FORMATION OF STEROID HORMONES IN VITRO BY DEVELOPING CORPORA LUTEA OF THE RABBIT

1977 ◽  
Vol 75 (3) ◽  
pp. 355-361 ◽  
Author(s):  
AKIRA SUZUKI ◽  
TAKAHIDE MORI ◽  
TOSHIO NISHIMURA

Rabbits were injected with human chorionic gonadotrophin (HCG), and slices of developing corpora lutea taken from the ovaries 15, 18, 24, 48, 72 and 96 h after injection were incubated with [1-14C]sodium acetate at 37 °C for 3 h. The incorporation of labelled acetate into ten steroids, including progestagens, androgens and oestrogens, was analysed. In the initial step of corpus luteal formation, the specific incorporation (incorporation of [1-14C]acetate/100 mg tissue) increased sharply. The major steroidal products were progesterone, 17-hydroxyprogesterone and 20α-hydroxypregn-4-en-3-one. Between 18 and 48 h, the increase in specific incorporation was more gradual than in the initial step. Although the pattern was also dominated by progestagens, a temporary increase in the incorporation of acetate into androgens and oestrogens was observed. In the final step, a sharp rise in the total incorporation (incorporation of [1-14C]acetate/corpus luteum) was found, whereas the specific incorporation increased only slightly. The principal steroids produced were progesterone, pregnenolone and 20α-hydroxypregn-4-en-3-one. Incorporation into C19 steroids declined markedly and that into C18 steroids could not be detected. This profile of steroidogenesis 96 h after injection of HCG was similar to that of the corpus luteum in pregnancy. Thus marked quantitative and qualitative changes have been demonstrated during the period of formation of corpora lutea in the rabbit.

1969 ◽  
Vol 62 (3) ◽  
pp. 449-460 ◽  
Author(s):  
P. Preumont ◽  
I. D. Cooke ◽  
K. J. Ryan

ABSTRACT In vitro incubations of isolated porcine pre-ovulatory follicles, corpora lutea, and minced whole pre- and post-ovulatory ovaries were undertaken in an attempt to elucidate the pathways of oestrogen biosynthesis and steroid metabolism in the various cellular compartments of the porcine ovary. The conversion of radioactive acetate to cholesterol, the conversions of pregnenolone and progesterone to 16-hydroxyprogesterone, 17-hydroxyprogesterone, oestrone and oestradiol and the conversion of pregnenolone to 17-hydroxypregnenolone were observed following incubation with pre-ovulatory follicles in vitro. No significant effect of human chorionic gonadotrophin (HCG) added in vitro on steroid metabolism could be demonstrated. The conversions of dehydroepiandrosterone to oestradiol and 16-hydroxydehydroepiandrosterone to 16-hydroxyandrostenedione were obtained with corpora lutea in vitro. Pre-ovulatory whole ovarian mince incubations converted progesterone to 16-hydroxyprogesterone and androstenedione to oestrone. When 16-hydroxyandrostenedione, 16-hydroxyprogesterone and 16-hydroxydehydroepiandrosterone were incubated as substrates and/or formed in these various studies, no evidence for the formation of oestriol or other 16-oxygenated oestrogens could be demonstrated, although non-16 hydroxylated neutral steroids were aromatized in paired experiments.


1984 ◽  
Vol 103 (1) ◽  
pp. 107-110 ◽  
Author(s):  
M. G. Hunter

ABSTRACT Human luteal tissue recovered from varying stages of the luteal phase was minced and incubated for 3 h and the effect of human chorionic gonadotrophin (hCG), prolactin and hCG + prolactin on progesterone and oestradiol production measured. While hCG generally enhanced both progesterone and oestradiol synthesis, prolactin alone at either 20 or 200 μg/l had no significant effect on steroidogenesis. When prolactin was added along with hCG in four of six corpora lutea, however, progesterone production significantly increased and in three of six corpora lutea oestradiol production was increased above that induced by hCG alone. It is concluded that prolactin may play some role in the control of steroidogenesis by the human corpus luteum. J. Endocr. (1984) 103, 107–110


1981 ◽  
Vol 91 (2) ◽  
pp. 197-203 ◽  
Author(s):  
M. C. RICHARDSON ◽  
G. M. MASSON

Cell suspensions were prepared from tissue samples of human corpora lutea obtained during the mid- and late-luteal phase of the menstrual cycle. Both oestradiol and progesterone production by dispersed cells were stimulated by similar concentrations of human chorionic gonadotrophin (hCG). As the degree of stimulation of production by hCG was greater for progesterone than for oestradiol (five- to tenfold compared with two- to threefold higher than basal production), the ratio of progesterone to oestradiol produced varied according to the level of trophic stimulation. A comparison of cell suspensions prepared from mid- and late-luteal phase corpora lutea, exposed to the same concentration of hCG (10 i.u./ml) in vitro, did not reveal a shift to oestradiol production in the late-luteal phase. Provision of additional testosterone during incubation raised the level of oestradiol production by dispersed luteal cells. At an optimum concentration of testosterone (1 μmol/l), oestradiol synthesis was not raised further in the presence of hCG or N6, O2-dibutyryl cyclic AMP, suggesting a lack of induction or activation of the aromatase system by gonadotrophin in short-term cultures. Basal and stimulated levels of progesterone production were not significantly impaired in the presence of testosterone.


