daily injections of 15 mg PGF_2α (prostaglandin-F_2α) for 4 or 5 days. These monkeys also showed a drop in blood P levels; moreover 5 of these monkeys had vaginal bleeding for 2–3 days starting either on day 19 or 20 of the cycle. This bleeding did not appear to be a normal physiological menstrual flow, since all of the monkeys commenced menstrual flow at the expected time. Four monkeys received daily injections of 10 mg P for 3 days. These monkeys also had normal cycle lengths in spite of the treatment. Finally 9 monkeys received daily injections of 20 μg E₂ for 3 days, and starting on the third day of E₂ treatment these monkeys also received injections of 15 mg PGF_2α for 4 or 5 days. Shortened cycle lengths were recorded in 8/9 monkeys in this group. Six monkeys had 22-day cycles, 2 monkeys had 24-day cycles and the remaining monkey had a cycle length of 26 days. Thus 8/9 monkeys had shortened luteal phases due to sequential treatment of E₂ and PGF_2α. The cycle lengths in all the treatment groups were normal subsequent to treatments. These results provide potentially useful information for further studies in the human as a method of contraception.     

Temporal and endocrine sequelae of aspirating follicular contents in rhesus monkeys

Aspiration of ovarian follicular contents in humans is a well-established procedure used to obtain oocytes for fertilization in vitro (IVF). However, the effects of aspiration on the menstrual cycle and resulting luteal function have been incompletely characterized. The present study was designed to investigate alterations in the temporal and endocrine characteristics of menstrual cycles following aspiration of contents of the dominant preovulatory follicle (DF) on day 10 of the cycle in normal rhesus monkeys. When aspiration was performed prior to the preovulatory surge of luteinizing hormone (LH), cycle length was extended to 38.6 ± 8.6 [15] (x days ± SD, [n]), as compared to 29.5 ± 5.7 [8] days when the surge occurred before the time of aspiration. Mean and maximal amounts of progesterone (P) in the luteal phase and the number of days in which P-values were > 1 ng/ml were significantly greater when aspiration was performed prior to the surge of LH than for aspiration after this event. Laparoscopic observations made in the midluteal phase in animals of the former group demonstrated that the corpus luteum (CL was derived from a follicle other than the original DF which had been aspirated on day 10 of the menstrual cycle; observations in the latter group of animals indicated that the CL was derived from the DF.     

News from the molecular frontier. Review of Paternity in Primates: Genetic Tests and Theories R. D. Martin,A. F. Dixson,E. J. Wickings eds. Karger, 1992, xi + 287 pp, $198.50

We have previously demonstrated that the exogenous administration of estradiol-17β (E₂) to rhesus monkeys induces atresia of the dominant preovulatory follicle (DF); and that this effect is mediated centrally, via the inhibition of follicle-stimulating hormone, and is also exerted directly at the level of the ovarian granulosa cell. We wished to investigate whether the local effect of E₂ is transduced through interaction with the nuclear receptor for estrogen, particularly in light of certain evidence that suggests a general lack of estrogen receptor (E-R) in the rhesus monkey ovary, except in the germinal epithelium. In the present study, we evaluated the presence of E-R by both autoradiographic and immunocytochemical techniques. Frozen sections of ovaries from rhesus females were incubated in experiment 1 with either ³H-E₂ or ¹²⁵I-E₂, in the presence or absence of excess, non-radioactive ligand or analogues diethylstilbestrol (DES) or the receptor antagonist 4-OH-tamoxifen (TAM). ³H-E₂ binding was most intense over functional corpora lutea, and was reduced to background with excess DES; label was also evident over antral follicles, Image analysis showed specific binding of ¹²⁵I-E₂ by ovaries. In experiment 2, cryostat sections were processed for immunocytochemical staining using the per-oxidase-anti-peroxidase (PAP) method and the H222 monoclonal antibody to the E-R. Intense, specific label was observed over nuclei of germinal epithelium, but, additionally, for the first time, to granulosa cells of antral follicles and other compartments of the ovary. In this paper, we report the first evidence for estrogen binding to rhesus monkey ovary; tins binding is specific, apparently receptor mediated, and corroborated independently by autoradiographic and immunocytochemical means. We herein provide substantial support for estrogen's dramatic effects being exerted directly at the level of the monkey ovary. © 1993 Wiley-Liss, Inc.     

17 β-hydroxysteroid dehydrogenase in monkey endometrium during the menstrual cycle and at the time of implantation

In order to further identify physiological similarities between 17β-hydroxysteroid dehydrogenase (HSD) in human and monkey endometrium, and to evaluate the role of estradiol-17β (E₂) oxidation to estrone (E₁) during periimplantation events, 30 rhesus monkeys were studied at different intervals of the nonfertile menstrual cycle (days 8, 12, 15, 18 and 24). Also, five pregnant monkeys provided endometrial tissue on day 24 of the fertile menstrual cycle, near the expected time of implantation. HSD activity in endometrium was low at midfollicular phase (day 8), increased to maximal levels (8-fold) during the periovulatory span (days 12 and 15),and was intermediate in mid to late luteal phase (days 18 and 24) in non-fertile menstrual cycles. In the absence of ovulation, HSD was low throughout. These enzyme data fit with a pattern of daily peripheral serum levels of E₂ and progesterone (P) and suggest that when the normal sequence of P follows elevated estrogens in late follicular phase, HSD activity is markedly enhanced in the early luteal phase. However, HSD activity in endometrium did not increase more in the fertile menstrual cycle, despite further elevations of serum P during rescue of the corpus luteum.     

Menopause in female rhesus monkeys

Fifteen female rhesus monkeys (Macaca mulatto), ranging in age from 8 to 34 years, were studied for one year to characterize the endocrine and menstrual changes associated with menopause in this species. Five monkeys were premenopausal; these younger monkeys, ages 8–11 years, menstruated and showed cyclic ovarian activity during the 12–month study period, as evidenced by menses and periodic elevations of serum estradiol (E₂) and luteinizing hormone (LH) concentrations. Four females, ages 24–26 years, were in transition to menopause. Two of these perimenopausal females menstruated and secreted E₂ and LH in a periodic fashion; the other two females showed elevated LH concentrations, consistently low E₂ levels, and no evidence of menstruation. Six females, ages 27–34 years, were clearly postmenopausal; LH concentrations were high, whereas E₂ concentrations were uniformly low. There was a significant inverse correlation between basal E₂ concentrations and age, and a significant positive correlation between age and LH concentrations across all 15 animals. Hormonal changes indicative of ovulation, when they occurred, were generally restricted to the winter and early spring months. Histological analysis of ovaries from four postmenopausal females revealed little or no evidence of active folliculogenesis. These data indicate that menopause in female rhesus monkeys does not occur until the second half of thethird decade of life. © 1995 Wiley-Liss, Inc.     

Effect of inhibition of estrogen synthesis during the luteal phase on function of the corpus luteum in rhesus monkeys

It has been hypothesized that estrogen synthesized by the corpus luteum initiates luteal regression during the nonfertile menstrual cycle in primates. To study the role of endogenous estrogens in functional regression of the monkey corpus luteum, we administered the aromatase inhibitor 1,4,6-androstatriene-3,17-dione (ATD) to rhesus monkeys during the luteal phase of the menstrual cycle. Twice-daily oral administration of ATD suppressed systemic and intraluteal estrogen levels by 80-90%. The midluteal phase rise in estradiol concentrations that occurs in rhesus monkeys was completely abolished by ATD treatment. Despite suppression of estrogen synthesis during the luteal phase, mean menstrual cycle length and length of the luteal phase were not different than in control monkeys treated with vehicle only. Progesterone levels were lower in the ATD-treated group on the second and third day of treatment, but did not differ from control levels during the remainder of the cycle. These data suggest that elevated estrogen synthesis during the luteal phase of the menstrual cycle is not a prerequisite for spontaneous luteolysis in rhesus monkeys.     