1982 ◽  
Vol 95 (1) ◽  
pp. 65-70 ◽  
Author(s):  
G. J. S. Tan ◽  
R. Tweedale ◽  
J. S. G. Biggs

The effects of oxytocin on dispersed luteal cells from human corpora lutea of the menstrual cycle were studied. Oxytocin at a concentration of 4 mi.u./ml produced a slight increase in basal progesterone production. However, higher oxytocin concentrations (400 and 800 mi.u./ml) markedly inhibited both basal and human chorionic gonadotrophin-induced progesterone production. These data provide evidence for an effect of oxytocin on the human corpus luteum. In view of the inhibitory action of oxytocin, increased secretion of this hormone may be important in the demise of the corpus luteum at the end of the menstrual cycle.


1983 ◽  
Vol 96 (3) ◽  
pp. 499-503 ◽  
Author(s):  
G. J. S. Tan ◽  
J. S. G. Biggs

The effects of prolactin on steroidogenesis were studied in dispersed luteal cells prepared from human corpora lutea of the menstrual cycle. Prolactin, at concentrations of 0·1–1000 ng/ml, had no effect on progesterone production by luteal cells during short-term incubation (3 h). However, in two out of five corpora lutea, higher concentrations of prolactin (100 and 1000 ng/ml) significantly reduced the oestradiol-17β production induced by human chorionic gonadotrophin (hCG; 10 i.u./ml); lower doses of prolactin had little effect. In the remaining corpora lutea, prolactin failed to affect either basal or hCG-induced production of oestradiol-17β. These results are discussed in relation to the mechanism by which prolactin influences human ovarian function.


1963 ◽  
Vol 42 (4) ◽  
pp. 509-513 ◽  
Author(s):  
D. Gospodarowicz ◽  
J. Legault-Démare

ABSTRACT Human chorionic gonadotrophin (HCG) and lactogenic hormone (LTH or prolactin) were found practically inactive on the incorporation of 14Cacetate into cholesterol of normal rat corpus luteum in vitro. On the contrary, when added simultaneously to the incubation medium, they increased by 90% the labeling of cholesterol. When pseudopregnancy corpora lutea were used, HCG alone stimulated to the same amount, but no stimulation was observed with LTH alone. These results show that the stimulation of cholesterol synthesis is produced by a synergic action of LTH and HCG, LTH being introduced either in vivo (pseudopregnancy) or in vitro.


2020 ◽  
Vol 32 (2) ◽  
pp. 177
Author(s):  
K. Hazano ◽  
S. Haneda ◽  
M. Matsui

In cattle, human chorionic gonadotrophin (hCG) is administered at Day 5 post-ovulation to improve fertility. This treatment can induce ovulation of the first-wave dominant follicle (W1DF), from which an accessory corpus luteum (CL) is generated. In addition, hCG has the effect of promoting CL development. It is possible that the locational relationship between the original and accessary CLs influences the effect of hCG on CL development, because the locational relationship of the CLs affects intraovarian blood flow. The present study aimed to clarify whether the locational relationship between the original and accessory CLs influences the effect of hCG on their development. Cross-bred beef heifers (Holstein×Japanese Black, n=56) were used for the present study. The oestrus cycle was synchronized using oestradiol benzoate (EB) and a controlled internal drug release (CIDR)-based program. Briefly, an administration of EB (2mg) with 9-day CIDR insertion was followed by administration of prostaglandin F2a analogue (PGF2a) on the day of CIDR removal, EB (1mg) 1 day after a PGF2a injection, and GnRH 12h after the second EB injection. At Day 5 post-ovulation, the locational relationship between the original CL and the W1DF was confirmed using transrectal ultrasonography (USG), and two groups were defined: ipsilateral group (IG; n=30), in which the CL and the W1DF are in the same ovary, and contralateral group (CG; n=26), in which the CL and the W1DF are in separate ovaries. Moreover, IG and CG were respectively subdivided into two groups, with or without hCG (1500IU) treatment (IG/hCG, n=15; IG without hCG, n=15, and CG/hCG, n=14; CG without hCG, n=12). The diameter and luteal tissue area (i.e. minus the cavity area) of the original CL and the accessory CL were examined at Days 5, 7, and 14, using USG. Two-way repeated-measures ANOVA was used to compare the diameter and luteal tissue area between IG/hCG and IG without hCG, and between CG/hCG and CG without hCG. In CG, the diameter (P<0.01) and luteal tissue area of the original CL (P<0.001) at Day 7 was increased by receiving hCG, while it did not change in IG. The diameter and luteal tissue area of the original CL at Day 14 were not affected by the administration of hCG in either CG or IG. Moreover, for the accessory CL, no difference of the diameter and luteal tissue area was observed between CG and IG. The present study showed that hCG treatment at Day 5 post-ovulation stimulate the growth of the original CL at Day 7, when the original CL and accessory CL are on contralateral sides. Our results suggest that the effect of administration of the hCG at Day 5 post-ovulation on the original CL development depends on the locational relationship between the original and accessory CL (IG or CG). The function of the CL affects the intrauterine environment for embryonic development. Therefore, it is necessary to investigate the effect of the hCG injection at Day 5 on the function of CL (i.e. plasma P4 concentration) in IG and CG, respectively.