Plasma estrogens,progesterone and chorionic gonadotropin in pregnant rhesus monkeys (Macaca mulatta) after ovariectomy

Maternal peripheral plasma concentrations of estrone (E₁), estradiol-17β (E₂), and progesterone (P) were determined in 4 rhesus monkey ovariectomized in early pregnancy (22–24 days). After ovariectomy, plasma concentrations of E₁ and E₂ were basal for 1 to 2 weeks. In contrast, slightly higher estrogen levels, which may be attributed to the ovaries, were found in intact pregnant monkeys. E₂ levels increased rapidly after this and exceeded those of E₁ until the 5th month of gestation. From that time until parturition, E₁ levels equaled or exceed those of E₂ in most instances. The pattern of P concentrations was similar to that observed in intact monkeys. Urinary chorionic gonadotropin (MCG) levels in ovariectomized monkeys were not significantly differen from those found in normal pregnancies. Thus, the patterns for circulating E_l, E₂ and P, as well as for the excretion of MCG,. after ovari-ectomy were remarkably similar to those found in intact, pregnant rhesus monkeys, indicating minimal ovarian influence.     

Dynamics of circulating concentrations of gonadotropins and ovarian hormones throughout the menstrual cycle in the bonnet monkey: role of inhibin A in the regulation of follicle‐stimulating hormone secretion

In higher primates, increased circulating follicle‐stimulating hormone (FSH) levels seen during late menstrual cycle and during menstruation has been suggested to be necessary for initiation of follicular growth, recruitment of follicles and eventually culminating in ovulation of a single follicle. With a view to establish the dynamics of circulating FSH secretion with that of inhibin A (INH A) and progesterone (P₄) secretions during the menstrual cycle, blood was collected daily from bonnet monkeys beginning day 1 of the menstrual cycle up to 35 days. Serum INH A levels were low during early follicular phase, increased significantly coinciding with the mid cycle luteinizing hormone (LH) surge to reach maximal levels during the mid luteal phase before declining at the late luteal phase, essentially paralleling the pattern of P₄ secretion seen throughout the luteal phase. Circulating FSH levels were low during early and mid luteal phases, but progressively increased during the late luteal phase and remained high for few days after the onset of menses. In another experiment, lutectomy performed during the mid luteal phase resulted in significant decrease in INH A concentration within 2 hr (58.3±2 vs. 27.3±3 pg/mL), and a 2‐ to 3‐fold rise in circulating FSH levels by 24 hr (0.20±0.02 vs. 0.53±0.14 ng/mL) that remained high until 48 hr postlutectomy. Systemic administration of Cetrorelix (150 µg/kg body weight), a gonadotropin releasing hormone receptor antagonist, at mid luteal phase in monkeys led to suppression of serum INH A and P₄ concentrations 24 hr post treatment, but circulating FSH levels did not change. Administration of exogenous LH, but not FSH, significantly increased INH A concentration. The results taken together suggest a tight coupling between LH and INH A secretion and that INH A is largely responsible for maintenance of low FSH concentration seen during the luteal phase. Am. J. Primatol. 71:817–824, 2009. © 2009 Wiley‐Liss, Inc.     

Effects of ovarian tissue reduction on the menstrual cycle: persistent normalcy after near-total oophorectomy

These experiments were designed to evaluate whether removal of approximately 95% visible ovarian tissue would interrupt the short- or long-term regulation of cyclic ovarian function. On cycle Days 2 4 (onset of menses = Day 1), the entire left ovary and approximately 90% of the right ovary were removed from three cycling cynomolgus monkeys. After approximately 95% ovariectomy, there was an acute elevation of follicle-stimulating hormone (FSH) and luteinizing hormone (LH), which lasted 11 +/- 2 days. A midcycle-like gonadotropin surge occurred 20 +/- 3 days following approximately 95% ovariectomy; the next menses occurred 19 +/- 1 days later. Follicular phase patterns of estradiol preceded the midcycle gonadotropin surge, and luteal phase progesterone levels indicated subsequent ovulation. Two of three monkeys resumed normal menstrual cyclicity in the following cycle with follicular phase, luteal phase, and menstrual cycle lengths similar to pretreatment levels. Histological examination of the ovarian remnant removed on Day 21 of the next cycle revealed a morphologically normal corpus luteum and many small follicles. A second group of 6 rhesus monkeys also underwent approximately 95% ovariectomy for long-term evaluation of menstrual cyclicity; typical 28-day menstrual cycle patterns were observed in 4 of the 6 monkeys for 5 mo, with 2 of these 3 animals maintaining regular menstrual cycles for 1 yr. In summary, our data suggest that normal ovarian function, i.e. recruitment, selection, and dominance of the ovulatory follicle, ovulation, and subsequent corpus luteum function, is maintained with only approximately 5% of functional ovarian tissue remaining.(ABSTRACT TRUNCATED AT 250 WORDS)     