1989 ◽  
Vol 120 (2) ◽  
pp. 325-330 ◽  
Author(s):  
J. Th. J. Uilenbroek ◽  
P. J. A. Woutersen ◽  
P. D. M. van der Vaart

ABSTRACT Corpora lutea could be identified under the dissection microscope up to 7 days after formation. They were isolated during the oestrous cycle and pseudopregnancy and the progesterone and 20α-OH-progesterone contents were compared with serum values of these steroids. The pattern of progesterone in serum resembled that found in the corpora lutea. However, the pattern of 20α-OH-progesterone concentrations in serum and corpora lutea were different. While 20α-OH-progesterone concentrations in the corpora lutea showed large variations during the cycle, changes in serum concentrations of 20α-OH-progesterone were relatively small. Measurement of hormone concentrations in isolated corpora lutea is therefore a sensitive method for studying corpus luteum activity. To study whether corpora lutea derived after ovulation of immature follicles showed deficient luteal activity, rats at dioestrus (2 days before pro-oestrus) were induced to ovulate by the injection of 10 IU human chorionic gonadotrophin (hCG) and subsequent luteal activity was studied by measuring hormone concentrations in the corpora lutea on day 5 of pseudopregnancy. Concentrations of progesterone, but not of 20α-OH-progesterone, in corpora lutea derived from follicles induced to ovulate at dioestrusday 1 were significantly lower than those in corpora lutea derived from follicles induced to ovulate at prooestrus. This difference was observed not only when pseudopregnancy was induced by cervical stimulation but also when it was induced by implantation of a pituitary gland under the kidney capsule. However, in the latter case, corpora lutea already present on the day of hCG injection also became activated. The present experiments demonstrate that by measuring hormone concentrations in isolated corpora lutea changes in luteal activity can be studied effectively. Moreover, it appears that corpora lutea derived from immature follicles contained less progesterone than those derived from fully mature follicles. Journal of Endocrinology (1989) 120, 325–330


2012 ◽  
Vol 24 (3) ◽  
pp. 472 ◽  
Author(s):  
D. Rizos ◽  
S. Scully ◽  
A. K. Kelly ◽  
A. D. Ealy ◽  
R. Moros ◽  
...  

The aim of the present study was to test the hypothesis that elevated concentrations of progesterone (P4) resulting from the induction of an accessory corpus luteum (CL) by human chorionic gonadotrophin (hCG) administration on Day 5 after oestrus would lead to advanced conceptus elongation on Day 14 following embryo transfer on Day 7. The oestrous cycles of cross-bred beef heifers were synchronised and animals were randomly assigned to receive either of two treatments: (1) intramuscular injection of 3000 IU hCG on Day 5 after oestrus (n = 14); or (2) intramuscular injection of saline on Day 5 after oestrus (n = 13). Ovaries were scanned daily by transrectal ultrasonography to assess CL development. Serum concentrations of P4 were determined from daily blood samples collected from the jugular vein. In vitro-produced bovine blastocysts were transferred to synchronised recipients on Day 7 after oestrus (n = 15 blastocysts per recipient). Heifers were killed on Day 14 after oestrus and the uterus was flushed to recover the embryos. Injection of hCG on Day 5 induced ovulation of the dominant follicle in all treated heifers and increased the total area of luteal tissue on the ovary, which was associated with a significant increase (P < 0.001) in serum concentrations of P4 from Day 7 to Day 14. Positive associations were detected between circulating P4 with CL area (within-day correlations ranging from r = 0.45 to r = 0.67) and total area of luteal tissue (within-day correlations ranging from r = 0.65 to r = 0.86) Administration of hCG did not affect the proportion of Day 14 conceptuses recovered. However, compared with the control group, hCG-treated heifers had increased conceptus length (3.91 ± 1.23 vs 5.57 ± 1.02 mm, respectively; P = 0.06), width (1.00 ± 0.06 vs 1.45 ± 0.05 mm, respectively; P = 0.002) and area (5.71 ± 0.97 vs 8.31 ± 0.83, respectively; P = 0.02). Although numerically greater, mean interferon-τ (IFNT) production in vitro did not differ significantly (P = 0.54) between embryos recovered from hCG-treated and control heifers. In contrast, there was a strong positive correlation between individual embryo length (r = 0.76; P < 0.001) and individual embryo area (r = 0.72; P < 0.001) and IFNT production. In conclusion, administration of hCG on Day 5 after oestrus resulted in the formation of an accessory CL and hypertrophy of the original CL, the result of which was an increase in P4 concentrations from Day 7 onwards. These elevated P4 concentrations were associated with an increased conceptus area. Furthermore, conceptus size was highly correlated with IFNT secretion in vitro.


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