Characterization of estrogen and progesterone receptors in monkey endometrium: methodology and effects of estradiol and/or progesterone on endometrium of castrate monkeys.

Appropriate methods for repeated surgical collection of endometrial tissue from rhesus monkeys, and characterization of cytosol and nuclear estrogen (E₂) and progesterone (P) receptors (R) are described. Tissue collection was made in the mid-luteal phase at abdominal fundal hysterotomy. Functional status of the ovaries was determined by visual inspection and RIA of E₂ and P in serum. Receptor assay procedures were devised permitting the measurement of total cytosol and nuclear receptor concentration. Sucrose density gradients of labelled cytosol were made and a 4S saturable binding component for ³H P and for ³H E₂ were found. Equilibrium dissociation constants of ³H E₂ and ³H R5020 were 2.1×10^?10M and 3.6 × 10^?9M, respectively. These binding characteristics are similar to those found in human endometrium and suggest that these surrogate primates have extensive utility in investigation of factors influencing E₂R and PR concentrations in endometrial tissue during the menstrual cycle and implantation. Simulated menstrual cycle were produced in 20 castrate monkeys by sequential treatment with estradiol and progesterone in silastic capsules. RIA of E₂ and P, and gonadotropins in peripheral serum provided assuredness of the hormonal status of each monkey under treatment. Cytosol and nuclear receptors for E₂ and P were measured in the endometrium after different intervals of the treatment. E₂ receptor (E₂R) levels were not changed during the estrogen sequence, but were lowered by progesterone therapy in both cytosol and nuclear components. Progesterone receptor (PR) synthesis in cytosol was induced by exogenous estrogen. The total concentration of PR decreased with the uptake of P by the cell; meanwhile, the ratio of cytosol to nuclear P receptors declined. These data suggest that this sequential estrogen-progesterone regimen induces the changes in E₂R and PR patterns in the endometrium of ovariectomized monkeys which occurs due to ovarian cyclicity in the normal menstrual cycle.     

Maternal estrogen and progesterone levels after hypophysectomy in early pregnancy and in term fetuses or newborn monkeys

Pregnant rhesus monkeys ($\text{Macaca}$$\text{mulatta}$) were hypophysectomized at 8–10 weeks gestation to determine effects on plasma levels of estrone (E₁), estradiol-17β (E₂) and progesterone (P). The first group of monkeys was subsequently fetectomized At 107–114 days. After hypophysectomy there was an initial decrease in maternal peripheral plasma E₂ followed by a rise to preoperative levels within 4–5 weeks. Plasma levels of e₁ and P were not markedly altered. After fetectomy, peripheral estrogen concentrations, especially E₂, declined markedly. In the second experimental series, we have examined the effects of maternal hypophysectomy on levels of E₁, E₂ and P either (1) in both mother and newborn baby or (2) in mother, term fetus and umbilical vein. Groups of hypophysectomized and intact pregnant monkeys (3 each) were delivered by cesarean section at the expected time of parturition. Other hypophysectomized and intact monkeys (2 each) delivered normally. E₂ levels were elevated significantly in plasma of hypophysectomized monkeys at the time of cesarean delivery and in newborn babies of hypophysectomized mothers shortly after parturition. Except for these differences, the maternal hypophysis apparently is not a major factor in the control of E₁, E₂ and P concentrations in pregnant rhesus monkeys.